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Sample records for alkalithermophile natranaerobius thermophilus

  1. Catabolic control of sugar metabolism in Streptococcus thermophilus

    NARCIS (Netherlands)

    Bogaard, van den P.T.C.

    2002-01-01

    Streptococcus thermophilus is used in many industrial dairy fermentations that require processing of milk at elevated temperatures. Its primary function is the rapid conversion of lactose to lactate while it also contributes to important sensory qualities. S. thermophilus strain CNRZ302 is unable to

  2. Genetic analysis of lipolytic activities in Thermus thermophilus HB27.

    Science.gov (United States)

    Leis, Benedikt; Angelov, Angel; Li, Haijuan; Liebl, Wolfgang

    2014-12-10

    The extremely thermophilic bacterium Thermus thermophilus HB27 displays lipolytic activity for the hydrolysis of triglycerides. In this study we performed a mutational in vivo analysis of esterases and lipases that confer growth on tributyrin. We interrupted 10 ORFs suspected to encode lipolytic enzymes. Two chromosomal loci were identified that resulted in reduced hydrolysis capabilities against tributyrin and various para-nitrophenyl acyl esters. By implementation of a convenient new one-step method which abstains from the use of selectable markers, a mutant strain with multiple scar-less deletions was constructed by sequentially deleting ORFs TT_C1787, TT_C0340, TT_C0341 and TT_C0904. The quadruple deletion mutant of T. thermophilus exhibited significantly lower lipolytic activity (approximately 25% residual activity compared to wild type strain) over a broad range of fatty acyl esters and had lost the ability to grow on agar plates containing tributyrin as the sole carbon source. Furthermore, we were able to determine the impact of each gene disruption on the lipolytic activity profile in this model organism and show that the esterase activity in T. thermophilus HB27 is due to a concerted action of several hydrolases having different substrate preferences and activities. The esterase-less T. thermophilus multi-deletion mutant from this study can be used as a screening and expression host for esterase genes from thermophiles or metagenomes. PMID:25102235

  3. Genetic analysis of lipolytic activities in Thermus thermophilus HB27.

    Science.gov (United States)

    Leis, Benedikt; Angelov, Angel; Li, Haijuan; Liebl, Wolfgang

    2014-12-10

    The extremely thermophilic bacterium Thermus thermophilus HB27 displays lipolytic activity for the hydrolysis of triglycerides. In this study we performed a mutational in vivo analysis of esterases and lipases that confer growth on tributyrin. We interrupted 10 ORFs suspected to encode lipolytic enzymes. Two chromosomal loci were identified that resulted in reduced hydrolysis capabilities against tributyrin and various para-nitrophenyl acyl esters. By implementation of a convenient new one-step method which abstains from the use of selectable markers, a mutant strain with multiple scar-less deletions was constructed by sequentially deleting ORFs TT_C1787, TT_C0340, TT_C0341 and TT_C0904. The quadruple deletion mutant of T. thermophilus exhibited significantly lower lipolytic activity (approximately 25% residual activity compared to wild type strain) over a broad range of fatty acyl esters and had lost the ability to grow on agar plates containing tributyrin as the sole carbon source. Furthermore, we were able to determine the impact of each gene disruption on the lipolytic activity profile in this model organism and show that the esterase activity in T. thermophilus HB27 is due to a concerted action of several hydrolases having different substrate preferences and activities. The esterase-less T. thermophilus multi-deletion mutant from this study can be used as a screening and expression host for esterase genes from thermophiles or metagenomes.

  4. New Insights into Various Production Characteristics of Streptococcus thermophilus Strains

    Science.gov (United States)

    Cui, Yanhua; Xu, Tingting; Qu, Xiaojun; Hu, Tong; Jiang, Xu; Zhao, Chunyu

    2016-01-01

    Streptococcus thermophilus is one of the most valuable homo-fermentative lactic acid bacteria, which, for a long time, has been widely used as a starter for the production of fermented dairy products. The key production characteristics of S. thermophilus, for example the production of extracellular polysaccharide, proteolytic enzymes and flavor substances as well as acidifying capacity etc., have an important effect on the quality of dairy products. The acidification capacity of the strains determines the manufacturing time and quality of dairy products. It depends on the sugar utilization ability of strains. The production of extracellular polysaccharide is beneficial for improving the texture of dairy products. Flavor substances increase the acceptability of dairy products. The proteolytic activity of the strain influences not only the absorption of the nitrogen source, but also the formation of flavor substances. Different strains have obvious differences in production characteristics via long-time evolution and adaptation to environment. Gaining new strains with novel and desirable characteristics is an important long-term goal for researchers and the fermenting industry. The understanding of the potential molecular mechanisms behind important characteristics of different strains will promote the screening and breeding of excellent strains. In this paper, key technological and functional properties of different S. thermophilus strains are discussed, including sugar metabolism, proteolytic system and amino acid metabolism, and polysaccharide and flavor substance biosynthesis. At the same time, diversity of genomes and plasmids of S. thermophilus are presented. Advances in research on key production characteristics and molecular levels of S. thermophilus will increase understanding of molecular mechanisms of different strains with different important characteristics, and improve the industrialization control level for fermented foods. PMID:27754312

  5. Bioprospecting of strains of Streptococcus thermophilus from Indian fermented milk products for folate production

    OpenAIRE

    Iyer, Ramya; Tomar, Sudhir Kumar; Mohanty, Ashok Kumar; Singh, Prashant; Singh, Rameshwar

    2011-01-01

    International audience This study deals with the bio-prospecting of folate producing strains of Streptococcus thermophilus isolated from milk and different fermented milk products of Indian origin. From a total of 500 randomly selected colonies isolated from 209 different samples, 117 isolates were identified as S. thermophilus by classical biochemical and molecular characterization. Frequency of incidence of S. thermophilus in the different samples of milk and milk products was variable w...

  6. Production of probiotic fresh white cheese using co-culture with Streptococcus thermophilus

    OpenAIRE

    Oktay Yerlikaya; Elif Ozer

    2014-01-01

    In this research, the probiotic Streptococcus thermophilus was inoculated into milk as co-culture to produce probiotic cheese. The effects of using Streptococcus thermophilus with other probiotic bacteria on cheese composition, and microbiological viability during 28 days of storage were investigated. Sensorial properties were determined only at 1st and 28th days of storage. The results showed that the use of Streptococcus thermophilus as co-culture in probiotic cheese production did not affe...

  7. Genome Signature Difference between Deinococcus radiodurans and Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Hiromi Nishida

    2012-01-01

    Full Text Available The extremely radioresistant bacteria of the genus Deinococcus and the extremely thermophilic bacteria of the genus Thermus belong to a common taxonomic group. Considering the distinct living environments of Deinococcus and Thermus, different genes would have been acquired through horizontal gene transfer after their divergence from a common ancestor. Their guanine-cytosine (GC contents are similar; however, we hypothesized that their genomic signatures would be different. Our findings indicated that the genomes of Deinococcus radiodurans and Thermus thermophilus have different tetranucleotide frequencies. This analysis showed that the genome signature of D. radiodurans is most similar to that of Pseudomonas aeruginosa, whereas the genome signature of T. thermophilus is most similar to that of Thermanaerovibrio acidaminovorans. This difference in genome signatures may be related to the different evolutionary backgrounds of the 2 genera after their divergence from a common ancestor.

  8. BlpC regulated bacteriocin production in Streptococcus thermophilus NRRL-B59671

    Science.gov (United States)

    Streptococcus thermophilus NRRL-B59671 was previously shown to produce a bacteriocin with anti-pediococcal activity, but the genetic components required for production were not characterized. Genome sequencing of several strains of S. thermophilus has identified a genetic locus which encodes a quor...

  9. Unidirectional reconstitution into detergent-destabilized liposomes of the purified lactose transport system of Streptococcus thermophilus

    NARCIS (Netherlands)

    Veenhoff, Liesbeth; Liang, Wei-Jun; Henderson, Peter J.F.; Leblanc, Gérard; Poolman, Bert

    1996-01-01

    The lactose transport protein (LacS) of Streptococcus thermophilus was amplified to levels as high as 8 and 30% of total membrane protein in Escherichia coli and S, thermophilus, respectively, In both organisms the protein was functional and the expression levels were highest with the streptococcal

  10. Viability and cholesterol uptake by Streptococcus thermophilus cultures in artificial git fluids

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    Małgorzata Ziarno

    2010-03-01

    Full Text Available Background. Streptococcus thermophilus is traditionally used in association with Lb. delbrueckii subsp. bulgaricus as a starter culture for the production of yoghurt. Some researchers have indicated that S. thermophilus may provide additional health benefits, for example it may reduce cholesterol levels. The aim of this study was to in vitro evaluate the cholesterol uptake and the viability of S. thermophilus isolates in artificial GIT environments. Material and methods. Twelve isolates of S. thermophilus were cultured in artificial gastric fluid (with pepsin added and in artificial duodenal fluid (with the enzyme complex added, and in M17 broth containing cholesterol at an initial concentration of 600 µg/mL, as well as in M17 broth without cholesterol. Immediately after the adding of bacteria inoculums and at the end of experiment, the concentration of cholesterol and the number of bacteria were measured. Results. S. thermophilus did not remove statistically significant amounts of cholesterol from artificial gastric fluid. The isolates showed the ability to uptake cholesterol from M17 broth and artificial duodenal fluid, and the degree of cholesterol uptake depended on the isolate. All isolates of S. thermophilus remove much more cholesterol from M17 broth than from artificial duodenal fluid. All S. thermophilus isolates had worse survival in artificial gastric or duodenal fluids than in M17 broth. Conclusions. The ability of S. thermophilus cells to survive in artificial gastric fluid and artificial duodenal fluid varied according to the isolates.

  11. Uptake of Branched-Chain Amino Acids by Streptococcus thermophilus

    OpenAIRE

    Akpemado, K. M.; Bracquart, P. A.

    1983-01-01

    The transport of branched-chain amino acids in Streptococcus thermophilus was energy dependent. The metabolic inhibitors of glycolysis and ATPase enzymes were active, but the proton-conducting uncouplers were not. Transport was optimal at temperatures of between 30 and 45°C and at pH 7.0 for the three amino acids leucine, valine, and isoleucine; a second peak existed at pH 5.0 with valine and isoleucine. By competition and kinetics studies, the branched-chain amino acids were found to share a...

  12. Preliminary X-ray crystallographic study of glucose dehydrogenase from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    The glucose dehydrogenase (GDH) protein from T. thermophilus HB8 was cloned, expressed, purified and crystallized. GDH crystals belong to space group P21 and diffract to 1.9 Å resolution. Thermus thermophilus is an aerobic chemoorganotroph that has been found to grow anaerobically in the presence of nitrate. Crystals of glucose dehydrogenase (GDH) from T. thermophilus HB8 belong to space group P21, with unit-cell parameters a = 36.90, b = 132.96, c = 60.78 Å, β = 97.2°. Preliminary studies and molecular-replacement calculations reveal that the asymmetric unit contains two monomers

  13. Fast induction of nisin resistance in Streptococcus thermophilus INIA 463 during growth in milk.

    Science.gov (United States)

    Garde, Sonia; Avila, Marta; Medina, Margarita; Nuñez, Manuel

    2004-11-01

    Streptococcus thermophilus INIA 463 became nisin-resistant after exposure in skim milk to subminimal inhibitory concentrations of nisin (1-3 IU/ml) for less than 2 h. Addition of 20 IU/ml caused a 4 log unit decrease in S. thermophilus counts of a culture not exposed previously to nisin, whereas no decrease was observed in the culture exposed to nisin for 2 h. Transfer of immunity genes as responsible for nisin resistance was discarded. The presence of extracellular or intracellular specific nisin-degrading enzymes was not detected in the nisin-resistant variant of S. thermophilus INIA 463. Nisin resistance was caused by the induction of a resistance mechanism. Transmission electron microscopy (TEM) revealed that the nisin-resistant variant of S. thermophilus INIA 463 had a thickened cell wall compared to the wild strain. Resistance to nisin was lost after one transfer (4 h growth) in nisin-free skim milk. PMID:15364470

  14. Mechanism of resistance to macrolide-lincosamide-streptogramin antibiotics in Streptococcus thermophilus

    Science.gov (United States)

    Resistance to macrolide-lincosamide-streptogramin (MLS) group antibiotics in the dairy bacterium Streptococcus thermophilus (ST) is documented but the mechanism of resistance has not been elucidated. MIC values for erythromycin (Erm), azithromycin (Azm), tylosin (Tyl), spiramycin (Spm), pristinamyci...

  15. Activation of Silent gal Genes in the lac-gal Regulon of Streptococcus thermophilus

    NARCIS (Netherlands)

    Vaughan, Elaine E.; Bogaard, Patrick T.C. van den; Catzeddu, Pasquale; Kuipers, Oscar P.; Vos, Willem M. de

    2001-01-01

    Streptococcus thermophilus strain CNRZ 302 is unable to ferment galactose, neither that generated intracellularly by lactose hydrolysis nor the free sugar. Nevertheless, sequence analysis and complementation studies with Escherichia coli demonstrated that strain CNRZ 302 contained structurally intac

  16. Cleavage of Phage DNA by the Streptococcus thermophilus CRISPR3-Cas System

    OpenAIRE

    Magadán, Alfonso H.; Marie-Ève Dupuis; Manuela Villion; Sylvain Moineau

    2012-01-01

    Streptococcus thermophilus, similar to other Bacteria and Archaea, has developed defense mechanisms to protect cells against invasion by foreign nucleic acids, such as virus infections and plasmid transformations. One defense system recently described in these organisms is the CRISPR-Cas system (Clustered Regularly Interspaced Short Palindromic Repeats loci coupled to CRISPR-associated genes). Two S. thermophilus CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been shown to actively blo...

  17. Effects of Argonaute on Gene Expression in Thermus thermophilus.

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    Daan C Swarts

    Full Text Available Eukaryotic Argonaute proteins mediate RNA-guided RNA interference, allowing both regulation of host gene expression and defense against invading mobile genetic elements. Recently, it has become evident that prokaryotic Argonaute homologs mediate DNA-guided DNA interference, and play a role in host defense. Argonaute of the bacterium Thermus thermophilus (TtAgo targets invading plasmid DNA during and after transformation. Using small interfering DNA guides, TtAgo can cleave single and double stranded DNAs. Although TtAgo additionally has been demonstrated to cleave RNA targets complementary to its DNA guide in vitro, RNA targeting by TtAgo has not been demonstrated in vivo.To investigate if TtAgo also has the potential to control RNA levels, we analyzed RNA-seq data derived from cultures of four T. thermophilus strain HB27 variants: wild type, TtAgo knockout (Δago, and either strain transformed with a plasmid. Additionally we determined the effect of TtAgo on expression of plasmid-encoded RNA and plasmid DNA levels.In the absence of exogenous DNA (plasmid, TtAgo presence or absence had no effect on gene expression levels. When plasmid DNA is present, TtAgo reduces plasmid DNA levels 4-fold, and a corresponding reduction of plasmid gene transcript levels was observed. We therefore conclude that TtAgo interferes with plasmid DNA, but not with plasmid-encoded RNA. Interestingly, TtAgo presence stimulates expression of specific endogenous genes, but only when exogenous plasmid DNA was present. Specifically, the presence of TtAgo directly or indirectly stimulates expression of CRISPR loci and associated genes, some of which are involved in CRISPR adaptation. This suggests that TtAgo-mediated interference with plasmid DNA stimulates CRISPR adaptation.

  18. Isolation and characterization of chromosomal promoters of Streptococcus salivarius subsp. thermophilus.

    Science.gov (United States)

    Slos, P; Bourquin, J C; Lemoine, Y; Mercenier, A

    1991-01-01

    A promoter probe vector, pTG244, was constructed with the aim of isolating transcription initiation signals from Streptococcus thermophilus (Streptococcus salivarius subsp. thermophilus). pTG244 is based on the Escherichia coli-streptococcus shuttle vector pTG222, into which the promoterless chloramphenicol acetyltransferase gene of Bacillus pumilus (cat-86) was cloned. Random Sau3A fragments from the S. thermophilus A054 chromosomal DNA were cloned upstream of the cat-86 gene by using E. coli as the host. The pool of recombinant plasmids were introduced into S. thermophilus and Lactococcus lactis subsp. lactis in order to search for promoter activity in these hosts. For S. thermophilus, it was necessary to first select erythromycin-resistant transformants and then to screen for chloramphenicol resistance among these. Direct selection of chloramphenicol-resistant clones was, however, possible in L. lactis subsp. lactis. Six fragments exhibiting promoter activity were characterized in S. thermophilus by measuring the levels of cat-86 transcription and/or chloramphenicol acetyltransferase specific activity. Three of the promoter-carrying fragments were sequenced. The 5' ends of their corresponding mRNAs were determined by S1 mapping and shown to correspond to a purine residue in all cases. Upstream from these potential transcription start points, sequences homologous to the E. coli sigma 70 and the Bacillus subtilis vegetative sigma 43 (or sigma A) consensus promoters were identified. Images PMID:1854195

  19. Isolation and characterization of Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus from plants in Bulgaria.

    Science.gov (United States)

    Michaylova, Michaela; Minkova, Svetlana; Kimura, Katsunori; Sasaki, Takashi; Isawa, Kakuhei

    2007-04-01

    One of the traditional ways of preparation of yogurt starter in Bulgaria is placing a branch of a particular plant species into boiled sheep's milk maintained at about 45 degrees C, which is further incubated until a dense coagulum is obtained. To investigate the possible origin of the yogurt starter bacteria, Lactobacillus delbrueckii ssp. bulgaricus (L. bulgaricus) and Streptococcus thermophilus (S. thermophilus), the traditional way of yogurt-starter preparation was followed. Hundreds of plant samples were collected from four regions in Bulgaria and incubated in sterile skim milk. The two target bacteria at low frequencies from the plant samples collected were successfully isolated. Phenotypic and genotypic characteristics of these bacterial isolates revealed that they were identified as L. bulgaricus and S. thermophilus. Twenty isolates of L. bulgaricus and S. thermophilus, respectively, were selected from the isolated strains and further characterized with regard to their performance in yogurt production. Organoleptic and physical properties of yogurt prepared using the isolated strains from plants were not significantly different from those prepared using commercial yogurt-starter strains. It was therefore suggested that L. bulgaricus and S. thermophilus strains widely used for commercial yogurt production could have originated from plants in Bulgaria. To our knowledge, this is the first report on the isolation and characterization of L. bulgaricus and S. thermophilus strains from plants.

  20. INSERTION OF A HETEROLOGOUS GENE CONSTRUCT INTO A NON-FUNCTIONAL ORF OF THE STREPTOCOCCUS THERMOPHILUS CHROMOSOME

    Science.gov (United States)

    A vector was constructed to allow for the integration of recombinant genes within the chromosome of Streptococcus thermophilus. The vector, pINTRS, contained a temperature sensitive origin of replication and an erythromycin resistance gene for initial selection in S. thermophilus. The region of th...

  1. Carbohydrate Utilization in Streptococcus thermophilus : Characterization of the Genes for Aldose 1-Epimerase (Mutarotase) and UDPglucose 4-Epimerase

    NARCIS (Netherlands)

    Poolman, Bert; Royer, Theresa J.; Mainzer, Stanley E.; Schmidt, Brian F.

    1990-01-01

    The complete nucleotide sequences of the genes encoding aldose 1-epimerase (mutarotase) (galM) and UDPglucose 4-epimerase (galE) and flanking regions of Streptococcus thermophilus have been determined. Both genes are located immediately upstream of the S. thermophilus lac operon. To facilitate the i

  2. Multi-site-specific 16S rRNA methyltransferase RsmF from Thermus thermophilus

    DEFF Research Database (Denmark)

    Demirci, Hasan; Larsen, Line H G; Hansen, Trine;

    2010-01-01

    Cells devote a significant effort toward the production of multiple modified nucleotides in rRNAs, which fine tune the ribosome function. Here, we report that two methyltransferases, RsmB and RsmF, are responsible for all four 5-methylcytidine (m(5)C) modifications in 16S rRNA of Thermus...... thermophilus. Like Escherichia coli RsmB, T. thermophilus RsmB produces m(5)C967. In contrast to E. coli RsmF, which introduces a single m(5)C1407 modification, T. thermophilus RsmF modifies three positions, generating m(5)C1400 and m(5)C1404 in addition to m(5)C1407. These three residues are clustered near...

  3. Therapeutic effect of Streptococcus thermophilus CRL 1190-fermented milk on chronic gastritis

    Institute of Scientific and Technical Information of China (English)

    Cecilia; Rodríguez; Marta; Medici; Fernanda; Mozzi; Graciela; Font; de; Valdez

    2010-01-01

    AIM: To investigate the potential therapeutic effect of exopolysaccharide (EPS)-producing Streptococcus thermophilus (S. thermophilus) CRL 1190 fermented milk on chronic gastritis in Balb/c mice. METHODS: Balb/c mice were fed with the fermented milk for 7 d after inducing gastritis with acetyl-salicylic acid (ASA, 400 mg/kg body weight per day for 10 d). Omeprazole was included in this study as a positive therapeutic control. The gastric in? ammatory activity was evaluated from gastric histology and in? amm...

  4. Streptococcus thermophilus, an emerging and promising tool for heterologous expression: Advantages and future trends.

    Science.gov (United States)

    Lecomte, Xavier; Gagnaire, Valérie; Lortal, Sylvie; Dary, Annie; Genay, Magali

    2016-02-01

    Streptococcus thermophilus is the second most used bacterium in dairy industry. It is daily consumed by millions of people through the worldwide consumption of yogurts, cheeses and fermented milks. S. thermophilus presents many features that make it a good candidate for the production of heterologous proteins. First, its ability to be naturally transformable allows obtaining swiftly and easily recombinant strains using various genetic tools available. Second, its Generally Recognised As Safe status and its ability to produce beneficial molecules or to liberate bioactive peptides from milk proteins open up the way for the development of new functional foods to maintain health and well-being of consumers. Finally, its ability to survive the intestinal passage and to be metabolically active in gastrointestinal tract allows considering S. thermophilus as a potential tool for delivering various biological molecules to the gastrointestinal tract. The aim of this review is therefore to take stock of various genetic tools which can be employed in S. thermophilus to produce heterologous proteins and to highlight the advantages and future trends of use of this bacterium as a heterologous expression host. PMID:26611164

  5. Prosthetic joint infection due to Lysobacter thermophilus diagnosed by 16S rRNA gene sequencing.

    Science.gov (United States)

    Dhawan, B; Sebastian, S; Malhotra, R; Kapil, A; Gautam, D

    2016-01-01

    We report the first case of prosthetic joint infection caused by Lysobacter thermophilus which was identified by 16S rRNA gene sequencing. Removal of prosthesis followed by antibiotic treatment resulted in good clinical outcome. This case illustrates the use of molecular diagnostics to detect uncommon organisms in suspected prosthetic infections.

  6. Inhibition of microbial adhesion to silicone rubber treated with biosurfactant from Streptococcus thermophilus A

    NARCIS (Netherlands)

    Rodrigues, L; van der Mei, H; Banat, IM; Teixeira, J; Oliveira, R

    2006-01-01

    Microbial adhesion of four bacterial and two yeast strains isolated from explanted voice prostheses to silicone rubber before and after conditioning with a biosurfactant obtained from the probiotic bacterium Streptococcus thermophilus A was investigated in a parallel plate flow chamber. The silicone

  7. Streptococcus thermophilus, an emerging and promising tool for heterologous expression: Advantages and future trends.

    Science.gov (United States)

    Lecomte, Xavier; Gagnaire, Valérie; Lortal, Sylvie; Dary, Annie; Genay, Magali

    2016-02-01

    Streptococcus thermophilus is the second most used bacterium in dairy industry. It is daily consumed by millions of people through the worldwide consumption of yogurts, cheeses and fermented milks. S. thermophilus presents many features that make it a good candidate for the production of heterologous proteins. First, its ability to be naturally transformable allows obtaining swiftly and easily recombinant strains using various genetic tools available. Second, its Generally Recognised As Safe status and its ability to produce beneficial molecules or to liberate bioactive peptides from milk proteins open up the way for the development of new functional foods to maintain health and well-being of consumers. Finally, its ability to survive the intestinal passage and to be metabolically active in gastrointestinal tract allows considering S. thermophilus as a potential tool for delivering various biological molecules to the gastrointestinal tract. The aim of this review is therefore to take stock of various genetic tools which can be employed in S. thermophilus to produce heterologous proteins and to highlight the advantages and future trends of use of this bacterium as a heterologous expression host.

  8. Streptococcus thermophilus and its biosurfactants inhibit adhesion by Candida spp. on silicone rubber

    NARCIS (Netherlands)

    Busscher, HJ; vanHoogmoed, CG; GeertsemaDoornbusch, GI; vanderKuijlBooij, M; vanderMei, HC

    1997-01-01

    The adhesion of yeasts, two Candida albicans and two Candida tropicalis strains isolated from naturally colonized voice prostheses, to silicone rubber with and without a salivary conditioning film in the absence and presence of adhering Streptococcus thermophilus B, a biosurfactant-releasing dairy i

  9. Molecular cloning, sequencing and expression in Escherichia coli cells Thermus thermophilus leucyl-tRNA synthetase

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    Kovalenko O. P.

    2011-12-01

    Full Text Available Aim. Cloning and sequencing of the T. thermophilus leucyl-tRNA synthetase (LeuRSTT followed by the creation of genetically engineered construct for protein expression in E.coli cells and its purification. Methods. Searching for the LeuRSTT gene was performed by Southern blot hybridization with chromosomal DNA, where digoxigenin-labeled PCR fragments of DNA were used as probes. Results. The gene of T. thermophilus HB27 leucyl-tRNA synthetase was cloned and sequenced. The open reading frame encodes a polypeptide chain of 878 amino acid residues in length (molecular mass 101 kDa. Comparison of the amino acid sequence of T. thermophilus LeuRS with that of the enzymes from other organisms showed that LeuRSTT was a part of the group of similar enzymes of prokaryotes, formed by the proteins of protobacteriae, rickettsia and mitochondria of eukaryotes. The resulting phylogenetic tree of LeuRSs reveals dichotomous branching into two lines: prokaryotic/eukaryotic mitochondrial and arhaeal/eukaryotic cytosolic proteins. Differences between prokaryotic and arhaeal branches of the LeuRSs phylogenetic tree are primarily due to the structure of two domains of the enzyme – the editing and the C-terminal. T. thermophilus LeuRS was expressed in E. coli cells by cloning the corresponding gene into pET29b vector. Conclusions. The cloned T. thermophilus leuS gene and expressed recombinant protein will be used for structural and functional studies on LeuRSTT, including X-ray analysis of the enzyme and its mutant forms in complex with different substrates

  10. PEDIOCIN PRODUCTION IN MILK BY PEDIOCOCCUS ACIDILACTICI IN CO-CULTURE WITH STREPTOCOCCUS THERMOPHILUS AND LACTOBACILLUS DELBRUECKII SUBSP. BULGARICUS

    Science.gov (United States)

    The production of pediocin in milk by Pediococcus acidilactici was evaluated in co-culture with the dairy fermentation cultures Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis subsp. lactis. The cultures were tested singly or in different combinations...

  11. Enzymatic fractionation of the antimicrobial peptides casocidin and isracidin by Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus

    Science.gov (United States)

    The cumulative effect of peptidase and protease activities associated with cells of Streptococcus thermophilus (ST) and Lactobacillus delbrueckii subsp. bulgaricus (LB) was evaluated on the milk-protein based antimicrobial peptides casocidin and isracidin. Reaction mixtures of casocidin or isracidin...

  12. Mixed-culture transcriptome analysis reveals the molecular basis of mixed-culture growth in Streptococcus thermophilus and Lactobacillus bulgaricus

    NARCIS (Netherlands)

    Sieuwerts, S.; Molenaar, D.; Hijum, van S.A.F.T.; Beerthuyzen, M.; Stevens, M.J.A.; Janssen, P.W.; Ingham, C.J.; Bok, de F.A.M.; Vos, de W.M.; Hylckama Vlieg, van J.E.T.

    2010-01-01

    Many food fermentations are performed using mixed cultures of lactic acid bacteria. Interactions between strains are of key importance for the performance of these fermentations. Yogurt fermentation by Streptococcus thermophilus and Lactobacillus bulgaricus (basonym, Lactobacillus delbrueckii subsp.

  13. Sequencing and transcriptional analysis of the streptococcus thermophilus histamine biosynthesis gene cluster: Factors that affect differential hdca expression

    OpenAIRE

    Calles-Enríquez, Marina; Hjort Eriksen, Benjamin; Skov Andersen, Pia; Rattray, F.; Johansen, Annette H.; Fernández García, María; Ladero Losada, Víctor Manuel; Álvarez González, Miguel Ángel

    2010-01-01

    Histamine, a toxic compound that is formed by the decarboxylation of histidine through the action of microbial decarboxylases, can accumulate in fermented food products. From a total of 69 Streptococcus thermophilus strains screened, two strains, CHCC1524 and CHCC6483, showed the capacity to produce histamine. The hdc clusters of S. thermophilus CHCC1524 and CHCC6483 were sequenced, and the factors that affect histamine biosynthesis and histidine-decarboxylating gene (hdcA) expression were st...

  14. Carbohydrate metabolism is essential for the colonization of Streptococcus thermophilus in the digestive tract of gnotobiotic rats.

    Directory of Open Access Journals (Sweden)

    Muriel Thomas

    Full Text Available Streptococcus thermophilus is the archetype of lactose-adapted bacterium and so far, its sugar metabolism has been mainly investigated in vitro. The objective of this work was to study the impact of lactose and lactose permease on S. thermophilus physiology in the gastrointestinal tract (GIT of gnotobiotic rats. We used rats mono-associated with LMD-9 strain and receiving 4.5% lactose. This model allowed the analysis of colonization curves of LMD-9, its metabolic profile, its production of lactate and its interaction with the colon epithelium. Lactose induced a rapid and high level of S. thermophilus in the GIT, where its activity led to 49 mM of intra-luminal L-lactate that was related to the induction of mono-carboxylic transporter mRNAs (SLC16A1 and SLC5A8 and p27(Kip1 cell cycle arrest protein in epithelial cells. In the presence of a continuous lactose supply, S. thermophilus recruited proteins involved in glycolysis and induced the metabolism of alternative sugars as sucrose, galactose, and glycogen. Moreover, inactivation of the lactose transporter, LacS, delayed S. thermophilus colonization. Our results show i/that lactose constitutes a limiting factor for colonization of S. thermophilus, ii/that activation of enzymes involved in carbohydrate metabolism constitutes the metabolic signature of S. thermophilus in the GIT, iii/that the production of lactate settles the dialogue with colon epithelium. We propose a metabolic model of management of carbohydrate resources by S. thermophilus in the GIT. Our results are in accord with the rationale that nutritional allegation via consumption of yogurt alleviates the symptoms of lactose intolerance.

  15. The ltp gene of temperate Streptococcus thermophilus phage TP-J34 confers superinfection exclusion to Streptococcus thermophilus and Lactococcus lactis

    International Nuclear Information System (INIS)

    The ltp gene, located within the lysogeny module of temperate Streptococcus thermophilus phage TP-J34, has been shown to be expressed in lysogenic strain S. thermophilus J34. It codes for a lipoprotein, as demonstrated by inhibition of cleavage of the signal sequence by globomycin. Exposure of Ltp on the surface of Lactococcus lactis protoplasts bearing a plasmid-encoded copy of ltp has been demonstrated by immunogold labeling and electron microscopy. Expression of ltp in prophage- and plasmid-cured S. thermophilus J34-6f interfered with TP-J34 infection. While plating efficiency was reduced by a factor of about 40 and lysis of strain J34-6f in liquid medium was delayed considerably, phage adsorption was not affected at all. Intracellular accumulation of phage DNA was shown to be inhibited by Ltp. This indicates interference of Ltp with infection at the stage of triggering DNA release and injection into the cell, indicating a role of Ltp in superinfection exclusion. Expression of ltp in L. lactis Bu2-60 showed that the same superinfection exclusion mechanism was strongly effective against phage P008, a member of the lactococcal 936 phage species: no plaque-formation was detectable with even 109 phage per ml applied, and lysis in liquid medium did not occur. In Lactococcus also, Ltp apparently inhibited phage DNA release and/or injection. Ltp appears to be a member of a family of small, secreted proteins with a 42 amino acids repeat structure encoded by genes of Gram-positive bacteria. Some of these homologous genes are part of the genomes of prophages

  16. Standardization of lyophilization medium for Streptococcus thermophilus subjected to viability escalation on freeze drying

    Directory of Open Access Journals (Sweden)

    Rohit Sharma

    2014-09-01

    Full Text Available The objective of the present study is to develop a lyophilization medium for Streptococcus thermophilus (NCIM 2904 as the industrial exploitation of this bacterium totally depends upon preservation and lyophilization protocols. Protective effect of 18 compounds were observed individually and in combinations with different sugars, sugar alcohols, polymers, protein concentrates and buffers. Among all the protectants tested, ammonium citrate (1% w/w, K2HPO4 (1% w/w and KH2PO4 (1% w/w provided lowest protection to these bacterial cells while 10% (w/w sodium caseinate, whey protein concentrate, sweet whey powder, and skim milk showed significant results in viability escalation. Survival in carbon sources like lactose, sucrose and maltodextrine was also favored maximally. Combination of sodium caseinate 10%, skim milk 5%, sucrose 5%, lactose 5% and mono sodium glutamate 1% in distilled water in ratio of 1:5 with S. thermophilus showed survival percentage of 96%.

  17. Physicochemical characteristics and bioactivities of the exopolysaccharide and its sulphated polymer from Streptococcus thermophilus GST-6.

    Science.gov (United States)

    Zhang, Jian; Cao, Yongqiang; Wang, Ji; Guo, Xialei; Zheng, Yi; Zhao, Wen; Mei, Xueyang; Guo, Ting; Yang, Zhennai

    2016-08-01

    Exopolysaccharide (EPS) produced by Streptococcus thermophilus GST-6 in skim milk was extracted and purified. The EPS was composed of glucose and galactose in a molar ratio of 1.80:1.03 with identical primary structure to the EPS from S. thermophilus ST1 reported previously. The purified EPS was sulphated at a sulphonation degree of 0.26±0.03, and presence of sulphate groups in the sulphated EPS (SEPS) was confirmed. Microstructural studies demonstrated a porous web with coarse surface for the EPS while the SEPS appeared as stacked flakes with relatively uniform shapes. Sulphonation of the EPS slightly decreased its degrading temperature from 234.6°C to 232.5°C. The DPPH, superoxide and hydroxyl radicals scavenging activities of the EPS were significantly (PSEPS also showed stronger inhibitory activity than the EPS against Eschericia coli, Salmonella typhimurium and Staphylococcus aureus. PMID:27112886

  18. Streptococcus thermophilus Biofilm Formation: A Remnant Trait of Ancestral Commensal Life?

    Directory of Open Access Journals (Sweden)

    Benoit Couvigny

    Full Text Available Microorganisms have a long history of use in food production and preservation. Their adaptation to food environments has profoundly modified their features, mainly through genomic flux. Streptococcus thermophilus, one of the most frequent starter culture organisms consumed daily by humans emerged recently from a commensal ancestor. As such, it is a useful model for genomic studies of bacterial domestication processes. Many streptococcal species form biofilms, a key feature of the major lifestyle of these bacteria in nature. However, few descriptions of S. thermophilus biofilms have been reported. An analysis of the ability of a representative collection of natural isolates to form biofilms revealed that S. thermophilus was a poor biofilm producer and that this characteristic was associated with an inability to attach firmly to surfaces. The identification of three biofilm-associated genes in the strain producing the most biofilms shed light on the reasons for the rarity of this trait in this species. These genes encode proteins involved in crucial stages of biofilm formation and are heterogeneously distributed between strains. One of the biofilm genes appears to have been acquired by horizontal transfer. The other two are located in loci presenting features of reductive evolution, and are absent from most of the strains analyzed. Their orthologs in commensal bacteria are involved in adhesion to host cells, suggesting that they are remnants of ancestral functions. The biofilm phenotype appears to be a commensal trait that has been lost during the genetic domestication of S. thermophilus, consistent with its adaptation to the milk environment and the selection of starter strains for dairy fermentations.

  19. The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli

    OpenAIRE

    Sapranauskas, Rimantas; Gasiunas, Giedrius; Fremaux, Christophe; Barrangou, Rodolphe; Horvath, Philippe; Siksnys, Virginijus

    2011-01-01

    The CRISPR/Cas adaptive immune system provides resistance against phages and plasmids in Archaea and Bacteria. CRISPR loci integrate short DNA sequences from invading genetic elements that provide small RNA-mediated interference in subsequent exposure to matching nucleic acids. In Streptococcus thermophilus, it was previously shown that the CRISPR1/Cas system can provide adaptive immunity against phages and plasmids by integrating novel spacers following exposure to these foreign genetic elem...

  20. Bioremediation potential of a newly isolate solvent tolerant strain Bacillus thermophilus PS11

    OpenAIRE

    PAYEL SARKAR; SHILPI GHOSH

    2012-01-01

    The increased generation of solvent waste has been stated as one of the most critical environmental problems. Though microbial bioremediation has been widely used for waste treatment but their application in solvent waste treatment is limited since the solvents have toxic effects on the microbial cells. A solvent tolerant strain of Bacillus thermophilus PS11 was isolated from soil by cyclohexane enrichment. Transmission electron micrograph of PS11 showed convoluted cell membrane and accumulat...

  1. Genetically Engineered Immunomodulatory Streptococcus thermophilus Strains Producing Antioxidant Enzymes Exhibit Enhanced Anti-Inflammatory Activities

    OpenAIRE

    del Carmen, Silvina; de Moreno de LeBlanc, Alejandra; Martin, Rebeca; Chain, Florian; Langella, Philippe; Bermúdez-Humarán, Luis G; LeBlanc, Jean Guy

    2014-01-01

    The aims of this study were to develop strains of lactic acid bacteria (LAB) having both immunomodulatory and antioxidant properties and to evaluate their anti-inflammatory effects both in vitro, in different cellular models, and in vivo, in a mouse model of colitis. Different Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains were cocultured with primary cultures of mononuclear cells. Analysis of the pro- and anti-inflammatory cytokines secreted by these cells...

  2. Genetically engineered immunomodulatory Streptococcus thermophilus strains producing antioxidant enzymes exhibit enhanced anti-inflammatory activities.

    Science.gov (United States)

    Del Carmen, Silvina; de Moreno de LeBlanc, Alejandra; Martin, Rebeca; Chain, Florian; Langella, Philippe; Bermúdez-Humarán, Luis G; LeBlanc, Jean Guy

    2014-02-01

    The aims of this study were to develop strains of lactic acid bacteria (LAB) having both immunomodulatory and antioxidant properties and to evaluate their anti-inflammatory effects both in vitro, in different cellular models, and in vivo, in a mouse model of colitis. Different Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains were cocultured with primary cultures of mononuclear cells. Analysis of the pro- and anti-inflammatory cytokines secreted by these cells after coincubation with candidate bacteria revealed that L. delbrueckii subsp. bulgaricus CRL 864 and S. thermophilus CRL 807 display the highest anti-inflammatory profiles in vitro. Moreover, these results were confirmed in vivo by the determination of the cytokine profiles in large intestine samples of mice fed with these strains. S. thermophilus CRL 807 was then transformed with two different plasmids harboring the genes encoding catalase (CAT) or superoxide dismutase (SOD) antioxidant enzymes, and the anti-inflammatory effects of recombinant streptococci were evaluated in a mouse model of colitis induced by trinitrobenzenesulfonic acid (TNBS). Our results showed a decrease in weight loss, lower liver microbial translocation, lower macroscopic and microscopic damage scores, and modulation of the cytokine production in the large intestines of mice treated with either CAT- or SOD-producing streptococci compared to those in mice treated with the wild-type strain or control mice without any treatment. Furthermore, the greatest anti-inflammatory activity was observed in mice receiving a mixture of both CAT- and SOD-producing streptococci. The addition of L. delbrueckii subsp. bulgaricus CRL 864 to this mixture did not improve their beneficial effects. These findings show that genetically engineering a candidate bacterium (e.g., S. thermophilus CRL 807) with intrinsic immunomodulatory properties by introducing a gene expressing an antioxidant enzyme enhances its anti

  3. Description of Tylenchorhynchus thermophilus n. sp. (Nematoda: Tylenchina) from Saltgrass in Death Valley, California

    OpenAIRE

    Golden, A. Morgan; Baldwin, James G.; Mundo-Ocampo, M.

    1995-01-01

    A stunt nematode, Tylenchorhynchus thermophilus n. sp., is described and illustrated from soil collected around roots of saltgrass (Distichlis spicata) in Death Valley, California. It is distinguished from the similar species, T. ewingi, T. mexicanus, and T. mashoodi, in having a longer female body, longer tail with more annules, and larger phasmids. Physical and chemical analysis of soil from saltgrass roots showed it to consist of 71% sand and possess high salinity (salt content of 0.51%) a...

  4. In vitro reconstitution of Cascade-mediated CRISPR immunity in Streptococcus thermophilus

    OpenAIRE

    Sinkunas, Tomas; Gasiunas, Giedrius; Waghmare, Sakharam P; Dickman, Mark J; Barrangou, Rodolphe; Horvath, Philippe; Siksnys, Virginijus

    2013-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR)-encoded immunity in Type I systems relies on the Cascade (CRISPR-associated complex for antiviral defence) ribonucleoprotein complex, which triggers foreign DNA degradation by an accessory Cas3 protein. To establish the mechanism for adaptive immunity provided by the Streptococcus thermophilus CRISPR4-Cas (CRISPR-associated) system (St-CRISPR4-Cas), we isolated an effector complex (St-Cascade) containing 61-nucleotide CRISPR R...

  5. Lateral Transfer of the Denitrification Pathway Genes among Thermus thermophilus Strains▿

    Science.gov (United States)

    Alvarez, Laura; Bricio, Carlos; José Gómez, Manuel; Berenguer, José

    2011-01-01

    Nitrate respiration is a common and strain-specific property in Thermus thermophilus encoded by the nitrate respiration conjugative element (NCE) that can be laterally transferred by conjugation. In contrast, nitrite respiration and further denitrification steps are restricted to a few isolates of this species. These later steps of the denitrification pathway are under the regulatory control of an NCE-encoded transcription factor, but nothing is known about their coding sequences or its putative genetic linkage to the NCE. In this study we examine the genetic linkage between nitrate and nitrite respiration through lateral gene transfer (LGT) assays and describe a cluster of genes encoding the nitrite-nitric oxide respiration in T. thermophilus PRQ25. We show that the whole denitrification pathway can be transferred from the denitrificant strain PRQ25 to an aerobic strain, HB27, and that the genes coding for nitrite and nitric oxide respiration are encoded near the NCE. Sequence data from the draft genome of PRQ25 confirmed these results and allowed us to describe the most compact nor-nir cluster known thus far and to demonstrate the expression and activities of the encoded enzymes in the HB27 denitrificant derivatives obtained by LGT. We conclude that this NCE nor-nir supercluster constitutes a whole denitrification island that can be spread by lateral transfer among Thermus thermophilus strains. PMID:21169443

  6. Small-scale analysis of exopolysaccharides from Streptococcus thermophilus grown in a semi-defined medium

    Directory of Open Access Journals (Sweden)

    Rådström Peter

    2001-09-01

    Full Text Available Abstract Background Exopolysaccharides (EPSs produced by lactic acid bacteria are important for the texture of fermented foods and have received a great deal of interest recently. However, the low production levels of EPSs in combination with the complex media used for growth of the bacteria have caused problems in the accurate analysis of the EPS. The purpose of this study was to find a growth medium for physiological studies of the lactic acid bacterium Streptococcus thermophilus, and to develop a simple method for qualitative and quantitative analysis of EPSs produced in this medium. Results A semi-defined polysaccharide medium was developed and evaluated on six strains of Streptococcus thermophilus. The EPSs were analysed using a novel protocol incorporating ultracentrifugation for the removal of interfering sugars, hydrolysis and analysis of the monomer composition by High Performance Anion-Exchange Chromatography with pulsed amperometric detection. The medium and analysis method allowed accurate quantification and monomer analysis of 0.5 ml samples of EPSs from tube cultures. Conclusions The presented medium should be useful for physiological studies of S. thermophilus, and, in combination with the method of analysis of EPS, will allow downscaling of physiological studies and screening for EPSs.

  7. Crystallization and preliminary crystallographic analysis of molybdenum-cofactor biosynthesis protein C from Thermus thermophilus

    International Nuclear Information System (INIS)

    The molybdenum-cofactor biosynthesis protein C from T. thermophilus has been crystallized in two different space groups, P21 and R32; the crystals diffracted to 1.9 and 1.75 Å resolution, respectively. The Gram-negative aerobic eubacterium Thermus thermophilus is an extremely important thermophilic microorganism that was originally isolated from a thermal vent environment in Japan. The molybdenum cofactor in this organism is considered to be an essential component required by enzymes that catalyze diverse key reactions in the global metabolism of carbon, nitrogen and sulfur. The molybdenum-cofactor biosynthesis protein C derived from T. thermophilus was crystallized in two different space groups. Crystals obtained using the first crystallization condition belong to the monoclinic space group P21, with unit-cell parameters a = 64.81, b = 109.84, c = 115.19 Å, β = 104.9°; the crystal diffracted to a resolution of 1.9 Å. The other crystal form belonged to space group R32, with unit-cell parameters a = b = 106.57, c = 59.25 Å, and diffracted to 1.75 Å resolution. Preliminary calculations reveal that the asymmetric unit contains 12 monomers and one monomer for the crystals belonging to space group P21 and R32, respectively

  8. Preparation of low galactose yogurt using cultures of Gal(+) Streptococcus thermophilus in combination with Lactobacillus delbrueckii ssp. bulgaricus.

    Science.gov (United States)

    Anbukkarasi, Kaliyaperumal; UmaMaheswari, Thiyagamoorthy; Hemalatha, Thiagarajan; Nanda, Dhiraj Kumar; Singh, Prashant; Singh, Rameshwar

    2014-09-01

    Streptococcus thermophilus is an important lactic starter used in the production of yogurt. Most strains of S. thermophilus are galactose negative (Gal(-)) and are able to metabolize only glucose portion of lactose and expel galactose into the medium. This metabolic defect leads to the accumulation of free galactose in yogurt, resulting in galactosemia among consumers. Hence there is an absolute need to develop low galactose yogurt. Therefore, in this study, three galactose positive (Gal(+)) S. thermophilus strains from National Collection of Dairy Cultures (NCDC) viz. NCDC 659 (AJM), NCDC 660 (JM1), NCDC 661 (KM3) and a reference galactose negative (Gal(-)) S. thermophilus NCDC 218 were used for preparation of low galactose yogurt. In milk fermented using S. thermophilus isolates alone, NCDC 659 released less galactose (0.27 %) followed by NCDC 661 (0.3 %) and NCDC 660 (0.45 %) after 10 h at 42 °C. Milk was fermented in combination with Gal(-) L. delbrueckii subsp. bulgaricus NCDC 04, in which NCDC 659 released least galactose upto 0.49 % followed by NCDC 661 (0.51 %) and NCDC 660 (0.60 %) than reference Gal(-) NCDC 218(0.79 %). Low galactose yogurt was prepared following standard procedure using Gal(+) S. thermophilus isolates and Gal(-) L. delbrueckii subsp. bulgaricus NCDC 04 in 1:1 ratio. Among which low galactose yogurt by NCDC 659 combination contained less galactose 0.37 % followed by NCDC 661 (0.51 %), NCDC 660 (0.65 %) and reference Gal(-) NCDC 218 (0.98 %) after 4 h of fermentation. This study clearly reveals that Gal(+) S. thermophilus isolates can be paired with Gal(-) L. delbrueckii subsp. bulgaricus for developing low galactose yogurt. PMID:25190881

  9. Preparation of low galactose yogurt using cultures of Gal(+) Streptococcus thermophilus in combination with Lactobacillus delbrueckii ssp. bulgaricus.

    Science.gov (United States)

    Anbukkarasi, Kaliyaperumal; UmaMaheswari, Thiyagamoorthy; Hemalatha, Thiagarajan; Nanda, Dhiraj Kumar; Singh, Prashant; Singh, Rameshwar

    2014-09-01

    Streptococcus thermophilus is an important lactic starter used in the production of yogurt. Most strains of S. thermophilus are galactose negative (Gal(-)) and are able to metabolize only glucose portion of lactose and expel galactose into the medium. This metabolic defect leads to the accumulation of free galactose in yogurt, resulting in galactosemia among consumers. Hence there is an absolute need to develop low galactose yogurt. Therefore, in this study, three galactose positive (Gal(+)) S. thermophilus strains from National Collection of Dairy Cultures (NCDC) viz. NCDC 659 (AJM), NCDC 660 (JM1), NCDC 661 (KM3) and a reference galactose negative (Gal(-)) S. thermophilus NCDC 218 were used for preparation of low galactose yogurt. In milk fermented using S. thermophilus isolates alone, NCDC 659 released less galactose (0.27 %) followed by NCDC 661 (0.3 %) and NCDC 660 (0.45 %) after 10 h at 42 °C. Milk was fermented in combination with Gal(-) L. delbrueckii subsp. bulgaricus NCDC 04, in which NCDC 659 released least galactose upto 0.49 % followed by NCDC 661 (0.51 %) and NCDC 660 (0.60 %) than reference Gal(-) NCDC 218(0.79 %). Low galactose yogurt was prepared following standard procedure using Gal(+) S. thermophilus isolates and Gal(-) L. delbrueckii subsp. bulgaricus NCDC 04 in 1:1 ratio. Among which low galactose yogurt by NCDC 659 combination contained less galactose 0.37 % followed by NCDC 661 (0.51 %), NCDC 660 (0.65 %) and reference Gal(-) NCDC 218 (0.98 %) after 4 h of fermentation. This study clearly reveals that Gal(+) S. thermophilus isolates can be paired with Gal(-) L. delbrueckii subsp. bulgaricus for developing low galactose yogurt.

  10. Cloning, expression and purification of D-Tyr-tRNATyr-deacylase from Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Rybak M. Yu.

    2015-06-01

    Full Text Available D-Tyr-tRNATyr-deacylase (DTD is a conservative enzyme, found in all domains of life, which ensures an additional checkpoint in the recycling of misaminoacylated D-Tyr-tRNATyr. DTD is capable of accelerating the hydrolysis of the ester linkage of D-Tyr-tRNATyr producing a free tRNA and D-tyrosine, thereby preventing an incorrect incorporation of D-amino acids into proteins. Deacylase distinguishes between D- and L-aminoacyl moieties and does not hydrolyze L-aminoacylated tRNA. The structural bases of this specificity and the mechanism of D-aminoacyl-tRNA hydrolysis are poorly understood. Aim. To clone D-Tyr-tRNATyr-deacylase from T. thermophilus (DTDTT, optimize the conditions for its expression in E.coli and develop an efficient purification procedure yielding the high quality enzyme suitable for the structural and functional studies. Methods. For amplification of DTD gene from T. thermophilus genomic DNA and its cloning into the pProEXHTb expression vector modern techniques were applied. Purification of the recombinant DTD protein was done with three types of column chromatography. His-tag was cleaved out from DTD by TEV protease. The cleavage was confirmed by Western blot analysis with anti-His-tag antibodies. Molecular weight of purified DTDTT was determined by the gel-filtration. Results. The expression construct pProEXHTb, containing DTD sequence from T. thermophilus, was obtained and successfully expressed in the BL21(DE3pLysS E.coli strain. The protein of interest was purified to homogeneity by the combination of affinity (Ni-NTA, anion-exchange (Q-Sepharose and size-exclusion (Superdex S 200 chromatographies. 2 mg of more than 90% pure recombinant DTD can be obtained from 1 L of bacterial culture. Molecular weight of purified DTD from T. thermophilus was determined to be 32 kDa, suggesting its dimeric structure. Conclusions. The pProEXHTb expression vector can be used for expression of DTD from T. thermophilus. The preparative amounts of

  11. Cell-wall proteinases PrtS and PrtB have a different role in Streptococcus thermophilus/Lactobacillus bulgaricus mixed cultures in milk.

    Science.gov (United States)

    Courtin, P; Monnet, V; Rul, F

    2002-11-01

    The manufacture of yoghurt relies on the simultaneous utilization of two starters: Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (Lb. bulgaricus). A protocooperation usually takes place between the two species, which often results in enhanced milk acidification and aroma formation compared to pure cultures. Cell-wall proteinases of Lactococcus lactis and lactobacilli have been shown to be essential to growth in milk in pure cultures. In this study, the role of proteinases PrtS from S. thermophilus and PrtB from Lb. bulgaricus in bacterial growth in milk was evaluated; a negative mutant for the prtS gene of S. thermophilus CNRZ 385 was constructed for this purpose. Pure cultures of S. thermophilus CNRZ 385 and its PrtS-negative mutant were made in milk as well as mixed cultures of S. thermophilus and Lb. bulgaricus: S. thermophilus CNRZ 385 or its PrtS-negative mutant was associated with several strains of Lb. bulgaricus, including a PrtB-negative strain. The pH and growth of bacterial populations of the resulting mixed cultures were followed, and the Lactobacillus strain was found to influence both the extent of the benefit of Lb. bulgaricus/S. thermophilus association on milk acidification and the magnitude of S. thermophilus population dominance at the end of fermentation. In all mixed cultures, the sequential growth of S. thermophilus then of Lb. bulgarius and finally of both bacteria was observed. Although proteinase PrtS was essential to S. thermophilus growth in milk in pure culture, it had no effect on bacterial growth and thus on the final pH of mixed cultures in the presence of PrtB. In contrast, proteinase PrtB was necessary for the growth of S. thermophilus, and its absence resulted in a higher final pH. From these results, a model of growth of both bacteria in mixed cultures in milk is proposed.

  12. Influence of different proteolytic strains of Streptococcus thermophilus in co-culture with Lactobacillus delbrueckii subsp. bulgaricus on the metabolite profile of set-yoghurt

    NARCIS (Netherlands)

    Settachaimongkon, S.; Nout, M.J.R.; Antunes Fernandes, E.C.; Hettinga, K.A.; Vervoort, J.J.M.; Hooijdonk, van A.C.M.; Zwietering, M.H.; Smid, E.J.; Valenberg, van H.J.F.

    2014-01-01

    Proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus is one of the key factors that determine the fermentation process and final quality of yoghurt. In this study, the interaction between different proteolytic strains of S. thermophilus and L. delbruec

  13. Metabolic network reconstruction, growth characterization and 13C-metabolic flux analysis of the extremophile Thermus thermophilus HB8.

    Science.gov (United States)

    Swarup, Aditi; Lu, Jing; DeWoody, Kathleen C; Antoniewicz, Maciek R

    2014-07-01

    Thermus thermophilus is an extremely thermophilic bacterium with significant biotechnological potential. In this work, we have characterized aerobic growth characteristics of T. thermophilus HB8 at temperatures between 50 and 85°C, constructed a metabolic network model of its central carbon metabolism and validated the model using (13)C-metabolic flux analysis ((13)C-MFA). First, cells were grown in batch cultures in custom constructed mini-bioreactors at different temperatures to determine optimal growth conditions. The optimal temperature for T. thermophilus grown on defined medium with glucose was 81°C. The maximum growth rate was 0.25h(-1). Between 50 and 81°C the growth rate increased by 7-fold and the temperature dependence was described well by an Arrhenius model with an activation energy of 47kJ/mol. Next, we performed a (13)C-labeling experiment with [1,2-(13)C] glucose as the tracer and calculated intracellular metabolic fluxes using (13)C-MFA. The results provided support for the constructed network model and highlighted several interesting characteristics of T. thermophilus metabolism. We found that T. thermophilus largely uses glycolysis and TCA cycle to produce biosynthetic precursors, ATP and reducing equivalents needed for cells growth. Consistent with its proposed metabolic network model, we did not detect any oxidative pentose phosphate pathway flux or Entner-Doudoroff pathway activity. The biomass precursors erythrose-4-phosphate and ribose-5-phosphate were produced via the non-oxidative pentose phosphate pathway, and largely via transketolase, with little contribution from transaldolase. The high biomass yield on glucose that was measured experimentally was also confirmed independently by (13)C-MFA. The results presented here provide a solid foundation for future studies of T. thermophilus and its metabolic engineering applications.

  14. Description of Tylenchorhynchus thermophilus n. sp. (Nematoda: Tylenchina) from Saltgrass in Death Valley, California.

    Science.gov (United States)

    Golden, A M; Baldwin, J G; Mundo-Ocampo, M

    1995-09-01

    A stunt nematode, Tylenchorhynchus thermophilus n. sp., is described and illustrated from soil collected around roots of saltgrass (Distichlis spicata) in Death Valley, California. It is distinguished from the similar species, T. ewingi, T. mexicanus, and T. mashoodi, in having a longer female body, longer tail with more annules, and larger phasmids. Physical and chemical analysis of soil from saltgrass roots showed it to consist of 71% sand and possess high salinity (salt content of 0.51%) and a pH of 9.3.

  15. Purification, Characterization, Gene Cloning, Sequencing, and Overexpression of Aminopeptidase N from Streptococcus thermophilus A

    OpenAIRE

    Chavagnat, Frederic; Casey, Michael G.; Meyer, Jacques

    1999-01-01

    The general aminopeptidase PepN from Streptococcus thermophilus A was purified to protein homogeneity by hydroxyapatite, anion-exchange, and gel filtration chromatographies. The PepN enzyme was estimated to be a monomer of 95 kDa, with maximal activity on N-Lys–7-amino-4-methylcoumarin at pH 7 and 37°C. It was strongly inhibited by metal chelating agents, suggesting that it is a metallopeptidase. The activity was greatly restored by the bivalent cations Co2+, Zn2+, and Mn2+. Except for prolin...

  16. Structure of a UPF0150-family protein from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    The crystal structure of the hypothetical protein TTHA0281 from T. thermophilus HB8 has been determined at 1.9 Å resolution. The TTHA0281 protein forms a homotetramer in which each monomer adopts an α-β-β-β-α fold. TTHA0281 is a hypothetical protein from Thermus thermophilus HB8 that belongs to an uncharacterized protein family, UPF0150, in the Pfam database and to COG1598 in the National Center for Biotechnology Information Database of Clusters of Orthologous Groups. The X-ray crystal structure of the protein was determined by a multiple-wavelength anomalous dispersion technique and was refined at 1.9 Å resolution to a final R factor of 18.5%. The TTHA0281 monomer adopts an α-β-β-β-α fold and forms a homotetramer. Based on the properties and functions of structural homologues of the TTHA0281 monomer, the TTHA0281 protein is speculated to be involved in RNA metabolism, including RNA binding and cleavage

  17. Structure of the stand-alone RAM-domain protein from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    The crystal structure of the stand-alone RAM domain from T. thermophilus HB8 has been determined at 2.4 Å resolution. The structure revealed that five dimers are arranged to form a ring. The stand-alone RAM (regulation of amino-acid metabolism) domain protein SraA from Thermus thermophilus HB8 (TTHA0845) was crystallized in the presence of zinc ions. The X-ray crystal structure was determined using a multiple-wavelength anomalous dispersion technique and was refined at 2.4 Å resolution to a final R factor of 25.0%. The monomeric structure is a βαββαβ fold and it dimerizes mainly through interactions between the antiparallel β-sheets. Furthermore, five SraA dimers form a ring with external and internal diameters of 70 and 20 Å, respectively. This decameric structure is unique compared with the octameric and dodecameric structures found for other stand-alone RAM-domain proteins and the C-terminal RAM domains of Lrp/AsnC-family proteins

  18. Structure of the minimized α/β-hydrolase fold protein from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    The crystal structure of the minimized α/β-hydrolase fold protein encoded by the gene TTHA1544 from T. thermophilus HB8 has been determined at 2.0 Å resolution. The gene encoding TTHA1544 is a singleton found in the Thermus thermophilus HB8 genome and encodes a 131-amino-acid protein. The crystal structure of TTHA1544 has been determined at 2.0 Å resolution by the single-wavelength anomalous dispersion method in order to elucidate its function. There are two molecules in the asymmetric unit. Each molecule consists of four α-helices and six β-strands, with the β-strands composing a central β-sheet. A structural homology search revealed that the overall structure of TTHA1544 resembles the α/β-hydrolase fold, although TTHA1544 lacks the catalytic residues of a hydrolase. These results suggest that TTHA1544 represents the minimized α/β-hydrolase fold and that an additional component would be required for its activity

  19. Lactose uptake driven by galactose efflux in Streptococcus thermophilus: Evidence for a galactose-lactose antiporter

    International Nuclear Information System (INIS)

    Galactose-nonfermenting (Gal-) Streptococcus thermophilus TS2 releases galactose into the extracellular medium when grown in medium containing excess lactose. Starved and de-energized Gal- cells, however, could be loaded with galactose to levels approximately equal to the extracellular concentration (0 to 50 mM). When loaded cells were separated from the medium and resuspended in fresh broth containing 5 mM lactose, galactose efflux occurred. De-energized, galactose-loaded cells, resuspended in buffer or medium, accumulated [14C]lactose at a greater rate and to significantly higher intracellular concentrations than unloaded cells. Uptake of lactose by loaded cells was inhibited more than that by unloaded cells in the presence of extracellular galactose, indicating that a galactose gradient was involved in the exchange system. When de-energized, galactose-loaded cells were resuspended in carbohydrate-free medium at pH 6.7, a proton motive force (Δp) of 86 to 90 mV was formed, whereas de-energized, nonloaded cells maintained a Δp of about 56 mV. However, uptake of lactose by loaded cells occurred when the proton motive force was abolished by the addition of an uncoupler or in the presence of a proton-translocating ATPase inhibitor. These results support the hypothesis that galactose efflux in Gal- S. thermophilus is electrogenic and that the exchange reaction (lactose uptake and galactose efflux) probably occurs via an antiporter system

  20. Production of gamma-aminobutyric acid by Streptococcus salivarius subsp. thermophilus Y2 under submerged fermentation.

    Science.gov (United States)

    Yang, S-Y; Lü, F-X; Lu, Z-X; Bie, X-M; Jiao, Y; Sun, L-J; Yu, B

    2008-04-01

    Gamma-aminobutyric acid (GABA), a major inhibitory neurotransmitter in the central nervous system, has several well-known physiological functions and has been applied to the production of many drugs and functional foods. The technology of GABA production via submerged fermentation by Streptococcus salivarius subsp. thermophilus Y2 was investigated in this paper. It indicated that the GABA production was related to the biochemical characteristics of glutamate decarboxylase (GAD) of S. salivarius subsp. thermophilus Y2. After 24 h of fermentation at 37 degrees C, which is the suitable culture conditions for GAD-production, then the culture condition were adjusted to the optimal temperature (40 degrees C) and pH (4.5) for the GAD reaction activity in biotransformation of cells and pyridoxal 5'-phosphate (0.02 mmol/l) were added to the broth at the 48 h, the GABA production was increased up to 1.76-fold, reaching 7984.75 +/- 293.33 mg/l. The strain shows great potential use as a starter for GABA-containing yoghurt, cheese and other functional fermented food productions. PMID:17514494

  1. Bioremediation potential of a newly isolate solvent tolerant strain Bacillus thermophilus PS11

    Directory of Open Access Journals (Sweden)

    PAYEL SARKAR

    2012-01-01

    Full Text Available The increased generation of solvent waste has been stated as one of the most critical environmental problems. Though microbial bioremediation has been widely used for waste treatment but their application in solvent waste treatment is limited since the solvents have toxic effects on the microbial cells. A solvent tolerant strain of Bacillus thermophilus PS11 was isolated from soil by cyclohexane enrichment. Transmission electron micrograph of PS11 showed convoluted cell membrane and accumulation of solvents in the cytoplasm, indicating the adaptation of the bacterial strain to the solvent after 48h of incubation. The strain was also capable of growing in presence of wide range of other hydrophobic solvents with log P-values below 3.5. The isolate could uptake 50 ng/ml of uranium in its initial 12h of growth, exhibiting both solvent tolerance and metal resistance property. This combination of solvent tolerance and metal resistance will make the isolated Bacillus thermophilus PS11 a potential tool for metal bioremediation in solvent rich wastewaters.

  2. The role of the PHP domain associated with DNA polymerase X from Thermus thermophilus HB8 in base excision repair.

    Science.gov (United States)

    Nakane, Shuhei; Nakagawa, Noriko; Kuramitsu, Seiki; Masui, Ryoji

    2012-11-01

    Base excision repair (BER) is one of the most commonly used DNA repair pathways involved in genome stability. X-family DNA polymerases (PolXs) play critical roles in BER, especially in filling single-nucleotide gaps. In addition to a polymerase core domain, bacterial PolXs have a polymerase and histidinol phosphatase (PHP) domain with phosphoesterase activity which is also required for BER. However, the role of the PHP domain of PolX in bacterial BER remains unresolved. We found that the PHP domain of Thermus thermophilus HB8 PolX (ttPolX) functions as two types of phosphoesterase in BER, including a 3'-phosphatase and an apurinic/apyrimidinic (AP) endonuclease. Experiments using T. thermophilus HB8 cell lysates revealed that the majority of the 3'-phosphatase and AP endonuclease activities are attributable to the another phosphoesterase in T. thermophilus HB8, endonuclease IV (ttEndoIV). However, ttPolX possesses significant 3'-phosphatase activity in ΔttendoIV cell lysate, indicating possible complementation. Our experiments also reveal that there are only two enzymes that display the 3'-phosphatase activity in the T. thermophilus HB8 cell, ttPolX and ttEndoIV. Furthermore, phenotypic analysis of ΔttpolX, ΔttendoIV, and ΔttpolX/ΔttendoIV using hydrogen peroxide and sodium nitrite supports the hypothesis that ttPolX functions as a backup for ttEndoIV in BER.

  3. Crystal structure of TTC0263, a thermophilic TPR protein from Thermus thermophilus HB27.

    Science.gov (United States)

    Lim, Hyosun; Kim, Kyunggon; Han, Dohyun; Oh, Jongkil; Kim, Youngsoo

    2007-08-31

    The hypothetical protein TTC0263 of Thermus thermophilus HB27 is a thermophilic tetratricopeptide repeat (TPR)-containing protein. In the present study, the TPR region (residues 26-230) was resolved at 2.5 A with R-factors of R/Rfree = 23.6%/28.6%. TTC0263 consists of 11 helices that form five TPR units. Uniquely, it contains one atypical "extended" TPR (eTPR) unit. This comprises extended helical residues near the loop region of TTC0263, such that the helical length of eTPR is longer than that of the canonical TPR sequence. In addition, the hybrid TPR domain of TTC0263 possesses oligomer-forming characteristics. TPR domains are generally involved in forming multi-subunit complexes by interacting with each other or with other subunit proteins. The dynamic structure of TTC0263 described here goes some way to explaining how TPR domains mediate the formation of multi-subunit complexes.

  4. Expression, purification and characterization of recombinant protein tyrosine phosphatase from Thermus thermophilus HB27

    Institute of Scientific and Technical Information of China (English)

    Yejing Wang; Fanguo Meng; Yingmei Zhang

    2009-01-01

    The low-molecular-weight protein tyrosine phospha-tases (PTPase) exist ubiquitously in prokaryotes and eukaryotes and play important roles in the regulation of physiological activities. We report here the expression, purification and characterization of an active and soluble PTPase from Thermus thermophilus HB27 in Escherichia coli. This PTPase has an optimum pH range of 2.8-4.8 when using p-nitrophenyl phos-phate as the substrate. The thermal inactivation results indicate a high thermal stability of this enzyme, with the optimum temperature of 75℃ for activity. It can be activated by Mn2+, Mg2+, Ca2+, Ba2+, and Ni2+, but inhibited by Zn2+, Cu2+, Cl-, and SO2-4. These results suggest that this heat-resistant PTPase may play impor-tant roles in vivo in the adaptation of the microorgan-ism to extreme temperatures and specific nutritional conditions.

  5. Identification of lactobacillus casei-BDI and Streptococcus thermophilus by fluorescence spectra

    Institute of Scientific and Technical Information of China (English)

    Hao Wu; Tuo Zhu; Wei Chen; Hao Zhang; Jianlei Yang

    2009-01-01

    Fluorescence analysis applied in the study of lactic acid bacteria (LAB) provides a new method and theory to study probiotics and realize the detection and identification of the strains.It is also possible to achieve automation and computerization.In this letter,the differences between the fluorescence spectra of lactobacillus casei-BDI (Lc-BDI) and Streptococcus thermophilus (St) are shown,and the second-order derivative spectra are used to further study the diversity of these two strains.According to the results,with thc excitation wavelengths of 285 and 340 nm,there are significant differences between them.The experiment is repeated for 6 times,showing good repetitiveness.

  6. Assembling the Streptococcus thermophilus clustered regularly interspaced short palindromic repeats (CRISPR) array for multiplex DNA targeting.

    Science.gov (United States)

    Guo, Lijun; Xu, Kun; Liu, Zhiyuan; Zhang, Cunfang; Xin, Ying; Zhang, Zhiying

    2015-06-01

    In addition to the advantages of scalable, affordable, and easy to engineer, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) technology is superior for multiplex targeting, which is laborious and inconvenient when achieved by cloning multiple gRNA expressing cassettes. Here, we report a simple CRISPR array assembling method which will facilitate multiplex targeting usage. First, the Streptococcus thermophilus CRISPR3/Cas locus was cloned. Second, different CRISPR arrays were assembled with different crRNA spacers. Transformation assays using different Escherichia coli strains demonstrated efficient plasmid DNA targeting, and we achieved targeting efficiency up to 95% with an assembled CRISPR array with three crRNA spacers.

  7. An efficient gene replacement and deletion system for an extreme thermophile, Thermus thermophilus.

    Science.gov (United States)

    Tamakoshi, M; Yaoi, T; Oshima, T; Yamagishi, A

    1999-04-15

    A Thermus thermophilus host strain of which the leuB gene was totally deleted was constructed from a delta pyrE strain by a two step method. First, the leuB gene was replaced with the pyrE gene. Second, the inserted pyrE gene was deleted by using 5-fluoroorotic acid. A plasmid vector with the leuB marker was constructed and the plasmid complemented the leuB deficiency of the host. When the leuB gene from Escherichia coli and its derivative encoding a stabilized enzyme were expressed with the host-vector system, their growth temperature reflected the stability of the enzyme. These results suggest that the gene replacement deletion method using the pyrE gene is useful for the construction of a reliable plasmid vector system and it can be applied to the selection of stabilized enzymes. PMID:10227171

  8. Reconstitution of translation from Thermus thermophilus reveals a minimal set of components sufficient for protein synthesis at high temperatures and functional conservation of modern and ancient translation components

    OpenAIRE

    Ying ZHOU; Asahara, Haruichi; Gaucher, Eric A.; Chong, Shaorong

    2012-01-01

    Thermus thermophilus is a thermophilic model organism distantly related to the mesophilic model organism E. coli. We reconstituted protein translation of Thermus thermophilus in vitro from purified ribosomes, transfer ribonucleic acids (tRNAs) and 33 recombinant proteins. This reconstituted system was fully functional, capable of translating natural messenger RNA (mRNA) into active full-length proteins at temperatures up to 65°C and with yields up to 60 μg/ml. Surprisingly, the synthesis of a...

  9. Correlation of Activities of the Enzymes α-Phosphoglucomutase, UDP-Galactose 4-Epimerase, and UDP-Glucose Pyrophosphorylase with Exopolysaccharide Biosynthesis by Streptococcus thermophilus LY03

    OpenAIRE

    Degeest, Bart; De Vuyst, Luc

    2000-01-01

    The effects of different carbohydrates or mixtures of carbohydrates as substrates on bacterial growth and exopolysaccharide (EPS) production were studied for the yoghurt starter culture Streptococcus thermophilus LY03. This strain produces two heteropolysaccharides with the same monomeric composition (galactose and glucose in the ratio 4:1) but with different molecular masses. Lactose and glucose were fermented by S. thermophilus LY03 only when they were used as sole energy and carbohydrate s...

  10. Biotic interactions at hydrothermal vents: Recruitment inhibition by the mussel Bathymodiolus thermophilus

    Science.gov (United States)

    Lenihan, H. S.; Mills, S. W.; Mullineaux, L. S.; Peterson, C. H.; Fisher, C. R.; Micheli, F.

    2008-12-01

    The structure and dynamics of marine communities are regulated in part by variation in recruitment. As in other ecosystems, recruitment at deep-sea hydrothermal vents is controlled by the interplay of propagule supply and behavior, gradients in physical-chemical conditions, and biotic interactions during pre- and post-settlement periods. Recent research along the East Pacific Rise indicates that inhibition of recently settled larvae by mobile predators (mainly limpets) influences patterns of recruitment and subsequent community succession. We conducted a manipulative experiment at the same sites (˜2510 m water depth) to test whether high-density assemblages of the mussel Bathymodiolus thermophilus also inhibit recruitment. In a preliminary study, recruitment of vent invertebrates within the faunal zone dominated by B. thermophilus was strikingly different at two sites, East Wall and Worm Hole. East Wall had high densities of mussels but very low total recruitment. In contrast, Worm Hole had few mussels but high recruitment. Using the submersible Alvin, we transplanted a large number of mussels from East Wall to Worm Hole and quantified recruitment on basalt blocks placed in three treatments: (1) naturally high densities of mussels at East Wall; (2) naturally low densities of mussels at Worm Hole; and (3) high densities of transplanted mussels at Worm Hole. After 11 months, a total of 24 taxa had recruited to the basalt blocks. Recruitment was 44-60% lower in the transplanted high-density mussel patch at Worm Hole and the natural high-density patch at East Wall than within the natural low-density patch at Worm Hole. Biotic processes that may have caused the pattern of recruitment observed included predation of larvae via water filtration by mussels, larval avoidance of superior competitors, interference competition, and enhanced predation by species within the mussel-bed community. Our results indicate that biotic interactions affecting recruitment must be

  11. Mixed-culture transcriptome analysis reveals the molecular basis of mixed-culture growth in Streptococcus thermophilus and Lactobacillus bulgaricus.

    Science.gov (United States)

    Sieuwerts, Sander; Molenaar, Douwe; van Hijum, Sacha A F T; Beerthuyzen, Marke; Stevens, Marc J A; Janssen, Patrick W M; Ingham, Colin J; de Bok, Frank A M; de Vos, Willem M; van Hylckama Vlieg, Johan E T

    2010-12-01

    Many food fermentations are performed using mixed cultures of lactic acid bacteria. Interactions between strains are of key importance for the performance of these fermentations. Yogurt fermentation by Streptococcus thermophilus and Lactobacillus bulgaricus (basonym, Lactobacillus delbrueckii subsp. bulgaricus) is one of the best-described mixed-culture fermentations. These species are believed to stimulate each other's growth by the exchange of metabolites such as folic acid and carbon dioxide. Recently, postgenomic studies revealed that an upregulation of biosynthesis pathways for nucleotides and sulfur-containing amino acids is part of the global physiological response to mixed-culture growth in S. thermophilus, but an in-depth molecular analysis of mixed-culture growth of both strains remains to be established. We report here the application of mixed-culture transcriptome profiling and a systematic analysis of the effect of interaction-related compounds on growth, which allowed us to unravel the molecular responses associated with batch mixed-culture growth in milk of S. thermophilus CNRZ1066 and L. bulgaricus ATCC BAA-365. The results indicate that interactions between these bacteria are primarily related to purine, amino acid, and long-chain fatty acid metabolism. The results support a model in which formic acid, folic acid, and fatty acids are provided by S. thermophilus. Proteolysis by L. bulgaricus supplies both strains with amino acids but is insufficient to meet the biosynthetic demands for sulfur and branched-chain amino acids, as becomes clear from the upregulation of genes associated with these amino acids in mixed culture. Moreover, genes involved in iron uptake in S. thermophilus are affected by mixed-culture growth, and genes coding for exopolysaccharide production were upregulated in both organisms in mixed culture compared to monocultures. The confirmation of previously identified responses in S. thermophilus using a different strain combination

  12. Molecular methods for identification of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus using methionine biosynthesis and 16S rRNA genes.

    Science.gov (United States)

    Cebeci, Aysun; Gürakan, G Candan

    2008-11-01

    Yoghurt and starter culture producers are still searching strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus to produce healthier yogurt with longer shelf life, better texture, taste and quality. However, selective identification of Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus from a mixed population using microbiological and biochemical methods is difficult, time consuming and may not be accurate. In this study, a quick, sensitive and accurate method is proposed to identify both Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus using PCR. The method is comprised of two parts. In the first part, methionine biosynthesis genes, known to be present in both species were partially amplified by designed primers (cysmet2F and cysmet2R). Partial amplification of the methionine biosynthesis gene which gives 700 bp fragment resulted in selective identification of Lb. bulgaricus and Strep. thermophilus. All 16 Lb. bulgaricus and 6 Strep. thermophilus isolates assessed by this method gave the expected amplification. On the other hand, further analysis of other closely related species with the same primers have indicated that the same product was also amplified in two more lactobacilli namely, Lb. delbrueckii subsp. lactis and Lb. helveticus species. Thus, in the second part of the method, further differentiation of Lb. delbrueckii subsp. bulgaricus and Strep. thermophilus from each other and these species was achieved using restriction analysis of 16S rRNA gene with EcoRI.

  13. Reduction of the off-flavor volatile generated by the yogurt starter culture including Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in soymilk.

    Science.gov (United States)

    Kaneko, Daisuke; Igarashi, Toshinori; Aoyama, Kenji

    2014-02-19

    Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus establish a symbiotic relationship in milk; however, S. thermophilus predominantly grows in soymilk. This study determined that excess diacetyl was notably generated mainly by S. thermophilus in soymilk, and this flavor compound created an unpleasant odor in fermented soymilk. The addition of l-valine to soymilk reduced the amount of diacetyl and increased the levels of acetoin during fermentation by S. thermophilus . In addition, it was found that the expression of the ilvC gene was repressed and that of the als and aldB genes was stimulated in S. thermophilus by l-valine. Sensory evaluations with the triangle difference test and a preference test showed that the soymilk fermented with l-valine was significantly preferred compared with that without l-valine. In this study, we successfully controlled the metabolic flux of S. thermophilus in soymilk and produced more favorable fermented soymilk without the use of genetically modified lactic acid bacteria strains.

  14. Properties of Streptococcus thermophilus fermented milk containing variable concentrations of Bifidobacterium longum and Lactobacillus acidophilus Propriedades de leites fermentados por Streptococcus thermophilus contendo concentrações variáveis de Bifidobacterium longum e Lactobacillus acidophilus

    Directory of Open Access Journals (Sweden)

    Patrícia Blumer Zacarchenco

    2006-09-01

    Full Text Available Sensory evaluation and analysis of pH, titratable acidity and microbial counts after 1, 7, 14 and 21 days of storage of five combinations of lyophilized Bifidobacterium longum and/or Lactobacillus acidophilus added to milk fermented with Streptococcus thermophilus were studied during storage at 4ºC. The taste and acidity sensory attributes were significantly (PForam estudadas as características sensoriais, de pH e acidez de cinco combinações de leites fermentados por Streptococcus thermophilus, adicionados de Bifidobacterium longum e/ou Lactobacillus acidophilus liofilizados. Os efeitos sobre as características sensoriais tornaram-se significantes (p < 0,05 com o aumento do tempo de estocagem. As combinações favoritas continham, inicialmente, Bif. longum e L. acidophilus (10(8 e 10(7ufc/mL, respectivamente e Bif. longum apenas (10(8ufc/mL. Estas combinações não diferiram significativamente do leite fermentado padrão, nem entre si. As notas mais baixas e os maiores valores de acidez titulável foram apresentados pelo leite fermentado por Streptococcus thermophilus contendo apenas L. acidophilus (10(8ufc/mL, concentração inicial. Durante 21 dias de estocagem a 4ºC, as contagens de células viáveis de Str. thermophilus não mudaram, as de Bif. longum mantiveram-se constantes ou reduziram em um ciclo logarítmico e, as de L. acidophilus reduziram de 1 a 2 ciclos logarítmicos. Não foi observada inibição da pós-acidificação resultante da presença de bifidobactéria ou L. acidophilus.

  15. Expression in E. coli and purification of Thermus thermophilus translation initiation factors IF1 and IF3.

    Science.gov (United States)

    Wolfrum, Alexandra; Brock, Stephan; Mac, Thi; Grillenbeck, Norbert

    2003-05-01

    The initiation of protein translation in bacteria requires in addition to mRNA, fMet-tRNA, and ribosomal subunits three protein factors, the initiation factor 1 (IF1), initiation factor 2 (IF2), and initiation factor 3 (IF3). The genes coding for IF1 and IF3 from Thermus thermophilus have been identified and cloned into pET expression vector and were expressed as soluble proteins in Escherichia coli. IF1 was purified by a DEAE-cellulose chromatography, followed by heat denaturation, chromatography on Hydroxylapatit, and gel permeation chromatography using Sephacryl 200HR. For the purification of IF3, a heat denaturation step is followed by anion-exchange chromatography on Q-Sepharose FF and gel permeation chromatography on Sephacryl 200HR. Using these procedures we obtained chromatographically pure and biologically active preparations of both T. thermophilus IF1 and IF3. PMID:12729721

  16. Identification and molecular modeling of a family 5 endocellulase from Thermus caldophilus GK24, a cellulolytic strain of Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Dae-Sil Lee

    2006-12-01

    Full Text Available The genome of T. caldophilus GK24 was recently sequenced and annotated as 14contigs, equivalent to 2.3 mega basepairs (Mbp of DNA. In the current study, we identifieda unique 13.7 kbp DNA sequence, which included the endocellulase gene of T. caldophilusGK24, which did not appear to be present in the complete genomic sequence of the closelyrelated species T. thermophilus HB27 and HB8. Congo-red staining revealed a uniquephenotype of cellulose degradation by strain GK24 that was distinct from other closelyrelated Thermus strains. The results showed that strain GK24 is an aerobic, thermophilic,cellulolytic eubacterium which belongs to the group T. thermophilus. In order to understandthe mechanism of production of cellobiose in T. caldophilus GK24, a three-dimensionalmodel of the endocellulase, TcCel5A, was generated based on known crystal structures.Using this model, we carried out a flexible cellotetraose docking study.

  17. Characterization and evolution of a family of integrative and potentially conjugative or mobilizable elements from Streptococcus thermophilus

    OpenAIRE

    Pavlovic, Guillaume; Burrus, Vincent; Toulmay, Alexandre; Choulet, Frédéric; Decaris, Bernard; Guédon, Gérard

    2004-01-01

    International audience The integrative and conjugative elements (ICEs) excise by site-specific recombination, self-transfer the resulting circular form by conjugation and integrate into the genome of the recipient bacterium. The 34.7-kb element from Streptococcus thermophilus CNRZ368, ICESt1, excises and integrates by site-specific recombination. This element also possesses a conjugation module distantly related to that of the conjugative transposon Tn916 from Enterococcus faecalis. Theref...

  18. Purification, properties, and sequence specificity of SslI, a new type II restriction endonuclease from Streptococcus salivarius subsp. thermophilus.

    Science.gov (United States)

    Benbadis, L; Garel, J R; Hartley, D L

    1991-01-01

    SslI, a type II restriction endonuclease, was purified from Streptococcus salivarius subsp. thermophilus strain BSN 45. SslI is an isoschizomer of BstNI. SslI activity was maximum at pH 8.8, 0 to 50 mM NaCl, 2 to 8 mM Mg2+, and 42 degrees C. Activity against phage DNA in vitro was demonstrated. Images PMID:1785940

  19. Differentiation of Enterococcus faecium from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains by PCR and dot-blot hybridisation.

    Science.gov (United States)

    Langa, S; Fernández, A; Martín, R; Reviriego, C; Marín, M L; Fernández, L; Rodríguez, J M

    2003-12-01

    Variations in length and sequence of the 16S/23S spacer region of Enterococcus faecium provided the basis for development of simple PCR and dot-blot hybridisation assays that enabled the differentiation of potentially probiotic Enterococcus faecium strains from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Such assays may be useful for differentiation of yoghurt starter cultures and enterococcal strains when they are simultaneously present in probiotic food products.

  20. Phage-Induced Expression of CRISPR-Associated Proteins is Revealed by Shotgun Proteomics in Streptococcus thermophilus

    Energy Technology Data Exchange (ETDEWEB)

    Young, Jacque C [ORNL; Dill, Brian [ORNL; Pan, Chongle [ORNL; Hettich, Robert {Bob} L [ORNL; Banfield, Jillian F. [University of California, Berkeley; Shah, Manesh B [ORNL; Fremaux, Christophe [Danisco France SAS; Horvath, Philippe [Danisco France SAS; Barrangou, Rodolphe [Danisco USA; Verberkmoes, Nathan C [ORNL

    2012-01-01

    The CRISPR/Cas system, comprised of clustered regularly interspaced short palindromic repeats along with their associated (Cas) proteins, protects bacteria and archaea from viral predation and invading nucleic acids. While the mechanism of action for this acquired immunity is currently under investigation, the response of Cas protein expression to phage infection has yet to be elucidated. In this study, we employed shotgun proteomics to measure the global proteome expression in a model system for studying the CRISPR/Cas response: infection of S. thermophilus DGCC7710 with phage 2972. Host and viral proteins were simultaneously measured following inoculation at two different multiplicities of infection and across various time points using two-dimensional liquid chromatography tandem mass spectroscopy. Thirty-seven out of forty predicted viral proteins were detected, including all proteins of the structural virome and viral effector proteins. In total, 1,013 of 2,079 predicted S. thermophilus proteins were detected, facilitating the monitoring of host protein synthesis changes in response to virus infection. Importantly, Cas proteins from all four CRISPR loci in the S. thermophilus DGCC7710 genome were detected, including loci previously thought to be inactive. Many Cas proteins were found to be constitutively expressed, but several demonstrated increased abundance during peak infection, including the Cas9 proteins from the CRISPR1 and CRISPR3 loci, which are key players in the interference phase of the CRISPR/Cas response. Altogether, these results provide novel insights into the proteomic response of S. thermophilus, specifically CRISPR-associated proteins, upon phage 2972 infection.

  1. Persistence of wild Streptococcus thermophilus strains on wooden vat and during the manufacture of a traditional Caciocavallo type cheese.

    Science.gov (United States)

    Settanni, L; Di Grigoli, A; Tornambé, G; Bellina, V; Francesca, N; Moschetti, G; Bonanno, A

    2012-04-01

    The present work was undertaken to evaluate the influence of the wooden dairy plant equipment on the microbiological characteristics of curd to be transformed into Caciocavallo Palermitano cheese. Traditional raw milk productions were performed concomitantly with standard cheese making trials carried out in stainless steel vat inoculated with a commercial starter. Milk from two different farms (A and B) was separately processed. The wooden vat was found to be a reservoir of lactic acid bacteria (LAB), while unwanted (spoilage and/or pathogenic) microorganisms were not hosted or were present at very low levels. All microbial groups were numerically different in bulk milks, showing higher levels for the farm B. LAB, especially thermophilic cocci, dominated the whole cheese making process of all productions. Undesired microorganisms decreased in number or disappeared during transformation, particularly after curd stretching. LAB were isolated from the wooden vat surface and from all dairy samples, subjected to phenotypic and genetic characterization and identification. Streptococcus thermophilus was the species found at the highest concentration in all samples analyzed and it also dominated the microbial community of the wooden vat. Fourteen other LAB species belonging to six genera (Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Streptococcus and Weissella) were also detected. All S. thermophilus isolates were genetically differentiated and a consortium of four strains persisted during the whole traditional production process. As confirmed by pH and the total acidity after the acidification step, indigenous S. thermophilus strains acted as a mixed starter culture.

  2. Cloning, expression, purification, crystallization and preliminary X-ray crystallographic study of molybdopterin synthase from Thermus thermophilus HB8

    International Nuclear Information System (INIS)

    The molybdopterin synthase from T. thermophilus HB8 was cloned, expressed, purified and crystallized. The crystals belong to space group P21 and diffracted to a resolution of 1.64 Å. Thermus thermophilus is a Gram-negative aerobic thermophilic eubacterium which can grow at temperatures ranging from 323 to 355 K. In addition to their importance in thermostability or adaptation strategies for survival at high temperatures, the thermostable enzymes in thermophilic organisms contribute to a wide range of biotechnological applications. The molybdenum cofactor in all three kingdoms consists of a tricyclic pyranopterin termed molybdopterin that bears the cis-dithiolene group responsible for molybdenum ligation. The crystals of molybdopterin synthase from T. thermophilus HB8 belong to the primitive monoclinic space group P21, with unit-cell parameters a = 33.94, b = 103.32, c = 59.59 Å, β = 101.3°. Preliminary studies and molecular-replacement calculations reveal the presence of three monomers in the asymmetric unit

  3. Specialized adaptation of a lactic acid bacterium to the milk environment: the comparative genomics of Streptococcus thermophilus LMD-9

    Science.gov (United States)

    2011-01-01

    Background Streptococcus thermophilus represents the only species among the streptococci that has “Generally Regarded As Safe” status and that plays an economically important role in the fermentation of yogurt and cheeses. We conducted comparative genome analysis of S. thermophilus LMD-9 to identify unique gene features as well as features that contribute to its adaptation to the dairy environment. In addition, we investigated the transcriptome response of LMD-9 during growth in milk in the presence of Lactobacillus delbrueckii ssp. bulgaricus, a companion culture in yogurt fermentation, and during lytic bacteriophage infection. Results The S. thermophilus LMD-9 genome is comprised of a 1.8 Mbp circular chromosome (39.1% GC; 1,834 predicted open reading frames) and two small cryptic plasmids. Genome comparison with the previously sequenced LMG 18311 and CNRZ1066 strains revealed 114 kb of LMD-9 specific chromosomal region, including genes that encode for histidine biosynthetic pathway, a cell surface proteinase, various host defense mechanisms and a phage remnant. Interestingly, also unique to LMD-9 are genes encoding for a putative mucus-binding protein, a peptide transporter, and exopolysaccharide biosynthetic proteins that have close orthologs in human intestinal microorganisms. LMD-9 harbors a large number of pseudogenes (13% of ORFeome), indicating that like LMG 18311 and CNRZ1066, LMD-9 has also undergone major reductive evolution, with the loss of carbohydrate metabolic genes and virulence genes found in their streptococcal counterparts. Functional genome distribution analysis of ORFeomes among streptococci showed that all three S. thermophilus strains formed a distinct functional cluster, further establishing their specialized adaptation to the nutrient-rich milk niche. An upregulation of CRISPR1 expression in LMD-9 during lytic bacteriophage DT1 infection suggests its protective role against phage invasion. When co-cultured with L. bulgaricus, LMD-9

  4. Structure of N-formylglycinamide ribonucleotide amidotransferase II (PurL) from Thermus thermophilus HB8

    Science.gov (United States)

    Suzuki, Sakiko; Yanai, Hisaaki; Kanagawa, Mayumi; Tamura, Satoko; Watanabe, Yuzo; Fuse, Kyotaro; Baba, Seiki; Sampei, Gen-ichi; Kawai, Gota

    2012-01-01

    The crystal structure of PurL from Thermus thermophilus HB8 (TtPurL; TTHA1519) was determined in complex with an adenine nucleotide, PO4 3− and Mg2+ at 2.35 Å resolution. TtPurL consists of 29 α-helices and 28 β-strands, and one loop is disordered. TtPurL consists of four domains, A1, A2, B1 and B2, and the structures of the A1–B1 and A2–B2 domains were almost identical to each other. Although the sequence identity between TtPurL and PurL from Thermotoga maritima (TmPurL) is higher than that between TtPurL and the PurL domain of the large PurL from Salmonella typhimurium (StPurL), the secondary structure of TtPurL is much more similar to that of StPurL than to that of TmPurL. PMID:22232163

  5. Crystal structures and ligand binding of PurM proteins from Thermus thermophilus and Geobacillus kaustophilus.

    Science.gov (United States)

    Kanagawa, Mayumi; Baba, Seiki; Watanabe, Yuzo; Nakagawa, Noriko; Ebihara, Akio; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Sampei, Gen-Ichi; Kawai, Gota

    2016-03-01

    Crystal structures of 5-aminoimidazole ribonucleotide (AIR) synthetase, also known as PurM, from Thermus thermophilus (Tt) and Geobacillus kaustophilus (Gk) were determined. For TtPurM, the maximum resolution was 2.2 Å and the space group was P21212 with four dimers in an asymmetric unit. For GkPurM, the maximum resolution was 2.2 Å and the space group was P21212 with one monomer in asymmetric unit. The biological unit is dimer for both TtPurM and GkPurM and the dimer structures were similar to previously determined structures of PurM in general. For TtPurM, ∼50 residues at the amino terminal were disordered in the crystal structure whereas, for GkPurM, the corresponding region covered the ATP-binding site forming an α helix in part, suggesting that the N-terminal region of PurM changes its conformation upon binding of ligands. FGAM binding site was predicted by the docking simulation followed by the MD simulation based on the SO4 (2-) binding site found in the crystal structure of TtPurM.

  6. Long-Range PCR Amplification of DNA by DNA Polymerase III Holoenzyme from Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Wendy Ribble

    2015-01-01

    Full Text Available DNA replication in bacteria is accomplished by a multicomponent replicase, the DNA polymerase III holoenzyme (pol III HE. The three essential components of the pol III HE are the α polymerase, the β sliding clamp processivity factor, and the DnaX clamp-loader complex. We report here the assembly of the functional holoenzyme from Thermus thermophilus (Tth, an extreme thermophile. The minimal holoenzyme capable of DNA synthesis consists of α, β and DnaX (τ and γ, δ and δ′ components of the clamp-loader complex. The proteins were each cloned and expressed in a native form. Each component of the system was purified extensively. The minimum holoenzyme from these five purified subunits reassembled is sufficient for rapid and processive DNA synthesis. In an isolated form the α polymerase was found to be unstable at temperatures above 65°C. We were able to increase the thermostability of the pol III HE to 98°C by addition and optimization of various buffers and cosolvents. In the optimized buffer system we show that a replicative polymerase apparatus, Tth pol III HE, is capable of rapid amplification of regions of DNA up to 15,000 base pairs in PCR reactions.

  7. Site-directed mutation of a laccase from Thermus thermophilus: Effect on the activity profile

    Directory of Open Access Journals (Sweden)

    Liu Xin

    2012-01-01

    Full Text Available A site-directed mutant R453T of a laccase from Thermus thermophilus HB27 (Tth-laccase was constructed in order to investigate the effect on laccase catalytic properties. The mutated gene was cloned and overexpressed in Escherichia coli. Nickel-affinity purification was achieved and followed by copper ion incorporation. The mature mutated enzyme was quantitatively equal to the wild type. A photometric assay based on the oxidation of the substrate 2,2-azino-bis-(3- ethylbenzthiazoline-6-sulfonate (ABTS was employed in comparison with the wild-type Tth-laccase on catalytic properties. The R453T mutant exhibited improvement in substrate affinity and specific activity at room temperature, whereas those parameters were not significantly influenced when the temperature increased up to 65°C or higher. The mutant had better catalytic activity than that of the wild type at acidic pH. Investigated by circular dichroism spectroscopy, the mutant Tth-laccase displayed similar profiles at low and high temperatures.

  8. Sequences spanning the leader-repeat junction mediate CRISPR adaptation to phage in Streptococcus thermophilus.

    Science.gov (United States)

    Wei, Yunzhou; Chesne, Megan T; Terns, Rebecca M; Terns, Michael P

    2015-02-18

    CRISPR-Cas systems are RNA-based immune systems that protect prokaryotes from invaders such as phages and plasmids. In adaptation, the initial phase of the immune response, short foreign DNA fragments are captured and integrated into host CRISPR loci to provide heritable defense against encountered foreign nucleic acids. Each CRISPR contains a ∼100-500 bp leader element that typically includes a transcription promoter, followed by an array of captured ∼35 bp sequences (spacers) sandwiched between copies of an identical ∼35 bp direct repeat sequence. New spacers are added immediately downstream of the leader. Here, we have analyzed adaptation to phage infection in Streptococcus thermophilus at the CRISPR1 locus to identify cis-acting elements essential for the process. We show that the leader and a single repeat of the CRISPR locus are sufficient for adaptation in this system. Moreover, we identified a leader sequence element capable of stimulating adaptation at a dormant repeat. We found that sequences within 10 bp of the site of integration, in both the leader and repeat of the CRISPR, are required for the process. Our results indicate that information at the CRISPR leader-repeat junction is critical for adaptation in this Type II-A system and likely other CRISPR-Cas systems.

  9. Expression and Characterization of Recombinant Thermostable Alkaline Phosphatase from a Novel Thermophilic Bacterium Thermus thermophilus XM

    Institute of Scientific and Technical Information of China (English)

    Jianbo LI; Limei XU; Feng YANG

    2007-01-01

    A gene (tap) encoding a thermostable alkaline phosphatase from the thermophilic bacterium Thermus thermophilus XM was cloned and sequenced. It is 1506 bp long and encodes a protein of 501 amino acid residues with a calculated molecular mass of 54.7 kDa. Comparison of the deduced amino acid sequence with other alkaline phosphatases showed that the regions in the vicinity of the phosphorylation site and metal binding sites are highly conserved. The recombinant thermostable alkaline phosphatase was expressed as a His6-tagged fusion protein in Escherichia coli and its enzymatic properties were characterized after purification. The pH and temperature optima for the recombinant thermostable alkaline phosphatases activity were pH 12 and 75 ℃. As expected, the enzyme displayed high thermostability, retaining more than 50% activity after incubating for 6 h at 80 ℃. Its catalytic function was accelerated in the presence of 0.1 mM Co2+, Fe2+, Mg2+, or Mn2+ but was strongly inhibited by 2.0 mM Fe2+. Under optimal conditions, the Michaelis constant (Km) for cleavage of p-nitrophenyl-phosphate was 0.034 mM. Although it has much in common with other alkaline phosphatases, the recombinant thermostable alkaline phosphatase possesses some unique features, such as high optimal pH and good thermostability.

  10. Independent in vitro assembly of all three major morphological parts of the 30S ribosomal subunit of Thermus thermophilus.

    Science.gov (United States)

    Agalarov, S C; Selivanova, O M; Zheleznyakova, E N; Zheleznaya, L A; Matvienko, N I; Spirin, A S

    1999-12-01

    Fragments of the 16S rRNA of Thermus thermophilus representing the 3' domain (nucleotides 890-1515) and the 5' domain (nucleotides 1-539) have been prepared by transcription in vitro. Incubation of these fragments with total 30S ribosomal proteins of T. thermophilus resulted in formation of specific RNPs. The particle assembled on the 3' RNA domain contained seven proteins corresponding to Escherichia coli ribosomal proteins S3, S7, S9, S10, S13, S14, and S19. All of them have previously been shown to interact with the 3' domain of the 16S RNA and to be localized in the head of the 30S ribosomal subunit. The particle formed on the 5' RNA domain contained five ribosomal proteins corresponding to E. coli proteins S4, S12, S17, S16, and S20. These proteins are known to be localized in the main part of the body of the 30S subunit. Both types of particle were compact and had sedimentation coefficients of 15.5 S and 13 S, respectively. Together with our recent demonstration of the reconstitution of the RNA particle representing the platform of the T. thermophilus 30S ribosomal subunit [Agalarov, S.C., Zheleznyakova, E.N., Selivanova, O.M., Zheleznaya, L.A., Matvienko, N.I., Vasiliev, V.D. & Spirin, A.S. (1998) Proc. Natl Acad. Sci. USA 95, 999-1003], these experiments establish that all three main structural lobes of the small ribosomal subunit can be reconstituted independently of each other and prepared in the individual state.

  11. A novel pheromone quorum-sensing system controls the development of natural competence in Streptococcus thermophilus and Streptococcus salivarius.

    Science.gov (United States)

    Fontaine, Laetitia; Boutry, Céline; de Frahan, Marie Henry; Delplace, Brigitte; Fremaux, Christophe; Horvath, Philippe; Boyaval, Patrick; Hols, Pascal

    2010-03-01

    In streptococcal species, the key step of competence development is the transcriptional induction of comX, which encodes the alternative sigma factor sigma(X), which positively regulates genes necessary for DNA transformation. In Streptococcus species belonging to the mitis and mutans groups, induction of comX relies on the activation of a three-component system consisting of a secreted pheromone, a histidine kinase, and a response regulator. In Streptococcus thermophilus, a species belonging to the salivarius group, the oligopeptide transporter Ami is essential for comX expression under competence-inducing conditions. This suggests a different regulation pathway of competence based on the production and reimportation of a signal peptide. The objective of our work was to identify the main actors involved in the early steps of comX induction in S. thermophilus LMD-9. Using a transcriptomic approach, four highly induced early competence operons were identified. Among them, we found a Rgg-like regulator (Ster_0316) associated with a nonannotated gene encoding a 24-amino-acid hydrophobic peptide (Shp0316). Through genetic deletions, we showed that these two genes are essential for comX induction. Moreover, addition to the medium of synthetic peptides derived from the C-terminal part of Shp0316 restored comX induction and transformation of a Shp0316-deficient strain. These peptides also induced competence in S. thermophilus and Streptococcus salivarius strains that are poorly transformable or not transformable. Altogether, our results show that Ster_0316 and Shp0316, renamed ComRS, are the two members of a novel quorum-sensing system responsible for comX induction in species from the salivarius group, which differs from the classical phosphorelay three-component system identified previously in streptococci. PMID:20023010

  12. A Novel Pheromone Quorum-Sensing System Controls the Development of Natural Competence in Streptococcus thermophilus and Streptococcus salivarius▿ †

    Science.gov (United States)

    Fontaine, Laetitia; Boutry, Céline; de Frahan, Marie Henry; Delplace, Brigitte; Fremaux, Christophe; Horvath, Philippe; Boyaval, Patrick; Hols, Pascal

    2010-01-01

    In streptococcal species, the key step of competence development is the transcriptional induction of comX, which encodes the alternative sigma factor σX, which positively regulates genes necessary for DNA transformation. In Streptococcus species belonging to the mitis and mutans groups, induction of comX relies on the activation of a three-component system consisting of a secreted pheromone, a histidine kinase, and a response regulator. In Streptococcus thermophilus, a species belonging to the salivarius group, the oligopeptide transporter Ami is essential for comX expression under competence-inducing conditions. This suggests a different regulation pathway of competence based on the production and reimportation of a signal peptide. The objective of our work was to identify the main actors involved in the early steps of comX induction in S. thermophilus LMD-9. Using a transcriptomic approach, four highly induced early competence operons were identified. Among them, we found a Rgg-like regulator (Ster_0316) associated with a nonannotated gene encoding a 24-amino-acid hydrophobic peptide (Shp0316). Through genetic deletions, we showed that these two genes are essential for comX induction. Moreover, addition to the medium of synthetic peptides derived from the C-terminal part of Shp0316 restored comX induction and transformation of a Shp0316-deficient strain. These peptides also induced competence in S. thermophilus and Streptococcus salivarius strains that are poorly transformable or not transformable. Altogether, our results show that Ster_0316 and Shp0316, renamed ComRS, are the two members of a novel quorum-sensing system responsible for comX induction in species from the salivarius group, which differs from the classical phosphorelay three-component system identified previously in streptococci. PMID:20023010

  13. Identification of novel esterase-active enzymes from hot environments by use of the host bacterium Thermus thermophilus

    Directory of Open Access Journals (Sweden)

    Benedikt eLeis

    2015-04-01

    Full Text Available Functional metagenomic screening strategies, which are independent of known sequence information, can lead to the identification of truly novel genes and enzymes. Since E. coli has been used exhaustively for this purpose as a host, it is important to establish alternative expression hosts and to use them for functional metagenomic screening for new enzymes. In this study we show that Thermus thermophilus HB27 is an excellent screening host and can be used as an alternative provider of truly novel biocatalysts. In a previous study we constructed the mutant strain BL03 that was no longer able to grow on defined minimal medium supplemented with tributyrin as the sole carbon source and could be used as a host to screen for metagenomic DNA fragments that could complement growth on tributyrin. Several thousand single fosmid clones from thermophilic metagenomic libraries from heated compost and hot spring water samples were subjected to a comparative screening for esterase activity in both T. thermophilus strain BL03 and E. coli EPI300. We scored a greater number of active clones in the thermophilic bacterium than in the mesophilic E. coli. From all clones functionally screened in E. coli, only two thermostable α/β-fold hydrolase enzymes with high amino acid sequence similarity to already characterized enzymes were identifiable. In contrast, five further fosmids were found that conferred lipolytic activities in T. thermophilus. Four open reading frames (ORFs were found which did not share significant similarity to known esterase enzymes. Two of the genes were expressed in both hosts and the novel thermophilic esterases, which based on their primary structures could not be assigned to known esterase or lipase families, were purified and preliminarily characterized. Our work underscores the benefit of using additional screening hosts other than E. coli for the identification of novel biocatalysts with industrial relevance.

  14. Quantitative analysis of histidine decarboxylase gene (hdcA) transcription and histamine production by Streptococcus thermophilus PRI60 under conditions relevant to cheese making.

    Science.gov (United States)

    Rossi, Franca; Gardini, Fausto; Rizzotti, Lucia; La Gioia, Federica; Tabanelli, Giulia; Torriani, Sandra

    2011-04-01

    This study evaluated the influence of parameters relevant for cheese making on histamine formation by Streptococcus thermophilus. Strains possessing a histidine decarboxylase (hdcA) gene represented 6% of the dairy isolates screened. The most histaminogenic, S. thermophilus PRI60, exhibited in skim milk a high basal level of expression of hdcA, upregulation in the presence of free histidine and salt, and repression after thermization. HdcA activity persisted in cell extracts, indicating that histamine might accumulate after cell lysis in cheese.

  15. Quantitative Analysis of Histidine Decarboxylase Gene (hdcA) Transcription and Histamine Production by Streptococcus thermophilus PRI60 under Conditions Relevant to Cheese Making▿†

    Science.gov (United States)

    Rossi, Franca; Gardini, Fausto; Rizzotti, Lucia; La Gioia, Federica; Tabanelli, Giulia; Torriani, Sandra

    2011-01-01

    This study evaluated the influence of parameters relevant for cheese making on histamine formation by Streptococcus thermophilus. Strains possessing a histidine decarboxylase (hdcA) gene represented 6% of the dairy isolates screened. The most histaminogenic, S. thermophilus PRI60, exhibited in skim milk a high basal level of expression of hdcA, upregulation in the presence of free histidine and salt, and repression after thermization. HdcA activity persisted in cell extracts, indicating that histamine might accumulate after cell lysis in cheese. PMID:21378060

  16. Röntgenstrukturanalyse der ba3 Cytochrom-c Oxidase aus Thermus thermophilus und ihres Substrates Cytochrom-c552

    OpenAIRE

    Than, Manuel E.

    2007-01-01

    Die dreidimensionalen Strukturen der ba3 Cytocrom-c Oxidase aus dem Eubakterium Thermus thermophilus sowie ihres Substrates Cytochrom-c552 wurden mit den Methoden der Röntgenstrukturanalyse bei einer Auflösung von 2,4 Å bzw. 1,28 Å aufgeklärt. Die Analyse dieser Strukturen ermöglichte das Verständnis der besonderen biochemischen Eigenschaften dieser beiden Proteine bezüglich Thermostabilität, Protonenpumpaktivität, Reaktionsmechanismus und Elektronenübertragung im Vergleich zu typischen Vert...

  17. The three major types of CRISPR-Cas systems function independently in CRISPR RNA biogenesis in Streptococcus thermophilus

    OpenAIRE

    Carte, Jason; Christopher, Ross T.; Smith, Justin T.; Olson, Sara; Barrangou, Rodolphe; Moineau, Sylvain; Glover, Claiborne V. C.; Graveley, Brenton R.; Terns, Rebecca M.; Terns, Michael P.

    2014-01-01

    CRISPR-Cas systems are small RNA-based immune systems that protect prokaryotes from invaders such as viruses and plasmids. We have investigated the features and biogenesis of the CRISPR (cr)RNAs in Streptococcus thermophilus (Sth) strain DGCC7710, which possesses four different CRISPR-Cas systems including representatives from the three major types of CRISPR-Cas systems. Our results indicate that the crRNAs from each CRISPR locus are specifically processed into divergent crRNA species by Cas ...

  18. High yield of mannosylglycerate production by upshock fermentation and bacterial milking of trehalose-deficient mutant Thermus thermophilus RQ-1

    OpenAIRE

    K Egorova; Grudieva, T.; Morinez, C.; Kube, J.; Santos, H.; Costa, M Da; Antranikian, G.

    2007-01-01

    Abstract A production process, using upshock fermentation and osmotic downshock, for the effective production/excretion of mannosylglycerate (MG) by the trehalose-deficient mutant of the strain Thermus thermophilus RQ-1 has been developed. In the first phase of fed-batch fermentation, the knockout mutant was grown at 70°C on a NaCl-free medium. After the culture reached the end of the exponential growth phase, upshift in temperature and NaCl concentration was applied. The temperature was inc...

  19. Zygote-mediated generation of genome-modified mice using Streptococcus thermophilus 1-derived CRISPR/Cas system.

    Science.gov (United States)

    Fujii, Wataru; Kakuta, Shigeru; Yoshioka, Shin; Kyuwa, Shigeru; Sugiura, Koji; Naito, Kunihiko

    2016-08-26

    Mammalian zygote-mediated genome-engineering by CRISPR/Cas is currently used for the generation of genome-modified animals. Here we report that a Streptococcus thermophilus-1 derived orthologous CRISPR/Cas system, which recognizes the 5'-NNAGAA sequence as a protospacer adjacent motif (PAM), is useful in mouse zygotes and is applicable for generating knockout mice (87.5%) and targeted knock-in mice (45.5%). The induced mutation could be inherited in the next generation. This novel CRISPR/Cas can expand the feasibility of the zygote-mediated generation of genome-modified animals that require an exact mutation design. PMID:27318086

  20. Structural comparison between the open and closed forms of citrate synthase from Thermus thermophilus HB8.

    Science.gov (United States)

    Kanamori, Eiji; Kawaguchi, Shin-Ichi; Kuramitsu, Seiki; Kouyama, Tsutomu; Murakami, Midori

    2015-01-01

    The crystal structures of citrate synthase from the thermophilic eubacteria Thermus thermophilus HB8 (TtCS) were determined for an open form at 1.5 Å resolution and for closed form at 2.3 Å resolution, respectively. In the absence of ligands TtCS in the open form was crystalized into a tetragonal form with a single subunit in the asymmetric unit. TtCS was also co-crystallized with citrate and coenzyme-A to form an orthorhombic crystal with two homodimers in the asymmetric unit. Citrate and CoA are found in the active site situated between the large domain and the small domain in all subunit whereas the complex shows two distinct closed conformations, the fully closed form and partially closed form. Structural comparisons are performed to describe conformational changes associated with binding of products of TtCS. Upon binding of citrate, basic residues in the active site move toward citrate and make a hydrogen bond network in the active site, inducing a large-scale rotation of the small domain relative to the large domain. CoA is sandwiched between the small and large domains and then the cysteamine tail is inserted into the active site with a cooperative rotation around mainchain dihedrals in the hinge region connecting helices M and N. According to this rotation these helices are extended to close the active site completely. The considerable flexibility and structural rearrangements in the hinge region are crucial for an ordered bibi reaction in catalysis for microbial CSs.

  1. Study on Streptococcus thermophilus isolated from Qula and associated characteristic of acetaldehyde and diacetyl in their fermented milk.

    Science.gov (United States)

    Zha, Musu; Yu, Jie; Zhang, Yong; Wang, Hongmei; Bai, Na; Qin, Yanting; Liangliang, De; Liu, Wenjun; Zhang, Heping; Bilige, Menghe

    2015-01-01

    In this study, the lactic acid bacterial population of Qula cheese from the Gansu and Sichuan provinces of China were isolated and identified. Eight strains of Streptococcus thermophilus were isolated, of which five strains were selected for further characterization based on their fermentation properties. The changes in a number of parameters, including titration acidity, pH, viable counts, PrtS protease activity and the production of acetaldehyde, diacetyl and organic acid, were monitored during fermentation and the storage of fermented milks produced by the respective strain. All of the strains displaying acidifying capacity and all five fermented milks maintained high viable counts of S. thermophilus from fermentation to storage. Our study found that the changes in the monitored parameters were strain-specific and varied considerably among the five tested strains. Fermented milks produced by strain IMAU80809 had the highest concentration of acetaldehyde and were most favorable in the sensory evaluation. This study confirms that Qula cheese is a good source for isolating novel lactic acid bacterial strains with different fermentation properties, which will be very useful for further development and industrialization of traditionally fermented dairy products. PMID:26018501

  2. Optimization and effect of dairy industrial waste as media components in the production of hyaluronic acid by Streptococcus thermophilus.

    Science.gov (United States)

    Mohan, Naresh; Balakrishnan, Rengesh; Sivaprakasam, Senthilkumar

    2016-08-17

    Hyaluronic acid (HA) production using a dairy industrial waste is a more cost-efficient strategy than using an expensive synthetic medium. In this study, we investigated the production of HA using Streptococcus thermophilus under shake flask conditions using dairy industrial waste as nutritional supplements, namely whey permeate (WP) and whey protein hydrolysate (WPH). Preliminary screening using Plackett-Burman design exhibited WP, WPH, initial pH, and inoculum size as significant factors influencing HA titer. Response surface methodology design of four factors was formulated at three levels for enhanced production of HA. Shake flask HA fermentation by S. thermophilus was performed under global optimized process conditions and the optimal HA titer (342.93 mg L(-1)) corroborates with Box-Behnken design prediction. The molecular weight of HA was elucidated as 9.22-9.46 kDa. The ultralow-molecular weight HA reported in this study has a potential role in drug and gene delivery applications. PMID:26681350

  3. Differential regulation of two closely related integrative and conjugative elements from Streptococcus thermophilus

    Directory of Open Access Journals (Sweden)

    Carraro Nicolas

    2011-10-01

    Full Text Available Abstract Background Two closely related ICEs, ICESt1 and ICESt3, have been identified in the lactic acid bacterium Streptococcus thermophilus. While their conjugation and recombination modules are almost identical (95% nucleotide identity and their regulation modules related, previous work has demonstrated that transconjugants carrying ICESt3 were generated at rate exceeding by a 1000 factor that of ICESt1. Results The functional regulation of ICESt1 and ICESt3 transcription, excision and replication were investigated under different conditions (exponential growth or stationary phase, DNA damage by exposition to mitomycin C. Analysis revealed an identical transcriptional organization of their recombination and conjugation modules (long unique transcript whereas the transcriptional organization of their regulation modules were found to be different (two operons in ICESt1 but only one in ICESt3 and to depend on the conditions (promoter specific of stationary phase in ICESt3. For both elements, stationary phase and DNA damage lead to the rise of transcript levels of the conjugation-recombination and regulation modules. Whatever the growth culture conditions, excision of ICESt1 was found to be lower than that of ICESt3, which is consistent with weaker transfer frequencies. Furthermore, for both elements, excision increases in stationary phase (8.9-fold for ICESt1 and 1.31-fold for ICESt3 and is strongly enhanced by DNA damage (38-fold for ICESt1 and 18-fold for ICESt3. Although ICEs are generally not described as replicative elements, the copy number of ICESt3 exhibited a sharp increase (9.6-fold after mitomycin C exposure of its harboring strain CNRZ385. This result was not observed when ICESt3 was introduced in a strain deriving ICESt1 host strain CNRZ368, deleted for this element. This finding suggests an impact of the host cell on ICE behavior. Conclusions All together, these results suggest a novel mechanism of regulation shared by ICESt1

  4. Impact of engineered Streptococcus thermophilus trains overexpressing glyA gene on folic acid and acetaldehyde production in fermented milk Impacto de linhagens de Streptococcus thermophilus com aumento da expressão do gene glyA na produção de ácido folico e acetaldeído em leite fermentado

    Directory of Open Access Journals (Sweden)

    Ana Carolina Sampaio Dória Chaves

    2003-11-01

    Full Text Available The typical yogurt flavor is caused by acetaldehyde produced through many different pathways by the yogurt starter bacteria L. bulgaricus and S. thermophilus. The attention was focused on one specific reaction for acetaldehyde and folic acid formation catalyzed by serine hydroxymethyltransferase (SHMT, encoded by the glyA gene. In S. thermophilus, this enzyme SHMT also plays the typical role of the enzyme threonine aldolase (TA that is the interconvertion of threonine into glycine and acetaldehyde. The behavior of engineered S. thermophilus strains in milk fermentation is described, folic acid and acetaldehyde production were measured and pH and counts were followed. The engineered S. thermophilus strains StA2305 and StB2305, have the glyA gene (encoding the enzyme serine hydroxymethyltransferase overexpressed. These engineered strains showed normal growth in milk when it was supplemented with Casitione. When they were used in milk fermentation it was observed an increase in folic acid and in acetaldehyde production by StA2305 and for StB2305 it was noticed a significative increase in folic acid formation.O acetaldeído, responsável pelo sabor e aroma característicos de iogurte, é produzido por diferentes vias metabólicas pelas bactérias lácticas: Streptococcus thermophilus (S. thermophilus e Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus. Neste trabalho, a atenção foi focada especificamente na reação para a formação de acetaldeído e de ácido fólico, catalisada pela enzima serina hidroximetil transferase (SHMT, codificada pelo gene glyA. A enzima SHMT catalisa diversas reações e, no caso da bactéria S. thermophilus, ela exerce também a atividade característica da enzima treonina aldolase (TA, definida como a interconversão do aminoácido treonina em glicina e acetaldeído. Foram construídas linhagens de S. thermophilus (StA2305 e StB2305 com super expressão do gene glyA. Estas linhagens modificadas apresentaram

  5. Electron transfer among the CuA-, heme b- and a3-centers of Thermus thermophilus cytochrome ba3

    DEFF Research Database (Denmark)

    Farver, Ole; Chen, Ying; Fee, James A;

    2006-01-01

    The 1-methyl-nicotinamide radical (MNA(*)), produced by pulse radiolysis has previously been shown to reduce the Cu(A)-site of cytochromes aa(3), a process followed by intramolecular electron transfer (ET) to the heme a but not to the heme a(3) [Farver, O., Grell, E., Ludwig, B., Michel, H. and...... Pecht, I. (2006) Rates and equilibrium of CuA to heme a electron transfer in Paracoccus denitrificans cytochrome c oxidase. Biophys. J. 90, 2131-2137]. Investigating this process in the cytochrome ba(3) of Thermus thermophilus (Tt), we now show that MNA(*) also reduces Cu(A) with a subsequent ET to the...... heme b and then to heme a(3), with first-order rate constants 11200 s(-1), and 770 s(-1), respectively. The results provide clear evidence for ET among the three spectroscopically distinguishable centers and indicate that the binuclear a(3)-Cu(B) center can be reduced in molecules containing a single...

  6. Complete genome sequence of Streptococcus thermophilus MN-BM-A01, a strain with high exopolysaccharides production.

    Science.gov (United States)

    Bai, Ying; Sun, Erna; Shi, Yudong; Jiang, Yunyun; Chen, Yun; Liu, Songling; Zhao, Liang; Zhang, Ming; Guo, Huiyuan; Zhang, Hao; Mu, Zhishen; Ren, Fazheng

    2016-04-20

    Streptococcus thermophilus MN-BM-A01 (ST MN-BM-A01) (CGMCC No. 11383) was a strain isolated from Yogurt Block in Gansu, China. The yogurt fermented with this strain has good flavor, acidity, and viscosity. Moreover, ST MN-BM-A01 could produce a high level of EPS which can confer the yogurt with improved rheological properties. We reported the complete genome sequence of ST MN-BM-A01 that contains 1,876,516bp encoding 1704 coding sequences (CDSs), 67 tRNA genes and 6 rRNA operons. The genomic sequence indicated that this strain included a 35.3-kb gene cluster involved in EPS biosynthesis. PMID:26956372

  7. Selection of an adhesive phenotype of Streptococcus salivarius subsp. thermophilus for use in fixed-bed reactors.

    Science.gov (United States)

    Ragout, A; Siñeriz, F; Kaul, R; Guoqiang, D; Mattiasson, B

    1996-09-01

    Streptococcus salivarius subsp. thermophilus was cultivated in a chemostat in order to obtain an adhesive phenotype of this strain. When the system was operated at low dilution rates (D biomass productivity (P = 4.1 g1(-1) h-1). On the other hand, when the culture was grown at dilution rates faster than 0.2 h-1, only the free suspended cells were present in the culture broth, and were washed out at velocities of about 1.0 h-1. The biomass productivity was consequently lower (P = 1.33 g1(-1) h-1) than in the previous case. The selected adhesive phenotype was grown on different glass beads and the possibility of lactate fermentation in a continuous and semicontinuous mode was demonstrated.

  8. Bifidobacterium breve and Streptococcus thermophilus secretion products enhance T helper 1 immune response and intestinal barrier in mice.

    Science.gov (United States)

    Ménard, Sandrine; Laharie, David; Asensio, Corinne; Vidal-Martinez, Teresita; Candalh, Céline; Rullier, Anne; Zerbib, Frank; Mégraud, Francis; Matysiak-Budnik, Tamara; Heyman, Martine

    2005-11-01

    Lactic acid bacteria or their secretion products can modulate immune responses differently in normal and inflammatory conditions. This comparative study analyzes the effect of oral administration of living lactic acid bacteria, or their conditioned media, on the epithelial and immune functions of colitis-prone C57BL/6 IL-10-deficient mice. Mice were untreated (control) or infected with Helicobacter hepaticus with or without oral treatment with living bacteria, Bifidobacterium breve C50 and Streptococcus thermophilus 065 (LB), or their culture-conditioned media (CM). Histology, cytokine mRNA, electrical resistance, and barrier capacity of colonic samples as well as cytokine secretion by mesenteric lymph node (MLN) cells were studied. Helicobacter hepaticus mice developed only mild colitis, which was not modified in LB or CM groups. In the CM (but not the LB) group, the colonic barrier was reinforced as compared to the other groups, as evidenced by decreased horseradish peroxidase (HRP) transcytosis and mannitol fluxes and increased electrical resistance. In MLN, the percentage of CD4+ and CD8+ T cells secreting IFNgamma was significantly higher in CM (2.06% and 1.98%, respectively) mice than in H. hepaticus (1.1% and 0.47%, P secretion by MLN cells was significantly higher in the CM group as compared to the other groups. In the absence of severe colitis, Bifidobacterium breve C50- and Streptococcus thermophilus 065-conditioned media can reinforce intestinal barrier capacity and stimulate Th1 immune response, highlighting the involvement of lactic acid bacteria-derived components in host defense.

  9. Comparative genomics of Thermus thermophilus and Deinococcus radiodurans: divergent routes of adaptation to thermophily and radiation resistance

    Directory of Open Access Journals (Sweden)

    Daly Michael J

    2005-10-01

    Full Text Available Abstract Background Thermus thermophilus and Deinococcus radiodurans belong to a distinct bacterial clade but have remarkably different phenotypes. T. thermophilus is a thermophile, which is relatively sensitive to ionizing radiation and desiccation, whereas D. radiodurans is a mesophile, which is highly radiation- and desiccation-resistant. Here we present an in-depth comparison of the genomes of these two related but differently adapted bacteria. Results By reconstructing the evolution of Thermus and Deinococcus after the divergence from their common ancestor, we demonstrate a high level of post-divergence gene flux in both lineages. Various aspects of the adaptation to high temperature in Thermus can be attributed to horizontal gene transfer from archaea and thermophilic bacteria; many of the horizontally transferred genes are located on the single megaplasmid of Thermus. In addition, the Thermus lineage has lost a set of genes that are still present in Deinococcus and many other mesophilic bacteria but are not common among thermophiles. By contrast, Deinococcus seems to have acquired numerous genes related to stress response systems from various bacteria. A comparison of the distribution of orthologous genes among the four partitions of the Deinococcus genome and the two partitions of the Thermus genome reveals homology between the Thermus megaplasmid (pTT27 and Deinococcus megaplasmid (DR177. Conclusion After the radiation from their common ancestor, the Thermus and Deinococcus lineages have taken divergent paths toward their distinct lifestyles. In addition to extensive gene loss, Thermus seems to have acquired numerous genes from thermophiles, which likely was the decisive contribution to its thermophilic adaptation. By contrast, Deinococcus lost few genes but seems to have acquired many bacterial genes that apparently enhanced its ability to survive different kinds of environmental stresses. Notwithstanding the accumulation of

  10. Growth and gas formation by Lactobacillus wasatchensis, a novel obligatory heterofermentative nonstarter lactic acid bacterium, in Cheddar-style cheese made using a Streptococcus thermophilus starter.

    Science.gov (United States)

    Ortakci, Fatih; Broadbent, Jeffery R; Oberg, Craig J; McMahon, Donald J

    2015-11-01

    A novel slow-growing, obligatory heterofermentative, nonstarter lactic acid bacterium (NSLAB), Lactobacillus wasatchensis WDC04, was studied for growth and gas production in Cheddar-style cheese made using Streptococcus thermophilus as the starter culture. Cheesemaking trials were conducted using S. thermophilus alone or in combination with Lb. wasatchensis deliberately added to cheese milk at a level of ~10(4) cfu/mL. Resulting cheeses were ripened at 6 or 12°C. At d 1, starter streptococcal numbers were similar in both cheeses (~10(9) cfu/g) and fast-growing NSLAB lactobacilli counts were below detectable levels (blowing in Cheddar-style cheeses, especially when the cheese is ripened at elevated temperature. PMID:26364109

  11. The extent of co-metabolism of glucose and galactose by L. lactis changes with the expression of the lacSZ operon from Streptococcus thermophilus

    DEFF Research Database (Denmark)

    Solem, Christian; Købmann, Brian Jensen; Jensen, Peter Ruhdal

    2008-01-01

    The lactose transporter and β-galactosidase from Streptococcus thermophilus, encoded by the lacSZ operon, were introduced into the lactose-negative strain Lactococcus lactis MG1363 and the expression of the lacSZ operon was modulated by substitution of the native promoter with randomized synthetic...... and only glucose was metabolized in glycolysis. Interestingly, strains with low expression of the operon showed a mixed acid metabolism and co-metabolism of galactose and glucose. The lactose flux increased gradually with increasing expression of the lacSZ operon until an optimum was observed...... promoters. A series of strains with various expression levels of lacSZ were examined for their fermentation of lactose. Strains with a high expression level were found to metabolize lactose in a similar manner to S. thermophilus, i.e. the galactose moiety of lactose was excreted to the growth medium...

  12. Co-utilization of glucose and xylose by evolved Thermus thermophilus LC113 strain elucidated by (13)C metabolic flux analysis and whole genome sequencing.

    Science.gov (United States)

    Cordova, Lauren T; Lu, Jing; Cipolla, Robert M; Sandoval, Nicholas R; Long, Christopher P; Antoniewicz, Maciek R

    2016-09-01

    We evolved Thermus thermophilus to efficiently co-utilize glucose and xylose, the two most abundant sugars in lignocellulosic biomass, at high temperatures without carbon catabolite repression. To generate the strain, T. thermophilus HB8 was first evolved on glucose to improve its growth characteristics, followed by evolution on xylose. The resulting strain, T. thermophilus LC113, was characterized in growth studies, by whole genome sequencing, and (13)C-metabolic flux analysis ((13)C-MFA) with [1,6-(13)C]glucose, [5-(13)C]xylose, and [1,6-(13)C]glucose+[5-(13)C]xylose as isotopic tracers. Compared to the starting strain, the evolved strain had an increased growth rate (~2-fold), increased biomass yield, increased tolerance to high temperatures up to 90°C, and gained the ability to grow on xylose in minimal medium. At the optimal growth temperature of 81°C, the maximum growth rate on glucose and xylose was 0.44 and 0.46h(-1), respectively. In medium containing glucose and xylose the strain efficiently co-utilized the two sugars. (13)C-MFA results provided insights into the metabolism of T. thermophilus LC113 that allows efficient co-utilization of glucose and xylose. Specifically, (13)C-MFA revealed that metabolic fluxes in the upper part of metabolism adjust flexibly to sugar availability, while fluxes in the lower part of metabolism remain relatively constant. Whole genome sequence analysis revealed two large structural changes that can help explain the physiology of the evolved strain: a duplication of a chromosome region that contains many sugar transporters, and a 5x multiplication of a region on the pVV8 plasmid that contains xylose isomerase and xylulokinase genes, the first two enzymes of xylose catabolism. Taken together, (13)C-MFA and genome sequence analysis provided complementary insights into the physiology of the evolved strain. PMID:27164561

  13. 嗜热链球菌的特性与应用研究进展%Research Progress on the Property and Application of Streptococcus thermophilus

    Institute of Scientific and Technical Information of China (English)

    田辉; 梁宏彰; 霍贵成; Evivie Smith Etareri

    2015-01-01

    嗜热链球菌是一种重要的工业用乳酸菌,广泛应用于发酵乳制品生产。近年来,对嗜热链球菌的研究已不仅仅停留于应用方面,而有关该菌基因组学等深层次特性机理的研究越来越多,对嗜热链球菌的研究和应用开发具有较好的参考价值。综述了嗜热链球菌的认定和应用、基因组学特征、发酵特性、噬菌体防御和共生机制等方面的研究进展,并展望了现代生物技术在嗜热链球菌特性研究中的应用前景。%Streptococcus thermophilus is one of the most important species of lactic acid bacteria in industry, and it is widely used in the process of producing fermented milk. This review discusses researches on the qualification and application of S. thermophilus, the properties of genome and fermentation, phage defense system, and symbiotic mechanism. In the end, the potential application of modern biological technologies in the study of S. thermophilus properties is prospected.

  14. Dairy Streptococcus thermophilus improves cell viability of Lactobacillus brevis NPS-QW-145 and its γ-aminobutyric acid biosynthesis ability in milk.

    Science.gov (United States)

    Wu, Qinglong; Law, Yee-Song; Shah, Nagendra P

    2015-08-06

    Most high γ-aminobutyric acid (GABA) producers are Lactobacillus brevis of plant origin, which may be not able to ferment milk well due to its poor proteolytic nature as evidenced by the absence of genes encoding extracellular proteinases in its genome. In the present study, two glutamic acid decarboxylase (GAD) genes, gadA and gadB, were found in high GABA-producing L. brevis NPS-QW-145. Co-culturing of this organism with conventional dairy starters was carried out to manufacture GABA-rich fermented milk. It was observed that all the selected strains of Streptococcus thermophilus, but not Lactobacillus delbrueckii subsp. bulgaricus, improved the viability of L. brevis NPS-QW-145 in milk. Only certain strains of S. thermophilus improved the gadA mRNA level in L. brevis NPS-QW-145, thus enhanced GABA biosynthesis by the latter. These results suggest that certain S. thermophilus strains are highly recommended to co-culture with high GABA producer for manufacturing GABA-rich fermented milk.

  15. Influence of different proteolytic strains of Streptococcus thermophilus in co-culture with Lactobacillus delbrueckii subsp. bulgaricus on the metabolite profile of set-yoghurt.

    Science.gov (United States)

    Settachaimongkon, Sarn; Nout, M J Robert; Antunes Fernandes, Elsa C; Hettinga, Kasper A; Vervoort, Jacques M; van Hooijdonk, Toon C M; Zwietering, Marcel H; Smid, Eddy J; van Valenberg, Hein J F

    2014-05-01

    Proto-cooperation between Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus is one of the key factors that determine the fermentation process and final quality of yoghurt. In this study, the interaction between different proteolytic strains of S. thermophilus and L. delbrueckii subsp. bulgaricus was investigated in terms of microbial growth, acidification and changes in the biochemical composition of milk during set-yoghurt fermentation. A complementary metabolomics approach was applied for global characterization of volatile and non-volatile polar metabolite profiles of yoghurt associated with proteolytic activity of the individual strains in the starter cultures. The results demonstrated that only non-proteolytic S. thermophilus (Prt-) strain performed proto-cooperation with L. delbrueckii subsp. bulgaricus. The proto-cooperation resulted in significant higher populations of the two species, faster milk acidification, significant abundance of aroma volatiles and non-volatile metabolites desirable for a good organoleptic quality of yoghurt. Headspace SPME-GC/MS and (1)H NMR resulted in the identification of 35 volatiles and 43 non-volatile polar metabolites, respectively. Furthermore, multivariate statistical analysis allows discriminating set-yoghurts fermented by different types of starter cultures according to their metabolite profiles. Our finding underlines that selection of suitable strain combinations in yoghurt starters is important for achieving the best technological performance regarding the quality of product.

  16. Models for the a subunits of the Thermus thermophilus V/A-ATPase and Saccharomyces cerevisiae V-ATPase enzymes by cryo-EM and evolutionary covariance.

    Science.gov (United States)

    Schep, Daniel G; Zhao, Jianhua; Rubinstein, John L

    2016-03-22

    Rotary ATPases couple ATP synthesis or hydrolysis to proton translocation across a membrane. However, understanding proton translocation has been hampered by a lack of structural information for the membrane-embedded a subunit. The V/A-ATPase from the eubacterium Thermus thermophilus is similar in structure to the eukaryotic V-ATPase but has a simpler subunit composition and functions in vivo to synthesize ATP rather than pump protons. We determined the T. thermophilus V/A-ATPase structure by cryo-EM at 6.4 Å resolution. Evolutionary covariance analysis allowed tracing of the a subunit sequence within the map, providing a complete model of the rotary ATPase. Comparing the membrane-embedded regions of the T. thermophilus V/A-ATPase and eukaryotic V-ATPase from Saccharomyces cerevisiae allowed identification of the α-helices that belong to the a subunit and revealed the existence of previously unknown subunits in the eukaryotic enzyme. Subsequent evolutionary covariance analysis enabled construction of a model of the a subunit in the S. cerevisae V-ATPase that explains numerous biochemical studies of that enzyme. Comparing the two a subunit structures determined here with a structure of the distantly related a subunit from the bovine F-type ATP synthase revealed a conserved pattern of residues, suggesting a common mechanism for proton transport in all rotary ATPases.

  17. Transcriptomic and metabolic responses of Staphylococcus aureus in mixed culture with Lactobacillus plantarum, Streptococcus thermophilus and Enterococcus durans in milk.

    Science.gov (United States)

    Zdenkova, Kamila; Alibayov, Babek; Karamonova, Ludmila; Purkrtova, Sabina; Karpiskova, Renata; Demnerova, Katerina

    2016-09-01

    Staphylococcus aureus is a major food-borne pathogen due to the production of enterotoxin and is particularly prevalent in contaminated milk and dairy products. The lactic acid bacteria (LAB) are widely used as biocontrol agents in fermented foods which can inhibit pathogenic flora. In our work, we investigated the influence of three strains of LAB (Lactobacillus plantarum, Streptococcus thermophilus and Enterococcus durans) on the relative expression of three enterotoxin genes (sea, sec, sell) and eight virulence and/or regulatory genes (sarA, saeS, codY, srrA, rot, hld/RNAIII, agrA/RNAII, sigB) in two S. aureus strains (MW2 and Sa1612) in TSB and reduced-fat milk (1.5 %) at 30 °C over a 24-h period. The tested LAB and S. aureus strains proved to be mutually non-competitive or only slightly competitive during co-cultivation. In addition, under the above-mentioned conditions, differential gene expression between the S. aureus MW2 and Sa1612 strains was well documented. S. aureus growth was changed in mixed culture with LAB; however, its effect on the repression of sea and sec expression correlated with production of these virulence factors. In comparison, the presence of LAB strains generally inhibited the expression of sec, sell, sarA, seaS, agrA/RNAII and hld/RNAIII genes. The effect of LAB strains presence on the expression of sea, codY, srrA, rot and sigB genes was medium, time, LAB and S. aureus strain specific. SEA and SEC production was significantly reduced in milk compared to TSB in pure culture. After the 24-h cultivation, S. aureus MW2 and Sa1612 SEC production was 187 and 331 times lower in milk compared to TSB, respectively (0.07 and 0.39 ng/mL in milk, versus 13.1 and 129.2 ng/mL in TSB, respectively). At the same time S. aureus MW2 and Sa1612 SEA production was 77 and 68 times lower in milk compared to TSB, respectively (0.99 and 0.17 ng/mL in milk, versus 76.4 and 11.5 ng/mL in TSB, respectively). This study has revealed new insights into the

  18. Transcriptomic and metabolic responses of Staphylococcus aureus in mixed culture with Lactobacillus plantarum, Streptococcus thermophilus and Enterococcus durans in milk.

    Science.gov (United States)

    Zdenkova, Kamila; Alibayov, Babek; Karamonova, Ludmila; Purkrtova, Sabina; Karpiskova, Renata; Demnerova, Katerina

    2016-09-01

    Staphylococcus aureus is a major food-borne pathogen due to the production of enterotoxin and is particularly prevalent in contaminated milk and dairy products. The lactic acid bacteria (LAB) are widely used as biocontrol agents in fermented foods which can inhibit pathogenic flora. In our work, we investigated the influence of three strains of LAB (Lactobacillus plantarum, Streptococcus thermophilus and Enterococcus durans) on the relative expression of three enterotoxin genes (sea, sec, sell) and eight virulence and/or regulatory genes (sarA, saeS, codY, srrA, rot, hld/RNAIII, agrA/RNAII, sigB) in two S. aureus strains (MW2 and Sa1612) in TSB and reduced-fat milk (1.5 %) at 30 °C over a 24-h period. The tested LAB and S. aureus strains proved to be mutually non-competitive or only slightly competitive during co-cultivation. In addition, under the above-mentioned conditions, differential gene expression between the S. aureus MW2 and Sa1612 strains was well documented. S. aureus growth was changed in mixed culture with LAB; however, its effect on the repression of sea and sec expression correlated with production of these virulence factors. In comparison, the presence of LAB strains generally inhibited the expression of sec, sell, sarA, seaS, agrA/RNAII and hld/RNAIII genes. The effect of LAB strains presence on the expression of sea, codY, srrA, rot and sigB genes was medium, time, LAB and S. aureus strain specific. SEA and SEC production was significantly reduced in milk compared to TSB in pure culture. After the 24-h cultivation, S. aureus MW2 and Sa1612 SEC production was 187 and 331 times lower in milk compared to TSB, respectively (0.07 and 0.39 ng/mL in milk, versus 13.1 and 129.2 ng/mL in TSB, respectively). At the same time S. aureus MW2 and Sa1612 SEA production was 77 and 68 times lower in milk compared to TSB, respectively (0.99 and 0.17 ng/mL in milk, versus 76.4 and 11.5 ng/mL in TSB, respectively). This study has revealed new insights into the

  19. Probiotic Leuconostoc mesenteroides ssp.cremoris and Streptococcus thermophilus induce IL-12 and IFN-γ production

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    AIM: To investigate the capacity of potentially probiotic strains from six bacterial genera to induce cytokine production alone or in combinations in order to identify potential enhancing or synergistic effects in order to select probiotic bacteria for in vivo purposes. METHODS: Cytokine production in human peripheral blood mononuclear cells (PBMC) in response to stimulation with eleven different potentially probiotic bacterial strains from Streptococcus, Lactobacillus, Bifidobacterium, Lactococcus, Leuconostoc and Propionibacterium genera was analysed. Production and mRNA expression of TNF-α, IL-12,IFN-γ and IL-10 were determined by ELISA and Northern blotting, respectively. RESULTS: All tested bacteria induced TNF-α production. The best inducers of Thl type cytokines IL-12 and IFN-γ were Streptococcus and Leuconostoc strains. All Bifidobacteriurn and Propionibacteriurn strains induced higher IL-10 production than other studied bacteria. Stimulation of PBNC with any bacterial combinations did not result in enhanced cytokine production suggesting that different bacteria whether gram-positive or gram negative compete with each other during host cell interactions. CONCLUSION: The probiotic S. thermophilus and Leuconostoc strains are more potent inducers of Thl type cytokines IL-12 and IFN-γ than the probiotic Lactobacillus strains. Bacterial combinations did not result in enhanced cytokine production.

  20. Electron paramagnetic resonance studies of the soluble CuA protein from the cytochrome ba3 of Thermus thermophilus.

    Science.gov (United States)

    Karpefors, M; Slutter, C E; Fee, J A; Aasa, R; Källebring, B; Larsson, S; Vänngård, T

    1996-11-01

    The electron paramagnetic resonance (EPR) spectrum of the binuclear CuA center in the water-soluble subunit II fragment from cytochrome ba3 of Thermus thermophilus was recorded at 3.93, 9.45, and 34.03 GHz, and the EPR parameters were determined by computer simulations. The frequency and M1 dependence of the linewidth was discussed in terms of g strain superimposed on a correlation between the A and g values. The g values were found to be gx = 1.996, gy = 2.011, gz = 2.187, and the two Cu ions contribute nearly equally to the hyperfine structure, with magnitude of Ax magnitude of approximately 15 G, magnitude of Ay magnitude = 29 G, and magnitude of Az magnitude of = 28.5 G (65Cu). Theoretical CNDO/S calculations, based on the x-ray structure of the Paracoccus denitrificans enzyme, yield a singly occupied antibonding orbital in which each Cu is pi*-bonded to one S and sigma*-bonded to the other. In contrast to the equal spin distribution suggested by the EPR simulations, the calculated contributions from the Cu ions differ by a factor of 2. However, only small changes in the ligand geometry are needed to reproduce the experimental results.

  1. Enzymatic fragmentation of the antimicrobial peptides casocidin and isracidin by Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus.

    Science.gov (United States)

    Somkuti, George A; Paul, Moushumi

    2010-06-01

    The cumulative effect of peptidase and protease activities associated with cells of Streptococcus thermophilus (ST) and Lactobacillus delbrueckii subsp. bulgaricus (LB) was evaluated on the milk protein-based antimicrobial peptides casocidin and isracidin. Reaction mixtures of casocidin or isracidin and nonproliferating mid-log cells of these essential yogurt starter cultures were individually incubated for up to 4 h at pH 4.5 and 7.0, and samples removed at various time points were analyzed by reverse phase-high performance liquid chromatography (RP-HPLC) and MALDI-TOF/TOF-MS. Both casocidin and isracidin remained largely unchanged following exposure to cell suspensions of ST or LB strains at pH 4.5. Casocidin was extensively degraded by both ST and LB strains at pH 7.0, whereas isracidin remained largely intact after incubation for 4 h with ST strains but was degraded by exposure to LB strains. The results showed the feasibility of using the bovine casein-based peptides casocidin and isracidin as food grade antimicrobial supplements to impart fermented dairy foods additional protection against bacterial contamination. The structural integrity and efficacy of these biodefensive peptides may be preserved by timing their addition near the end of the fermentation of yogurt-like dairy foods (at or below pH 4.5), when conditions for bacterial proteolytic activity become unfavorable.

  2. Pulse Radiolysis Studies of Temperature Dependent Electron Transfers among Redox Centers in ba(3)-Cytochrome c Oxidase from Thermus thermophilus

    DEFF Research Database (Denmark)

    Farver, Ole; Wherland, Scot; Antholine, William E;

    2010-01-01

    The functioning of cytochrome c oxidases involves orchestration of long-range electron transfer (ET) events among the four redox active metal centers. We report the temperature dependence of electron transfer from the Cu(A)(r) site to the low-spin heme-(a)b(o) site, i.e., Cu(A)(r) + heme......-a(b)(o) → Cu(A)(o) + heme-a(b)(r) in three structurally characterized enzymes: A-type aa(3) from Paracoccus denitrificans (PDB code 3HB3 ) and bovine heart tissue (PDB code 2ZXW ), and the B-type ba(3) from T. thermophilus (PDB codes 1EHK and 1XME ). k,T data sets were obtained with the use of pulse radiolysis...... in cytochrome ba(3) had no effect on the rate of this reaction whereas the II-Met160Leu Cu(A)-mutation was slower by an amount corresponding to a decreased driving force of ∼0.06 eV. The structures support the presence of a common, electron-conducting "wire" between Cu(A) and heme-a(b). The transfer...

  3. Inactivation and unfolding of protein tyrosine phosphatase from Thermus thermophilus HB27 during urea and guanidine hydrochloride denaturation.

    Directory of Open Access Journals (Sweden)

    Yejing Wang

    Full Text Available The effects of urea and guanidine hydrochloride (GdnHCl on the activity, conformation and unfolding process of protein tyrosine phosphatase (PTPase, a thermostable low molecular weight protein from Thermus thermophilus HB27, have been studied. Enzymatic activity assays showed both urea and GdnHCl resulted in the inactivation of PTPase in a concentration and time-dependent manner. Inactivation kinetics analysis suggested that the inactivation of PTPase induced by urea and GdnHCl were both monophasic and reversible processes, and the effects of urea and GdnHCl on PTPase were similar to that of mixed-type reversible inhibitors. Far-ultraviolet (UV circular dichroism (CD, Tryptophan and 1-anilinonaphthalene -8-sulfonic acid (ANS fluorescence spectral analyses indicated the existence of a partially active and an inactive molten globule-like intermediate during the unfolding processes induced by urea and GdnHCl, respectively. Based on the sequence alignment and the homolog Tt1001 protein structure, we discussed the possible conformational transitions of PTPase induced by urea and GdnHCl and compared the conformations of these unfolding intermediates with the transient states in bovine PTPase and its complex structures in detail. Our results may be able to provide some valuable clues to reveal the relationship between the structure and enzymatic activity, and the unfolding pathway and mechanism of PTPase.

  4. Gene cloning and molecular characterization of the Talaromyces thermophilus lipase catalyzed efficient hydrolysis and synthesis of esters.

    Science.gov (United States)

    Romdhane, Ines Belhaj-Ben; Frikha, Fakher; Maalej-Achouri, Inès; Gargouri, Ali; Belghith, Hafedh

    2012-02-15

    A genomic bank from Talaromyces thermophilus fungus was constructed and screened using a previously isolated fragment lipase gene as probe. From several clones isolated, the nucleotide sequence of the lipase gene (TTL gene) was completed and sequenced. The TTL coding gene consists of an open reading frame (ORF) of 1083bp encoding a protein of 269 Aa with an estimated molecular mass of 30kDa. The TTL belongs to the same gene family as Thermomyces lanuginosus lipase (TLL, Lipolase®), a well known lipase with multiple applications. The promoter sequence of the TTL gene showed the conservation of known consensus sequences PacC, CreA, Hap2-3-4 and the existence of a particular sequence like the binding sites of Oleate Response Element (ORE) and Fatty acids Responsis Element (FARE) which are similar to that already found to be specific of lipolytic genes in Candida and Fusarium, respectively. Northern blot analysis showed that the TTL expression was much higher on wheat bran than on olive oil as sole carbon source. Compared to the Lipolase®, this enzyme was found to be more efficient for the hydrolysis and the synthesis of esters; and its synthetic efficiency even reached 91.6% from Waste Cooking Oil triglycerides.

  5. Deferribacter thermophilus gen. nov., sp. nov., a novel thermophilic manganese- and iron-reducing bacterium isolated from a petroleum reservoir.

    Science.gov (United States)

    Greene, A C; Patel, B K; Sheehy, A J

    1997-04-01

    A thermophilic anaerobic bacterium, designated strain BMAT (T = type strain), was isolated from the production water of Beatrice oil field in the North Sea (United Kingdom). The cells were straight to bent rods (1 to 5 by 0.3 to 0.5 microns) which stained gram negative. Strain BMAT obtained energy from the reduction of manganese (IV), iron(III), and nitrate in the presence of yeast extract, peptone, Casamino Acids, tryptone, hydrogen, malate, acetate, citrate, pyruvate, lactate, succinate, and valerate. The isolate grew optimally at 60 degrees C (temperature range for growth, 50 to 65 degrees C) and in the presence of 2% (wt/vol) NaCl (NaCl range for growth, 0 to 5% [wt/vol]). The DNA base composition was 34 mol% G + C. Phylogenetic analyses of the 16S rRNA gene indicated that strain BMAT is a member of the domain Bacteria. The closest known bacterium is the moderate thermophile Flexistipes sinusarabici (similarity value, 88%). Strain BMAT possesses phenotypic and phylogenetic traits that do not allow its classification as a member of any previously described genus; therefore, we propose that this isolate should be described as a member of a novel species of a new genus, Deferribacter thermophilus gen. nov., sp. nov.

  6. Galactose and Lactose Genes from the Galactose-Positive Bacterium Streptococcus salivarius and the Phylogenetically Related Galactose-Negative Bacterium Streptococcus thermophilus: Organization, Sequence, Transcription, and Activity of the gal Gene Products

    Science.gov (United States)

    Vaillancourt, Katy; Moineau, Sylvain; Frenette, Michel; Lessard, Christian; Vadeboncoeur, Christian

    2002-01-01

    Streptococcus salivarius is a lactose- and galactose-positive bacterium that is phylogenetically closely related to Streptococcus thermophilus, a bacterium that metabolizes lactose but not galactose. In this paper, we report a comparative characterization of the S. salivarius and S. thermophilus gal-lac gene clusters. The clusters have the same organization with the order galR (codes for a transcriptional regulator and is transcribed in the opposite direction), galK (galactokinase), galT (galactose-1-P uridylyltransferase), galE (UDP-glucose 4-epimerase), galM (galactose mutarotase), lacS (lactose transporter), and lacZ (β-galactosidase). An analysis of the nucleotide sequence as well as Northern blotting and primer extension experiments revealed the presence of four promoters located upstream from galR, the gal operon, galM, and the lac operon of S. salivarius. Putative promoters with virtually identical nucleotide sequences were found at the same positions in the S. thermophilus gal-lac gene cluster. An additional putative internal promoter at the 3′ end of galT was found in S. thermophilus but not in S. salivarius. The results clearly indicated that the gal-lac gene cluster was efficiently transcribed in both species. The Shine-Dalgarno sequences of galT and galE were identical in both species, whereas the ribosome binding site of S. thermophilus galK differed from that of S. salivarius by two nucleotides, suggesting that the S. thermophilus galK gene might be poorly translated. This was confirmed by measurements of enzyme activities. PMID:11790749

  7. Biotransformation of aflatoxin B1 and aflatoxin G1 in peanut meal by anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Chen, Yujie; Kong, Qing; Chi, Chen; Shan, Shihua; Guan, Bin

    2015-10-15

    The purpose of this study was to explore the ability of anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to biotransform aflatoxins in peanut meal. The pH of the peanut meal was adjusted above 10, and then heated for 10 min at 100 °C, 115 °C and 121 °C. The S. thermophilus and L. delbrueckii subsp. bulgaricus were precultured together in MRS broth for 48 h at 37 °C. The heated peanut meal was mixed with precultured MRS broth containing 7.0×10(8) CFU/mL of S. thermophilus and 3.0×10(3) CFU/mL of L. delbrueckii subsp. bulgaricus with the ratio of 1 to 1 (weight to volume) and incubated in anaerobic jars at 37 °C for 3 days. The aflatoxin content in the peanut meal samples was determined by HPLC. The results showed that the peanut meal contained mainly aflatoxin B1 (AFB1) (10.5±0.64 μg/kg) and aflatoxin G1 (AFG1) (18.7±0.55 μg/kg). When heat treatment was combined with anaerobic solid fermentation, the biotransformation rate of aflatoxins in peanut meal could attain 100%. The cytotoxicity of fermented peanut meal to L929 mouse connective tissue fibroblast cells was determined by MTT assay and no significant toxicity was observed in the fermented peanut meal. Furthermore, heat treatment and anaerobic solid fermentation did not change the amino acid concentrations and profile in peanut meal.

  8. Influence of different carbon sources on exopolysaccharide production by Lactobacillus delbrueckii subsp. bulgaricus (B3, G12 and Streptococcus thermophilus (W22

    Directory of Open Access Journals (Sweden)

    Zehra Nur Yuksekdag

    2008-06-01

    Full Text Available Exopolysaccharides (EPSs production was studied by Lactobacillus delbrueckii subsp. bulgaricus (B3, G12 and Streptococcus thermophilus (W22 in the medium containing various carbon sources (glucose, fructose, sucrose or lactose. For all the strains, glucose was the most efficient carbon source and B3, G12 and W22 strains produced 211, 175 and 120 EPS mg/L respectively. Also, the influence of different concentrations of glucose (5,10,15,20,25,30 g/L on EPS production and growth was studied. The results indicated that EPS production and growth were stimulated by the high glucose concentration (30 g/L.

  9. Quantitative analysis of the lactic acid and acetaldehyde produced by Streptococcus thermophilus and Lactobacillus bulgaricus strains isolated from traditional Turkish yogurts using HPLC.

    Science.gov (United States)

    Gezginc, Y; Topcal, F; Comertpay, S; Akyol, I

    2015-03-01

    The present study was conducted to evaluate the lactic acid- and acetaldehyde-producing abilities of lactic acid bacterial species isolated from traditionally manufactured Turkish yogurts using HPLC. The lactic acid bacterial species purified from the yogurts were the 2 most widely used species in industrial yogurt production: Streptococcus thermophilus and Lactobacillus bulgaricus. These bacteria have the ability to ferment hexose sugars homofermentatively to generate lactic acid and some carbonyl compounds, such as acetaldehyde through pyruvate metabolism. The levels of the compounds produced during fermentation influence the texture and the flavor of the yogurt and are themselves influenced by the chemical composition of the milk, processing conditions, and the metabolic activity of the starter culture. In the study, morphological, biochemical, and molecular characteristics were employed to identify the bacteria obtained from homemade yogurts produced in different regions of Turkey. A collection of 91 Strep. thermophilus and 35 L. bulgaricus strains were investigated for their lactic acid- and acetaldehyde-formation capabilities in various media such as cow milk, LM17 agar, and aerobic-anaerobic SM17 agar or de Man, Rogosa, and Sharpe agar. The amounts of the metabolites generated by each strain in all conditions were quantified by HPLC. The levels were found to vary depending on the species, the strain, and the growth conditions used. Whereas lactic acid production ranged between 0 and 77.9 mg/kg for Strep. thermophilus strains, it ranged from 0 to 103.5 mg/kg for L. bulgaricus. Correspondingly, the ability to generate acetaldehyde ranged from 0 to 105.9 mg/kg in Strep. thermophilus and from 0 to 126.9 mg/kg in L. bulgaricus. Our study constitutes the first attempt to determine characteristics of the wild strains isolated from traditional Turkish yogurts, and the approach presented here, which reveals the differences in metabolite production abilities of the

  10. Molecular identification of differentially regulated genes in the hydrothermal-vent species Bathymodiolus thermophilus and Paralvinella pandorae in response to temperature

    Directory of Open Access Journals (Sweden)

    Shillito Bruce

    2009-05-01

    Full Text Available Abstract Background Hydrothermal vents and cold seeps represent oases of life in the deep-sea environment, but are also characterized by challenging physical and chemical conditions. The effect of temperature fluctuations on vent organisms in their habitat has not been well explored, in particular at a molecular level, most gene expression studies being conducted on coastal marine species. In order to better understand the response of hydrothermal organisms to different temperature regimes, differentially expressed genes (obtained by a subtractive suppression hybridization approach were identified in the mussel Bathymodiolus thermophilus and the annelid Paralvinella pandorae irlandei to characterize the physiological processes involved when animals are subjected to long term exposure (2 days at two contrasting temperatures (10° versus 20°C, while maintained at in situ pressures. To avoid a potential effect of pressure, the experimental animals were initially thermally acclimated for 24 hours in a pressurized vessel. Results For each species, we produced two subtractive cDNA libraries (forward and reverse from sets of deep-sea mussels and annelids exposed together to a thermal challenge under pressure. RNA extracted from the gills, adductor muscle, mantle and foot tissue were used for B. thermophilus. For the annelid model, whole animals (small individuals were used. For each of the four libraries, we sequenced 200 clones, resulting in 78 and 83 unique sequences in mussels and annelids (about 20% of the sequencing effort, respectively, with only half of them corresponding to known genes. Real-time PCR was used to validate differentially expressed genes identified in the corresponding libraries. Strong expression variations have been observed for some specific genes such as the intracellular hemoglobin, the nidogen protein, and Rab7 in P. pandorae, and the SPARC protein, cyclophilin, foot protein and adhesive plaque protein in B. thermophilus

  11. ns-μs Time-Resolved Step-Scan FTIR of ba3 Oxidoreductase from Thermus thermophilus: Protonic Connectivity of w941-w946-w927

    Science.gov (United States)

    Nicolaides, Antonis; Soulimane, Tewfik; Varotsis, Constantinos

    2016-01-01

    Time-resolved step-scan FTIR spectroscopy has been employed to probe the dynamics of the ba3 oxidoreductase from Thermus thermophilus in the ns-μs time range and in the pH/pD 6–9 range. The data revealed a pH/pD sensitivity of the D372 residue and of the ring-A propionate of heme a3. Based on the observed transient changes a model in which the protonic connectivity of w941-w946-927 to the D372 and the ring-A propionate of heme a3 is described. PMID:27690021

  12. A highly conserved DNA replication module from Streptococcus thermophilus phages is similar in sequence and topology to a module from Lactococcus lactis phages.

    Science.gov (United States)

    Desiere, F; Lucchini, S; Bruttin, A; Zwahlen, M C; Brüssow, H

    1997-08-01

    A highly conserved DNA region extending over 5 kb was observed in Streptococcus thermophilus bacteriophages. Comparative sequencing of one temperate and 26 virulent phages demonstrated in the most extreme case an 18% aa difference for a predicted protein, while the majority of the phages showed fewer, if any aa changes. The relative degree of aa conservation was not homogeneous over the DNA segment investigated. Sequence analysis of the conserved segment revealed genes possibly involved in DNA transactions. Three predicted proteins (orf 233, 443, and 382 gene product (gp)) showed nucleoside triphosphate binding motifs. Orf 443 gp showed in addition a DEAH box motif, characteristically found in a subgroup of helicases, and a variant zinc finger motif known from a phage T7 helicase/primase. Tree analysis classified orf 443 gp as a distant member of the helicase superfamily. Orf 382 gp showed similarity to putative plasmid DNA primases. Downstream of orf 382 a noncoding repeat region was identified that showed similarity to a putative minus origin from a cryptic S. thermophilus plasmid. Four predicted proteins showed not only high degrees of aa identity (34 to 63%) with proteins from Lactococcus lactis phages, but their genes showed a similar topological organization. We interpret this as evidence for a horizontal gene transfer event between phages of the two bacterial genera in the distant past. PMID:9268169

  13. Purification, crystallization and preliminary X-ray diffraction of SecDF, a translocon-associated membrane protein, from Thermus thermophilus

    Energy Technology Data Exchange (ETDEWEB)

    Tsukazaki, Tomoya; Mori, Hiroyuki [Institute for Virus Research, Kyoto University, Kyoto 606-8507 (Japan); Fukai, Shuya; Numata, Tomoyuki [Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Kanagawa 226-8501 (Japan); Perederina, Anna [Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Schools of Medicine and Dentistry, Birmingham, Alabama 35294 (United States); Adachi, Hiroaki [SOSHO Inc., 7-7-15-208 Asagi, Saito, Ibaraki, Osaka 567-0085 (Japan); Department of Electrical, Electronic and Information Engineering, Osaka University, Osaka 565-0871 (Japan); CREST, JST, Saitama 332-0012 (Japan); Matsumura, Hiroyoshi [SOSHO Inc., 7-7-15-208 Asagi, Saito, Ibaraki, Osaka 567-0085 (Japan); CREST, JST, Saitama 332-0012 (Japan); Department of Materials Chemistry, Osaka University, Osaka 565-0871 (Japan); Takano, Kazufumi [SOSHO Inc., 7-7-15-208 Asagi, Saito, Ibaraki, Osaka 567-0085 (Japan); CREST, JST, Saitama 332-0012 (Japan); Department of Material and Life Science, Osaka University, Osaka 565-0871 (Japan); Murakami, Satoshi [SOSHO Inc., 7-7-15-208 Asagi, Saito, Ibaraki, Osaka 567-0085 (Japan); CREST, JST, Saitama 332-0012 (Japan); PRESTO, JST, Saitama 332-0012 (Japan); Institute of Scientific and Industrial Research, Osaka University, Osaka 567-0047 (Japan); Inoue, Tsuyoshi [SOSHO Inc., 7-7-15-208 Asagi, Saito, Ibaraki, Osaka 567-0085 (Japan); CREST, JST, Saitama 332-0012 (Japan); Department of Materials Chemistry, Osaka University, Osaka 565-0871 (Japan); Mori, Yusuke; Sasaki, Takatomo [SOSHO Inc., 7-7-15-208 Asagi, Saito, Ibaraki, Osaka 567-0085 (Japan); Department of Electrical, Electronic and Information Engineering, Osaka University, Osaka 565-0871 (Japan); CREST, JST, Saitama 332-0012 (Japan); Vassylyev, Dmitry G. [Department of Biochemistry and Molecular Genetics, University of Alabama at Birmingham, Schools of Medicine and Dentistry, Birmingham, Alabama 35294 (United States); Nureki, Osamu, E-mail: onureki@bio.titech.ac.jp [Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Kanagawa 226-8501 (Japan); PRESTO, JST, Saitama 332-0012 (Japan); Ito, Koreaki, E-mail: onureki@bio.titech.ac.jp [Institute for Virus Research, Kyoto University, Kyoto 606-8507 (Japan); CREST, JST, Saitama 332-0012 (Japan)

    2006-04-01

    SecDF is a multi-path membrane protein required for efficient protein translocation and integration via translocon. Purification and crystallization of T. thermophilus SecDF have been achieved by exploiting unique crystallization techniques that allowed the collection of a 3.74 Å data set. Thermus thermophilus has a multi-path membrane protein, TSecDF, as a single-chain homologue of Escherichia coli SecD and SecF, which form a translocon-associated complex required for efficient preprotein translocation and membrane-protein integration. Here, the cloning, expression in E. coli, purification and crystallization of TSecDF are reported. Overproduced TSecDF was solubilized with dodecylmaltoside, chromatographically purified and crystallized by vapour diffusion in the presence of polyethylene glycol. The crystals yielded a maximum resolution of 4.2 Å upon X-ray irradiation, revealing that they belonged to space group P4{sub 3}2{sub 1}2. Attempts were made to improve the diffraction quality of the crystals by combinations of micro-stirring, laser-light irradiation and dehydration, which led to the eventual collection of complete data sets at 3.74 Å resolution and preliminary success in the single-wavelength anomalous dispersion analysis. These results provide information that is essential for the determination of the three-dimensional structure of this important membrane component of the protein-translocation machinery.

  14. Potential role of Thermus thermophilus and T. oshimai in high rates of nitrous oxide (N2O) production in ∼80 °C hot springs in the US Great Basin.

    Science.gov (United States)

    Hedlund, B P; McDonald, A I; Lam, J; Dodsworth, J A; Brown, J R; Hungate, B A

    2011-11-01

    Ambient nitrous oxide (N(2)O) emissions from Great Boiling Spring (GBS) in the US Great Basin depended on temperature, with the highest flux, 67.8 ± 2.6 μmol N(2)O-N m(-2) day(-1) , occurring in the large source pool at 82 °C. This rate of N(2)O production contrasted with negligible production from nearby soils and was similar to rates from soils and sediments impacted with agricultural fertilizers. To investigate the source of N(2)O, a variety of approaches were used to enrich and isolate heterotrophic micro-organisms, and isolates were screened for nitrate reduction ability. Nitrate-respiring isolates were identified by 16S rRNA gene sequencing as Thermus thermophilus (31 isolates) and T. oshimai (three isolates). All isolates reduced nitrate to N(2)O but not to dinitrogen and were unable to grow with N(2)O as a terminal electron acceptor. Representative T. thermophilus and T. oshimai strains contained genes with 96-98% and 93% DNA identity, respectively, to the nitrate reductase catalytic subunit gene (narG) of T. thermophilus HB8. These data implicate T. thermophilus and T. oshimai in high flux of N(2)O in GBS and raise questions about the genetic basis of the incomplete denitrification pathway in these organisms and on the fate of biogenic N(2)O in geothermal environments.

  15. Water-soluble, recombinant CuA-domain of the cytochrome ba3 subunit II from Thermus thermophilus.

    Science.gov (United States)

    Slutter, C E; Sanders, D; Wittung, P; Malmström, B G; Aasa, R; Richards, J H; Gray, H B; Fee, J A

    1996-03-19

    Recently, the genes of cytochrome ba3 from thermus thermophilus [Keightley, J.A., et al. (1995) J. Biol. Chem. 270, 20345-20358], a homolog of the heme-copper oxidase family, have been cloned. We report here expression of a truncated gene, encoding the copper A (CuA) domain of cytochrome ba3, that is regulated by a T7 RNA polymerase promoter in Escherichia coli. The CuA-containing domain is purified in high yields as a water-soluble, thermostable, purple-colored protein. Copper analysis by chemical assay, mass spectrometry, X-ray fluorescence, and EPR spin quantification show that this protein contains two copper ions bound in a mixed-valence state, indicating that the CuA site in cytochrome ba3, is a binuclear center. The absorption spectrum of the CuA site, free of the heme interference in cytochrome ba3, is similar to the spectra of other soluble fragments from the aa3-type oxidase of Parachccus denitrificans [Lappalainen, P., et al. (1993) J. Biol Chem. 268, 26416-26421] and the caa3-type oxidase of Bacillus subtilis [von Wachenfeldt, C. et al. (1994) FEBS Lett. 340, 109-113]. There are intense bands at 480 nm (3100 M(-1) cm(-1)) and 530 nm (3200 M(-1) cm(-1)), a band in the near -IR centered at 790 nm (1900 M(-1) cn(-1)), and a weaker band at 363 nm (1300M(-1) cm(-1)). The visible CD spectrum shows a positive-going band at 460 nm and a negative-going band at 527 nm, the opposite signs of which may result from the binuclear nature of the site. The secondary structure prediction from the far-UV CD spectrum indicates that this domain is predominantly beta-sheet, in agreement with the recent X-ray structure reported for the complete P. denitrificans cytochrome aa3 molecule [Iwata, S., et al. (1995) Nature 376, 660-669] and the engineered, purple CyoA protein [Wilmanns, M., et al. (1996) Proc. Natl Acad. Sci. U.S.A. 92, 11955-11959]. However, the thermostability of the fragment described here (Tm approximately 80 degrees C) and the stable binding of copper over a

  16. Functional expression of a penicillin acylase from the extreme thermophile Thermus thermophilus HB27 in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Torres Leticia L

    2012-08-01

    Full Text Available Abstract Background Penicillin acylases (PACs are enzymes of industrial relevance in the manufacture of β-lactam antibiotics. Development of a PAC with a longer half-life under the reaction conditions used is essential for the improvement of the operational stability of the process. A gene encoding a homologue to Escherichia coli PAC was found in the genome of the thermophilic bacterium Thermus thermophilus (Tth HB27. Because of the nature of this PAC and its complex maturation that is crucial to reach its functional heterodimeric final conformation, the overexpression of this enzyme in a heterologous mesophilic host was a challenge. Here we describe the purification and characterization of the PAC protein from Tth HB27 overexpressed in Escherichia coli. Results Fusions to a superfolder green fluorescent protein and differential membrane solubilization assays indicated that the native enzyme remains attached through its amino-terminal end to the outer side of the cytoplasmic membrane of Tth cells. In order to overexpress this PAC in E. coli cells, a variant of the protein devoid of its membrane anchoring segment was constructed. The effect of the co-expression of chaperones and calcium supplementation of the culture medium was investigated. The total production of PAC was enhanced by the presence of DnaK/J and GrpE and even more by trigger factor and GroEL/ES. In addition, 10 mM calcium markedly improved both PAC specific and volumetric activities. Recombinant PAC was affinity-purified and proper maturation of the protein was confirmed by SDS-PAGE and MALDI-TOF analysis of the subunits. The recombinant protein was tested for activity towards several penicillins, cephalosporins and homoserine lactones. Hydrophobic acyl-chain penicillins were preferred over the rest of the substrates. Penicillin K (octanoyl penicillin was the best substrate, with the highest specificity constant value (16.12 mM-1.seg-1. The optimum pH was aprox. 4 and the optimum

  17. High resolution structure of the ba3 cytochrome c oxidase from Thermus thermophilus in a lipidic environment.

    Directory of Open Access Journals (Sweden)

    Theresa Tiefenbrunn

    Full Text Available The fundamental chemistry underpinning aerobic life on Earth involves reduction of dioxygen to water with concomitant proton translocation. This process is catalyzed by members of the heme-copper oxidase (HCO superfamily. Despite the availability of crystal structures for all types of HCO, the mode of action for this enzyme is not understood at the atomic level, namely how vectorial H(+ and e(- transport are coupled. Toward addressing this problem, we report wild type and A120F mutant structures of the ba(3-type cytochrome c oxidase from Thermus thermophilus at 1.8 Å resolution. The enzyme has been crystallized from the lipidic cubic phase, which mimics the biological membrane environment. The structures reveal 20 ordered lipid molecules that occupy binding sites on the protein surface or mediate crystal packing interfaces. The interior of the protein encloses 53 water molecules, including 3 trapped in the designated K-path of proton transfer and 8 in a cluster seen also in A-type enzymes that likely functions in egress of product water and proton translocation. The hydrophobic O(2-uptake channel, connecting the active site to the lipid bilayer, contains a single water molecule nearest the Cu(B atom but otherwise exhibits no residual electron density. The active site contains strong electron density for a pair of bonded atoms bridging the heme Fe(a3 and Cu(B atoms that is best modeled as peroxide. The structure of ba(3-oxidase reveals new information about the positioning of the enzyme within the membrane and the nature of its interactions with lipid molecules. The atomic resolution details provide insight into the mechanisms of electron transfer, oxygen diffusion into the active site, reduction of oxygen to water, and pumping of protons across the membrane. The development of a robust system for production of ba(3-oxidase crystals diffracting to high resolution, together with an established expression system for generating mutants, opens the

  18. 东太平洋海隆热液区嗜热深海偏顶蛤(Bathymodiolus thermophilus)的形态和分布特征%Characteristics of morphology and distribution about Bathymodiolus thermophilus from hydrothermal area of the EPR

    Institute of Scientific and Technical Information of China (English)

    王建佳; 林荣澄; 黄丁勇; 刘昕明; 牛文涛

    2013-01-01

    在2009-2010年的中国大洋第21航次环球科学考察中, “大洋一号”考察船对东太平洋海隆1.2°N周边新发现的“宝石山”热液区进行了综合调查,并利用我国自主研发的电视抓斗采集到了贻贝类样品,经形态学研究鉴定为嗜热深海偏顶蛤Bathymodiolus thermophilus,Kenk&Wilson,1985,这是此物种在该区域分布的新纪录.文章对样品进行了详细描述,并与其它海区发现的B.thermophilus进行了形态学上的比较和讨论.%During the 21st ocean survey of China during 2009-2010,the vessel " DA YANG YI HAO" made a comprehensive investigation in the hydrothermal area of the new discovered 'Precious Stone Mountain' surrounding 1.2 °N of the EPR,and it acquired some specimens of Mytilidae by use of the independently researched and developed TV-Grab.After morphological researches,we identified these specimens as Bathymodiolus thermophilus,Kenk & Wilson,1985.This is new record in the regional distribution of B.thermophilus.The article described the samples,and carried out some morphological comparations and discussions with this species found from other areas.

  19. EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), 2014. Scientific Opinion on the substantiation of a health claim related to beta-galactosidase from Streptococcus thermophilus and reduction of gastrointestinal discomfort pursuant to Article 14 of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    deliver an opinion on the scientific substantiation of a health claim related to beta-galactosidase from Streptococcus thermophilus and reduction of gastrointestinal discomfort. According to the applicant, the food constituent which is the subject of the claim is beta-galactosidase from Streptococcus...... thermophilus. Streptococcus thermophilus is added to infant formulae in the production process for fermentation purposes and the live organisms are inactivated after the fermentation process. The Panel considers that the food constituent, beta-galactosidase from Streptococcus thermophilus, under the conditions...... of use proposed by the applicant, is sufficiently characterised. The claimed effect proposed by the applicant is “reduction of gastro-intestinal discomfort”. The target population proposed by the applicant is infants and young children. The Panel considers that reduction of gastrointestinal...

  20. Interaction of Thermus thermophilus, ArsC enzyme and gold nanoparticles naked-eye assays speciation between As(III) and As(V)

    Science.gov (United States)

    Politi, Jane; Spadavecchia, Jolanda; Fiorentino, Gabriella; Antonucci, Immacolata; Casale, Sandra; De Stefano, Luca

    2015-10-01

    The thermophilic bacterium Thermus thermophilus HB27 encodes chromosomal arsenate reductase (TtArsC), the enzyme responsible for resistance to the harmful effects of arsenic. We report on adsorption of TtArsC onto gold nanoparticles for naked-eye monitoring of biomolecular interaction between the enzyme and arsenic species. Synthesis of hybrid biological-metallic nanoparticles has been characterized by transmission electron microscopy (TEM), ultraviolet-visible (UV-vis), dynamic light scattering (DLS) and phase modulated infrared reflection absorption (PM-IRRAS) spectroscopies. Molecular interactions have been monitored by UV-vis and Fourier transform-surface plasmon resonance (FT-SPR). Due to the nanoparticles’ aggregation on exposure to metal salts, pentavalent and trivalent arsenic solutions can be clearly distinguished by naked-eye assay, even at 85 μM concentration. Moreover, the assay shows partial selectivity against other heavy metals.

  1. Mutations in conserved helix 69 of 23S rRNA of Thermus thermophilus that affect capreomycin resistance but not posttranscriptional modifications

    DEFF Research Database (Denmark)

    Monshupanee, Tanakarn; Gregory, Steven T; Douthwaite, Stephen;

    2008-01-01

    for resistance to the tuberactinomycin antibiotic capreomycin. Two base substitutions, A1913U and mU1915G, and a single base deletion, DeltamU1915, were identified in helix 69 of 23S rRNA, a structural element that forms part of an interribosomal subunit bridge with the decoding center of 16S rRNA, the site...... to capreomycin. Finally, none of the mutations in helix 69 interferes with methylation at C1920 or with pseudouridylation at positions 1911 and 1917. We conclude that the resistance phenotype is a consequence of structural changes introduced by the mutations....... of previously reported capreomycin resistance base substitutions. Capreomycin resistance in other bacteria has been shown to result from inactivation of the TlyA methyltransferase which 2'-O methylates C1920 of 23S rRNA. Inactivation of the tlyA gene in T. thermophilus does not affect its sensitivity...

  2. Sugar Utilization and Acid Production by Free and Entrapped Cells of Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, and Lactococcus lactis subsp. lactis in a Whey Permeate Medium

    Science.gov (United States)

    Audet, Pascal; Paquin, Celine; Lacroix, Christophe

    1989-01-01

    Cells of Streptococcus salivarius subsp. thermophilus and Lactococcus lactis subsp. lactis entrapped in k-carrageenan-locust bean gum gel performed similarly to free cells in the conversion of lactose to lactic acid. Bead diameter influenced the fermentation rate. Cells entrapped in smaller beads (0.5 to 1.0 mm) showed higher release rates, higher lactose, glucose, and formic acid utilization, higher galactose accumulation, and higher lactic acid production than did cells entrapped in larger beads (1.0 to 2.0 mm). Values for smaller beads were comparable with those for free cells. Immobilization affected the fermentation rate of lactic acid bacteria, especially Lactobacillus delbrueckii subsp. bulgaricus. Entrapped cells of L. delbrueckii subsp. bulgaricus demonstrated a lower lactic acid production than did free cells in batch fermentation. The kinetics of the production of formic and pyruvic acids by L. lactis subsp. lactis and S. salivarius subsp. thermophilus are presented. PMID:16347822

  3. Study on the effect of melamine on the growth of Streptococcus thermophilus%三聚氰胺对嗜热链球菌生长影响的研究

    Institute of Scientific and Technical Information of China (English)

    郭丽琼; 田雪琴; 苏霞; 吴厚玖; 曹秋旭; 董文博

    2012-01-01

    通过在MRS液体培养基里分别添加0‰、0.25‰、0.5‰、1‰、2‰、3‰及饱和的三聚氰胺,接种嗜热链球菌,在42℃下培养48h,通过测定培养液的pH变化和计数细菌总数来考察三聚氰胺对嗜热链球菌生长的影响。结果表明:嗜热链球菌不能以三聚氰胺为氮源物质而加以降解利用,三聚氰胺在水溶解度范围内对嗜热链球菌的生长没有促进作用,只有微弱的抑制效果。%MRS liquid mediums added respectively with 0‰,0.25‰,0.5‰,1‰,2‰,3‰ and saturation concentration melamine were used to cultivate Streptococcus thermophilus.After 48h at 42℃ in the culture medium,the impaction of melamine on the growth of Streptococcus thermophilus was tested through measuring the pH of the medium and calculating the final total number of bacteria.The experimental results showed that melamine couldn't be used and degraded by Streptococcus thermophilus as a nitrogen and melamine within the water solubility couldn't promote the growth of Streptococcus thermophilus while it had a little inhibition effect on the growth of it.

  4. Structure of a conserved hypothetical protein, TTHA0849 from Thermus thermophilus HB8, at 2.4 Å resolution: a putative member of the StAR-related lipid-transfer (START) domain superfamily

    International Nuclear Information System (INIS)

    The crystal structure of a conserved hypothetical protein, TTHA0849 from T. thermophilus HB8, has been determined at 2.4 Å resolution. The compact α+β structure shows the typical folding of the steroidogenic acute regulatory-related lipid-transfer (START) domain. The crystal structure of a conserved hypothetical protein, TTHA0849 from Thermus thermophilus HB8, has been determined at 2.4 Å resolution as a part of a structural and functional genomics project on T. thermophilus HB8. The main-chain folding shows a compact α+β motif, forming a hydrophobic cavity in the molecule. A structural similarity search reveals that it resembles those steroidogenic acute regulatory proteins that contain the lipid-transfer (START) domain, even though TTHA0849 shows comparatively weak sequence identity to polyketide cyclases. However, the size of the ligand-binding cavity is distinctly smaller than other START domain-containing proteins, suggesting that it catalyses the transfer of smaller ligand molecules

  5. Use of the dynamic gastro-intestinal model TIM to explore the survival of the yogurt bacterium Streptococcus thermophilus and the metabolic activities induced in the simulated human gut.

    Science.gov (United States)

    Uriot, Ophélie; Galia, Wessam; Awussi, Ahoefa Ablavi; Perrin, Clarisse; Denis, Sylvain; Chalancon, Sandrine; Lorson, Emilie; Poirson, Chantal; Junjua, Maira; Le Roux, Yves; Alric, Monique; Dary, Annie; Blanquet-Diot, Stéphanie; Roussel, Yvonne

    2016-02-01

    Streptococcus thermophilus, a lactic acid bacterium used to produce yogurts and cheeses is more and more considered for its potential probiotic properties. This implies that additional information should be obtained regarding its survival and metabolic activity in the human Gastro-Intestinal Tract (GIT). In this study, we screened 30 S. thermophilus strains for urease, small heat shock protein, and amino-acid decarboxylase functions which may play a role in survival in the upper part of the GIT. The survival kinetics of 4 strains was investigated using the TIM, a physiologically relevant in vitro dynamic gastric and small intestinal model. The three strains LMD9, PB18O and EBLST20 showed significantly higher survival than CNRZ21 in all digestive compartments of the TIM, which may be related to the presence of urease and heat shock protein functions. When LMD9 bacterial cells were delivered in a fermented milk formula, a significant improvement of survival in the TIM was observed compared to non-fermented milk. With the RIVET (Recombinase In Vivo Expression Technology) method applied to the LMD9 strain, a promoter located upstream of hisS, responsible for the histidyl-transfer RNA synthesis, was found to be specifically activated in the artificial stomach. The data generated on S. thermophilus survival and its adaptation capacities to the digestive tract are essential to establish a list of biomarkers useful for the selection of probiotic strains.

  6. Characteristics of third-generation glucose biosensors based on Corynascus thermophilus cellobiose dehydrogenase immobilized on commercially available screen-printed electrodes working under physiological conditions.

    Science.gov (United States)

    Zafar, Muhammad Nadeem; Safina, Gulnara; Ludwig, Roland; Gorton, Lo

    2012-06-01

    In this article, we describe a third-generation amperometric glucose biosensor working under physiological conditions. This glucose biosensor consists of a recently discovered cellobiose dehydrogenase from the ascomycete Corynascus thermophilus (CtCDH) immobilized on different commercially available screen-printed electrodes made of carbon (SPCEs), carboxyl-functionalized single-walled carbon nanotubes (SPCE-SWCNTs), or multiwalled carbon nanotubes (SPCE-MWCNTs) by simple physical adsorption or a combination of adsorption followed by cross-linking using poly(ethyleneglycol) (400) diglycidyl ether (PEGDGE) or glutaraldehyde (GA). The CtCDH-based third-generation glucose biosensor has a linear range between 0.025 and 30 mM and a detection limit of 10 μM glucose. Biosensors based on SWCNTs showed a higher sensitivity and catalytic response than the ones functionalized with MWCNTs and the SPCEs. A drastic increase in response was observed for all three electrodes when the adsorbed enzyme was cross-linked with PEGDGE or GA. The operational stability of the biosensor was tested for 7 h by repeated injections of 50 mM glucose, and only a slight decrease in the electrochemical response was found. The selectivity of the CtCDH-based biosensor was tested on other potentially interfering carbohydrates such as mannose, galactose, sucrose, and fucose that might be present in blood. No significant analytical response from any of these compounds was observed. PMID:22381371

  7. Comparison of the backbone dynamics of wild-type Hydrogenobacter thermophilus cytochrome c{sub 552} and its b-type variant

    Energy Technology Data Exchange (ETDEWEB)

    Tozawa, Kaeko; Ferguson, Stuart J.; Redfield, Christina, E-mail: christina.redfield@bioch.ox.ac.uk [University of Oxford, Department of Biochemistry (United Kingdom); Smith, Lorna J., E-mail: lorna.smith@chem.ox.ac.uk [University of Oxford, Department of Chemistry (United Kingdom)

    2015-06-15

    Cytochrome c{sub 552} from the thermophilic bacterium Hydrogenobacter thermophilus is a typical c-type cytochrome which binds heme covalently via two thioether bonds between the two heme vinyl groups and two cysteine thiol groups in a CXXCH sequence motif. This protein was converted to a b-type cytochrome by substitution of the two cysteine residues by alanines (Tomlinson and Ferguson in Proc Natl Acad Sci USA 97:5156–5160, 2000a). To probe the significance of the covalent attachment of the heme in the c-type protein, {sup 15}N relaxation and hydrogen exchange studies have been performed for the wild-type and b-type proteins. The two variants share very similar backbone dynamic properties, both proteins showing high {sup 15}N order parameters in the four main helices, with reduced values in an exposed loop region (residues 18–21), and at the C-terminal residue Lys80. Some subtle changes in chemical shift and hydrogen exchange protection are seen between the wild-type and b-type variant proteins, not only for residues at and neighbouring the mutation sites, but also for some residues in the heme binding pocket. Overall, the results suggest that the main role of the covalent linkages between the heme group and the protein chain must be to increase the stability of the protein.

  8. Characterization of DNA polymerase X from Thermus thermophilus HB8 reveals the POLXc and PHP domains are both required for 3'-5' exonuclease activity.

    Science.gov (United States)

    Nakane, Shuhei; Nakagawa, Noriko; Kuramitsu, Seiki; Masui, Ryoji

    2009-04-01

    The X-family DNA polymerases (PolXs) comprise a highly conserved DNA polymerase family found in all kingdoms. Mammalian PolXs are known to be involved in several DNA-processing pathways including repair, but the cellular functions of bacterial PolXs are less known. Many bacterial PolXs have a polymerase and histidinol phosphatase (PHP) domain at their C-termini in addition to a PolX core (POLXc) domain, and possess 3'-5' exonuclease activity. Although both domains are highly conserved in bacteria, their molecular functions, especially for a PHP domain, are unknown. We found Thermus thermophilus HB8 PolX (ttPolX) has Mg(2+)/Mn(2+)-dependent DNA/RNA polymerase, Mn(2+)-dependent 3'-5' exonuclease and DNA-binding activities. We identified the domains of ttPolX by limited proteolysis and characterized their biochemical activities. The POLXc domain was responsible for the polymerase and DNA-binding activities but exonuclease activity was not detected for either domain. However, the POLXc and PHP domains interacted with each other and a mixture of the two domains had Mn(2+)-dependent 3'-5' exonuclease activity. Moreover, site-directed mutagenesis revealed catalytically important residues in the PHP domain for the 3'-5' exonuclease activity. Our findings provide a molecular insight into the functional domain organization of bacterial PolXs, especially the requirement of the PHP domain for 3'-5' exonuclease activity.

  9. Improved Growth of Lactobacillus bulgaricus and Streptococcus thermophilus as well as Increased Antioxidant Activity by Biotransforming Litchi Pericarp Polysaccharide with Aspergillus awamori

    Directory of Open Access Journals (Sweden)

    Sen Lin

    2013-01-01

    Full Text Available This study was conducted to increase the bioactivity of litchi pericarp polysaccharides (LPPs biotransformed by Aspergillus awamori. Compared to the non-A. awamori-fermented LPP, the growth effects of A. awamori-fermented LPP on Lactobacillus bulgaricus and Streptococcus thermophilus were four and two times higher after 3 days of fermentation, respectively. Increased 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and DNA protection activity of litchi pericarp polysaccharides were also achieved after A. awamori fermentation. Moreover, the relative content of glucose and arabinose in LPP after fermentation decreased from 58.82% to 22.60% and from 18.82% to 10.09%, respectively, with a concomitant increase in the relative contents of galactose, rhamnose, xylose, and mannose. Furthermore, lower molecular weight polysaccharides were obtained after A. awamori fermentation. It can be concluded that A. awamori was effective in biotransforming LPP into a bioactive mixture with lower molecular weight polysaccharides and higher antioxidant activity and relative galactose content.

  10. Comparative Analysis of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) of Streptococcus thermophilus St-I and its Bacteriophage-Insensitive Mutants (BIM) Derivatives.

    Science.gov (United States)

    Li, Wan; Bian, Xin; Evivie, Smith Etareri; Huo, Gui-Cheng

    2016-09-01

    The CRISPR-Cas (CRISPR together with CRISPR-associated proteins) modules are the adaptive immune system, acting as an adaptive and heritable immune system in bacteria and archaea. CRISPR-based immunity acts by integrating short virus sequences in the cell's CRISPR locus, allowing the cell to remember, recognize, and clear infections. In this study, the homology of CRISPRs sequence in BIMs (bacteriophage-insensitive mutants) of Streptococcus thermophilus St-I were analyzed. Secondary structures of the repeats and the PAMs (protospacer-associated motif) of each CRISPR locus were also predicted. Results showed that CRISPR1 has 27 repeat-spacer units, 5 of them had duplicates; CRISPR2 has one repeat-spacer unit; CRISPR3 has 28 repeat-spacer units. Only BIM1 had a new spacer acquisition in CRISPR3, while BIM2 and BIM3 had no new spacers' insertion, thus indicating that while most CRISPR1 were more active than CRISPR3, new spacer acquisition occurred just in CRSPR3 in some situations. These findings will help establish the foundation for the study of CRSPR-Cas systems in lactic acid bacteria.

  11. A Flexible Domain-Domain Hinge Promotes an Induced-fit Dominant Mechanism for the Loading of Guide-DNA into Argonaute Protein in Thermus thermophilus

    KAUST Repository

    Zhu, Lizhe

    2016-02-24

    Argonaute proteins (Ago) are core components of the RNA Induced Silencing Complex (RISC) that load and utilize small guide nucleic acids to silence mRNAs or cleave foreign DNAs. Despite the essential role of Ago in gene regulation and defense against virus, the molecular mechanism of guide-strand loading into Ago remains unclear. We explore such a mechanism in the bacterium Thermus thermophilus Ago (TtAgo), via a computational approach combining molecular dynamics, bias-exchange metadynamics, and protein-DNA docking. We show that apo TtAgo adopts multiple closed states that are unable to accommodate guide-DNA. Conformations able to accommodate the guide are beyond the reach of thermal fluctuations from the closed states. These results suggest an induced-fit dominant mechanism for guide-strand loading in TtAgo, drastically different from the two-step mechanism for human Ago 2 (hAgo2) identified in our previous study. Such a difference between TtAgo and hAgo2 is found to mainly originate from the distinct rigidity of their L1-PAZ hinge. Further comparison among known Ago structures from various species indicates that the L1-PAZ hinge may be flexible in general for prokaryotic Agos but rigid for eukaryotic Agos. © 2016 American Chemical Society.

  12. Comparative Analysis of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) of Streptococcus thermophilus St-I and its Bacteriophage-Insensitive Mutants (BIM) Derivatives.

    Science.gov (United States)

    Li, Wan; Bian, Xin; Evivie, Smith Etareri; Huo, Gui-Cheng

    2016-09-01

    The CRISPR-Cas (CRISPR together with CRISPR-associated proteins) modules are the adaptive immune system, acting as an adaptive and heritable immune system in bacteria and archaea. CRISPR-based immunity acts by integrating short virus sequences in the cell's CRISPR locus, allowing the cell to remember, recognize, and clear infections. In this study, the homology of CRISPRs sequence in BIMs (bacteriophage-insensitive mutants) of Streptococcus thermophilus St-I were analyzed. Secondary structures of the repeats and the PAMs (protospacer-associated motif) of each CRISPR locus were also predicted. Results showed that CRISPR1 has 27 repeat-spacer units, 5 of them had duplicates; CRISPR2 has one repeat-spacer unit; CRISPR3 has 28 repeat-spacer units. Only BIM1 had a new spacer acquisition in CRISPR3, while BIM2 and BIM3 had no new spacers' insertion, thus indicating that while most CRISPR1 were more active than CRISPR3, new spacer acquisition occurred just in CRSPR3 in some situations. These findings will help establish the foundation for the study of CRSPR-Cas systems in lactic acid bacteria. PMID:27378131

  13. Structural basis for the hydrolysis of ATP by a nucleotide binding subunit of an amino acid ABC transporter from Thermus thermophilus.

    Science.gov (United States)

    Devi, Seenivasan Karthiga; Chichili, Vishnu Priyanka Reddy; Jeyakanthan, J; Velmurugan, D; Sivaraman, J

    2015-06-01

    ATP-binding cassette (ABC) transporters are a major family of small molecule transporter proteins, and their deregulation is associated with several diseases, including cancer. Here, we report the crystal structure of the nucleotide binding domain (NBD) of an amino acid ABC transporter from Thermus thermophilus (TTHA1159) in its apo form and as a complex with ADP along with functional studies. TTHA1159 is a putative arginine ABC transporter. The apo-TTHA1159 was crystallized in dimeric form, a hitherto unreported form of an apo NBD. Structural comparison of the apo and ADP-Mg(2+) complexes revealed that Phe14 of TTHA1159 undergoes a significant conformational change to accommodate ADP, and that the bound ADP interacts with the P-loop (Gly40-Thr45). Modeling of ATP-Mg(2+):TTHA1159 complex revealed that Gln86 and Glu164 are involved in water-mediated hydrogen bonding contacts and Asp163 in Mg(2+) ion-mediated hydrogen bonding contacts with the γ-phosphate of ATP, consistent with the findings of other ABC transporters. Mutational studies confirmed the necessity of each of these residues, and a comparison of the apo/ADP Mg(2+):TTHA1159 with its ATP-complex model suggests the likelihood of a key conformational change to the Gln86 side chain for ATP hydrolysis. PMID:25916755

  14. Improved Helicobacter pylori Eradication Rate of Tailored Triple Therapy by Adding Lactobacillus delbrueckii and Streptococcus thermophilus in Northeast Region of Thailand: A Prospective Randomized Controlled Clinical Trial

    Directory of Open Access Journals (Sweden)

    Taweesak Tongtawee

    2015-01-01

    Full Text Available Background and Aim. To evaluate the effect of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus to Helicobacter pylori eradication in different periods of therapeutic protocol. Methods. Infected patients were randomized to one-week tailored triple therapy (esomeprazole 20 mg bid, clarithromycin 500 mg bid/metronidazole 400 mg tid if clarithromycin resistant, and amoxicillin 1000 mg bid with placebo (group 1, n=100; one week of pretreatment with probiotics (group 2, n=100; and one week of pretreatment with probiotic followed by one week of the same probiotics after treatment (group 3, n=100. Result. PP analysis involved 292 patients, 98 in group 1, 97 in group 2, and 97 in group 3. Successful eradication was observed in 229 patients; by PP analysis, the eradication rates were significantly higher (P<0.01, 95% CI; 0.71–0.97 in group 2 and group 3 than group 1. ITT analysis eradication rates were significantly higher in group 2 and group 3 than group 1 (P<0.01 95% CI; 0.72–0.87, and there is no significant difference between the three groups (P=0.32 in terms of adverse events. Conclusion. Adding probiotics before or before and after tailored treatment can improve Helicobacter pylori eradication rates. This trial is registered with Thai Clinical Trials Registry number: TCTR20141209001.

  15. Discovery and characterization of RecA protein of thermophilic bacterium Thermus thermophilus MAT72 phage Tt72 that increases specificity of a PCR-based DNA amplification.

    Science.gov (United States)

    Stefanska, Aleksandra; Kaczorowska, Anna-Karina; Plotka, Magdalena; Fridjonsson, Olafur H; Hreggvidsson, Gudmundur O; Hjorleifsdottir, Sigridur; Kristjansson, Jakob K; Dabrowski, Slawomir; Kaczorowski, Tadeusz

    2014-07-20

    The recA gene of newly discovered Thermus thermophilus MAT72 phage Tt72 (Myoviridae) was cloned and overexpressed in Escherichia coli. The 1020-bp gene codes for a 339-amino-acid polypeptide with an Mr of 38,155 which shows 38.7% positional identity to the E. coli RecA protein. When expressed in E. coli, the Tt72 recA gene did not confer the ability to complement the ultraviolet light (254nm) sensitivity of an E. coli recA mutant. Tt72 RecA protein has been purified with good yield to catalytic and electrophoretic homogeneity using a three-step chromatography procedure. Biochemical characterization indicated that the protein can pair and promote ATP-dependent strand exchange reaction resulting in formation of a heteroduplex DNA at 60°C under conditions otherwise optimal for E. coli RecA. When the Tt72 RecA protein was included in a standard PCR-based DNA amplification reaction, the specificity of the PCR assays was significantly improved by eliminating non-specific products.

  16. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium logum LA 10, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing

    DEFF Research Database (Denmark)

    Tetens, Inge

    substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and reducing intestinal discomfort. The food that is the subject of the health claim is a combination of B. longum LA 101, L...... and effect relationship cannot be established between the consumption of a combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103, and S. thermophilus LA 104 and reducing intestinal discomfort. © European Food Safety Authority, 2013...

  17. On the Thermus thermophilus HB8 potential pathogenicity triggered from rhamnolipids secretion: morphological alterations and cytotoxicity induced on fibroblastic cell line.

    Science.gov (United States)

    Pantazaki, A A; Choli-Papadopoulou, T

    2012-05-01

    A limited number of bacterial strains usually grown under nutrient limitation secrete rhamnolipids (RLs), which are recorded as virulence factors that are implicated in the pathogenicity of a microorganism. The non-pathogenic T. thermophilus HB8 produces extracellular rhamnolipids (TthRLs) under defined cultivation conditions using sunflower seed oil and sodium gluconate as carbon sources. In particular, the secreted TthRLs have been isolated, purified and identified with ATR-FTIR. Their effects on the cells' viability were examined when they were supplemented in a culture of human skin fibroblasts. Purified TthRLs triggered a sequence of rapid and pronounced morphological alterations characterized by transformation of fibroblast shape from polygonal to fusiform; retraction with cytoplasm condensation, rounding up, distortion of nuclei and loss of lamellar processes, and finally disruption of membrane. The addition of TthRLs in the cultured fibroblasts caused cytotoxicity, in contrast to that of rhamnose that stimulated viability, as it was assessed by MTT test. These results revealed that among the constituents of RLs that are implicated in the cytotoxicity, it has to be attributed to the lipidic chain variation and not to the carbohydrate part. TthRLs cytotoxicity on fibroblasts is comparable, and provoked similar effects, to that caused by saponin white, a known surfactant. TthRLs secretion might be a crucial point for the transformation of a non-pathogenic bacterium to a pathogenic one under certain environmental conditions favoring their secretion. RLs secretion in the microorganism's world might be a general route for the passage in the pathogenicity to ensure their survival under nutrient limitation conditions. PMID:21611776

  18. Deciphering the Dynamics of Non-Covalent Interactions Affecting Thermal Stability of a Protein: Molecular Dynamics Study on Point Mutant of Thermus thermophilus Isopropylmalate Dehydrogenase.

    Science.gov (United States)

    Sharma, Reetu; Sastry, G Narahari

    2015-01-01

    Thermus thermophilius isopropylmalate dehydrogenase catalyzes oxidative decarboxylation and dehydrogenation of isopropylmalate. Substitution of leucine to alanine at position 172 enhances the thermal stability among the known point mutants. Exploring the dynamic properties of non-covalent interactions such as saltbridges, hydrogen bonds and hydrophobic interactions to explain thermal stability of a protein is interesting in its own right. In this study dynamic changes in the non-covalent interactions are studied to decipher the deterministic features of thermal stability of a protein considering a case study of a point mutant in Thermus thermophilus isopropylmalate dehydrogenase. A total of four molecular dynamic simulations of 0.2 μs were carried out on wild type and mutant's functional dimers at 300 K and 337 K. Higher thermal stability of the mutant as compared to wild type is revealed by root mean square deviation, root mean square fluctuations and Cα-Cα distance with an increase in temperature from 300 K to 337 K. Most of the regions of wild type fluctuate higher than the corresponding regions of mutant with an increase in temperature. Cα-Cα distance analysis suggests that long distance networks are significantly affected in wild type as compared to the mutant. Short lived contacts are higher in wild type, while long lived contacts are lost at 337 K. The mutant forms less hydrogen bonds with water as compared to wild type at 337 K. In contrast to wild type, the mutant shows significant increase in unique saltbridges, hydrogen bonds and hydrophobic contacts at 337 K. The current study indicates that there is a strong inter-dependence of thermal stability on the way in which non-covalent interactions reorganize, and it is rewarding to explore this connection in single mutant studies.

  19. Deciphering the Dynamics of Non-Covalent Interactions Affecting Thermal Stability of a Protein: Molecular Dynamics Study on Point Mutant of Thermus thermophilus Isopropylmalate Dehydrogenase.

    Directory of Open Access Journals (Sweden)

    Reetu Sharma

    Full Text Available Thermus thermophilius isopropylmalate dehydrogenase catalyzes oxidative decarboxylation and dehydrogenation of isopropylmalate. Substitution of leucine to alanine at position 172 enhances the thermal stability among the known point mutants. Exploring the dynamic properties of non-covalent interactions such as saltbridges, hydrogen bonds and hydrophobic interactions to explain thermal stability of a protein is interesting in its own right. In this study dynamic changes in the non-covalent interactions are studied to decipher the deterministic features of thermal stability of a protein considering a case study of a point mutant in Thermus thermophilus isopropylmalate dehydrogenase. A total of four molecular dynamic simulations of 0.2 μs were carried out on wild type and mutant's functional dimers at 300 K and 337 K. Higher thermal stability of the mutant as compared to wild type is revealed by root mean square deviation, root mean square fluctuations and Cα-Cα distance with an increase in temperature from 300 K to 337 K. Most of the regions of wild type fluctuate higher than the corresponding regions of mutant with an increase in temperature. Cα-Cα distance analysis suggests that long distance networks are significantly affected in wild type as compared to the mutant. Short lived contacts are higher in wild type, while long lived contacts are lost at 337 K. The mutant forms less hydrogen bonds with water as compared to wild type at 337 K. In contrast to wild type, the mutant shows significant increase in unique saltbridges, hydrogen bonds and hydrophobic contacts at 337 K. The current study indicates that there is a strong inter-dependence of thermal stability on the way in which non-covalent interactions reorganize, and it is rewarding to explore this connection in single mutant studies.

  20. Crystal structures of a subunit of the formylglycinamide ribonucleotide amidotransferase, PurS, from Thermus thermophilus , Sulfolobus tokodaii and Methanocaldococcus jannaschii

    Energy Technology Data Exchange (ETDEWEB)

    Watanabe, Yuzo; Yanai, Hisaaki; Kanagawa, Mayumi; Suzuki, Sakiko; Tamura, Satoko; Okada, Kiyoshi; Baba, Seiki; Kumasaka, Takashi; Agari, Yoshihiro; Chen, Lirong; Fu, Zheng-Qing; Chrzas, John; Wang, Bi-Cheng; Nakagawa, Noriko; Ebihara, Akio; Masui, Ryoji; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Sampei, Gen-ichi; Kawai, Gota

    2016-07-27

    The crystal structures of a subunit of the formylglycinamide ribonucleotide amidotransferase, PurS, fromThermus thermophilus,Sulfolobus tokodaiiandMethanocaldococcus jannaschiiwere determined and their structural characteristics were analyzed. For PurS fromT. thermophilus, two structures were determined using two crystals that were grown in different conditions. The four structures in the dimeric form were almost identical to one another despite their relatively low sequence identities. This is also true for all PurS structures determined to date. A few residues were conserved among PurSs and these are located at the interaction site with PurL and PurQ, the other subunits of the formylglycinamide ribonucleotide amidotransferase. Molecular-dynamics simulations of the PurS dimer as well as a model of the complex of the PurS dimer, PurL and PurQ suggest that PurS plays some role in the catalysis of the enzyme by its bending motion.

  1. Development of a pentaplex PCR assay for the simultaneous detection of Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, L. delbrueckii subsp. lactis, L. helveticus, L. fermentum in whey starter for Grana Padano cheese.

    Science.gov (United States)

    Cremonesi, Paola; Vanoni, Laura; Morandi, Stefano; Silvetti, Tiziana; Castiglioni, Bianca; Brasca, Milena

    2011-03-30

    A pentaplex PCR assay for the rapid, selective and simultaneous detection of Lactobacillus helveticus, L. delbrueckii subsp. lactis, L. delbrueckii subsp. bulgaricus, Streptococcus thermophilus, and L. fermentum, was developed. The target sequences were a group of genes coding for beta-galactosidase production (S. thermophilus and L. delbrueckii subsp. bulgaricus), for cell-enveloped associated proteinase synthesis (L. helveticus), for dipeptide transport system production (L. delbrueckii subsp. lactis) and for arginine-ornithine antiporter protein production (L. fermentum). The analytical specificity of the assay was evaluated with 5 reference strains and 140 lactic acid bacterial strains derived from raw milk cheeses and belonging to the Lactobacillus, Streptococcus, Lactococcus and Enterococcus genera. The identification limit for each target strain was 10(3)CFU/ml. This new molecular assay was used to investigate the LAB population by direct extraction of DNA from the 12 whey cultures for Grana Padano. The pentaplex PCR assay revealed a good correspondence with microbiological analyses and allowed to identify even minor LAB community members which, can be out-competed in vitro by numerically more abundant microbial species.

  2. Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR.

    Science.gov (United States)

    Pega, J; Rizzo, S; Pérez, C D; Rossetti, L; Díaz, G; Ruzal, S M; Nanni, M; Descalzo, A M

    2016-09-01

    The quality of functional food products designed for the prevention of degenerative diseases can be affected by the incorporation of bioactive compounds. In many types of cheese, the performance of starter microorganisms is critical for optimal elaboration and for providing potential probiotic benefits. Phytosterols are plant lipophilic triterpenes that have been used for the design of functional dairy products because of their ability to lower serum cholesterol levels in humans. However, their effect on the starter culture behavior during cheesemaking has not yet been studied. Here, we followed DNA and RNA kinetics of the bacterium Streptococcus thermophilus, an extensively used dairy starter with probiotic potential, during industrial production of a functional, semi-soft, reduced-fat cheese containing phytosterol esters and alpha-tocopherol as bioactive compounds. For this purpose, real-time quantitative PCR (qPCR) and reverse transcription-qPCR (RT-qPCR) assays were optimized and applied to samples obtained during the manufacture and ripening of functional and control cheeses. An experimental set-up was used to evaluate the detection threshold of free nucleic acids for extraction protocols based on pelleted microorganisms. To our knowledge, this straight-forward approach provides the first experimental evidence indicating that DNA is not a reliable marker of cell integrity, whereas RNA may constitute a more accurate molecular signature to estimate both bacterial viability and metabolic activity. Compositional analysis revealed that the bioactive molecules were effectively incorporated into the cheese matrix, at levels considered optimal to exert their biological action. The starter S. thermophilus was detected by qPCR and RT-qPCR during cheese production at the industrial level, from at least 30min after its inoculation until 81days of ripening, supporting the possible role of this species in shaping organoleptic profiles. We also showed for the first time that

  3. Effect of the addition of phytosterols and tocopherols on Streptococcus thermophilus robustness during industrial manufacture and ripening of a functional cheese as evaluated by qPCR and RT-qPCR.

    Science.gov (United States)

    Pega, J; Rizzo, S; Pérez, C D; Rossetti, L; Díaz, G; Ruzal, S M; Nanni, M; Descalzo, A M

    2016-09-01

    The quality of functional food products designed for the prevention of degenerative diseases can be affected by the incorporation of bioactive compounds. In many types of cheese, the performance of starter microorganisms is critical for optimal elaboration and for providing potential probiotic benefits. Phytosterols are plant lipophilic triterpenes that have been used for the design of functional dairy products because of their ability to lower serum cholesterol levels in humans. However, their effect on the starter culture behavior during cheesemaking has not yet been studied. Here, we followed DNA and RNA kinetics of the bacterium Streptococcus thermophilus, an extensively used dairy starter with probiotic potential, during industrial production of a functional, semi-soft, reduced-fat cheese containing phytosterol esters and alpha-tocopherol as bioactive compounds. For this purpose, real-time quantitative PCR (qPCR) and reverse transcription-qPCR (RT-qPCR) assays were optimized and applied to samples obtained during the manufacture and ripening of functional and control cheeses. An experimental set-up was used to evaluate the detection threshold of free nucleic acids for extraction protocols based on pelleted microorganisms. To our knowledge, this straight-forward approach provides the first experimental evidence indicating that DNA is not a reliable marker of cell integrity, whereas RNA may constitute a more accurate molecular signature to estimate both bacterial viability and metabolic activity. Compositional analysis revealed that the bioactive molecules were effectively incorporated into the cheese matrix, at levels considered optimal to exert their biological action. The starter S. thermophilus was detected by qPCR and RT-qPCR during cheese production at the industrial level, from at least 30min after its inoculation until 81days of ripening, supporting the possible role of this species in shaping organoleptic profiles. We also showed for the first time that

  4. Optimization of exopolysaccharide production conditions fermented by Streptococcus thermophilus from Tibetan kefir%藏灵菇中嗜热链球菌高产胞外多糖发酵条件的优化

    Institute of Scientific and Technical Information of China (English)

    刘慧; 张红星; 代娟

    2007-01-01

    利用从藏灵菇中筛选的产胞外多糖(EPS)的嗜热链球菌(Streptococcus thermophilus)研究提高多糖产量的环境因素.针对影响胞外多糖合成的4个因素,采用4因素3水平L9(34)正交试验,确定了高产胞外多糖优化发酵条件,即发酵温度为32℃,发酵时间为16h,培养基起始pH值为6.5,接种量为3%.在此优化条件下,胞外多糖的产量是优化前的2.6倍.

  5. A novel mode of ferric ion coordination by the periplasmic ferric ion-binding subunit FbpA of an ABC-type iron transporter from Thermus thermophilus HB8.

    Science.gov (United States)

    Wang, Shipeng; Ogata, Misaki; Horita, Shoichiro; Ohtsuka, Jun; Nagata, Koji; Tanokura, Masaru

    2014-01-01

    Crystal structures of FbpA, the periplasmic ferric ion-binding protein of an iron-uptake ABC transporter, from Thermus thermophilus HB8 (TtFbpA) have been solved in apo and ferric ion-bound forms at 1.8 and 1.7 Å resolution, respectively. The latter crystal structure shows that the bound ferric ion forms a novel six-coordinated complex with three tyrosine side chains, two bicarbonates and a water molecule in the metal-binding site. The results of gel-filtration chromatography and dynamic light scattering show that TtFbpA exists as a monomer in solution regardless of ferric ion binding and that TtFbpA adopts a more compact conformation in the ferric ion-bound state than in the apo state in solution.

  6. Multi-frequency EPR evidence for a binuclear CuA center in cytochrome c oxidase: studies with a 63Cu- and 65Cu-enriched, soluble domain of the cytochrome ba3 subunit II from Thermus thermophilus.

    Science.gov (United States)

    Fee, J A; Sanders, D; Slutter, C E; Doan, P E; Aasa, R; Karpefors, M; Vänngård, T

    1995-07-01

    We have recorded multi-frequency EPR spectra of 63Cu- and 65Cu-labeled, water-soluble CuA-protein from the cytochrome ba3 of T. thermophilus. The spectrum taken at the highest frequency (34.03 GHz) shows no hyperfine structure and is nominally axial with apparent gz approximately 2.18 and gxy approximately 2.00. The spectrum taken at the lowest frequency (3.93 GHz) shows a rich hyperfine structure. Analyses of the spectra show that the observed splitting arises from an interaction of the unpaired electron with two Cu nuclei and support the notion that CuA is a mixed-valent [Cu(II)/Cu(I)] complex in which the unpaired electronic spin is distributed evenly over the two Cu ions.

  7. 保加利亚乳杆菌和嗜热链球菌发酵紫红薯酸牛奶%Purple sweet potatoes yoghurt fermented by Lactobacillus bulgaricus and Streptococcus thermophilus

    Institute of Scientific and Technical Information of China (English)

    卢兆芸; 陈海婴; 彭冬英; 段学辉; 胡泽红

    2011-01-01

    研究以鲜牛奶和紫红薯为主要原料,保加利亚乳杆菌(Lactobacillus bulgaricus)与嗜热链球菌(Streptococcus thermophilus)为发酵剂,木糖醇为甜味剂,进行了发酵条件的平行和L9(34)正交试验.结果表明,保加利亚乳杆菌与嗜热链球菌混合量比1∶1(数量比),接种量5%,紫红薯添加量15%,木糖醇10%,果胶0.1%,明胶0.1%,CMC 0.1%,变性淀粉0.5%,45℃,发酵4h,制得的紫红薯酸牛奶不仅保留纯酸牛奶的芳香和口感,又具备紫红薯的营养和色泽,是新型紫红薯营养酸奶发酵的优化工艺.%A new type of yoghurt, a clabber mile with the purple sweet potatoes fermented by Lactobadllus bulgaricus and Streptococcus ther-mophilus was studied. The orthogonal experiments of L9(34)results showed that, the optimize fermentation conditions and additive compositions of purple sweet potatoes yoghurt were: purple sweet potatoes 15%, sweeter xylitol 10%, inoculate strain 5%, 45 ℃, fermentation time 4 hours. The dairy products fermented with best nutrition and flavor of yoghurt, and keep the natural colour of purple sweet potatoes.

  8. 益生菌Lactobacillus plantarum P8与Streptococcus thermophilus混合发酵对发酵乳品质的影响%Effect of probiotic Lactobacillus plantarum P8 and Streptococcus thermophilus on the quality of fermented milk

    Institute of Scientific and Technical Information of China (English)

    何秋雯; 王水泉; 鲍雅静; 礼贺; 王记成; 张和平; 陈卫

    2012-01-01

    将益生菌Lactobacillus plantarum P8以不同接种比例分别与Streptococcus thermophilus ND03、ND05和ND07复配发酵牛乳,并以S.thermophilus单菌发酵为对照.于0h、发酵结束(pH=4.5)及4℃冷藏24 h,分别测定发酵乳样品的发酵时间,及pH值、黏度、脱水收缩率和活菌数并进行感官鉴评.结果表明,L.plantarum P8与S.thermophilus ND03复配,发酵时间显著高于其他组,各组样品黏度和脱水收缩率差异不显著;L.plantarum P8与S.thermophilus ND05分别以5×106 mL-1接种1∶1复配时,样品中L.plantarum P8的活菌数为5.26×107mL-1,酸度适宜,无乳清析出,有良好的黏度,产品风味较好,将该组合应用于发酵乳生产,较可行.%L. plantarutn P8 was mixed respectively with S. thermophillus ND03, ND05 and ND07 with different proportion for bovine milk fermentation with single S. thermophillus fermented milk as the control. At the baseline (0 h), the end of fermentation (pH=4.5) and storage (4℃ for 24 h) fermentation time, pH, viscosity, syneresis, and sensory characteristics of yogurts as well as the bacterial viable counts were evaluated. The results indicated that fermention time of milk fermented by combined L. plantarum P8 and S. thermophilus ND03 was significantly longer than other groups(P<0.05=,while viscosity and syneresis had no significant differences(P>0.05). Moreover, the fermented milk showed great quality with fine viscosity, moderate acidity, no whey separating, good spinnablility and nice flavor while combined L. plantarum P8 and S. thermophilus ND05 with 1:1 ratio(5×l06 mL-1 each) for fermentation, suggesting that this combination could be applied for yoghurt production.

  9. Biofilm-forming Capabilities of Lactobacillus bulgaricus and Streptococcus thermophilus%保加利亚乳杆菌和嗜热链球菌生物膜形成研究

    Institute of Scientific and Technical Information of China (English)

    王坤; 闫颖娟; 姜梅; 陈晓红; 李伟; 董明盛

    2011-01-01

    The biofilm-forming capabilities of Streptococcus thermophilus and Lactobacillus bulgaricus on different carriers(coconut,stainless steel mesh,plastic sheet,glass sheet or ceramic parts) was studied by using placed slice and plate culture count methods.The results showed that stainless steel mesh and coconut were suitable carriers for biofilm formation.During 7 days,the CFU(colony-forming units) of the single-species biofilm on coconut and stainless steel mesh initially reached 1 × 107 CFU/cm2 and then was kept at 3.2 × 106 CFU/cm2.The mixed-species biofilm was stable at approximately 1 × 107 CFU/cm2 in continuous seven days.In addition,scanning electron microscope was used to observe mixed-species biofilm formed by S.thermophilus and L.bulgaricus,and a typical mixed-species biofilm was observed on coconut and stainless steel mesh.%采用置片法和菌落计数法对嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus)在不同载体表面(椰果粒、不锈钢网布、塑料片、陶瓷片和玻璃片)上形成生物膜的能力进行研究。结果表明:椰果粒和不锈钢网布是适宜乳酸菌单菌生物膜形成的载体,培养7d椰果粒和不锈钢网布上的单菌生物膜初始菌密度可达107CFU/cm2,并且在后续培养中能稳定在3.2×106CFU/cm2左右;混菌生物膜菌密度连续7d稳定在1×107CFU/cm2左右。利用扫描电镜对S.thermophilus和L.bulgaricus形成的混菌生物膜进行观察,结果表明在椰果粒和不锈钢网布表面上S.thermophilus和L.bulgaricus形成典型的混菌生物膜结构。

  10. Optimize culture conditions of streptococcus thermophilus for fermentation producing nisin by response surface methodology%响应面法优化 Nisin 发酵的培养条件

    Institute of Scientific and Technical Information of China (English)

    贺小贤; 郑茂奎; 刘欢; 彭光杰

    2014-01-01

    In order to improve fermentation titer unit of Nisin ,the MRS was used as the fer‐mentation medium by streptococcus thermophilus and titer unit of Nisin to Staphylococcus aureus which is used as indicator bacteria serve as detection index of titer unit of Nisin .On the basis of single factor experiment ,optimization of fermentation initial pH ,inoculation a‐mount of Streptococcus thermophilus and corn steep liquor content for producing Nisin by u‐sing response surface analysis methodology .The experiment results showed that the optimal fermentation condition was initial pH 6 .60 ,inoculation amount of 4 .80% and corn steep liq‐uor content of 25 .60% ,and corn steep liquor content is the maximum factor for affecting the fermentation titer unit of Nisin and inoculation is the minimal factor .Under these conditions , the Nisin titer unit was increased by 18 .07% and reached up to 1 602 .77 ± 6 .38 IU/mL which is consistent with the predicted value .These show that it is effective for optimizing culture conditions of Nisin by response surface methodology .%为了提高嗜热乳链球菌发酵生产Nisin的效价,以M RS为发酵培养基,以金黄色葡萄球菌为效价检测指示菌,以Nisin效价为考察指标,在单因素实验的基础上,采用响应面分析法优化发酵初始pH、接种量以及玉米浆添加量.结果表明影响Nisin效价的最大因素是玉米浆添加量,其次是发酵初始p H ,接种量影响最小.最佳初始p H6.60,接种量4.80%,玉米浆添加量2560%,在此条件下 Nisin 的效价可达1602.77±6.38 IU/mL ,较优化前提高了18.07%,其实验结果与模型预测值拟合良好,说明通过响应面试验设计对Nisin发酵条件的优化是有效的.

  11. Preparation of Gamma-Aminobutyric Acid Using Streptococcus salivarius subsp.thermophilus Y-2 Cells%唾液链球菌嗜热亚种Y-2细胞转化法制备γ-氨基丁酸

    Institute of Scientific and Technical Information of China (English)

    杨胜远; 陆兆新; 吕凤霞; 别小妹

    2011-01-01

    研究反应pH值、反应温度、重金属盐、表面活性剂、底物浓度、菌体质最浓度和磷酸吡哆醛添加量对Streptococcus salivarius subsp.thermophilus Y-2细胞转化法生产γ-氨基丁酸的影响.获得反应体系的最佳组成为:湿菌体25g/L、BaCl2 40mmol/L、Triton X-100体积分数0.02%、L-谷氨酸单钠盐(L-monosodium glutamate,MSG)47.5g/L和L-谷氨酸(L-glutamic acid,L-Glu)90.0g/L.该体系在40℃、pH4.5和搅拌速度100r/min的最适转化条件下进行反应72h,转化液GABA产量达到了(87.16±4.33)g/L,细胞平均生产力为(48.42±2.41)mg/(h·g),摩尔转化率为(97.60±4.71)%.

  12. Enhanced Heavy Metal Tolerance and Accumulation by Transgenic Sugar Beets Expressing Streptococcus thermophilus StGCS-GS in the Presence of Cd, Zn and Cu Alone or in Combination.

    Science.gov (United States)

    Liu, Dali; An, Zhigang; Mao, Zijun; Ma, Longbiao; Lu, Zhenqiang

    2015-01-01

    Phytoremediation is a promising means of ameliorating heavy metal pollution through the use of transgenic plants as artificial hyperaccumulators. A novel Streptococcus thermophilus γ-glutamylcysteine synthetase-glutathione synthetase (StGCS-GS) that synthesizes glutathione (GSH) with limited feedback inhibition was overexpressed in sugar beet (Beta vulgaris L.), yielding three transgenic lines (s2, s4 and s5) with enhanced tolerance to different concentrations of cadmium, zinc and copper, as indicated by their increased biomass, root length and relative growth compared with wild-type plants. Transgenic sugar beets accumulated more Cd, Zn and Cu ions in shoots than wild-type, as well as higher GSH and phytochelatin (PC) levels under different heavy metal stresses. This enhanced heavy metal tolerance and increased accumulation were likely due to the increased expression of StGCS-GS and consequent overproduction of both GSH and PC. Furthermore, when multiple heavy metal ions were present at the same time, transgenic sugar beets overexpressing StGCS-GS resisted two or three of the metal combinations (50 μM Cd-Zn, Cd-Cu, Zn-Cu and Cd-Zn-Cu), with greater absorption in shoots. Additionally, there was no obvious competition between metals. Overall, the results demonstrate the explicit role of StGCS-GS in enhancing Cd, Zn and Cu tolerance and accumulation in transgenic sugar beet, which may represent a highly promising new tool for phytoremediation.

  13. Enhanced Heavy Metal Tolerance and Accumulation by Transgenic Sugar Beets Expressing Streptococcus thermophilus StGCS-GS in the Presence of Cd, Zn and Cu Alone or in Combination.

    Directory of Open Access Journals (Sweden)

    Dali Liu

    Full Text Available Phytoremediation is a promising means of ameliorating heavy metal pollution through the use of transgenic plants as artificial hyperaccumulators. A novel Streptococcus thermophilus γ-glutamylcysteine synthetase-glutathione synthetase (StGCS-GS that synthesizes glutathione (GSH with limited feedback inhibition was overexpressed in sugar beet (Beta vulgaris L., yielding three transgenic lines (s2, s4 and s5 with enhanced tolerance to different concentrations of cadmium, zinc and copper, as indicated by their increased biomass, root length and relative growth compared with wild-type plants. Transgenic sugar beets accumulated more Cd, Zn and Cu ions in shoots than wild-type, as well as higher GSH and phytochelatin (PC levels under different heavy metal stresses. This enhanced heavy metal tolerance and increased accumulation were likely due to the increased expression of StGCS-GS and consequent overproduction of both GSH and PC. Furthermore, when multiple heavy metal ions were present at the same time, transgenic sugar beets overexpressing StGCS-GS resisted two or three of the metal combinations (50 μM Cd-Zn, Cd-Cu, Zn-Cu and Cd-Zn-Cu, with greater absorption in shoots. Additionally, there was no obvious competition between metals. Overall, the results demonstrate the explicit role of StGCS-GS in enhancing Cd, Zn and Cu tolerance and accumulation in transgenic sugar beet, which may represent a highly promising new tool for phytoremediation.

  14. Enhanced Heavy Metal Tolerance and Accumulation by Transgenic Sugar Beets Expressing Streptococcus thermophilus StGCS-GS in the Presence of Cd, Zn and Cu Alone or in Combination.

    Science.gov (United States)

    Liu, Dali; An, Zhigang; Mao, Zijun; Ma, Longbiao; Lu, Zhenqiang

    2015-01-01

    Phytoremediation is a promising means of ameliorating heavy metal pollution through the use of transgenic plants as artificial hyperaccumulators. A novel Streptococcus thermophilus γ-glutamylcysteine synthetase-glutathione synthetase (StGCS-GS) that synthesizes glutathione (GSH) with limited feedback inhibition was overexpressed in sugar beet (Beta vulgaris L.), yielding three transgenic lines (s2, s4 and s5) with enhanced tolerance to different concentrations of cadmium, zinc and copper, as indicated by their increased biomass, root length and relative growth compared with wild-type plants. Transgenic sugar beets accumulated more Cd, Zn and Cu ions in shoots than wild-type, as well as higher GSH and phytochelatin (PC) levels under different heavy metal stresses. This enhanced heavy metal tolerance and increased accumulation were likely due to the increased expression of StGCS-GS and consequent overproduction of both GSH and PC. Furthermore, when multiple heavy metal ions were present at the same time, transgenic sugar beets overexpressing StGCS-GS resisted two or three of the metal combinations (50 μM Cd-Zn, Cd-Cu, Zn-Cu and Cd-Zn-Cu), with greater absorption in shoots. Additionally, there was no obvious competition between metals. Overall, the results demonstrate the explicit role of StGCS-GS in enhancing Cd, Zn and Cu tolerance and accumulation in transgenic sugar beet, which may represent a highly promising new tool for phytoremediation. PMID:26057126

  15. Obtención de ácido láctico por fermentación de almidón de ñame espino mediante el Lactobacillus delbrueckii ssp. bulgaricus y el Streptococcus thermophilus para su uso en la producción de ácido poliláctico

    OpenAIRE

    Tejada Tovar, Ronald Eduardo

    2015-01-01

    Resumen: El propósito del presente trabajo fue evaluar la producción de ácido láctico obtenido por la fermentación del almidón de ñame espino Discorea rotundata mediante la acción de la mezcla de Lactobacillus delbrueckii ssp. bulgaricus y el Streptococcus thermophilus, para su posible uso en la obtención de APL; estableciendo el proceso de obtención y caracterizando el producto obtenido mediante análisis de sus propiedades fisicoquímicas y funcionales; a este se le realizó un seguimiento med...

  16. Cloning and functional verification of codY promoter from Streptococcus thermophilus%嗜热链球菌codY基因启动子的克隆及功能验证

    Institute of Scientific and Technical Information of China (English)

    刘芳; 都立辉; 王道营; 徐为民; 霍贵成

    2013-01-01

    The experiment aimed to study the function of codY gene promoter. The promoter was amplified from Streptococcus thermophilus KLDS 3.0503 genome and inserted into the promoter probe vector pNZ273 to construct a recom-binant plasmid named pNZ273/codY. The plasmid p3. 8501-1 in KLDS3. 8501 was sequenced and its genetic information was analyzed. The replicated fragment of plasmid p3. 8501-1 was ligated together with the fragments including chloramphen-icol-resistance gene, codY gene promoter and gusA gene in plasmid pNZ273/codY.The ligated product was electrotrans-formed into KLDS3.0503 to construct a recombinant plasmid named pSTl. It was found that both in Lactococcus lactis and Streptococcus thermophilus, the codY gene promoter could switch on the gusA gene expression, and p-glucuronidase activity could be detected only when the strains grew to the logarithmic growth phase. These results indicated that the codY gene promoter could functioned as an inducible promoter because of its self-regulation mechanism.%为研究转录调控蛋白codY基因启动子的功能,首先从嗜热链球菌KLDS3.0503中克隆出codY基因启动子,将该启动子连入启动子探针型载体pNZ273的gusA基因上游,获得重组质粒pNZ273/codY.通过分析嗜热链球菌KLDS3.8501携带的质粒p3.8501-1的全序列后,将p3.8501-1质粒的复制区段与pNZ273/codY质粒中氯霉素抗性基因、codY基因启动子和gusA基因的片断连接到一起,电转化入嗜热链球菌KLDS3.0503中,获得重组质粒pST1.在乳酸乳球菌和嗜热链球菌中,该启动子均能启动下游gusA基因的表达,并且当菌株生长到对数生长期时,才检测到β-葡糖苷酸酶的活性.这说明codY启动子凭借自身的调控机制可以起到诱导型启动子的作用,并能控制外源基因的表达.

  17. Effect of High Hydrostatic Pressure on Streptococcus Thermophilus Bacteriophages%超高压对嗜热链球菌噬菌体灭活效果的研究

    Institute of Scientific and Technical Information of China (English)

    曲明椿; 曹玉敏; 董鹏; 陈芳; 胡小松

    2012-01-01

    The objective of this study is investigate the effect of high hydrostatic pressure (HHP) on Streptococcus thermophilus bacteriophages. Three strains of Streptococcus thermophilus bacteriophage were subjected to different pressure, treatment time with different initial bacteriophage load. The traditional heat treatment was used for comparison, and the structural changes of treated bactcriophages were observed to study the inactivation mechanism of HHP. The results showed that 400 MPa didn' t have obvious inactivation effect. At 600 MPa, all bacteriophages were completely inactivated after 10~20 min. For bacteriophage ALQ13.2 and DTI, the lower the initial load, the bigger the reduction achieved. However, Abc2 was independent on initial concentration. The traditional heat treatment at 72 ℃ for 15s didn't have obvious inaclivalion effect. Increasing time to 5 min inactivated most bacteriophages. There is no relevance between pressure resistance and heat resistance of bactenophages, ALQ13.2 was the most pressure-resistant while DT1 was the most heat-resistant. The result of electron microscope showed that the bacteriophage structure had various changes after HHP treatment.%为研究超高压(HHP)对嗜热链球菌噬菌体的灭活效果,以3株嗜热链球菌噬菌体为对象,在不同压力、处理时间及初始菌数条件下进行超高压处理,并与传统热处理进行比较;通过观察处理前、后噬菌体结构的变化,探明超高压灭活机理.试验结果表明,压力400 MPa下无明显灭活效果;压力600 MPa处理10~20 min,可将3株噬菌体全部杀灭.噬菌体ALQ13.2和DTI随初始菌数的下降,灭活效果逐渐增强;Abc2则无显著性差异.72 ℃、15 s的传统热处理对噬菌体无明显灭活作用,而时间延长至5min,可将大部分噬菌体杀灭.3株噬菌体的耐压性与耐热性无相关性,其中ALQ13.2最耐压,DTI最耐热.电镜结果显示超高压处理前、后噬菌体的结构发生多种变化.

  18. Overexpression of a novel thermostable and chloride-tolerant laccase from Thermus thermophilus SG0.5JP17-16 in Pichia pastoris and its application in synthetic dye decolorization.

    Directory of Open Access Journals (Sweden)

    Huiping Liu

    Full Text Available Laccases have been used for the decolorization and detoxification of synthetic dyes due to their ability to oxidize a wide variety of dyes with water as the sole byproduct. A putative laccase gene (LacTT from Thermus thermophilus SG0.5JP17-16 was screened using the genome mining approach, and it was highly expressed in Pichia pastoris, yielding a high laccase activity of 6130 U/L in a 10-L fermentor. The LacTT open reading frame encoded a protein of 466 amino acid residues with four putative Cu-binding regions. The optimal pH of the recombinant LacTT was 4.5, 6.0, 7.5 and 8.0 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonic acid (ABTS, syringaldazine (SGZ, guaiacol, and 2,6-dimethoxyphenol (2,6-DMP as the substrate, respectively. The optimal temperature of LacTT was 90°C with guaiacol as the substrate. LacTT was highly stable at pH 4.0-11.0 and thermostable at 40°C-90°C, confirming that it is a pH-stable and thermostable laccase. Furthermore, LacTT also exhibited high tolerance to halides such as NaCl, NaBr and NaF, and decolorized 100%, 94%, 94% and 73% of Congo Red, Reactive Black B and Reactive Black WNN, and Remazol Brilliant Blue R, respectively. Interestingly, addition of high concentration of NaCl increased the RBBR decolorization efficiency of LacTT. These results suggest that LacTT is a good candidate for industrial applications such as dyestuff processing and degradation of dyes in textile wastewaters.

  19. Overexpression of a Novel Thermostable and Chloride-Tolerant Laccase from Thermus thermophilus SG0.5JP17-16 in Pichia pastoris and Its Application in Synthetic Dye Decolorization

    Science.gov (United States)

    Liu, Huiping; Cheng, Yu; Du, Bing; Tong, Chaofan; Liang, Shuli; Han, Shuangyan; Zheng, Suiping; Lin, Ying

    2015-01-01

    Laccases have been used for the decolorization and detoxification of synthetic dyes due to their ability to oxidize a wide variety of dyes with water as the sole byproduct. A putative laccase gene (LacTT) from Thermus thermophilus SG0.5JP17-16 was screened using the genome mining approach, and it was highly expressed in Pichia pastoris, yielding a high laccase activity of 6130 U/L in a 10-L fermentor. The LacTT open reading frame encoded a protein of 466 amino acid residues with four putative Cu-binding regions. The optimal pH of the recombinant LacTT was 4.5, 6.0, 7.5 and 8.0 with 2,2'-azino-bis(3-ethylbenzothazoline-6-sulfonic acid) (ABTS), syringaldazine (SGZ), guaiacol, and 2,6-dimethoxyphenol (2,6-DMP) as the substrate, respectively. The optimal temperature of LacTT was 90°C with guaiacol as the substrate. LacTT was highly stable at pH 4.0–11.0 and thermostable at 40°C–90°C, confirming that it is a pH-stable and thermostable laccase. Furthermore, LacTT also exhibited high tolerance to halides such as NaCl, NaBr and NaF, and decolorized 100%, 94%, 94% and 73% of Congo Red, Reactive Black B and Reactive Black WNN, and Remazol Brilliant Blue R, respectively. Interestingly, addition of high concentration of NaCl increased the RBBR decolorization efficiency of LacTT. These results suggest that LacTT is a good candidate for industrial applications such as dyestuff processing and degradation of dyes in textile wastewaters. PMID:25790466

  20. 固定化唾液链球菌生产γ-氨基丁酸的研究%PRUDUCTION OF γ-AMINOBUTYRIC ACID BY IMMOBILIZED Streptococcus salivarius subsp.thermophilus Y-2

    Institute of Scientific and Technical Information of China (English)

    焦阳; 汪建敏; 杨胜远; 周利娟; 姚正颖; 吕凤霞; 邹晓葵; 陆兆新

    2009-01-01

    以唾液链球茵嗜热亚种(Streptococcus salivarius subsp.thermophilus Y-2)为供试菌株,考察了卡拉胶、明胶和海藻酸钙等材料将此菌株固定化的效果,并通过比较固定化细胞的谷氨酸脱羧酶(glutamatedecarboxylase,GAD)活性及γ-氨基丁酸的产量和载体机械强度,确定了海藻酸钙作为固定化细胞的适宜载体.优化后得到的最适固定化条件(W/V)为:海藻酸钠2%,CaCl_2 14%,茵体25%,凝胶平均颗粒直径1.64 mm,在此条件下测得固定化细胞的GAD活性为游离细胞的1.2倍.细胞多批次应用稳定性试验证明:固定化细胞较游离细胞有着更稳定的GAD活性,反复使用60 h后,固定化细胞GAD活性仍能保持其初始活性的90%以上,γ-氨基丁酸的积累量达到7.97 g/L.

  1. N7-Methylguanine at position 46 (m7G46) in tRNA from Thermus thermophilus is required for cell viability at high temperatures through a tRNA modification network.

    Science.gov (United States)

    Tomikawa, Chie; Yokogawa, Takashi; Kanai, Tamotsu; Hori, Hiroyuki

    2010-01-01

    N(7)-methylguanine at position 46 (m(7)G46) in tRNA is produced by tRNA (m(7)G46) methyltransferase (TrmB). To clarify the role of this modification, we made a trmB gene disruptant (DeltatrmB) of Thermus thermophilus, an extreme thermophilic eubacterium. The absence of TrmB activity in cell extract from the DeltatrmB strain and the lack of the m(7)G46 modification in tRNA(Phe) were confirmed by enzyme assay, nucleoside analysis and RNA sequencing. When the DeltatrmB strain was cultured at high temperatures, several modified nucleotides in tRNA were hypo-modified in addition to the lack of the m(7)G46 modification. Assays with tRNA modification enzymes revealed hypo-modifications of Gm18 and m(1)G37, suggesting that the m(7)G46 positively affects their formations. Although the lack of the m(7)G46 modification and the hypo-modifications do not affect the Phe charging activity of tRNA(Phe), they cause a decrease in melting temperature of class I tRNA and degradation of tRNA(Phe) and tRNA(Ile). (35)S-Met incorporation into proteins revealed that protein synthesis in DeltatrmB cells is depressed above 70 degrees C. At 80 degrees C, the DeltatrmB strain exhibits a severe growth defect. Thus, the m(7)G46 modification is required for cell viability at high temperatures via a tRNA modification network, in which the m(7)G46 modification supports introduction of other modifications.

  2. Effect of ABF Bio + malt extract on the growth of Streptococcus thermophilus%双麦麦精对嗜热链球菌生长影响的研究

    Institute of Scientific and Technical Information of China (English)

    赵爽; 刘少伟; 陈如琪; 吕宝龙; 卢艳花; 费楠

    2012-01-01

    主要研究双麦麦精对嗜热链球菌生长过程的影响。通过在营养成分缺省的培养基中添加双麦麦精,采用分光光度法测定菌液吸光值(OD600),根据OD值确定对应的嗜热链球菌的活菌数,从而分析双麦麦精对嗜热链球菌生长情况的影响。利用正交实验,通过直接计数法确定双麦麦精与其他营养成分的最佳比例。结果表明,双麦麦精对嗜热链球菌的生长有明显的促进作用,可为嗜热链球菌提供充足的碳源和生长因子。双麦麦精的添加量为16g/L时效果最佳。分析比较吸光值法和直接计数法测得的生长曲线,结果表明两种方法具有良好的相似性,其中在对数期有强相关性。%The effects of ABF Bio + malt extract on the main nutrition constituent of the Streptococcus thermophiles in its growth progress by adding ABF Bio + malt in the incomplete nutrient medium were studied.The number of viable cells in Streptococcus thermophiles counted according to its Bacilli absorbance was used to analyse the effect on Streptococcus thermophiles brought by ABF Bio + malt extract.The optimal proportion of ABF Bio + malt extract and other nutrients was determined by orthogonal experiment with direct counting method.The results showed that ABF Bio + malt extract played an important role of the augmentor the growth of streptococcus thermophilus as well as a provider of sufficient carbon source and growth factors for it.The ABF Bio + malt extract produced the best result when its additive amount reached 16g/L.After analysis and comparison of the growth curves which were measured by absorb value method and the direct counting method respectively, the curves were proved to be of good similarity, especially in the logarithmic phase.

  3. 乳酸杆菌及嗜热链球菌脉冲场凝胶电泳分子分型方法建立及应用%Development of pulsed field gel electrophoresis and application for characterization and identification of Lactobacillus and Streptococcus thermophilus

    Institute of Scientific and Technical Information of China (English)

    董银苹; 崔生辉; 于红霞; 李凤琴

    2011-01-01

    目的 建立乳酸杆菌及嗜热链球菌的PFGE分子分型方法,并对北京市售酸奶中分离的乳酸杆菌及嗜热链球菌进行分子分型.方法 选取ApaⅠ、NotⅠ、sfiⅠ、XbaⅠ和SmaⅠ共5种PFGE分析中常用的限制性内切酶,对从北京市售酸奶中分离到的52株乳酸杆菌、嗜热链球菌以及相应的标准菌株进行酶切,优化PFGE限制性内切酶种类及电泳条件,并用优化出的实验条件对菌株进行分子分型,同时进行聚类分析,与生化鉴定及16s rRNA基因鉴定结果进行对比分析.结果 限制性内切酶NotⅠ对保加利亚乳酸杆菌、发酵乳酸杆菌和德氏乳酸杆菌的酶切效果较好,而限制性内切酶Apa Ⅰ对嗜热链球菌、嗜酸乳酸杆菌及干酪乳酸杆菌的酶切效果较好.24株保加利亚乳酸杆菌被分为8个PFGE型,15株嗜热链球菌被分为8个PFGE型,7株嗜酸乳酸杆菌被分为3个PFGE型,2株德氏乳酸杆菌分属于2个不同的PFGE型.结论 建立的PFGE方法分析结果与生化鉴定及16s rRNA基因鉴定结果高度符合,所建方法适用于乳酸杆菌及嗜热链球菌的分子分型.%Objective To develop a pulsed field gel electrophoresis (PFGE) method for molecular typing of Lactobacillus and Streptococcus thermophilus (S.thermophilus) and to apply it in identification and characterization of both bacteria isolated from yoghourt collected from Beijing supermarket.Methods The five most useful restriction enzymes includingApa Ⅰ,Not ],Sfi Ⅰ,Xba Ⅰ and Sma Ⅰ were chosen to cut DNA of 52 strains of Lactobacillus,S.thermophilus as well as associated standard bacteria strains.The endonucleases and electrophoresis conditions for PFGE analysis were optimized and applied in molecular typing of Lactobacillus and S.thermophilus isolates.Cluster analysis based on the PFGE data was conducted.The identification results of PFGE were compared with those obtained in biochemical and 16s ribosomal RNA PCR identification tests

  4. EFSA NDA Panel (EFSA Panel on Dietetic Products, Nutrition and Allergies), 2014. Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and

    DEFF Research Database (Denmark)

    Tetens, Inge

    to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improvement of bowel function by increasing stool frequency. The food that is the subject of the health claim is a combination of four bacterial...... effect proposed by the applicant is "improves stool frequency". The Panel considers that improvement of bowel function by increasing stool frequency, provided that it does not result in diarrhoea, is a beneficial physiological effect. The Panel considers that the human study provided for the...... substantiation of the claim did not find an increase in stool frequency following consumption of a combination of the bacterial strains which is the subject of the claim. The Panel concludes that a cause and effect relationship has not been established between the consumption of a combination of B. longum LA 101...

  5. Purification and Structural Characterization of Exopolysaccharide Produced by Streptococcus thermophilus CH9%嗜热链球菌CH9胞外多糖的分离纯化及结构分析

    Institute of Scientific and Technical Information of China (English)

    张新源; 刘会平; 曹春玲; 季玲

    2012-01-01

    Crude exopolysaccharides (EPS) isolated from the fermentation supernatant of Streptococcus thermophilus CH9 were fractionated by DEAE-52 column chromatography into EPS1, EPS2 and EPS3. Sephadex G-100 column chromatography was used to further ffactionate the three factions to harvest their respective major components named as EPSla, EPS2a and EPS3a, respectively. EPSla was a neutral polysaccharide with a molecular weight of 1.8×10^6 U, and EPS2a and EPS3a were acidic polysaccharides with a molecular weight of 1.06×10^6u and 1.05 × 10^6 u, respectively. Agarose gel electrophoresis analysis indicated that all these three polysaccharides consisted of a single component. UV and IR spectroscopic analyses showed that all of them had characteristic adsorption peaks of polysaccharide. EPS2a and EPS3a might be polysaccharide- protein complexes. Congo red test revealed that EPSla rather than EPS2a and EPS3a had a three-stranded helix structure. Beta elimination test demonstrated that EPS la and EPS3a were -O-linked, but EPS2a was a -N-linked polysaccharide.%对嗜热链球菌CH9胞外多糖进行分离纯化,经DEAE-52离子交换柱及SephadexG-100凝胶柱得到单一组分,利用高效液相色谱(HPLC)测其分子质量,结果表明:EPSla为中性多糖,分子质量为1.8×10^6u;EPS2a、EPS3a为酸性多糖,分子质量分别为1.06×10^6、1.05X10^u。经琼脂糖凝胶电泳)及SephadexG-100凝胶柱鉴定3种糖均为单一组分。紫外及红外光谱扫描结果显示:3种糖均具有多糖的特征吸收峰,且EPS2a、EPS3a可能为糖与蛋白的复合物。刚果红实验表明:EPSla具有三股螺旋结构,而其他两种糖并未发现此现象。卢消去实验证明:EPSla、EPS3a为—O-连接,EPS2a为-N-连接。

  6. Effects of inoculum ratios of Lactobacillus delbrueckii subsp.bulgaricus and Streptococcus thermophilus on the quality of probiotic fermented milk by Bifidobacterium lactis%Lactobacillus delbrueckii subsp.bulgaricus和Streptococcus thermophilus接种比例对Bifidobacterium lactis益生菌发酵乳品质的影响

    Institute of Scientific and Technical Information of China (English)

    李慧; 白梅; 王记成; 魏爱彬; 孔亚楠; 张和平; 孙天松

    2012-01-01

    Different ratios (1:1, 1:10, 1:100, 1:1000) of Lactobacillus delbnteckii subsp. Bulgaricus ND02 (LB-ND02) and Streptococcus thermophilus ND03 (ST-ND03) plus with Bifidobacterium lactis V9 (B. Lactis V9, 2.0X107 g"1) were used as starter cultures for milk fermentation. Some indexs of prepared probiotic fermented milk were determined for evaluation for effect of different ratios of inoculum on the quality of probiotic fermented milk during the fermentation and storage. The results showed the fermentation time was significandy prolonged, viable numbers of B. Lactis V9 was significantly increased .After the fermentation followed by a 28 d storage, the survival of B. Lactis V9 were significandy different and the postacidifition was weakened as the ratio of LB-ND02 decreased. . It is suggested that different ratios of LB-ND02 and ST-ND03 significandy influence the fermentation time, viable numbers of B. Lactis V9, postacidifition and visicosity during the fermentation and storage.%将Lactobacillus delbrueckii subsp.bulgaricus ND02(LB-ND02)和Streptococcus thermophilus ND03(ST-ND03)按1∶1、1∶10、1∶100、1∶1000接种于脱脂乳中,同时接入益生菌Bifidobacterium lactis V9(B.lactis V9,接种量为2.0×107 g-1),于42℃进行发酵.通过对发酵及贮藏过程中发酵乳指标的测定,评价LB-ND02和ST-ND03的接种比例对发酵乳品质的影响.结果表明,随着LB-ND02接种比例减小,凝乳时间显著延长,B.lactis V9活菌数显著提高.4 ℃贮藏28 d后,随LB-ND02接种比例减小,B.lactis V9存活率差异显著,后酸化也显著减弱.研究发现,LB-ND02和ST-ND03的接种比例,显著影响发酵乳的发酵时间、B.lactis V9活菌数、后酸化及黏度.

  7. Fermentation Performance and Resistance of Biofilm Formed by Mixed Bacterial Strains of Lactobacillus bulgaricus and Streptococcus thermophilus%嗜热链球菌和保加利亚乳杆菌混菌生物膜发酵特性及其抗逆性

    Institute of Scientific and Technical Information of China (English)

    玛依诺·木图拉; 王坤; 陈晓红; 姜梅; 李伟; 董明盛

    2013-01-01

    利用扫描电子显微镜对嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus)在不锈钢网布及椰果粒表面形成的混菌生物膜进行观察,并对不锈钢网布表面混菌生物膜发酵特性及其抗逆性进行研究.结果表明:Streptococcus thermophilus和Lactobacillus bulgaricus可在两种载体表面形成典型的混菌生物膜结构,且不锈钢网布表面混菌生物膜的发酵液菌体密度变化曲线与细菌S型生长曲线相似,但在停滞期和对数期不明显,发酵液pH值呈现先下降后趋于平稳的趋势,终止pH值接近4.0;此外,不锈钢网布表面混菌生物膜比游离乳酸菌具有更强的抗性,并且可在15d内稳定地释放较高浓度的游离乳酸菌.

  8. 唾液链球菌嗜热亚种Y-2产谷氨酸脱羧酶的影响因子确立%Ascertainment of Factors Affecting Glutamate Decarboxylase Production by Streptococcus Salivarius ssp.thermophilus Y-2

    Institute of Scientific and Technical Information of China (English)

    杨胜远; 陆兆新; 余勃; 林谦; 焦阳; 别小妹; 吕凤霞

    2008-01-01

    从产酶和细胞生长较好的MRS培养基出发,对Streptococcus salivarius ssp.thermophilus Y-2产谷氨酸脱羧酶(glutamate decarboxylase,GAD)的影响因子进行探讨,结果当培养基组成和培养条件为蛋白胨15g/L、牛肉膏12.5g/L、蔗糖12.5g/L、柠檬酸二铵2.0g/L、乙酸钠5.0g/L、K2HPO4 2.0g/L、CaCl2 2.0 g/L、Tween 80 1.0ml、pH7.0、接种量2%(V/V)、发酵温度37℃、发酵时间12h时,较有利于菌株Y-2产GAD.Plackett-Burman设计法研究表明培养基初始pH值和K2HPO4为影响菌株Y-2产GAD的主要影响因素.经对菌株Y-2产GAD影响因素的筛选,新获得的培养基在组成上与MRS培养基相比已发生显著变化,GAD活力提高了1.3倍.

  9. Identification of PHA loci in Thermus thermophilus HB8 genome

    OpenAIRE

    Papi, R. M.; Mimikakou, G. E.; A. A. Pantazaki; Kyriakidis, D A

    2008-01-01

    Abstract Poster Presentation Journal URL: http://www3.interscience.wiley.com/journal/119877016/tocgroup Polyhydroxyalkanoic acids (PHAs) are carbon and energy storage polymers synthesized by most eubacteria and archea in response to environmental conditions. The PHA biosynthetic bacteria are classified into four groups based on the organization of the pha gene locus and the structure-function properties of the PHA synthases. Synthases are the key enzymes in the biosynthesis of PHA and c...

  10. Detection and transfer of the glutamate decarboxylase gene in Streptococcus thermophilus

    Science.gov (United States)

    GABA (gamma-aminobutyric acid) is generated from glutamate by the action of glutamic acid decarboxylase (GAD) and characterized by hypotensive, diuretic and tranquilizing effects in humans and animals. The production of GABA by lactic acid starter bacteria would enhance the functionality of fermen...

  11. Cloning, expression and purification of D-Tyr-tRNATyr-deacylase from Thermus thermophilus

    OpenAIRE

    Rybak M. Yu.; Kovalenko O. P.; Kryklyvyi I. A.; Tukalo M. A.

    2015-01-01

    D-Tyr-tRNATyr-deacylase (DTD) is a conservative enzyme, found in all domains of life, which ensures an additional checkpoint in the recycling of misaminoacylated D-Tyr-tRNATyr. DTD is capable of accelerating the hydrolysis of the ester linkage of D-Tyr-tRNATyr producing a free tRNA and D-tyrosine, thereby preventing an incorrect incorporation of D-amino acids into proteins. Deacylase distinguishes between D- and L-aminoacyl moieties and does not hydrolyze L-aminoacylated tRNA. The structural ...

  12. Probiotic Leuconostoc mesenteroides ssp. cremoris and Streptococcus thermophilus induce IL-12 and IFN-γ production

    OpenAIRE

    Kekkonen, Riina A; Kajasto, Elina; Miettinen, Minja; Veckman, Ville; Korpela, Riitta; Julkunen, Ilkka

    2008-01-01

    AIM: To investigate the capacity of potentially probiotic strains from six bacterial genera to induce cytokine production alone or in combinations in order to identify potential enhancing or synergistic effects in order to select probiotic bacteria for in vivo purposes.

  13. Sequences spanning the leader-repeat junction mediate CRISPR adaptation to phage in Streptococcus thermophilus

    OpenAIRE

    Wei, Yunzhou; Chesne, Megan T.; Terns, Rebecca M.; Terns, Michael P.

    2015-01-01

    CRISPR-Cas systems are RNA-based immune systems that protect prokaryotes from invaders such as phages and plasmids. In adaptation, the initial phase of the immune response, short foreign DNA fragments are captured and integrated into host CRISPR loci to provide heritable defense against encountered foreign nucleic acids. Each CRISPR contains a ∼100–500 bp leader element that typically includes a transcription promoter, followed by an array of captured ∼35 bp sequences (spacers) sandwiched bet...

  14. Purification and characterization of thermophilin T, a novel bacteriocin produced by Streptococcus thermophilus ACA-DC 0040

    NARCIS (Netherlands)

    Aktypis, A.; Kalantzopoulos, G.; Huis Veld,in't J.H.J.; Brink, B. ten

    1998-01-01

    ACA-DC 0040 produced an antimicrobial agent, which was named thermophilin T, active against several lactic acid bacteria strains of different species and food spoilage bacteria, such as Clostridium sporogenes C22/10 and Cl. tyrobutyricum NCDO-1754. The crude antimicrobial compound is sensitive to pr

  15. Sequencing and transcriptional analysis of the Streptococcus thermophilus histamine biosynthesis gene cluster: factors that affect differential hdcA expression

    DEFF Research Database (Denmark)

    Calles-Enríquez, Marina; Hjort, Benjamin Benn; Andersen, Pia Skov;

    2010-01-01

    expression levels correlated with high histamine levels. Limited expression was evident during the lag and exponential growth phases. Low-temperature (4 degrees C) incubation of milk inoculated with a histamine-producing strain showed lower levels of histamine than did inoculated milk kept at 42 degrees C...

  16. Isolation of Nisin-resistant Streptococcus thermophilus%Nisin抗性嗜热链球菌的筛选

    Institute of Scientific and Technical Information of China (English)

    焦世耀; 张兰威

    2005-01-01

    采用两倍稀释法在脱脂乳培养基和乳清培养基中测定了nisin对嗜热链球菌9的最小抑菌浓度(MIC).在脱脂乳培养基中逐渐增加nisin的浓度,驯化嗜热链球菌9,直到其抗性达到300IU/ml.经生产性能鉴定和遗传性状实验,获得两株适宜做酸奶发酵剂的nisin抗性嗜热链球菌9.31和9.4.

  17. New biotechnological perspectives of a NADH oxidase variant from Thermus thermophilus HB27 as NAD+-recycling enzyme

    Directory of Open Access Journals (Sweden)

    Rocha-Martín Javier

    2011-11-01

    Full Text Available Abstract Background The number of biotransformations that use nicotinamide recycling systems is exponentially growing. For this reason one of the current challenges in biocatalysis is to develop and optimize more simple and efficient cofactor recycling systems. One promising approach to regenerate NAD+ pools is the use of NADH-oxidases that reduce oxygen to hydrogen peroxide while oxidizing NADH to NAD+. This class of enzymes may be applied to asymmetric reduction of prochiral substrates in order to obtain enantiopure compounds. Results The NADH-oxidase (NOX presented here is a flavoenzyme which needs exogenous FAD or FMN to reach its maximum velocity. Interestingly, this enzyme is 6-fold hyperactivated by incubation at high temperatures (80°C under limiting concentrations of flavin cofactor, a change that remains stable even at low temperatures (37°C. The hyperactivated form presented a high specific activity (37.5 U/mg at low temperatures despite isolation from a thermophile source. Immobilization of NOX onto agarose activated with glyoxyl groups yielded the most stable enzyme preparation (6-fold more stable than the hyperactivated soluble enzyme. The immobilized derivative was able to be reactivated under physiological conditions after inactivation by high solvent concentrations. The inactivation/reactivation cycle could be repeated at least three times, recovering full NOX activity in all cases after the reactivation step. This immobilized catalyst is presented as a recycling partner for a thermophile alcohol dehydrogenase in order to perform the kinetic resolution secondary alcohols. Conclusion We have designed, developed and characterized a heterogeneous and robust biocatalyst which has been used as recycling partner in the kinetic resolution of rac-1-phenylethanol. The high stability along with its capability to be reactivated makes this biocatalyst highly re-useable for cofactor recycling in redox biotransformations.

  18. Purification and characterization of thermostable serine proteases encoded by the genes ttha0099 and ttha01320 from Thermus thermophilus HB8.

    Science.gov (United States)

    Li, Hui; Sun, Yajie; Jiao, Xue; Wang, Honglin; Zhu, Hu

    2016-07-01

    As an important class of proteases, serine proteases are required to show high activity under diverse conditions, especially at high temperatures. In the current study, two serine proteases SP348 and SP404 were analyzed by different bioinformatics tools. Both proteins are comprised of a trypsin domain and a PDZ domain, and belong to the trypsin family of proteases. The proteins were successfully expressed with Trx-tags as soluble proteins in the specialized Escherichia coli Rosetta-gami B(DE3)pLysS strain. A simple three-step purification protocol involving heat treatment, Ni-NTA purification and gel filtration was adopted to purify SP404. The molecular weight of recombinant SP404 was about 64 kDa. According to the circular dichroism spectroscopy analysis, SP404 is thermostable at 70 °C with alpha-helix, beta-sheet and random coil contents of about 8, 22 and 70 %, respectively. Our findings may broaden the range of microorganism-derived proteases and have a wide potential for industrial and fundamental studies. PMID:27215206

  19. 嗜热链球菌抗噬菌体菌株的选育%Selection of bacteriophage-resistant mutants of Streptococcus thermophilus

    Institute of Scientific and Technical Information of China (English)

    冷一非; 赵述淼; 朱辉; 梁运祥

    2010-01-01

    研究以酸奶生产菌株嗜热链球菌S2为出发菌株.采用自然选育的方法筛选出其抗噬菌体突变株.获得4株生产性能较好的抗噬菌株KS21、KS22、KS23、KS24,溶源性检测其为非溶源菌.对4株抗性菌株进行20次传代和酸奶发酵实验,结果表明抗性稳定,其中KS23发酵酸奶产酸达109°T,凝乳时间与出发菌株相当.细胞全蛋白SDS-PAGE电泳分析显示突变菌株与出发菌株在某些蛋白表达上有所差异,可能是抗性产生的原因.

  20. Impact of process parameters on the sourdough microbiota, selection of suitable starter strains, and description of the novel yeast Cryptococcus thermophilus sp. nov.

    OpenAIRE

    Vogelmann, Stephanie Anke

    2013-01-01

    The microbiota of a ripe sourdough consists of lactic acid bacteria (LAB), especially of the genus Lactobacillus, and yeasts. Their composition is influenced by the interplay of species or strains, the kind of substrate as well as the process parameters temperature, dough yield, redox potential, refreshment time, and number of propagation steps (Hammes and Gänzle, 1997). As taste and quality of sourdough breads are mainly influenced by the fermentation microbiota, intense research has been fo...

  1. Detection and Homology Analysis of CRISPR in Streptococcus thermophilus%嗜热链球菌中CRISPR序列的检测与同源性分析

    Institute of Scientific and Technical Information of China (English)

    邓凯波; 霍贵成

    2013-01-01

    为探明内蒙古的8株嗜热链球菌的CRISPR,用PCR方法扩增了其CRISPR序列,并采用生物信息学方法分析和预测了重复序列(direct repeat,DR)的同源性及二级结构.结果表明:除S4只含有一个CRISPR结构外,其余7株嗜热链球菌均检测出3条CRISPR序列,远高于CRISPR database公布细菌中具有该结构的比例(46.4%),且分别具特异性的重复序列(DR1~DR3); CRISPR最长达2853bp,最短仅101bp.对DR的二级结构预测发现,DR1~DR3均能形成回文结构,但茎环大小会有差异;在同源性比对中发现,除多数同属或同种外,供试DR还与个别远缘菌种具有高同源性,这种现象证明了供试菌株的CRISPR序列同样存在的水平基因转移,并可能存在其他的进化进程.

  2. Detection of CRSPR-Cas system in Streptococcus thermophilus%嗜热链球菌CRISPR-Cas系统的检测

    Institute of Scientific and Technical Information of China (English)

    李婉; 梁宏彰; 张丹青; 王娜娜; 唐雅茹; 李柏良; 霍贵成

    2016-01-01

    [目的]CRISPR-Cas系统为嗜热链球菌抵抗噬菌体等外源基因元件提供获得性免疫,分析NCBI中已公开发表全基因组序列的9株嗜热链球菌所含CRISPR-Cas系统的数目和类型,对实验室相应菌株的CRISPR-Cas系统进行检测.[方法]利用生物信息学方法对NCBI中9株已测序嗜热链球菌所含CRISPR-Cas系统进行分析,根据其Cas基因序列设计引物,对实验室嗜热链球菌菌株的Cas基因进行扩增、测序,分析实验室6株嗜热链球菌的CRISPR-Cas系统情况.[结果]9株标准菌株均含不同数目的CRISPR-Cas系统,其类型主要为Ⅱ-A型、Ⅲ-A型和Ⅰ-E型,各类型的标志Cas基因高度保守.6株供试菌中,S4仅含Cas9基因,其它5株均含有Cas9基因、Ca1O基因和Cas9*基因,79和KLDS3.0207还含有Cas3基因.[结论]可根据标准菌株高度保守的Cas基因设计引物,预测未知嗜热链球菌所含CRISPR-Cas系统的数目和类型.S4仅含1个Ⅱ-A型CRISPR-Cas系统,其它5株均含有2个Ⅱ-A型CRISPR-Cas系统和1个Ⅲ-A型CRISPR-Cas系统,此外,79和KLDS3.0207均含有1个Ⅰ-E型CRISPR-Cas系统.

  3. Molecular identification of differentially regulated genes in the hydrothermal-vent species Bathymodiolus thermophilus and Paralvinella pandorae in response to temperature

    OpenAIRE

    Shillito Bruce; Jollivet Didier; Boutet Isabelle; Moraga Dario; Tanguy Arnaud

    2009-01-01

    Abstract Background Hydrothermal vents and cold seeps represent oases of life in the deep-sea environment, but are also characterized by challenging physical and chemical conditions. The effect of temperature fluctuations on vent organisms in their habitat has not been well explored, in particular at a molecular level, most gene expression studies being conducted on coastal marine species. In order to better understand the response of hydrothermal organisms to different temperature regimes, d...

  4. Elektronentransfer zwischen Komplex III und IV der Atmungskette von Paracoccus denitrificans und Thermus thermophilus : funktionelle und kinetische Charakterisierung der Interaktionen anhand von löslichen Fragmenten

    OpenAIRE

    Janzon, Julia

    2007-01-01

    Adenosintriphosphat (ATP) als universelles Energieäquivalent der Zelle wird durch die oxidative Phosphorylierung synthetisiert, bei der Elektronen entlang des elektrochemischen Gefälles der Atmungskette über verschiedene Redoxkomplexe transferiert und durch die chemiosmotische Kopplung Protonen über die Membran gepumpt werden. Der Protonengradient wird dann von der ATP-Synthase genutzt, um ADP zu ATP zu phosphorylieren. Zentraler Redoxkomplex der Atmungskette vieler Pro- und Eukaryonten ist d...

  5. Effects of strain properties and proportion of Streptococcus thermophilus to Lactobacillus delbrueckii subsp.bulgaricus on the quality of yoghurt%菌株性能及球杆菌比例对酸奶品质的影响

    Institute of Scientific and Technical Information of China (English)

    杜昭平; 刘鑫; 马成杰; 华宝珍; 马爱民

    2011-01-01

    Fermentation properties of individual strains and combination fermentation of S.t S1,S.t R1(Streptococcus thermophilus)and L.b L1(Lactobacillus delbrueckii subsp.bulgaricus)with different proportion were performed.The differences of fermentation time,viscosity,syneresis,postacidification and sensory evaluation of yoghurt with different combinations were analyzed.The results indicated that the growth rate of S.t S1 was slower than that of S.t R1,and the acidic rate was similar between each other,but the exopolysaccharide production and yoghurt viscosity of S.t S1 were better than that of S.t R1.The above mentioned properties of strain L.b L1 were inferior to that of S.t S1 and S.t R1.The yoghurt combinatively fermented by S.t S1 and L.b L1 with proportion of 100:1 was better than other combinations,the viscosity and syneresis of which was 0.549Pa·s and 2.5mL/10g respectively,and the acidity rose only 12°T after 15 days at 4℃.The properties and combination modes of strains are with significant influence on the quality of yoghurt.%分析测定了从某直投式发酵剂中分离的两株嗜热链球菌(Streptococcus thermophilus)S.t S1、S.t R1和一株保加利亚乳杆菌(Lactobacillus delbrueckii subsp.bulgaricus)L.bL1的发酵性能,并将球菌与杆菌以不同比例组合发酵,比较酸奶发酵时间、粘度、乳清析出、后酸化和感官评价差异。结果表明,嗜热链球菌S.tS1生长速度略慢于S.tR1,酸化速度相当,但S.tS1的胞外多糖产量和发酵乳粘度明显高于S.tR1,保加利亚乳杆菌L.bL1的上述性

  6. The physiological functions of capsule polysaccharide produced by streptococcus salivarius subsp. Thermophilus%唾液链球菌嗜热亚种荚膜多糖生理功能特性研究

    Institute of Scientific and Technical Information of China (English)

    顾瑞霞; 刘爱萍; 程涛; 骆承庠

    2000-01-01

    通过对唾液链球菌嗜热亚种的研究发现,细胞固有发酵活性、抵抗外界不良环境能力与荚膜无关.破坏唾液链球菌嗜热亚种LCX2001的荚膜,会使发酵速度加快,但同时对外界不良环境的抵抗力下降.这些研究结果表明,有无荚膜,或荚膜大小对同一亚种所表现出来的不同生理特性没有决定作用;破坏荚膜,会影响细胞体与外界物质交换能力,对细胞体许多生理功能有影响;荚膜起到阻止外界物质的进入细胞和细胞内产生的乳酸向外界释放的作用.

  7. Study on Flavour Compounds Analysis in Milk by Coculturing with Streptococcus salivarius subsp.thermophilus%唾液链球菌嗜热亚种发酵乳风味成分分析

    Institute of Scientific and Technical Information of China (English)

    王伟军; 李延华; 张兰威; 于俊林

    2008-01-01

    本实验使用同时蒸馏萃取-气谱-质谱(SDE-GC-MS)技术对5株唾液链球菌嗜热亚种S.t-9、S.t-17YA、S.t-1703CA、S.t-1703D和S.t-03发酵乳中风味物质进行了定性及定量分析,分别鉴定出24种、20种、13种、20种、21种风味物质,共涉及6大类:酸类化合物、酯类化合物、醇类化合物、羰基化合物、芳环和杂环化合物,35种物质.另外,5株菌代谢主要风味物质2,3.丁二酮的能力大小为:S.t-1703D>S.t-1703CA>S.t-17YA>S.t-03>S.t-9,除S.t-9菌代谢2,3-丁二酮量较低外,其它4种菌代谢丁二酮的能力差异不大.

  8. KINETICS OF EXOPOLYSACCHARIDE BY STREPTOCOCCUS SALIVARIUS SUBSP. THERMOPHILUS LCX2001 FERMENTATION FOR PRODUCTION OF EXOPOLYSACCHARIDE%唾液链球菌嗜热亚种LCX2001胞外多糖分批发酵动力学

    Institute of Scientific and Technical Information of China (English)

    顾瑞霞; 刘爱萍; 骆承庠

    2001-01-01

    对唾液链球菌嗜热亚种LCンX2001生物合成胞外多糖(EPS)的动力学进行了研究,基于Logistic-equation方程和Luedeking-piret方程,得到了描述发酵过程的动力学模型和模型参数.该模型反映了细胞生长、乳酸生成、EPS生物合成和基质消耗之间的关系,模型的拟合结果与实验值吻合较好,误差小于10%.

  9. Optimization of Carrot Yoghurt Production by Fermentation of Immobilized Lactobacillus delbrueckii subsp bulgaricus and Streptococcus salivarius subsp thermophilus%固定化乳酸菌发酵胡萝卜酸奶工艺研究

    Institute of Scientific and Technical Information of China (English)

    吴定; 孙嘉文; 姚明兰; 路桂红

    2010-01-01

    通过固定化乳酸菌发酵胡萝卜酸奶,建立新的胡萝卜酸奶发酵工艺.经过单因素试验和正交试验,确定固定化乳酸菌发酵胡萝卜酸奶的配方和发酵工艺.胡萝卜酸奶的配方:奶粉9g、胡萝卜13g、白砂糖8g、蒸馏水100mL.胡萝卜酸奶发酵工艺:固定化德氏乳杆菌保加利亚亚种与固定化唾液链球菌嗜热亚种的比为2:1接种发酵液(菌种的总添加量为6g/100mL)、在37℃发酵培养7h、然后4℃冷藏12h.结果表明,采用固定化乳酸菌新技术可以发酵生产营养丰富、口感良好的胡萝卜酸奶.

  10. Scientific Opinion on the safety and efficacy of Probiotic LACTINA® (Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus bulgaricus, Lactobacillus lactis, Streptococcus thermophilus and Enterococcus faecium for chickens for fattening and piglets

    Directory of Open Access Journals (Sweden)

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP

    2013-04-01

    Full Text Available Probiotic LACTINA® is a feed additive consisting of six strains of lactic acid bacteria. In 2007 the European Food Safety Authority was requested to evaluate the safety and efficacy of the additive when used as a zootechnical additive (functional group: gut flora stabilisers in diets for chickens for fattening, piglets and pigs for fattening. At that time the FEEDAP Panel was able to conclude only on the safety for the environment. EFSA is now requested to assess the safety for the target animals and consumers, and the efficacy of the additive when added in diets for chickens for fattening at 5 x 108 - 9 x 109 CFU/kg feed and for piglets at 9 x 108 - 1 x 1010 CFU/kg feed. The identity of the six strains was established. As no antibiotic resistance of concern was detected and as five of the strains qualify for the Qualified Presumption of Safety approach to safety assessment, their use is presumed safe for target species and consumers. The Enterococcus faecium strain is not a recognised pathogen for animals and as the additive did not adversely affect growth of chickens or piglets, Probiotic LACTINA® is considered safe for the target species up to 1 x 1010 CFU/kg feed. The E. faecium strain does not contain marker genes typical of hospital-associated isolates responsible for clinical infections. The metabolic pathways of E. faecium are well known. Since no other harmful metabolites are expected and the additive does not contain excipients of concern, it is considered safe for consumers. Probiotic LACTINA® showed a potential to increase the body weight of suckling piglets when used at approximately 5 x 109 CFU/kg feed. The Panel could not conclude on the efficacy of Probiotic LACTINA® when used in diets for chickens for fattening or weaned piglets.

  11. Study on co-culture of Yeast and Streptococcus.thermophilus with Apple Pomace%苹果渣制备饲料酵母和嗜热链球菌

    Institute of Scientific and Technical Information of China (English)

    李瑛; 吕嘉枥

    2011-01-01

    With apple pomace-based materials, separate and mixed culture of yeast and Streptococcus thermophilusiS. T) was studied. The respective growth curves of them under different temperature were discussed, and on this basis, the feasibili-ty of the mixed culture was studied. The results indicated that : in separate fermentation the total number of viable cells was up to 4. 32× 108 cfu/ml, in staged constant-temperature-fermentation (first 4% yeast inoculated and cultured at 35℃ for 2 d, then 2% S. T inoculated) the total number of viable was up to 6. 53 X 108 cfu/ml, and in staged variable-temperature-fermenta-tion(first 4% yeast inoculated and cultured at 32℃ for 2 d, then up to 37℃ , 2% of S. T inoculated) the total number of viable cells was up to 7. 21× 108 cfu/ml, there were no major differences. Thus, the use of apple pomace mixed culture of yeasts and S. T was indeed feasible.%以苹果渣为主料,分别单独和混合培养饲料酵母菌和嗜热链球菌.探讨了各自在不同温度下的生长曲线,并在此基础上研究了混合培养的可行性.结果显示:单独发酵活菌总数可达4.32×108 cfu/ml,分段恒温发酵(35℃下培养,先接入4%酵母菌培养2d,再接入2%嗜热链球菌)活菌总数可达6.53×108 cfu/ml,分段变温培养(32℃下先接入4%酵母菌培养2d,再将温度调制37℃接入2%嗜热链球菌)活菌总数可达7.21×108 cfu/ml,没有较大差异.由此可见,利用苹果渣混合培养酵母菌和嗜热链球菌确实可行.

  12. Draft Genome Sequence of Lactobacillus delbrueckii subsp. bulgaricus LBB.B5

    Science.gov (United States)

    Hajo, Karima; Lenoci, Leonardo; Bron, Peter A.; Dijkstra, Annereinou; Alkema, Wynand; Wels, Michiel; Siezen, Roland J.; Minkova, Svetlana; van Hijum, Sacha A. F. T.

    2016-01-01

    Lactobacillus delbrueckii subsp. bulgaricus LBB.B5 originates from homemade Bulgarian yogurt and was selected for its ability to form a strong association with Streptococcus thermophilus. The genome sequence will facilitate elucidating the genetic background behind the contribution of LBB.B5 to the taste and aroma of yogurt and its exceptional protocooperation with S. thermophilus.

  13. 一株嗜热菌的分离、鉴定及海藻糖合成酶基因的克隆%Isolation and Identification of a Thermophile Bacterium and Cloning of Trehalose Synthase Gene

    Institute of Scientific and Technical Information of China (English)

    王慧荣; 韦宇拓; 黄日波

    2011-01-01

    从广西贺州市80℃热泉分离了一株嗜热菌TTH,生长温度范围在55℃~85℃,最适生长温度70℃左右,最高生长温度85℃,G+C物质的量分数为67.6%.对菌株TTH从形态、培养条件、16S rRNA基因序列和系统发育分析方面进行了研究,表明该菌株与3株嗜热栖热菌(Thermus thermophilus)的同源性达到98%,初步鉴定是嗜热栖热菌(Thermus thermophilus).克隆了该菌的海藻糖合成酶基因,它与菌株Thermus thermophilus RQ-1,Thermus caldophilus,Thermus thermophilus HB8,Thermus thermophilus的同源性达到87%.%A thermophile strain, designated as TTH, was isolated from 80℃ hot spring at Hezhou county in Guangxi province. It could grow at temperatures from 55℃ to 85℃, with the optimal temperature 70℃, the highest temperature 85℃ , and with 67. 6% molar content of the G + C. The researches which were based on the TTH morphological , culture conditions, 16S rDNA sequence, as well as the phylogenetic analysis show that the strain TTH has 98% sequence identity with three kinds of Thennus thermophilus. Therefore the strain is tentatively identified as Thermits thermophilus. Trehalose synthase gene of TTH was cloned and analyzed. It has 87% identity to the treS gene from Thermits thermophilus RQ-1, Thermits caldophilus, Thermits thermophilus HB8 and Thermits thermophilus.

  14. Antibiotic resistance of lactic acid bacteria isolated from Chinese yogurts.

    Science.gov (United States)

    Zhou, N; Zhang, J X; Fan, M T; Wang, J; Guo, G; Wei, X Y

    2012-09-01

    The aim of this study was to evaluate the susceptibility of 43 strains of lactic acid bacteria, isolated from Chinese yogurts made in different geographical areas, to 11 antibiotics (ampicillin, penicillin G, roxithromycin, chloramphenicol, tetracycline, chlortetracycline, lincomycin, kanamycin, streptomycin, neomycin, and gentamycin). The 43 isolates (18 Lactobacillus bulgaricus and 25 Streptococcus thermophilus) were identified at species level and were typed by random amplified polymorphic DNA analysis. Thirty-five genotypically different strains were detected and their antimicrobial resistance to 11 antibiotics was determined using the agar dilution method. Widespread resistance to ampicillin, chloramphenicol, chlortetracycline, tetracyclines, lincomycin, streptomycin, neomycin, and gentamycin was found among the 35 strains tested. All of the Strep. thermophilus strains tested were susceptible to penicillin G and roxithromycin, whereas 23.5 and 64.7% of Lb. bulgaricus strains, respectively, were resistant. All of the Strep. thermophilus and Lb. bulgaricus strains were found to be resistant to kanamycin. The presence of the corresponding resistance genes in the resistant isolates was investigated through PCR, with the following genes detected: tet(M) in 1 Lb. bulgaricus and 2 Strep. thermophilus isolates, ant(6) in 2 Lb. bulgaricus and 2 Strep. thermophilus isolates, and aph(3')-IIIa in 5 Lb. bulgaricus and 2 Strep. thermophilus isolates. The main threat associated with these bacteria is that they may transfer resistance genes to pathogenic bacteria, which has been a major cause of concern to human and animal health. To our knowledge, the aph(3')-IIIa and ant(6) genes were found in Lb. bulgaricus and Strep. thermophilus for the first time. Further investigations are required to analyze whether the genes identified in Lb. bulgaricus and Strep. thermophilus isolates might be horizontally transferred to other species.

  15. Diversity, Activity, and Evolution of CRISPR Loci in Streptococcus thermophilus▿ †

    OpenAIRE

    Horvath, Philippe; Romero, Dennis A.; Coûté-Monvoisin, Anne-Claire; Richards, Melissa; Deveau, Hélène; Moineau, Sylvain; Boyaval, Patrick; Fremaux, Christophe; Barrangou, Rodolphe

    2007-01-01

    Clustered regularly interspaced short palindromic repeats (CRISPR) are hypervariable loci widely distributed in prokaryotes that provide acquired immunity against foreign genetic elements. Here, we characterize a novel Streptococcus thermophilus locus, CRISPR3, and experimentally demonstrate its ability to integrate novel spacers in response to bacteriophage. Also, we analyze CRISPR diversity and activity across three distinct CRISPR loci in several S. thermophilus strains. We show that both ...

  16. 唾液链球菌嗜热亚种LCX2001胞外多糖合成条件的研究%Study on Exopolysaccharide Synthesis by Streptococcus salivarius subsp.thermophilus LCX2001 Grown in Chemically Defined Medium

    Institute of Scientific and Technical Information of China (English)

    顾瑞霞; 刘爱萍; 程涛; 骆承庠

    2000-01-01

    从众多的用于乳酸菌生长的合成和半合成培养基(MRS、M17、ATP、SL、Elliker、SDM),选择了ATP作为唾液链球菌嗜热亚种LCX2001胞外多糖产生的基本培养基,在此基础上开发了LCX合成培养基,并对碳源、培养基的初始pH、发酵温度和发酵时间进行了探讨.该菌生物合成EPS的工艺条件为:葡萄糖和/或乳糖是该菌株合成EPS的合适的碳源,添加量为20g/L,培养基的初始pH值为6.5,发酵温度为35℃,发酵时间为20h.

  17. Fermentation Characteristics of Lactobacillus delbrueckii subsp, bulgaricus Co-cultured with Different Streptococcus thermophilus Strains%不同发酵特性的嗜热链球菌与保加利亚乳杆菌共发酵的特性

    Institute of Scientific and Technical Information of China (English)

    李莎; 马成杰; 徐志平; 徐爱才; 华宝珍

    2015-01-01

    为探究具有不同优良性状的嗜热链球菌株与保加利亚乳杆菌株共发酵的特性,用产酸快的嗜热链球菌StCH-1菌株和产黏好的嗜热链球菌StTA040菌株与保加利亚乳杆菌Lb0925B菌株组合,测定其组合菌株的发酵性能.通过实验发现组合菌株发酵速率均比单菌株发酵速率快,其中含有嗜热链球菌StCH-1的组合菌株发酵速率较快,而含有嗜热链球菌StTA040的组合菌株的胞外多糖产量较高,发酵乳的黏度较高,持水力较强;三组分发酵剂的发酵速率快,发酵乳在贮藏期间黏度高,持水力强,通过感官分析得出三组分发酵剂制得的发酵乳的口感和风味最佳.

  18. Production of Synbiotic Yogurt-Like Using Indigenous Lactic Acid Bacteria as Functional Food

    Directory of Open Access Journals (Sweden)

    M. Astawan

    2012-04-01

    Full Text Available Yoghurt is a product of fermented milk using Lactobacillus bulgaricus and Streptococcus thermophilus as culture starter. Indigenous probiotic lactic acid bacteria, Lactobacillus plantarum 2C12 or Lactobacillus acidophilus 2B4, were applied in the making of functional synbiotic yoghurt-like with 5% of fructo-oligosaccharide (FOS as a prebiotic source. The aim of this study was to determine the best formula of functional synbiotic yoghurt-like among four formulas: F1 (L. bulgaricus + S. thermophilus, F2 (L. bulgaricus + S. thermophilus + L. plantarum 2C12, F3 (L. bulgaricus+ S. thermophilus + L. acidophilus 2B4, and F4 (L. bulgaricus + S. thermophilus + L. plantarum 2C12 + L. acidophilus 2B4 to be choosen and followed detection of it’s flavor to improve the product quality and consumer acceptance. The results showed that the F3 synbiotic yogurt made from mixed culture L. bulgaricus, S. thermophilus, and L. acidophilus 2B4 had the highest antibacterial effect against Enteropathogenic Escherichia coli (EPEC. Addition of 1.75% natural corn starch as a stabilizer produced optimum improvement in yoghurt consistency and minimize whey separation. Result of sensory evaluation indicated that the yoghurt with addition of 1% strawberry flavor and 0.1% vanilla flavor were ranked at first and second. Yoghurts were still good to be consumed after 15 d storage period at the refrigeration temperature (10 oC.

  19. Thermostabilization of desulfurization enzymes from Rhodococcos sp. IGTS8. Final technical report

    Energy Technology Data Exchange (ETDEWEB)

    John J. Kilbane II

    2000-12-15

    The objective of this project was to develop thermophilic cultures capable of expressing the desulfurization (dsz) operon of Rhodococcus sp. IGTS8. The approaches taken in this project included the development of plasmid and integrative expression vectors that function well in Thermus thermophilus, the cloning of Rhodococcus dsz genes in Thermus expression vectors, and the isolation of bacterial cultures that express the dsz operon at thermophilic temperatures. This project has resulted in the development of plasmid and integrative expression vectors for use in T. thermophilus. The dsz genes have been expressed at moderately thermophilic temperatures (52 C) in Mycobacterium phlei and at temperatures as high as 72 C in T. thermophilus. The tools and methods developed in this project will be generally useful for the expression of heterologous genes in Thermus. Key developments in the project have been the isolation of a Mycobacterium phlei culture capable of expressing the desulfurization operon at 52 C, development of plasmid and integrative expression vectors for Thermus thermophilus, and the development of a host-vector system based on the malate dehydrogenase gene that allows plasmids to be stably maintained in T. thermophilus and provides a convenient reporter gene for the accurate quantification of gene expression. Publications have been prepared regarding each of these topics; these preprints are included.

  20. Antibacterial Activity of Some Lactic Acid Bacteria Isolated from an Algerian Dairy Product

    Directory of Open Access Journals (Sweden)

    Abdelkader Mezaini

    2009-01-01

    Full Text Available In the present study, the antibacterial effect of 20 lactic acid bacteria isolates from a traditional cheese was investigated. 6 isolates showed antibacterial effect against Gram positive bacteria. Streptococcus thermophilus T2 strain showed the wide inhibitory spectrum against the Gram positive bacteria. Growth and bacteriocin production profiles showed that the maximal bacteriocin production, by S. thermophilus T2 cells, was measured by the end of the late-log phase (90 AU ml−1 with a bacteriocine production rate of 9.3 (AU ml−1 h−1. In addition, our findings showed that the bacteriocin, produced by S. thermophilus T2, was stable over a wide pH range (4–8; this indicates that such bacteriocin may be useful in acidic as well as nonacidic food. This preliminarily work shows the potential application of autochthonous lactic acid bacteria to improve safety of traditional fermented food.

  1. Effect of feed supplements on the occurrence of coccidia oocysts in the digestive tract of pheasants

    Directory of Open Access Journals (Sweden)

    Luboš Zábranský

    2016-05-01

    Full Text Available The occurrence of coccidiosis is an enormous problem of pheasant farming. The aim of this study is to show, that this disease can be cured by unconventional dietary supplements. Dietary supplements which were used in this study were as follows: prebiotics (Ascophyllum nodosum, probiotics (Lactobacillus thermophilus and homeopathics. Four groups of pheasants were made - three experimental groups and one control group. All groups were consisted of 12 individuals. The pheasant Phasianus colchicus was used in this experiment. Dietary supplements were served for 30days. The favourable effect of homeopatics (P < 0.05 and Lactobacillus thermophilus (P < 0.05 on the intestinal tract of pheasant was statistical significant.

  2. Probiotinių mikroorganizmų įtaka pieno rūgšties izomerų kiekiui jogurte

    OpenAIRE

    Garmienė, G.; Kulikauskienė, M.; Saikauskienė, V.

    2005-01-01

    Fermentiniu metodu tirtas pieno rūgšties izomerų kiekis jogurte. Nustatyta, kad jogurto gamyboje kartu vartojant tradicinių pieno rūgšties bakterijų Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus ir probiotinių kultūrų Bifidobacterium, Lactobacillus acidophilus, Lactobacillus casei subsp. casei mišinius bei didinant santykinį S. thermophilus kiekį bendroje tradicinių jogurto kultūrų masėje, galima produktui suteikti ne tik pageidaujamas kokybines savybes, bet ir sumaž...

  3. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA) Scientific Opinion on the substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus

    DEFF Research Database (Denmark)

    Tetens, Inge

    substantiation of a health claim related to a combination of Bifidobacterium longum LA 101, Lactobacillus helveticus LA 102, Lactococcus lactis LA 103 and Streptococcus thermophilus LA 104 and improves stools frequency. The food that is the subject of the health claim is a combination of B. longum LA 101, L...... and effect relationship cannot be established between the consumption of a combination of B. longum LA 101, L. helveticus LA 102, L. lactis LA 103 and S. thermophilus LA 104 and improves stool frequency. © European Food Safety Authority, 2013...

  4. An intermolecular binding mechanism involving multiple LysM domains mediates carbohydrate recognition by an endopeptidase

    DEFF Research Database (Denmark)

    Wong, Jaslyn E M M; Midtgaard, Søren Roi; Gysel, Kira;

    2015-01-01

    of multiple LysM domains in substrate binding has so far lacked support from high-resolution structures of ligand-bound complexes. Here, a structural study of the Thermus thermophilus NlpC/P60 endopeptidase containing two LysM domains is presented. The crystal structure and small-angle X-ray scattering...

  5. Influence of ions on growth and production of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus NCFB 2772

    NARCIS (Netherlands)

    Grobben, G.J.; Boels, I.C.; Sikkema, J.; Smith, M.R.; Bont, de J.A.M.

    2000-01-01

    Several lactic acid bacteria produce exopolysaccharides (EPS), either attached to the cell wall or excreted into the environment as slime material. EPS produced by Lactobacillus delbrueckii subsp. bulgaricus (Lb. bulgaricus) and Streptococcus thermophilus play an important role in improving the text

  6. ANTIMICROBIAL ACTIVITY AND PHOTOCHEMICAL SCREENING OF TINOSPORA CORDIFOLIA AND EUPHORBIA HIRTA

    OpenAIRE

    Amit Sandhu; Neha bhardwaj; Vikas Menon; Ruby Gupta

    2013-01-01

    The present study deals with the antimicrobial activity and phytochemical screening of the two medicinal plants, Tinospora cordifolia and Euphorbia hirta those are commonly available in India. Results of antimicrobial activity revealed that these medicinal plant extracts were very effective against Serratia marcescens, E. coli, Streptococcus thermophilus, Fusarium oxysporium, Aspergillus niger while these extracts showed very less inhibition against Trichoderma reesei. Phytochemical analysis ...

  7. STUDY OF THE COMBINED EFFECTS OF ARABINOGALACTAN AND LACTULOSE ON THE DEVELOPMENT OF THE STARTER CULTURES FOR YOGHURT Исследование совместного влияния арабиногалактана и лактулозы на развитие закваски для йогурта

    Directory of Open Access Journals (Sweden)

    Zavezenova I. V.

    2013-05-01

    Full Text Available In this article, we present the results of the research of influence of polysaccharides and prebiotics on physical, chemical, rheological, microbiological and organoleptic indicators of a sour-milk product on the basis of ferment starting cultures of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus. The optimum dose of polysaccharides and prebiotics adding into a sour-milk product is proved

  8. Thermophilin 13, a nontypical antilisterial poration complex bacteriocin, that functions without a receptor

    NARCIS (Netherlands)

    Marciset, O.; Jeronimus-Stratingh, C.M; Mollet, B.; Poolman, B.

    1997-01-01

    A novel broad host range antimicrobial substance, Thermophilin 13, has been isolated and purified from the growth medium of Streptococcus thermophilus. Thermophilin 13 is composed of the antibacterial peptide ThmA (M-r of 5776) and the enhancing factor ThmB (M-r of 3910); the latter peptide increase

  9. Response surface optimization of the medium components for the production of biosurfactants by probiotic bacteria

    NARCIS (Netherlands)

    Rodrigues, L; Teixeira, J; Oliveira, R; van der Mei, HC

    2006-01-01

    Optimization of the medium for biosurfactants production by probiotic bacteria (Lactococcus lactis 53 and Streptococcus thermophilus A) was carried out using response surface methodology. Both biosurfactants were proved to be growth-associated, thus the desired response selected for the optimization

  10. Examination of middle concentrations of oxytetracycline residues in cow milk after intrauterine application

    OpenAIRE

    Vuković V.; Kasalica A.; Vićentijević M.

    2011-01-01

    After intrauterine application of oxytetracycline in a dose of 2.0 g per cow, excretion of residues in milk was monitored using Resasurine test with Str. thermophilus and specific quantitative procedure (Vukovic, 1999). Mean concentrations of oxytetracycline residues in milk samples, the total quantity per milking and the total quantity per cow were monitored.

  11. Production of Volatile Compounds in Reconstituted Milk Reduced-Fat Cheese and the Physicochemical Properties as Affected by Exopolysaccharide-Producing Strain

    Directory of Open Access Journals (Sweden)

    Weijun Wang

    2012-12-01

    Full Text Available The application of the exopolysaccharide-producing strains for improving the texture and technical properties of reduced-fat cheese looks very promising. Streptococcus thermophilus TM11 was evaluated for production of reduced-fat cheese using reconstituted milk powder (CRMP. The physicochemical analysis of fresh and stored cheeses showed that this strain slightly increased moisture content resulting in cheese with higher yield and lower protein content compared to the direct acidified cheese. The volatiles of cheese were determined by SPME and GC equipped with a mass spectrometer. The results indicated that the major compounds included aldehydes, ketones and acids, whereas, alcohols and branched-chain aldehydes that contribute to exciting and harsh flavors were not found in CRMP. By the textural profile analysis, we found the cheese made with S. thermophilus TM11 had lower cohesiveness, resilience and higher adhesiveness than the direct acidified cheese, and had similar hardness. Further, S. thermophilus TM11 greatly changed the protein matrix with more opened cavities according to observation by scanning electron microscopy. Consequently, use of S. thermophilus TM11 could endow CRMP with the novel and suitable flavor properties and improved texture quality.

  12. Identification of lactic acid bacteria isolated from Tarhana, a traditional Turkish fermented food

    DEFF Research Database (Denmark)

    Sengun, Ilkin Yucel; Nielsen, Dennis Sandris; Karapinar, Mehmet;

    2009-01-01

    to constitute 27% of the isolates, 19% were identified as Streptococcus thermophilus, 19% as Lactobacillus fermentum, 12% as Enterococcus faecium, 7% as Pediococcus pentosaceus, 5% as Leuconostoc pseudomesenteroides, 4% as Weissella cibaria, 2% as Lactobacillus plantarum, 2% as Lactobacillus delbrueckii spp...

  13. Biochemical Characteristics and Viability of Probiotic and Yogurt Bacteria in Yogurt during the Fermentation and Refrigerated Storage

    Directory of Open Access Journals (Sweden)

    F Sarvari

    2014-09-01

    Full Text Available This research aimed to investigate the viability of probiotic bacteria (Lactobacillus acidophilus LA-5 and Bifidobacterium lactis BB-12 and yogurt bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus in yogurt during the fermentation, immediately after fermentation and during refrigerated storage (21 d, 4˚C. Also the biochemical characteristics of milk as affected by the commercial 4-strain mixed starter culture were investigated. Storage time affected the viability of all bacterial species. The concentration of lactic acid during the fermentation increased in parallel with the titrable acidity, and the concentration of acetic acid was proportional to the viability of Bifidobacterium lactis. The acetaldehyde level was decreased in the yogurt from day 0 up to the end of the storage. Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus were multiplied considerably during the fermentation. Streptococcus thermophilus could maintain its viability to the highest level, but Lactobacillus delbrueckii ssp. bulgaricus lost its viability rapidly during the cold storage compared to Streptococcus thermophilus. The multiplication and viability of probiotic bacteria were also influenced by the associative strains and species of yogurt organisms. Bifidobacteria counts were satisfactory. The loss of viability for bifidobacteria was gradual and steady during the storage, and they showed good stability during the storage as compared to Lactobacillus acidophilus.

  14. Effects of cultivation conditions on folate production by lactic acid bacteria

    NARCIS (Netherlands)

    Sybesma, W.; Starrenburg, M.; Tijsseling, L.; Hoefnagel, M.H.N.; Hugenholtz, J.

    2003-01-01

    A variety of lactic acid bacteria were screened for their ability to produce folate intracellularly and/or extracellularly. Lactococcus lactis, Streptococcus thermophilus, and Leuconostoc spp. all produced folate, while most Lactobacillus spp., with the exception of Lactobacillus plantarum, were not

  15. Crystal structures at 2.5 Angstrom resolution of seryl-tRNA synthetase complexed with two analogs of seryl adenylate

    DEFF Research Database (Denmark)

    Belrhali, H.; Yaremchuk, A.; Tukalo, M.;

    1994-01-01

    Crystal structures of seryl-tRNA synthetase from Thermus thermophilus complexed with two different analogs of seryl adenylate have been determined at 2.5 Angstrom resolution. The first complex is between the enzyme and seryl-hydroxamate-AMP (adenosine monophosphate), produced enzymatically...

  16. Nisin-induced Expression of Pediocin in Dairy Lactic Acid Bacteria

    Science.gov (United States)

    To test if a single vector, nisin-controlled expression (NICE) system could be used to regulate expression of the pediocin operon in Streptococcus thermophilus, Lactococcus lactis subsp. lactis and Lactobacillus casei, the intact pediocin operon was cloned into pMSP3535 immediately down stream of th...

  17. Genomics of Streptococcus salivarius, a major human commensal.

    Science.gov (United States)

    Delorme, Christine; Abraham, Anne-Laure; Renault, Pierre; Guédon, Eric

    2015-07-01

    The salivarius group of streptococci is of particular importance for humans. This group consists of three genetically similar species, Streptococcus salivarius, Streptococcus vestibularis and Streptococcus thermophilus. S. salivarius and S. vestibularis are commensal organisms that may occasionally cause opportunistic infections in humans, whereas S. thermophilus is a food bacterium widely used in dairy production. We developed Multilocus sequence typing (MLST) and comparative genomic analysis to confirm the clear separation of these three species. These analyses also identified a subgroup of four strains, with a core genome diverging by about 10%, in terms of its nucleotide sequence, from that of S. salivarius sensu stricto. S. thermophilus species displays a low level of nucleotide variability, due to its recent emergence with the development of agriculture. By contrast, nucleotide variability is high in the other two species of the salivarius group, reflecting their long-standing association with humans. The species of the salivarius group have genome sizes ranging from the smallest (∼ 1.7 Mb for S. thermophilus) to the largest (∼ 2.3 Mb for S. salivarius) among streptococci, reflecting genome reduction linked to a narrow, nutritionally rich environment for S. thermophilus, and natural, more competitive niches for the other two species. Analyses of genomic content have indicated that the core genes of S. salivarius account for about two thirds of the genome, indicating considerable variability of gene content and differences in potential adaptive features. Furthermore, we showed that the genome of this species is exceptionally rich in genes encoding surface factors, glycosyltransferases and response regulators. Evidence of widespread genetic exchanges was obtained, probably involving a natural competence system and the presence of diverse mobile elements. However, although the S. salivarius strains studied were isolated from several human body-related sites

  18. 益生菌豆乳发酵剂的筛选%Screening of probiotic strain of soymilk fermentation starter culture

    Institute of Scientific and Technical Information of China (English)

    包艳; 苏芳; 董喜梅; 姚国强; 王水泉; 张和平

    2011-01-01

    To produce fermented soy milk with favorable taste and sensory properties, L. Plantarum IMAU10156 and L. Plantarum IMAU60042 was repectively combined with L. Acidophilum S21-1, S. Thermophilus S21-2, B. Animalis L01-1 for soy milk fermnetation. The initial inoculation were lxl07 cfu/mL and then subjected to fermentation at 42 'C followed with a 4 °C storage for evaluation of viable counts, pH value, TA, viscosity, syneresis and sensory assessment (Id storage). The results showed that fermented soy milk in the L. Plantarum IMAU10156 and S. Thermophilus S21-2, L. Plantarum IMAU60042 and S. Thermophilus S21-2 group was of better fermented characteristic and pleasing aroma of beans, moderate sweet and great appearance plus with 6h short fermentation time, indicating that it is a promising potential starter cuture for probiotic soy milk fermentation.%为生产出具有良好风味和感官特性的发酵豆乳,选用Lacobacillus plantarum IMAU10156和L.IMAU60042分别与Lacbobacillus acidophilum S21-1,Streptococcus thermophilus S21-2,Bifidobacterium animalis L01-1复合发酵豆乳,接种量分别为1×107 mL-1,于42 °C发酵和4℃贮藏期间测定菌落总数、pH值、滴定酸度、黏度及脱水收缩性指标,并在贮藏1d进行感官鉴评.结果表明,L.plantarum IMAU10156与S.thermophilus S21-2,L.plantarum IMAU60042与S.thermophilus S21-2复合具有良好的发酵特性,在6h即可到达发酵终点,发酵豆乳产品具有纯正的豆乳发酵香味、酸甜适口、组织状态良好等特点,是两组具有开发价值的益生菌豆乳发酵剂.

  19. Sequence of the hyperplastic genome of the naturally competent Thermus scotoductus SA-01

    Directory of Open Access Journals (Sweden)

    Gounder Kamini

    2011-11-01

    Full Text Available Abstract Background Many strains of Thermus have been isolated from hot environments around the world. Thermus scotoductus SA-01 was isolated from fissure water collected 3.2 km below surface in a South African gold mine. The isolate is capable of dissimilatory iron reduction, growth with oxygen and nitrate as terminal electron acceptors and the ability to reduce a variety of metal ions, including gold, chromate and uranium, was demonstrated. The genomes from two different Thermus thermophilus strains have been completed. This paper represents the completed genome from a second Thermus species - T. scotoductus. Results The genome of Thermus scotoductus SA-01 consists of a chromosome of 2,346,803 bp and a small plasmid which, together are about 11% larger than the Thermus thermophilus genomes. The T. thermophilus megaplasmid genes are part of the T. scotoductus chromosome and extensive rearrangement, deletion of nonessential genes and acquisition of gene islands have occurred, leading to a loss of synteny between the chromosomes of T. scotoductus and T. thermophilus. At least nine large inserts of which seven were identified as alien, were found, the most remarkable being a denitrification cluster and two operons relating to the metabolism of phenolics which appear to have been acquired from Meiothermus ruber. The majority of acquired genes are from closely related species of the Deinococcus-Thermus group, and many of the remaining genes are from microorganisms with a thermophilic or hyperthermophilic lifestyle. The natural competence of Thermus scotoductus was confirmed experimentally as expected as most of the proteins of the natural transformation system of Thermus thermophilus are present. Analysis of the metabolic capabilities revealed an extensive energy metabolism with many aerobic and anaerobic respiratory options. An abundance of sensor histidine kinases, response regulators and transporters for a wide variety of compounds are indicative

  20. Bacteriological quality of freshly processed broiler chickens as affected by carcass pretreatment and gamma irradiation

    International Nuclear Information System (INIS)

    Chicken carcasses dipped in whey fermented by Streptococcus thermophilus, lactic acid solution or water and irradiated at 2.5 kGy by 60Co were evaluated for bacteriological quality on day 1, day-3 and at 3-day intervals for an 18-day storage (4 degrees C) period. Unirradiated carcasses treated similarly were used as control. Gram negative bacteria, Yersinia and Campylobacter counts were significantly (p0.01) lower in irradiated samples, but no significant (p0.05) differences were observed ammong the dipping solutions. Salmonellae were completely eliminated in irradiated samples. Whey fermented by S. thermophilus reduced the proportion of Salmonella contaminated carcasses from 67% to 20%. As evidenced by the bacterial counts the shelf-life was found to be 15 days for irradiated carcasses compared to about 6 days for the unirradiated samples

  1. Antibiotic susceptibility of different lactic acid bacteria strains.

    Science.gov (United States)

    Karapetkov, N; Georgieva, R; Rumyan, N; Karaivanova, E

    2011-12-01

    Five lactic acid bacteria (LAB) strains belonging to species Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis and Streptococcus thermophilus were tested for their susceptibility to 27 antibiotics. The minimum inhibitory concentrations of each antimicrobial were determined using a microdilution test. Among the strains a high susceptibility was detected for most of the cell-wall synthesis inhibitors (penicillins, cefoxitin and vancomycin) and resistance toward inhibitors of DNA synthesis (trimethoprim/sulfonamides and fluoroquinolones). Generally, the Lactobacillus strains were inhibited by antibiotics such as chloramphenicol, erythromycin and tetracycline at breakpoint levels lower or equal to the levels defined by the European Food Safety Authority. Despite the very similar profile of S. thermophilus LC201 to lactobacilli, the detection of resistance toward erythromycin necessitates the performance of additional tests in order to prove the absence of transferable resistance genes.

  2. 羊胎水解液乳酸菌发酵菌种的选择

    Institute of Scientific and Technical Information of China (English)

    苏晔; 魏泓; 等

    2002-01-01

    通过研究嗜酸乳杆菌(Lactobacillus acidophilus)1.1854;乳酸乳球菌(Lactobacillus)11454;短乳杆菌(Lactobacillus brevis)1.12;干酪乳杆菌干酪亚种(Lactobacillus casei)1.62;嗜热链球菌(Lactobacillus Streptococcus thermophilus)1.1855;植物乳杆菌9Lactobacillus plantarum)6种菌种对羊胎水解液菌发酵活力、双菌发酵活力、活菌数及风味的影响,确定了乳酸菌发酵羊水解液的最适菌种组合为嗜酸乳杆菌(Lactobacillus acidophilus)1.1854与嗜热链球菌(Lactobacillus Streptococ-cus thermophilus)1.1855。

  3. Evolution of Lysine Biosynthesis in the Phylum Deinococcus-Thermus

    Directory of Open Access Journals (Sweden)

    Hiromi Nishida

    2012-01-01

    Full Text Available Thermus thermophilus biosynthesizes lysine through the α-aminoadipate (AAA pathway: this observation was the first discovery of lysine biosynthesis through the AAA pathway in archaea and bacteria. Genes homologous to the T. thermophilus lysine biosynthetic genes are widely distributed in bacteria of the Deinococcus-Thermus phylum. Our phylogenetic analyses strongly suggest that a common ancestor of the Deinococcus-Thermus phylum had the ancestral genes for bacterial lysine biosynthesis through the AAA pathway. In addition, our findings suggest that the ancestor lacked genes for lysine biosynthesis through the diaminopimelate (DAP pathway. Interestingly, Deinococcus proteolyticus does not have the genes for lysine biosynthesis through the AAA pathway but does have the genes for lysine biosynthesis through the DAP pathway. Phylogenetic analyses of D. proteolyticus lysine biosynthetic genes showed that the key gene cluster for the DAP pathway was transferred horizontally from a phylogenetically distant organism.

  4. Structural basis of transcription activation.

    Science.gov (United States)

    Feng, Yu; Zhang, Yu; Ebright, Richard H

    2016-06-10

    Class II transcription activators function by binding to a DNA site overlapping a core promoter and stimulating isomerization of an initial RNA polymerase (RNAP)-promoter closed complex into a catalytically competent RNAP-promoter open complex. Here, we report a 4.4 angstrom crystal structure of an intact bacterial class II transcription activation complex. The structure comprises Thermus thermophilus transcription activator protein TTHB099 (TAP) [homolog of Escherichia coli catabolite activator protein (CAP)], T. thermophilus RNAP σ(A) holoenzyme, a class II TAP-dependent promoter, and a ribotetranucleotide primer. The structure reveals the interactions between RNAP holoenzyme and DNA responsible for transcription initiation and reveals the interactions between TAP and RNAP holoenzyme responsible for transcription activation. The structure indicates that TAP stimulates isomerization through simple, adhesive, stabilizing protein-protein interactions with RNAP holoenzyme. PMID:27284196

  5. Concentric-flow electrokinetic injector enables serial crystallography of ribosome and photosystem II.

    Science.gov (United States)

    Sierra, Raymond G; Gati, Cornelius; Laksmono, Hartawan; Dao, E Han; Gul, Sheraz; Fuller, Franklin; Kern, Jan; Chatterjee, Ruchira; Ibrahim, Mohamed; Brewster, Aaron S; Young, Iris D; Michels-Clark, Tara; Aquila, Andrew; Liang, Mengning; Hunter, Mark S; Koglin, Jason E; Boutet, Sébastien; Junco, Elia A; Hayes, Brandon; Bogan, Michael J; Hampton, Christina Y; Puglisi, Elisabetta V; Sauter, Nicholas K; Stan, Claudiu A; Zouni, Athina; Yano, Junko; Yachandra, Vittal K; Soltis, S Michael; Puglisi, Joseph D; DeMirci, Hasan

    2016-01-01

    We describe a concentric-flow electrokinetic injector for efficiently delivering microcrystals for serial femtosecond X-ray crystallography analysis that enables studies of challenging biological systems in their unadulterated mother liquor. We used the injector to analyze microcrystals of Geobacillus stearothermophilus thermolysin (2.2-Å structure), Thermosynechococcus elongatus photosystem II (<3-Å diffraction) and Thermus thermophilus small ribosomal subunit bound to the antibiotic paromomycin at ambient temperature (3.4-Å structure).

  6. Signature Lipids and Stable Carbon Isotope Analyses of Octopus Spring Hyperthermophilic Communities Compared with Those of Aquificales Representatives

    OpenAIRE

    Jahnke, Linda L.; Eder, Wolfgang; Huber, Robert; Hope, Janet M.; Hinrichs, Kai-Uwe; Hayes, John M.; Des Marais, David J.; Cady, Sherry L.; Summons, Roger E.

    2001-01-01

    The molecular and isotopic compositions of lipid biomarkers of cultured Aquificales genera have been used to study the community and trophic structure of the hyperthermophilic pink streamers and vent biofilm from Octopus Spring. Thermocrinis ruber, Thermocrinis sp. strain HI 11/12, Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus, and Aquifex aeolicus all contained glycerol-ether phospholipids as well as acyl glycerides. The n-C20:1 and cy-C21 fatty acids dominated all of the Aquificales...

  7. Phylogenetic analysis of the hyperthermophilic pink filament community in Octopus Spring, Yellowstone National Park

    Energy Technology Data Exchange (ETDEWEB)

    Reysenbach, A.L.; Wickham, G.S.; Pace, N.R. [Indiana Univ., Bloomington, IN (United States)

    1994-06-01

    This study uses a molecular phylogenetic approach to characterize the pink filament community at the outflow of Octopus Spring in Yellowstone National Park. The temperature range of the spring is from 84 to 88 C. The authors show that the pink filaments are most closely related to the hydrogen-oxidizing bacterium Aquifex pyrophilus and a close relative Hydrogenobacter thermophilus. 38 refs., 4 figs., 1 tab.

  8. Phosphorylation of Streptococcus salivarius Lactose Permease (LacS) by HPr(His∼P) and HPr(Ser-P)(His∼P) and Effects on Growth

    Science.gov (United States)

    Lessard, Christian; Cochu, Armelle; Lemay, Jean-Dominique; Roy, Denis; Vaillancourt, Katy; Frenette, Michel; Moineau, Sylvain; Vadeboncoeur, Christian

    2003-01-01

    The oral bacterium Streptococcus salivarius takes up lactose via a transporter called LacS that shares 95% identity with the LacS from Streptococcus thermophilus, a phylogenetically closely related organism. S. thermophilus releases galactose into the medium during growth on lactose. Expulsion of galactose is mediated via LacS and stimulated by phosphorylation of the transporter by HPr(His∼P), a phosphocarrier of the phosphoenolpyruvate:sugar phosphotransferase transport system (PTS). Unlike S. thermophilus, S. salivarius grew on lactose without expelling galactose and took up galactose and lactose concomitantly when it is grown in a medium containing both sugars. Analysis of the C-terminal end of S. salivarius LacS revealed a IIA-like domain (IIALacS) almost identical to the IIA domain of S. thermophilus LacS. Experiments performed with purified proteins showed that S. salivarius IIALacS was reversibly phosphorylated on a histidine residue at position 552 not only by HPr(His∼P) but also by HPr(Ser-P)(His∼P), a doubly phosphorylated form of HPr present in large amounts in rapidly growing S. salivarius cells. Two other major S. salivarius PTS proteins, IIABLMan and IIABHMan, were unable to phosphorylate IIALacS. The effect of LacS phosphorylation on growth was studied with strain G71, an S. salivarius enzyme I-negative mutant that cannot synthesize HPr(His∼P) or HPr(Ser-P)(His∼P). These results indicated that (i) the wild-type and mutant strains had identical generation times on lactose, (ii) neither strain expelled galactose during growth on lactose, (iii) both strains metabolized lactose and galactose concomitantly when grown in a medium containing both sugars, and (iv) the growth of the mutant was slightly reduced on galactose. PMID:14617640

  9. Probiotic activity of Lactobacillus delbrueckii subsp. bulgaricus in the oral cavity

    OpenAIRE

    Stamatova, Iva

    2010-01-01

    Yogurt consumption has been related to longevity of some populations living on the Balkans. Yogurt starter L. delbrueckii subsp. bulgaricus and Str. thermophilus have been recognized as probiotics with verified beneficial health effects. The oral cavity emerges as a arget for probiotic applications. Probiotics have demonstrated promising results in controlling dental diseases and oral yeast infections. However, L. bulgaricus despite its broad availability in dairy products has not been evalua...

  10. Cas9 function and host genome sampling in Type II-A CRISPR–Cas adaptation

    OpenAIRE

    Wei, Yunzhou; Terns, Rebecca M.; Terns, Michael P.

    2015-01-01

    Wei et al. found that Cas9, previously identified as the nuclease responsible for ultimate invader destruction, is also essential for adaptation in Streptococcus thermophilus. Cas9 nuclease activity is dispensable for adaptation. Wei et al. also revealed extensive, unbiased acquisition of the self-targeting host genome sequence by the CRISPR–Cas system that is masked in the presence of active target destruction.

  11. Competition mechanisms of lactic acid bacteria and bifidobacteria: Fermentative metabolism and colonization

    OpenAIRE

    Tabasco, Raquel; Fernández de Palencia, P.; Fontecha, F. Javier; Peláez, Carmen; Requena, Teresa

    2014-01-01

    Enzyme activities (α- and β-glucosidases, α- and β-galactosidases and β-fructofuranosidase) and organic acid production of four strains of lactic acid bacteria (LAB; Streptococcus thermophilus STY-31, Lactobacillus delbrueckii subsp. bulgaricus LBY-27, Lactobacillus casei LC-01 and Lactobacillus acidophilus LA-5) and Bifidobacterium lactis BB-12 were tested on milk and MRS fermentation broth with glucose, lactose or fructooligosaccharides (FOS) as carbon source. The highest β-galactosidase ac...

  12. Response surface optimization of the medium components for the production of biosurfactants by probiotic bacteria

    OpenAIRE

    L. R. Rodrigues; J.A. Teixeira; Oliveira, Rosário; Mei, Henny van der

    2006-01-01

    Optimization of the medium for biosurfactants production by probiotic bacteria (Lactococcus lactis 53 and Streptococcus thermophilus A) was carried out using response surface methodology. Both biosurfactants were proved to be growth-associated, thus the desired response selected for the optimization was the biomass concentration. The selected factors based on MRS medium for L. lactis 53 growth were peptone, meat extract, yeast extract, lactose, ammonium citrate and KH2PO4. For S. thermophi...

  13. Hazelnut milk fermentation using probiotic Lactobacillus rhamnosus GG and inulin

    OpenAIRE

    Bernat Pérez, Neus; Cháfer Nácher, María Teresa; Chiralt Boix, Mª Amparo; González Martínez, María Consuelo

    2014-01-01

    Following the consumer demand of healthy vegetable products due to their interesting nutritional profiles and potential functionalities, the fermentation process of hazelnut milk with Lactobacillus rhamnosus GG and S.thermophilus was studied. The effect of different factors (glucose, inulin and inoculum contents) was analysed to ensure sufficient probiotic survivals in a minimum time. The shelf life of the optimised product was characterised in terms of its main physicochemical and quality pa...

  14. Bacteriocin production by strain Lactobacillus delbrueckii ssp. bulgaricus BB18 during continuous prefermentation of yogurt starter culture and subsequent batch coagulation of milk.

    Science.gov (United States)

    Simova, E D; Beshkova, D M; Angelov, M P; Dimitrov, Zh P

    2008-06-01

    By screening for bacteriocin-producing lactic acid bacteria of 1,428 strains isolated from authentic Bulgarian dairy products, Lb. bulgaricus BB18 strain obtained from kefir grain was selected. Out of 11 yogurt starters containing Lb. bulgaricus BB18 and S. thermophilus strains resistant to bacteriocin secreted by Lb. bulgaricus BB18 a yogurt culture (S. thermophilus 11A+Lb. bulgaricus BB18) with high growth and bacteriocinogenic activity in milk was selected. Continuous (pH-stat 5.7) prefermentation processes were carried out in milk at 37 degrees C in a 2l MBR bioreactor (MBR AG, Zurich, Switzerland) with an IMCS controller for agitation speed, temperature, dissolved oxygen, CO2 and pH. Prefermented milk with pH 5.7 coagulated in a thermostat at 37 degrees C until pH 4.8-4.9. S. thermophilus 11A and Lb. bulgaricus BB18 grew independently in a continuous mode at similar and sufficiently high-dilution rates (D=1.83 h(-1)-S. thermophilus 11A; D=1.80 h(-1)-Lb. bulgaricus BB18). The yogurt cultures developed in a stream at a high-dilution rate (D=2.03-2.28 h(-1)). The progress of both processes (growth and bacteriocin production) depended on the initial ratio between the two microorganisms. The continuous prefermentation process promoted conditions for efficient fermentation and bacteriocinogenesis of the starter culture during the batch process: strong reduction of the times for bacteriocin production and coagulation of milk (to 4.5-5.0 h); high cell productivity (lactobacilli-4x10(12) CFU ml(-1), streptococci-6x10(12) CFU ml(-1)); high productivity of bacteriocins (4,500 BU ml(-1))-1.7 times higher than the bacteriocinogenic activity of the batch starter culture.

  15. Selection of folate-producing lactic acid bacteria for improving fermented goat milk

    OpenAIRE

    Sanna, Maria Giovanna; Mangia, Nicoletta Pasqualina; Giovanni GARAU; Murgia, Marco Ambrogio; Massa, Tomasina G.; Franco, Mario Andrea; Deiana, Pietrino

    2005-01-01

    Goat milk is a complete food but its low level of folic acid diminisches its nutritional efficacy. In this study, Streptococcus thermophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lb. delbrueckii subsp. lactis and Lb. helveticus strains were selected for folate production in goat milk to improve its quality. A suitable HPLC method was developed to directly determine both total folate and its biologically active derivatives such as 5-methyl-tetrahydrofolate (5- CH3-H4-PteGlu), tetrah...

  16. Evolución de la proteólisis durante la maduración de quesos Danbo elaborados con distintos cultivos iniciadores

    Directory of Open Access Journals (Sweden)

    Luis Alberto Panizzolo

    2011-12-01

    Full Text Available La determinación de nitrógeno soluble a pH 4,6 (NNC, nitrógeno soluble en ácido tricloroacético (12 % (NTCA y nitrógeno soluble en ácido fosfotúngstico (5 % (NPTA se utiliza como índice de maduración de quesos, ya que proporciona información adecuada de la extensión global de la proteólisis. La formación de péptidos y aminoácidos durante la maduración del queso contribuye directamente al desarrollo del sabor y textura del queso, de allí la importancia de conocer si el uso de distintos cultivos iniciadores genera diferencias en el desarrollo de la proteólisis. En este trabajo se estudiaron los cambios proteolíticos durante la maduración de quesos Danbo elaborados con cultivos iniciadores que difieren en la proporción de sus microorganismos componentes: Lote A con: 60 % Streptococcus thermophilus subsp. thermophilus – 40 % (Lactococcus lactis subsp. lactis + Lactococcus lactis subsp. cremoris y Lote B con: 50 % Streptococcus thermophilus subsp. thermophilus – 50 % (Lactococcus lactis subsp. lactis y Lactococcus lactis subsp. cremoris. Se analizaron quesos con 0, 15, 30 y 45 días de maduración y se determinó contenido de humedad, pH, contenido de nitrógeno en queso y contenido de nitrógeno de las distintas fracciones nitrogenadas. El lote elaborado con un mayor porcentaje de cultivo mesófilo (Lactococcus lactis subsp. lactis y Lactococcus lactis subsp. cremoris, lote B, fue el que presentó mayor proteólisis primaria y mayor formación de oligopéptidos y aminoácidos libres.

  17. Regulatory RNAs in the Less Studied Streptococcal Species: from Nomenclature to Identification

    Directory of Open Access Journals (Sweden)

    Mohamed Amine Zorgani

    2016-07-01

    Full Text Available Streptococcal species are Gram-positive bacteria involved in severe and invasive diseases in humans and animals. Although this group includes different pathogenic species involved in life-threatening infections for humans, it also includes beneficial species, such as Streptococcus thermophilus, which is used in yogurt production. In bacteria virulence factors are controlled by various regulatory networks including regulatory RNAs. For clearness and to develop logical thinking, we start this review with a revision of regulatory RNAs nomenclature. Previous reviews are mostly dealing with Streptococcus pyogenes and Streptococcus pneumoniae regulatory RNAs. We especially focused our analysis on regulatory RNAs in Streptococcus agalactiae, Streptococcus mutans, Streptococcus thermophilus and other less studied Streptococcus species. Although S. agalactiae RNome remains largely unknown, sRNAs (small RNAs are supposed to mediate regulation during environmental adaptation and host infection. In the case of S. mutans, sRNAs are suggested to be involved in competence regulation, carbohydrate metabolism and Toxin-Antitoxin systems. A new category of miRNA-size small RNAs (msRNAs was also identified for the first time in this species. The analysis of S. thermophilus sRNome shows that many sRNAs are associated to the bacterial immune system known as CRISPR-Cas system. Only few of the other different Streptococcus species have been the subject of studies pointed toward the characterization of regulatory RNAs. Finally, understanding bacterial sRNome can constitute one step forward to the elaboration of new strategies in therapy such as substitution of antibiotics in the management of S. agalactiae neonatal infections, prevention of S. mutans dental caries or use of S. thermophilus CRISPR-Cas system in genome editing applications.

  18. Regulatory RNAs in the Less Studied Streptococcal Species: From Nomenclature to Identification

    Science.gov (United States)

    Zorgani, Mohamed A.; Quentin, Roland; Lartigue, Marie-Frédérique

    2016-01-01

    Streptococcal species are Gram-positive bacteria involved in severe and invasive diseases in humans and animals. Although, this group includes different pathogenic species involved in life-threatening infections for humans, it also includes beneficial species, such as Streptococcus thermophilus, which is used in yogurt production. In bacteria virulence factors are controlled by various regulatory networks including regulatory RNAs. For clearness and to develop logical thinking, we start this review with a revision of regulatory RNAs nomenclature. Previous reviews are mostly dealing with Streptococcus pyogenes and Streptococcus pneumoniae regulatory RNAs. We especially focused our analysis on regulatory RNAs in Streptococcus agalactiae, Streptococcus mutans, Streptococcus thermophilus and other less studied Streptococcus species. Although, S. agalactiae RNome remains largely unknown, sRNAs (small RNAs) are supposed to mediate regulation during environmental adaptation and host infection. In the case of S. mutans, sRNAs are suggested to be involved in competence regulation, carbohydrate metabolism, and Toxin–Antitoxin systems. A new category of miRNA-size small RNAs (msRNAs) was also identified for the first time in this species. The analysis of S. thermophilus sRNome shows that many sRNAs are associated to the bacterial immune system known as CRISPR-Cas system. Only few of the other different Streptococcus species have been the subject of studies pointed toward the characterization of regulatory RNAs. Finally, understanding bacterial sRNome can constitute one step forward to the elaboration of new strategies in therapy such as substitution of antibiotics in the management of S. agalactiae neonatal infections, prevention of S. mutans dental caries or use of S. thermophilus CRISPR-Cas system in genome editing applications. PMID:27507970

  19. Determination of reactive oxygen generated from natural medicines and their antibacterial activity

    Directory of Open Access Journals (Sweden)

    Noriko Tajima

    2016-08-01

    Galla chinensis generated 2.4×10−4 mol/L hydrogen peroxide from a 1 mg/mL solution. In bacterial growth tests, Galla chinensis extract had antibacterial activity against Escherichia coli, Staphylococcus aureus, Bacteroides thetaiotaomicron, Campylobacter sputorum biovar sputorum, Streptococcus salivarius thermophilus, Lactobacillus casei, and Bifidobacterium longum infantis. This antibacterial activity was decreased by the addition of catalase. It revealed that hydrogen peroxide which Galla chinensis produced participated in antibacterial activity.

  20. Production optimization of probiotic soft cheese made from goat's and cow's milk

    OpenAIRE

    Ida Drgalić; Ljubica Tratnik; Rajka Božanić; Sandra Koruga

    2006-01-01

    The aim of this study was to determine optimal rennet share and fat content in milk for probiotic soft cheeseproduction made from goat's and cow's milk using DVS mixed probiotic culture ABT-4 (Lactobacillus acidophilus, Bifidobacterium spp. and Streptococcus thermophilus), with desirable sensory properties, which will be acceptable by consumers. The best sensory scores had samples of probiotic soft cheese produced in laboratory conditions from milk with 1% of milk fat, pasteurized at 65°C/30 ...

  1. In vitro Selektion kohlenhydratbindender Proteine

    OpenAIRE

    Talke, Anja

    2010-01-01

    The work presented here addresses the question whether an improved and generally accepted carbohydrate binding domain can be derived from a naturally occurring putative carbohydrate binding domain. For this purpose the surface layer homology (SLH) domain was selected because it is widely distributed in gram-positive bacteria. The SLH domains of five different strains of bacteria (B. sphaericus, B. stearothermophilus, T. thermophilus, E. acidaminophilum, T. kivui) were isolated. Four out of th...

  2. Scarce Evidence of Yogurt Lactic Acid Bacteria in Human Feces after Daily Yogurt Consumption by Healthy Volunteers

    OpenAIRE

    del Campo, Rosa; Bravo, Daniel; Cantón, Rafael; Ruiz-Garbajosa, Patricia; García-Albiach, Raimundo; Montesi-Libois, Alejandra; Yuste, Francisco-Javier; Abraira, Victor; Baquero, Fernando

    2005-01-01

    In a double-blind prospective study including 114 healthy young volunteers, the presence in human feces of the yogurt organisms Lactobacillus delbrueckii and Streptococcus thermophilus after repeated yogurt consumption (15 days) was analyzed by culture, specific PCR, and DNA hybridization of total fecal DNA. Detection of yogurt lactic acid bacteria in total fecal DNA by bacterial culture and PCR assay was consistently negative. DNA compatible with yogurt bacteria was found by hybridization ex...

  3. Use of native gels to measure protein binding to SSB.

    Science.gov (United States)

    Inoue, Jin; Mikawa, Tsutomu

    2012-01-01

    We describe a procedure to detect protein binding to SSB by polyacrylamide gel electrophoresis under non-denaturing conditions. As an example, we show the interaction of Thermus thermophilus (Tth) SSB with its cognate RecO protein. The interaction is detected as decay of the band corresponding to SSB by addition of RecO. We also demonstrate analysis of the RecO-RecR interaction as another example of this method. PMID:22976186

  4. Développement d'un milieu sélectif pour le dénombrement des bifidobactéries dans les laits fermentés

    OpenAIRE

    Bonaparte, Christine; Klein, Günter; Kneifel, Wolfgang; Reuter, Gerhard

    2001-01-01

    International audience Development of a selective culture medium for the enumeration of bifidobacteria in fermented milks. The use of selective culture media is necessary to demonstrate the presence of bifidobacteria in European fermented milks, since Lactobacillus (L.) acidophilus, L. delbrueckii ssp. bulgaricus and Streptococcus (S.) thermophilus usually dominate the flora in these products. But a fully selective medium suppressing these microorganisms does not exist as yet. Therefore, t...

  5. Diversity of thermophilic bacteria in raw, pasteurized and selectively-cultured milk, as assessed by culturing, PCR-DGGE and pyrosequencing.

    Science.gov (United States)

    Delgado, Susana; Rachid, Caio T C C; Fernández, Elena; Rychlik, Tomasz; Alegría, Angel; Peixoto, Raquel S; Mayo, Baltasar

    2013-10-01

    Thermophilic lactic acid bacteria (LAB) species, such as Streptococcus thermophilus, Lactobacillus delbrueckii and Lactobacillus helveticus, enjoy worldwide economic importance as dairy starters. To assess the diversity of thermophilic bacteria in milk, milk samples were enriched in thermophilic organisms through a stepwise procedure which included pasteurization of milk at 63 °C for 30 min (PM samples) and pasteurization followed by incubation at 42 °C for 24 h (IPM samples). The microbial composition of these samples was analyzed by culture-dependent (at 42 °C) and culture-independent (PCR-DGGE and pyrosequencing of 16S rRNA gene amplicons) microbial techniques. The results were then compared to those obtained for their corresponding starting raw milk counterparts (RM samples). Twenty different species were scored by culturing among 352 isolates purified from the counting plates and identified by molecular methods. Mesophilic LAB species (Lactococcus lactis, Lactococcus garvieae) were dominant (87% of the isolates) among the RM samples. However, S. thermophilus and Lb. delbrueckii were found to be the dominant recoverable organisms in both PM and IPM samples. The DGGE profiles of RM and PM samples were found to be very similar; the most prominent bands belonging to Lactococcus, Leuconostoc and Streptococcus species. In contrast, just three DGGE bands were obtained for IPM samples, two of which were assigned to S. thermophilus. The pyrosequencing results scored 95 operational taxonomic units (OTUs) at 3% sequence divergence in an RM sample, while only 13 were encountered in two IPM samples. This technique identified Leuconostoc citreum as the dominant microorganism in the RM sample, while S. thermophilus constituted more than 98% of the reads in the IPM samples. The procedure followed in this study allowed to estimate the bacterial diversity in milk and afford a suitable strategy for the isolation of new thermophilic LAB strains, among which adequate

  6. A selected core microbiome drives the early stages of three popular italian cheese manufactures.

    Directory of Open Access Journals (Sweden)

    Francesca De Filippis

    Full Text Available Mozzarella (M, Grana Padano (GP and Parmigiano Reggiano (PR are three of the most important traditional Italian cheeses. In the three cheese manufactures the initial fermentation is carried out by adding natural whey cultures (NWCs according to a back-slopping procedure. In this study, NWCs and the corresponding curds from M, GP and PR manufactures were analyzed by culture-independent pyrosequencing of the amplified V1-V3 regions of the 16S rRNA gene, in order to provide insights into the microbiota involved in the curd acidification. Moreover, culture-independent high-throughput sequencing of lacS gene amplicons was carried out to evaluate the biodiversity occurring within the S. thermophilus species. Beta diversity analysis showed a species-based differentiation between GP-PR and M manufactures indicating differences between the preparations. Nevertheless, all the samples shared a naturally-selected core microbiome, that is involved in the curd acidification. Type-level variability within S. thermophilus species was also found and twenty-eight lacS gene sequence types were identified. Although lacS gene did not prove variable enough within S. thermophilus species to be used for quantitative biotype monitoring, the possibility of using non rRNA targets for quantitative biotype identification in food was highlighted.

  7. Improvement of the Texture of Yogurt by Use of Exopolysaccharide Producing Lactic Acid Bacteria

    Directory of Open Access Journals (Sweden)

    Xue Han

    2016-01-01

    Full Text Available 19 Streptococcus thermophilus with high exopolysaccharide production were isolated from traditional Chinese fermented dairy products. The exopolysaccharide and viscosity of milk fermented by these 19 isolates were assayed. The strains of Streptococcus thermophilus zlw TM11 were selected because its fermented milk had the highest exopolysaccharide content (380 mg/L and viscosity (7716 mpa/s. Then Streptococcus thermophilus zlw TM11 was combined with Lactobacillus delbrueckii subsp. bulgaricus 3 4.5 and the combination was named SH-1. The quality of the yogurt fermented by SH-1 and two commercial starter cultures (YO-MIX 465, YF-L711 were compared. It was shown that the exopolysaccharide content of yogurt fermented by SH-1 was similar to that of yogurt fermented by YF-L711 and significantly higher than YO-MIX 465 (p<0.05. In addition, the yogurt fermented by SH-1 had the lowest syneresis (8.5% and better texture and sensory than the samples fermented by YO-MIX 465 and YF-L711. It manifested that the selected higher exopolysaccharide production starter SH-1 could be used as yogurt starter and reduce the amount of adding stabilizer, which can compare with the imported commercial starter culture.

  8. Improvement of the Texture of Yogurt by Use of Exopolysaccharide Producing Lactic Acid Bacteria

    Science.gov (United States)

    Han, Xue; Yang, Zhe; Jing, Xueping; Yu, Peng; Zhang, Yingchun

    2016-01-01

    19 Streptococcus thermophilus with high exopolysaccharide production were isolated from traditional Chinese fermented dairy products. The exopolysaccharide and viscosity of milk fermented by these 19 isolates were assayed. The strains of Streptococcus thermophilus zlw TM11 were selected because its fermented milk had the highest exopolysaccharide content (380 mg/L) and viscosity (7716 mpa/s). Then Streptococcus thermophilus zlw TM11 was combined with Lactobacillus delbrueckii subsp. bulgaricus 3 4.5 and the combination was named SH-1. The quality of the yogurt fermented by SH-1 and two commercial starter cultures (YO-MIX 465, YF-L711) were compared. It was shown that the exopolysaccharide content of yogurt fermented by SH-1 was similar to that of yogurt fermented by YF-L711 and significantly higher than YO-MIX 465 (p syneresis (8.5%) and better texture and sensory than the samples fermented by YO-MIX 465 and YF-L711. It manifested that the selected higher exopolysaccharide production starter SH-1 could be used as yogurt starter and reduce the amount of adding stabilizer, which can compare with the imported commercial starter culture. PMID:27294135

  9. STUDI PENGARUH PENGGUNAAN BIFIDOBAKTERIA TERHADAP FLAVOR YOUGHURT [Study on the Effect of the Use of Bifidobacteria on Flavor of Yoghurt

    Directory of Open Access Journals (Sweden)

    Anton Apriyantono4

    2005-04-01

    Full Text Available This experiment was carried out to study the effect of bifidobacteria on flavor of yoghurt. Parameters measured in the experiment were acidity, pH, viscosity, volatile composition, sensory acceptance and intensity of yoghurt sensory attributes.Results of the experiment indicated that the use of bifidobacteria in mixture of yoghurt culture was able to increase the levels of acidity and viscosity of yoghurt. The highest acidity and viscosity was found in yoghurt prepared by Lactobacillus bulgaricus and bifidobacteria mixture, and also by Lactobacillus bulgaricus, Streptococcus thermophilus and bifidobacteria mixture cultures. The major classes of volatile component identified were acids, alcohols, ketones and aldehydes. The major component identified were octanoic acid, acetoin and octadecanal. It was found that there was no significant difference in sensory acceptance of the panelist for colour, aroma and taste of yoghurt prepared by the various combination of cultures. However, consistency of yoghurt prepared by S. thermophilus with or without addition of bifidobacteria, was less compared to that of others. Yoghurt prepared by single culture of S. thermophilus showed higher intensity of bitter and syneresis. The use of bifidobacteria in the cultures mixture decrease the intensity of bitter and syneresis of the yoghurt.

  10. PETROLEUM BIOREFINING FOR POLLUTION PREVENTION

    Energy Technology Data Exchange (ETDEWEB)

    John J. Kilbane II

    2002-03-01

    The objective of this project was to isolate and characterize thermophilic bacterial cultures that can be used for the selective removal of nitrogen, sulfur, and/or metals in the biorefining of petroleum. The project was completed on schedule and no major difficulties were encountered. Significant progress was made on multiple topics relevant to the development of a petroleum biorefining process capable of operating at thermophilic temperatures. New cultures capable of selectively cleaving C-N or C-S bonds in molecules relevant to petroleum were obtained, and the genes encoding the enzymes for these unique biochemical reactions were cloned and sequenced. Genetic tools were developed that enable the use of Thermus thermophilus as a host to express any gene of interest, and information was obtained regarding the optimum conditions for the growth of T. thermophilus. The development of a practical biorefining process still requires further research and the future research needs identified in this project include the development of new enzymes and pathways for the selective cleavage of C-N or C-S bonds that have higher specific activities, increased substrate range, and are capable of functioning at thermophilic temperatures. Additionally, there is a need for process engineering research to determine the maximum yield of biomass and cloned gene products that can be obtained in fed-batch cultures using T. thermophilus, and to determine the best configuration for a process employing biocatalysts to treat petroleum.

  11. Antimicrobial potential of commercial silver nanoparticles and the characterization of their physical properties toward probiotic bacteria isolated from fermented milk products.

    Science.gov (United States)

    Mikiciuk, Jakub; Mikiciuk, Ewa; Wrońska, Anna; Szterk, Arkadiusz

    2016-01-01

    The application of nanotechnology in the agriculture and food sector is relatively recent compared to its usage in drug delivery or pharmaceuticals. Therefore, this paper presents a study of the effect of silver nanoparticles on probiotic bacteria based on the example of Lactobacillus acidophilus LA-5, Bifidobacterium animalis subsp. lactis BB-12 and Streptococcus thermophilus ST-Y31 isolated from fermented milk products. Probiotic bacteria are one of the most crucial groups of bacteria for the food industry, because of their claimed health-promoting properties. Studies have shown that the type and concentration of silver nanoparticle solutions have a significant impact on the tested probiotic bacteria which are profitable for the digestive system. In the presence of all tested silver nanoparticles, St. thermophilus ST-Y31 growth was inhibited significantly by the dilution method as opposed to the disk-diffusion method. Both the disk-diffusion and the dilution methods showed no significant differences between L. acidophilus LA-5 and B. animalis subsp. lactis BB-12. The concentrations 2 μg mL(-1) and 0.25 μg mL(-1) had the highest antibacterial activity and statistically significant impacts on the tested probiotic strains. To our knowledge, this is the first report on potential antimicrobial effect of nanosilver against the health-promoting probiotic bacteria L. acidophilus LA-5, B. animalis subsp. lactis BB-12 and St. thermophilus ST-Y31 isolated from fermented milk products. PMID:26766747

  12. Fermentation of aqueous plant seed extracts by lactic acid bacteria

    Energy Technology Data Exchange (ETDEWEB)

    Schafner, D.W.; Beuchat, R.L.

    1986-05-01

    The effects of lactic acid bacterial fermentation on chemical and physical changes in aqueous extracts of cowpea (Vigna unguiculata), peanut (Arachis hypogea), soybean (Glycine max), and sorghum (Sorghum vulgare) were studied. The bacteria investigated were Lactobacillus helveticus, L. delbrueckii, L. casei, L. bulgaricus, L. acidophilus, and Streptococcus thermophilus. Organisms were inoculated individually into all of the seed extracts; L. bulgaricus and S. thermophilus were also evaluated together as inocula for fermenting the legume extracts. During fermentation, bacterial population and changes in titratable acidity, pH, viscosity, and color were measured over a 72 h period at 37 degrees C. Maximum bacterial populations, titratable acidity, pH, and viscosity varied depending upon the type of extract and bacterial strain. The maximum population of each organism was influenced by fermentable carbohydrates, which, in turn, influenced acid production and change in pH. Change in viscosity was correlated with the amount of protein and titratable acidity of products. Color was affected by pasteurization treatment and fermentation as well as the source of extract. In the extracts inoculated simultaneously with L. bulgaricus and S. thermophilus, a synergistic effect resulted in increased bacterial populations, titratable acidity, and viscosity, and decreased pH in all the legume extracts when compared to the extracts fermented with either of these organisms individually. Fermented extracts offer potential as substitutes for cultured dairy products. 24 references.

  13. Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology

    Science.gov (United States)

    Yoo, Mi-Young; Lim, Sang-Dong

    2016-01-01

    Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products. PMID:27433115

  14. Improvement of the Texture of Yogurt by Use of Exopolysaccharide Producing Lactic Acid Bacteria.

    Science.gov (United States)

    Han, Xue; Yang, Zhe; Jing, Xueping; Yu, Peng; Zhang, Yingchun; Yi, Huaxi; Zhang, Lanwei

    2016-01-01

    19 Streptococcus thermophilus with high exopolysaccharide production were isolated from traditional Chinese fermented dairy products. The exopolysaccharide and viscosity of milk fermented by these 19 isolates were assayed. The strains of Streptococcus thermophilus zlw TM11 were selected because its fermented milk had the highest exopolysaccharide content (380 mg/L) and viscosity (7716 mpa/s). Then Streptococcus thermophilus zlw TM11 was combined with Lactobacillus delbrueckii subsp. bulgaricus 3 4.5 and the combination was named SH-1. The quality of the yogurt fermented by SH-1 and two commercial starter cultures (YO-MIX 465, YF-L711) were compared. It was shown that the exopolysaccharide content of yogurt fermented by SH-1 was similar to that of yogurt fermented by YF-L711 and significantly higher than YO-MIX 465 (p yogurt fermented by SH-1 had the lowest syneresis (8.5%) and better texture and sensory than the samples fermented by YO-MIX 465 and YF-L711. It manifested that the selected higher exopolysaccharide production starter SH-1 could be used as yogurt starter and reduce the amount of adding stabilizer, which can compare with the imported commercial starter culture. PMID:27294135

  15. Benzoic Acid Production with Respect to Starter Culture and Incubation Temperature during Yogurt Fermentation using Response Surface Methodology.

    Science.gov (United States)

    Yu, Hyung-Seok; Lee, Na-Kyoung; Jeon, Hye-Lin; Eom, Su Jin; Yoo, Mi-Young; Lim, Sang-Dong; Paik, Hyun-Dong

    2016-01-01

    Benzoic acid is occasionally used as a raw material supplement in food products and is sometimes generated during the fermentation process. In this study, the production of naturally occurring yogurt preservatives was investigated for various starter cultures and incubation temperatures, and considered food regulations. Streptococcus thermophilus, Lactobacillus acidophilus, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus reuteri, Lactobacillus plantarum, Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium bifidum, Bifidobacterium infantis, and Bifidobacterium breve were used as yogurt starter cultures in commercial starters. Among these strains, L. rhamnosus and L. paracasei showed the highest production of benzoic acid. Therefore, the use of L. rhamnosus, L. paracasei, S. thermophilus, and different incubation temperatures were examined to optimize benzoic acid production. Response surface methodology (RSM) based on a central composite design was performed for various incubation temperatures (35-44℃) and starter culture inoculum ratios (0-0.04%) in a commercial range of dairy fermentation processes. The optimum conditions were 0.04% L. rhamnosus, 0.01% L. paracasei, 0.02% S. thermophilus, and 38.12℃, and the predicted and estimated concentrations of benzoic acid were 13.31 and 13.94 mg/kg, respectively. These conditions maximized naturally occurring benzoic acid production during the yogurt fermentation process, and the observed production levels satisfied regulatory guidelines for benzoic acid in dairy products. PMID:27433115

  16. Physicochemical Characteristics and Antioxidant Capacity in Yogurt Fortified with Red Ginseng Extract.

    Science.gov (United States)

    Jung, Jieun; Paik, Hyun-Dong; Yoon, Hyun Joo; Jang, Hye Ji; Jeewanthi, Renda Kankanamge Chaturika; Jee, Hee-Sook; Li, Xiang; Lee, Na-Kyoung; Lee, Si-Kyung

    2016-01-01

    The objective of this study was to investigate characteristics and functionality of yogurt applied red ginseng extract. Yogurts added with red ginseng extract (0.5, 1, 1.5, and 2%) were produced using Lactobacillus acidophilus and Streptococcus thermophilus and stored at refrigerated temperature. During fermentation, pH was decreased whereas titratable aicidity and viable cell counts of L. acidophilus and S. thermophilus were increased. The composition of yogurt samples was measured on day 1, an increase of red ginseng extract content in yogurt resulted in an increase in lactose, protein, total solids, and ash content, whereas fat and moisture content decreased. The pH value and cell counts of L. acidophilus and S. thermophilus were declined, however titratable acidity was increased during storage period. The antioxidant capacity was measured as diverse methods. During refrigerated storage time, the value of antioxidant effect was decreased, however, yogurt fortified with red ginseng extract had higher capacity than plain yogurt. The antioxidant effect was improved in proportion to concentration of red ginseng extract. These data suggests that red ginseng extract could affect to reduce fermentation time of yogurt and enhance antioxidant capacity. PMID:27433113

  17. Microbial evolution of traditional mountain cheese and characterization of early fermentation cocci for selection of autochtonous dairy starter strains.

    Science.gov (United States)

    Carafa, Ilaria; Clementi, Francesca; Tuohy, Kieran; Franciosi, Elena

    2016-02-01

    The microbial population of Traditional Mountain (TM) cheese was investigated and characterized for the selection of cocci suitable for developing new starter cultures. Samples of milk, curd and cheese at different ripening times were enumerated in selective culture media and 640 colonies were isolated from curd and cheese after 24 h of ripening. The Lactic Acid Bacteria (LAB) isolated from M17 were clustered into 231 biotypes by RAPD-PCR analysis and identified as Lactococcus lactis, Streptococcus thermophilus and Enterococcus faecalis. Forty percent of enterococci showed the in vitro ability to inhibit raw milk resident coliforms, but they were excluded as possible starters due to the presence of associated risk factors. All lactococci and streptococci were tested for their technological properties; 4 Lc. lactis subsp. lactis and 2 Sc. thermophilus which were fast acidifiers and did not produce unpleasant flavours were subjected to the freeze-drying stability test. Lc. lactis subsp. lactis biotype 68 and Sc. thermophilus biotype 93 showed the best technological properties and may be appropriate for cheese production. This work gave evidence of the high biodiversity of TM-cheese autochthonous biotypes which could be used as starter cultures for the improvement of TM-cheese technology. PMID:26678135

  18. Microbial community dynamics in thermophilic undefined milk starter cultures.

    Science.gov (United States)

    Parente, Eugenio; Guidone, Angela; Matera, Attilio; De Filippis, Francesca; Mauriello, Gianluigi; Ricciardi, Annamaria

    2016-01-18

    Model undefined thermophilic starter cultures were produced from raw milk of nine pasta-filata cheesemaking plants using a selective procedure based on pasteurization and incubation at high temperature with the objective of studying the microbial community dynamics and the variability in performances under repeated (7-13) reproduction cycles with backslopping. The traditional culture-dependent approach, based on random isolation and molecular characterization of isolates was coupled to the determination of pH and the evaluation of the ability to produce acid and fermentation metabolites. Moreover, a culture-independent approach based on amplicon-targeted next-generation sequencing was employed. The microbial diversity was evaluated by 16S rRNA gene sequencing (V1-V3 regions), while the microdiversity of Streptococcus thermophilus populations was explored by using novel approach based on sequencing of partial amplicons of the phosphoserine phosphatase gene (serB). In addition, the occurrence of bacteriophages was evaluated by qPCR and by multiplex PCR. Although it was relatively easy to select for a community dominated by thermophilic lactic acid bacteria (LAB) within a single reproduction cycle, final pH, LAB populations and acid production activity fluctuated over reproduction cycles. Both culture-dependent and -independent methods showed that the cultures were dominated by either S. thermophilus or Lactobacillus delbrueckii subsp. lactis or by both species. Nevertheless, subdominant mesophilic species, including lactococci and spoilage organisms, persisted at low levels. A limited number of serB sequence types (ST) were present in S. thermophilus populations. L. delbrueckii and Lactococcus lactis bacteriophages were below the detection limit of the method used and high titres of cos type S. thermophilus bacteriophages were detected in only two cases. In one case a high titre of bacteriophages was concurrent with a S. thermophilus biotype shift in the culture

  19. 益生菌Lactobacillus casei Zhang对凝固型发酵乳感官品质的影响%Effects of Probiotic Lactobacillus casei Zhang on Sensory Properties of Set Fermented Milk

    Institute of Scientific and Technical Information of China (English)

    王记成; 郭壮; 其木格苏都; 张和平; 孟和毕力格

    2013-01-01

    [目的]评价益生菌Lactobacillus casei Zhang(L.casei Zhang)对凝固型发酵乳感官品质的影响.[方法]将Streptococcus thermophilus(S.thermophilus)与益生菌L.caseiZhang复配发酵,发酵结束(pH4.5)于4℃冷藏1d或28d,并对发酵乳进行感官鉴评试验.[结果]益生菌L.casei Zhang与S.thermophilus 复配发酵可赋予产品更好的细腻性、更为浓郁的奶油味和乙醛风味,从而提升产品的风味总体得分、气味总体得分及产品总得分.4℃贮藏7d时益生菌发酵乳具有最好的气味和风味感官特征,而贮藏14d时具有最好的表观和质地感官特征.[结论]益生菌L.casei Zhang可提高发酵乳制品的感官品质,同时贮藏时间对发酵乳感官特性产生影响.%[Objective] The objective of this study is to evaluate the effects of the probiotic Lactobacillus casei Zhang on the sensory properties of set fermented milk.[Method] Probiotic L.casei Zhang was inoculated together with the S.thermophilus strains into pasteurized milk for fermentation.The fermentation was ended when the pH value of fermented milk reached 4.5.Then,sensory properties of the products were determined after storage at 4℃ for 1 d or 28 d.[Result] Sensory analysis showed that the score of glossiness,acetaldehyde,buttery,acceptance of flavor and odor attributes of the fermented milks produced by probiotic L.casei Zhang and S.thermophilus were higher than those produced by single starter culture.Meanwhile,the acceptance of flavor and odor attributes of all of the fermented milks decreased after 7 days,and the highest sensory score of the acceptance of texture and appearance attributes were observed after 14 days of refrigerated storage.[Conclusion] Probiotic L.casei Zhang could improve the overall sensory quality of fermented milk,meanwhile the storage time had effects on some sensory characteristics of fermented milk.

  20. Influence of stevioside and rebaudioside A on fermentation bacterium in food%甜菊甙及A3甙对食品发酵菌的影响研究

    Institute of Scientific and Technical Information of China (English)

    钱丽丽; 范雪峰; 孙丽建; 张丽萍

    2011-01-01

    试验主要研究了甜菊总甙与A3甙这两种甜味剂对嗜热链球菌、植物乳杆菌和酿酒酵母的影响,受试菌选择以食品常用发酵菌为研究对象,在MRS和YPD培养基中添加不同量的甜菊甙和A3甙,测定各受试菌在甜菊甙和A3甙不同浓度下的生长曲线,同时做对照试验.受试菌的生长曲线测定方法采用比浊法,测定波长植物乳杆菌、嗜热链球菌为600nm,酿酒酵母为560nm.结果表明甜菊甙及A3甙对植物乳杆菌和酿酒酵母的生长影响作用不明显,对嗜热链球菌表现为双重影响,在低浓度0.01%~0.1%的甜菊甙和A3甙添加组对该菌生长有促进作用,而在甜菊甙和A3甙浓度0.5%~1%时则表现抑制生长作用,甜菊甙和A3甙之间对嗜热链球菌生长曲线影响无显著差异.通过研究,初步探究了发酵食品中添加这两种甜味剂对三种发酵菌生长情况的影响.%This paper studied how stevioside and rebaudiodside A - two kinds of food sweetener affect the growth of streptococcus thermophilus, lactobacillus plantarum and saccharomyces cerevisiae. Common food fermentation bacterium was selected in the study. Stevioside and rebaudiodside A with different amount were added to MRS broth and YPD broth cultured bacterium, and the growth of bacteria under different concentration was compared with the controlled trial. The determination method is turbidimetry at the wavelength of 600nm for streptococcus thermophilus, lactobacillus,and 560nm for saccharomyces cerevisiae. The results indicated that stevioside and rebaudiodside A have not significantly affected the growth of lactobacillus plantarum and saccharomyces cerevisiae; however, they showed two different effects on streptococcus thermophilus growth: at low concentration (0. 01% ~ 0. 5 % ), it promoted the growth of streptococcus thermophilus, while at higher concentration of more than 0. 5%, it inhibited the grow of streptococcus thermophilus. There was no obvious

  1. 益生菌Lactobacillus casei Zhang对凝固型发酵乳质构和挥发性风味物质的影响%Effects of Probiotic Lactobacillus casei Zhang on the Texture Profile and Volatile Compounds of Set Yogurt

    Institute of Scientific and Technical Information of China (English)

    其木格苏都; 郭壮; 王记成; 孟和毕力格; 张和平

    2013-01-01

    [目的]评价益生菌Lactobacillus casei Zhang(L.casei Zhang)货架期内活菌数的变化及其对发酵乳物理特性和挥发性风味物质的影响.[方法]将2株Streptococcus thermophilus(S.thermophilus)分别与益生菌L.casei Zhang复配发酵,发酵结束(pH 4.5)于4℃冷藏28 d,并对发酵乳各指标进行测定.[结果]4℃贮藏28 d时,发酵乳中益生菌L.casei Zhang活菌数没有显著性变化(P<0.05),且数量保持在108CFU/g以上;益生菌L.casei Zhang与S.thermophilus复配发酵有利于提高3-羟基-2-丁酮和己酸等挥发性风味物质的含量,同时短期贮藏过程中(1-7 d)可赋予产品较高的黏度、硬度、脆度、胶性和耐嚼性,随着贮藏时间的延长其对产品物理特性的影响随发酵用S.thermophilus菌株的不同而呈现不同的变化.[结论]4℃贮藏28 d过程中,益生菌L.casei Zhang具有非常好的稳定性,同时其与S.thermophillus复配发酵可赋予产品较好的质构和风味.%[Objective] The viability of the probiotic Lactobacillus casei Zhang during the product sbelf-life and its effects on the texture profile and the production of volatile compounds of fermented milk were studied.[Method] L.casei Zhang was inoculated together with either of the two Streptococcus thermophilus strains into pasteurized milk for fermentation,the fermentation was ended when the pH value of fermented milk reached 4.5,then,specific indexes of the products were determined after storage at 4℃C for 28 d.[Result] The viable count ofL.casei Zhang was highly stable (P<0.05),and was more than 108CFU/g aRer storage at 4℃ for 28 d.Furthermore,by simultaneous inoculating L.casei Zhang and the selected strains ofS.thermophilus into the milk at a certain proportion,it was beneficial to improve the content of 3-hydroxyl-2-butanone and hexylic acid in products; this method also enhanced the viscosity,hardness,fracturability,gumminess and chewiness of the products during short-term storage (1-7 d

  2. Study on anti-moulds of fruits and vegetables by lactic acid bacteria%乳酸菌对果蔬中霉菌拮抗作用的研究

    Institute of Scientific and Technical Information of China (English)

    白凤翎; 宋闪闪; 吕欣然; 石金鑫; 李莹; 励建荣

    2013-01-01

    应用保加利亚乳杆菌(Lactobacillus bulgaricus)、嗜热链球菌(Streptococcus thermophilus)、乳酸乳球菌(Lactococcus lactis)和嗜酸乳杆菌(Lactobacillus acidophilus)4种乳酸菌对从梨、黄瓜、葡萄等果蔬中分离的6种霉菌进行拮抗作用研究.短横线软琼脂拮抗孢子法结果表明,保加利亚乳杆菌、嗜热链球菌、嗜酸乳杆菌对6种霉菌生长平均抑制率分别为67.59%、69.37%和70.18%.菌饼法研究表明,保加利亚乳杆菌、嗜热链球菌、嗜酸乳杆菌对根霉A和D以及曲霉E都具有较强的拮抗作用,其中嗜热链球菌和嗜酸乳杆菌对霉菌D的抑制率分别为63.16%和57.17%,保加利亚乳杆菌对霉菌E抑制率为72.20%.保加利亚乳杆菌对霉菌E拮抗作用最强,嗜热链球菌和嗜酸乳杆菌对霉菌D的生长也具有较强的抑制作用,且在7d内拮抗作用稳定,乳酸乳球菌对所有霉菌没有拮抗作用.%The antagonism to moulds isolating mouldy pear,cucumber and grape by lactic acid bacteria such as Lactobacillus bulgaricus,Streptococcus thermophilus,Lactococcus lactis and Lactobacillus acidophilus were carried out.The results showed that the anti-mould rates of L.bulgaricus,S.thermophilus and L.acidophilus on the 6 mould strains were 67.59%,69.37% and 70.18% using dash spores method,respectively.By the bacteria-cake method,the results showed that L.bulgaricus,S.thermophilus and L.acidophilus have strong antagonistic activity to the Rhizopus A,D and the Aspergillus E,the anti-mould rate of S.thermophilus and L.acidophilus to the Rhizopus D were 63.16% and 57.17%,respectively,the anti-mould rates of L.bulgaricus to the Aspergillus E was 72.20%.It was the strongest antagonistic activities to the mould E by L.bulgaricus,and S.thermophilus and L.acidophilus have obvious anti-mould activity to the mould D,and the anti-mould activity was stability in the 7 days,and the anti-mould activity of L.lactis was of no effect on the 6

  3. A culture-dependent survey of thermophilic bacteria from hot springs in Xiamen area in China

    Institute of Scientific and Technical Information of China (English)

    YANG Bo; OUYANG Jianping; AO Jingqun; CHEN Xinhua

    2009-01-01

    Microbes are believed to play important roles in ecosystem function in many environments. The hot springs of Xiamen Island are close to the Xiamen Sea, and may have some characteristics different from those of inland hot springs. Microbes living in the hot springs of Xiamen may have new characteristics. However, little is known about microbial communities of hot springs close to the Xiamen Sea. A cuhure-dependent survey of microbial population in the Xiamen hot springs was pcrformed by using an approach combining total cellular protein profile identification and 16S rRNA gene sequencing. A total of 328 isolates of bacteria were obtained from liquid and sediment samples from the Xiamen hot springs, including neutrophilie thermophilic bacteria and moderately thermophilic acidophiles. Neutrophilic thermophilic bacteria, which grow at a temperature range of 55-90℃ including Rhodothermus marinus (Strain 1) , Thermus thermophilus (Strain 2), Thermus thiopara (Strain 3) , Geobacillus stearothermophilus(Strain 4) , Geobacillus thermoleovorans (Strain 5) , and Pseudomonas pseudoal-caligenes (Strain 6), were recovered by 2216E plates. Moderately thermophilic acidophiles, which can grow at temperatures above 50℃ and a pH range of 1. 8-3.5 such as Alicyclobacillus acidoterrestris (Strain 8) , Sul-fobacillus acidophilus (Strain 9), and Sulfobacillus thermosulfidooxidans (Strain 10), were isolated on selective solid medium containing sulfur and Fe2+. Among these strains, Rhodothermus marinus, Thermus thermophilus and Geobacillus stearothermophilus are not only thermophilcs, but also halophiles. One bacterium strain (Strain 6) shared 99% nucleotide sequence homology with Pseudomonas pseudoalcaligenes on the 16S rRNA gene se-quence, but was quite different from Pseudomonas pseudoalcaligenes in biological characteristics, suggesting that it may represent a novel thermophilic species. Results indicated that various species of neutrophilic thermophiles and moderately thermophilic

  4. A computational investigation on the connection between dynamics properties of ribosomal proteins and ribosome assembly.

    Directory of Open Access Journals (Sweden)

    Brittany Burton

    Full Text Available Assembly of the ribosome from its protein and RNA constituents has been studied extensively over the past 50 years, and experimental evidence suggests that prokaryotic ribosomal proteins undergo conformational changes during assembly. However, to date, no studies have attempted to elucidate these conformational changes. The present work utilizes computational methods to analyze protein dynamics and to investigate the linkage between dynamics and binding of these proteins during the assembly of the ribosome. Ribosomal proteins are known to be positively charged and we find the percentage of positive residues in r-proteins to be about twice that of the average protein: Lys+Arg is 18.7% for E. coli and 21.2% for T. thermophilus. Also, positive residues constitute a large proportion of RNA contacting residues: 39% for E. coli and 46% for T. thermophilus. This affirms the known importance of charge-charge interactions in the assembly of the ribosome. We studied the dynamics of three primary proteins from E. coli and T. thermophilus 30S subunits that bind early in the assembly (S15, S17, and S20 with atomic molecular dynamic simulations, followed by a study of all r-proteins using elastic network models. Molecular dynamics simulations show that solvent-exposed proteins (S15 and S17 tend to adopt more stable solution conformations than an RNA-embedded protein (S20. We also find protein residues that contact the 16S rRNA are generally more mobile in comparison with the other residues. This is because there is a larger proportion of contacting residues located in flexible loop regions. By the use of elastic network models, which are computationally more efficient, we show that this trend holds for most of the 30S r-proteins.

  5. The evaluation of mixtures of yeast and potato extracts in growth media for biomass production of lactic cultures.

    Science.gov (United States)

    Gaudreau, H; Renard, N; Champagne, C P; Van Horn, D

    2002-07-01

    The effectiveness of yeast extracts (YE) and potato extracts (PE) to promote growth of seven lactic cultures was evaluated by automated spectrophotometry (AS). Two aspects of the growth curve were analysed: (1) maximum biomass obtained (using ODmax) and (2) highest specific growth rate mu(max)) Eleven lots from the same PE-manufacturing process were examined for lot-to-lot variability. The ODmax values of three of the seven strains were significantly affected by lot source, but mu(max) was not significantly affected. The growth of bacteria was systematically lower in base medium containing 100% PE than in base medium containing 100% YE for both ODmax or mu(max) data, which could be related to the lower content in nitrogen-based compounds in PE. In AS assays, highest OD values for Lactobacillus casei EQ28, Lactobacillus rhamnosus R-011, Lactobacillus plantarum EQ12, and Streptococcus thermophilus R-083 were obtained with a mixture of PE and YE. Fermentations (2 L) were also carried out to determine the accuracy of AS to predict biomass levels obtained under fermentation trials. In these fermentations, replacement of 50% YE with PE was shown to enable good growth of S. thermophilus. With L. rhamnosus R-011, a high correlation (R2 = 0.95) was found between ODmax data obtained in the AS assays and that of the 2-L bioreactor when the same growth medium was used for both series of fermentations. However, AS was not as efficient when industrial media were used for the bioreactor assays. The relationship was still good for ODmax between AS data and that of the bioreactor data with L. rhamnosus R-011 in industrial LBS medium (R2 = 0.87), but was very poor with the S. thermophilus R-083 on Rosell #43 industrial medium (R2 = 0.33). Since PE cost 40% less than YE, there are strong economic advantages in considering such a partial replacement of YE by PE. PMID:12224561

  6. Effect of bacteria proportion on the fermentation of goat yoghurt with probiotic culture

    Directory of Open Access Journals (Sweden)

    Guowei Shu

    2015-12-01

    Full Text Available Background. Goat milk production in Shaanxi province is dominant in China, but the product is mainly in- fant formula and adult milk powder; product homogeneity is serious and has no goat yoghurt with probiotic culture. Methods. The effect of bacteria proportion (1:3:1, 1:2:1, 1:1:1, 2:1:1, 3:1:1 on pH, acidity, and viable counts and sensory evaluation of goat milk fermented by probiotics including L. acidophilus, B. bifidum  or L. casei besides, S. thermophilus and L. bulgaricus for developing AB-goat yoghurt and BC-goat yoghurt was investigated. Results. The optimum bacteria proportion of L. acidophilus : B. bifidum : S. thermophilus and L. bulgaricus for AB-goat yoghurt and B. bifidum : L. casei : S. thermophilus and L. bulgaricus for BC-goat yoghurt were both 2:1:1. The pH, acidity, the viable counts of L. acidophilus and B. bifidum, the total viable counts were respectively 4.60, 7.73 (g/L, 3.50×107 cfu/mL, 3.40×107 cfu/mL and 2.30×109 cfu/mL in AB-goat yoghurt. The pH, acidity, the viable counts of B. bifidum and L. casei, the total viable counts were respectively  4.61, 8.16 (g/L, 7.60×107 cfu/mL, 5.60×107 cfu/mL and 2.04×109 cfu/mL in BC-goat yoghurt. Conclusion. The bacteria proportion had a significant effect on fermentation of AB- and BC-goat yoghurt, the results are beneficial for developing AB-goat yoghurt and BC-goat yoghurt.

  7. Supplementation with fruit and okara soybean by-products and amaranth flour increases the folate production by starter and probiotic cultures.

    Science.gov (United States)

    Albuquerque, Marcela Albuquerque Cavalcanti de; Bedani, Raquel; Vieira, Antônio Diogo Silva; LeBlanc, Jean Guy; Saad, Susana Marta Isay

    2016-11-01

    The ability of two starter cultures (Streptococcus (S.) thermophilus ST-M6 and St. thermophilus TA-40) and eleven probiotic cultures (St. thermophilus TH-4, Lactobacillus (Lb.) acidophilus LA-5, Lb. fermentum PCC, Lb. reuteri RC-14, Lb. paracasei subsp. paracasei, Lb. casei 431, Lb. paracasei subsp. paracasei F19, Lb. rhamnosus GR-1, and Lb. rhamnosus LGG, Bifidobacterium (B.) animalis subsp. lactis BB-12, B. longum subsp. longum BB-46, and B. longum subsp. infantis BB-02) to produce folate in a modified MRS broth (mMRS) supplemented with different fruit (passion fruit, acerola, orange, and mango) and okara soybean by-products and amaranth flour was investigated. Initially, the folate content of each vegetable substrate was determined: passion fruit by-product showed the lowest folate content (8±2ng/mL) and okara the highest (457±22ng/mL). When the orange by-product and amaranth flour were added to mMRS, all strains were able to increase folate production after 24h of fermentation. B. longum subsp infantis BB-02 produced the highest concentrations (1223±116ng/mL) in amaranth flour. Okara was the substrate that had the lowest impact on the folate production by all strains evaluated. Lb. acidophilus LA-5 (297±36ng/mL) and B. animalis subsp. lactis BB-12 (237±23ng/mL) were also able to produce folate after growth in mMRS containing acerola and orange by-products, respectively. The results of this study demonstrate that folate production is not only strain-dependent but also influenced by the addition of different substrates in the growth media. PMID:27442847

  8. Development of a fermented ice-cream as influenced by in situ exopolysaccharide production: Rheological, molecular, microstructural and sensory characterization.

    Science.gov (United States)

    Dertli, Enes; Toker, Omer S; Durak, M Zeki; Yilmaz, Mustafa T; Tatlısu, Nevruz Berna; Sagdic, Osman; Cankurt, Hasan

    2016-01-20

    This study aimed to investigate the role of in situ exopolysaccharide (EPS) production by EPS(+)Streptococcus thermophilus strains on physicochemical, rheological, molecular, microstructural and sensory properties of ice cream in order to develop a fermented and consequently functional ice-cream in which no stabilizers would be required in ice-cream production. For this purpose, the effect of EPS producing strains (control, strain 1, strain 2 and mixture) and fermentation conditions (fermentation temperature; 32, 37 and 42 °C and time; 2, 3 and 4h) on pH, S. thermophilus count, EPS amount, consistency coefficient (K), and apparent viscosity (η50) were investigated and optimized using single and multiple response optimization tools of response surface methodology. Optimization analyses indicated that functional ice-cream should be fermented with strain 1 or strain mixture at 40-42 °C for 4h in order to produce the most viscous ice-cream with maximum EPS content. Optimization analysis results also revealed that strain specific conditions appeared to be more effective factor on in situ EPS production amount, K and η50 parameters than did fermentation temperature and time. The rheological analysis of the ice-cream produced by EPS(+) strains revealed its high viscous and pseudoplastic non-Newtonian fluid behavior, which demonstrates potential of S. thermophilus EPS as thickening and gelling agent in dairy industry. FTIR analysis proved that the EPS in ice-cream corresponded to a typical EPS, as revealed by the presence of carboxyl, hydroxyl and amide groups with additional α-glycosidic linkages. SEM studies demonstrated that it had a web-like compact microstructure with pores in ice-cream, revealing its application possibility in dairy products to improve their rheological properties. PMID:26572373

  9. Revisiting the Structures of Several Antibiotics Bound to the Bacterial Ribosome

    Energy Technology Data Exchange (ETDEWEB)

    D Bulkley; C Innis; G Blaha; T Steitz

    2011-12-31

    The increasing prevalence of antibiotic-resistant pathogens reinforces the need for structures of antibiotic-ribosome complexes that are accurate enough to enable the rational design of novel ribosome-targeting therapeutics. Structures of many antibiotics in complex with both archaeal and eubacterial ribosomes have been determined, yet discrepancies between several of these models have raised the question of whether these differences arise from species-specific variations or from experimental problems. Our structure of chloramphenicol in complex with the 70S ribosome from Thermus thermophilus suggests a model for chloramphenicol bound to the large subunit of the bacterial ribosome that is radically different from the prevailing model. Further, our structures of the macrolide antibiotics erythromycin and azithromycin in complex with a bacterial ribosome are indistinguishable from those determined of complexes with the 50S subunit of Haloarcula marismortui, but differ significantly from the models that have been published for 50S subunit complexes of the eubacterium Deinococcus radiodurans. Our structure of the antibiotic telithromycin bound to the T. thermophilus ribosome reveals a lactone ring with a conformation similar to that observed in the H. marismortui and D. radiodurans complexes. However, the alkyl-aryl moiety is oriented differently in all three organisms, and the contacts observed with the T. thermophilus ribosome are consistent with biochemical studies performed on the Escherichia coli ribosome. Thus, our results support a mode of macrolide binding that is largely conserved across species, suggesting that the quality and interpretation of electron density, rather than species specificity, may be responsible for many of the discrepancies between the models.

  10. Revisiting the structures of several antibiotics bound to the bacterial ribosome

    Energy Technology Data Exchange (ETDEWEB)

    Bulkley, David; Innis, C. Axel; Blaha, Gregor; Steitz, Thomas A. (Yale)

    2010-10-08

    The increasing prevalence of antibiotic-resistant pathogens reinforces the need for structures of antibiotic-ribosome complexes that are accurate enough to enable the rational design of novel ribosome-targeting therapeutics. Structures of many antibiotics in complex with both archaeal and eubacterial ribosomes have been determined, yet discrepancies between several of these models have raised the question of whether these differences arise from species-specific variations or from experimental problems. Our structure of chloramphenicol in complex with the 70S ribosome from Thermus thermophilus suggests a model for chloramphenicol bound to the large subunit of the bacterial ribosome that is radically different from the prevailing model. Further, our structures of the macrolide antibiotics erythromycin and azithromycin in complex with a bacterial ribosome are indistinguishable from those determined of complexes with the 50S subunit of Haloarcula marismortui, but differ significantly from the models that have been published for 50S subunit complexes of the eubacterium Deinococcus radiodurans. Our structure of the antibiotic telithromycin bound to the T. thermophilus ribosome reveals a lactone ring with a conformation similar to that observed in the H. marismortui and D. radiodurans complexes. However, the alkyl-aryl moiety is oriented differently in all three organisms, and the contacts observed with the T. thermophilus ribosome are consistent with biochemical studies performed on the Escherichia coli ribosome. Thus, our results support a mode of macrolide binding that is largely conserved across species, suggesting that the quality and interpretation of electron density, rather than species specificity, may be responsible for many of the discrepancies between the models.

  11. Adenosine Triphosphate Stimulates Aquifex aeolicus MutL Endonuclease Activity

    OpenAIRE

    Jerome Mauris; Thomas C Evans

    2009-01-01

    BACKGROUND: Human PMS2 (hPMS2) homologues act to nick 5' and 3' to misincorporated nucleotides during mismatch repair in organisms that lack MutH. Mn(++) was previously found to stimulate the endonuclease activity of these homologues. ATP was required for the nicking activity of hPMS2 and yPMS1, but was reported to inhibit bacterial MutL proteins from Thermus thermophilus and Aquifex aeolicus that displayed homology to hPMS2. Mutational analysis has identified the DQHA(X)(2)E(X)(4)E motif pre...

  12. Röntgenstrukturanalyse von LytB aus Aquifex aeolicus und Plasmodium falciparum

    OpenAIRE

    Rekittke, Ingo

    2010-01-01

    Der MEP-Weg stellt auf Grund seiner besonderen Eigenschaften ein interessantes Ziel für die de novo Entwicklung antimikrobialer und herbizider Wirkstoffe dar. Mit Ausnahme der letzten beiden Enzyme GcpE und LytB ist der MEP-Weg schon strukturell geklärt. Um diese Lücke zu schliessen, wurden in dieser Arbeit Kristallisationsexperimente mit Aquifex aeolicus und Plasmodium falciparum LytB und Thermus thermophilus GcpE durchgeführt. Durch die im Rahmen dieser Arbeit durchgeführten Experimente ...

  13. Growth of ABT-5 bacteria culture in whey supplemented with whey protein concentrate

    OpenAIRE

    Bojan Matijević; Rajka Božanić; Ljubica Tratnik

    2010-01-01

    The objective of this research was to decrease the duration of whey fermentation and to analyze the impact of whey protein concentrates (WPC) addition on growth and activity as well as on survival of mixed ABT-5 culture bacteria (Lactobacillus acidophilus, Bifidobacterium spp. and Streptococcus thermophilus) during 28 days of fermented whey cold storage (5 °C). Whey fermentation was observed at 37 °C with and without 1.5 and 3 % WPC addition. Whey fermentation with mixed ABT-5 bacterial cultu...

  14. 不同添加剂对冷藏期酸奶品质和活菌数的影响%The effects of different additives on the quality and active cell count of sour milk during the period of cold storage

    Institute of Scientific and Technical Information of China (English)

    韩向红; 黄吟; 王锐萍; 何滨

    2003-01-01

    该试验采用嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus balgaricus)作为混合菌剂对乳液及分别加入不同添加剂的乳液进行酸奶发酵,发现所添加的添加剂均可促进产酸并提高冷藏期酸奶的品质和活菌数,其中以胡萝卜汁效果最好.

  15. ПОКАЗАТЕЛИ КАЧЕСТВА СМЕСИ КИСЛОМОЛОЧНОГО МОРОЖЕНОГО

    Directory of Open Access Journals (Sweden)

    РЯБЦЕВА С.А.

    2013-01-01

    Full Text Available Исследование было проведено путем сквашивания смеси кисломолочного мороженого различными комбинациями культур: L. acidophilus, St. thermophilus, L. casei+ St. thermophilus, L. bulgaricus+ St. thermophilus, Lac. spp + St. thermophilus, симбиотическая закваска на основе кефирных грибков. Бактериальные закваски вносили в смесь в количестве 5%, после чего экспериментальные образцы подвергались физико-химической и органолептической оценки. Среди представленных образцов высокую оценку по показателям качества получила смесь содержащая 5% L. acidophilus.

  16. ПОКАЗАТЕЛИ КАЧЕСТВА СМЕСИ КИСЛОМОЛОЧНОГО МОРОЖЕНОГО

    OpenAIRE

    Ахмедова, В.; Рябцева, С.

    2013-01-01

    Исследование было проведено путем сквашивания смеси кисломолочного мороженого различными комбинациями культур: L. acidophilus, St. thermophilus, L. casei+ St. thermophilus, L. bulgaricus+ St. thermophilus, Lac. spp + St. thermophilus, симбиотическая закваска на основе кефирных грибков. Бактериальные закваски вносили в смесь в количестве 5%, после чего экспериментальные образцы подвергались физико-химической и органолептической оценки. Среди представленных образцов высокую оценку по показателя...

  17. Antimutagenic properties of lactic acid-cultured milk on chemical and fecal mutagens

    Energy Technology Data Exchange (ETDEWEB)

    Hosono, A.; Kashina, T.; Kada, T.

    1986-09-01

    The antimutagenic properties of milk cultured with Lactobacillus bulgaricus and Streptococcus thermophilus were examined using streptomycin-dependent strains of Salmonella in an in vitro assay system. The mutagens utilized for testing included 2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide, 4-nitroquinoline-N-oxide, and fecal mutagenic extracts from cats, monkeys, dogs and other mammals. Both types of cultured milk exhibited antimutagenic activity on all mutagens used. Antimutagenic activities of the cultured milks with 2-(2-furyl)-3-(5-nitro-2-furyl) acrylamide and 4-nitroquinoline-N-oxide increased with incubation time but were thermolabile beyond 55/sup 0/C for 10 min.

  18. Research on development of Bifidobacterium adolescentis-containing solidified yoghurt and selective medium for strains%青春双歧杆菌凝固型酸奶研制及选择性培养基比较

    Institute of Scientific and Technical Information of China (English)

    刘韩; 关宏; 杨文钦; 张琪; 韩明宇; 刘吉成

    2011-01-01

    以鲜牛奶、白砂糖为主要原料,以青春双歧杆菌、保加利亚乳杆菌、嗜热链球菌为发酵剂,制成营养保健的青春双歧杆菌凝固型酸奶.通过试验确定了青春双歧杆菌凝固型酸奶最佳工艺为:白砂糖8%,接种量3%,保加利亚乳杆菌:嗜热链球菌:青春双歧杆菌=1:1:4,发酵4h.结合MRS培养基和LM-MRS培养基可以对青春双歧杆菌凝固型酸奶中三种乳酸分别计数.青春双歧杆凝固型酸奶中保加利亚乳杆菌活菌数为8.0x108 cfu.mL-1,嗜热链球菌活茵数为8.5x108cfu·mL-1,青春双歧杆菌活茵数为2.0x106 cfu·mL-1.%The nutritional and healthy solidified yoghurt containing Bifidobacterium adolescentic has been made, using fresh milk and sugar as the main raw materials and using Bifidobacterium adolescentic, Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus as starter cultures. The optimum technological condition of solidified yoghurt of Bifidobacterium adolescentic was determined: they are respectively 8% of sugar, 3% of inoculum and three lactic acid bacteria which are Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus and Bifidobacterium adoles centic and the percentage of these three lactic acid bacteria is 1:1:4, the time of fermentation is 4 h. The number of Lactobacillus delbrueckii subsp. bulgaricus, Streptococcus thermophilus and Bifidobacterium adolescentic could be counted in solidified Bifidobacterium adolescentic. Solidified yoghurt contain the number of viable Lactobacillus delbrueckii subsp. bulgaricus is 8.0 × 108 cfu· mL-1, the number of viable Streptococcus thermophilus is 8.5× 108 cfu· mL-1, the number of viable Bifidobacterium adolescentic is 2.0×106 cfu·mL-1.

  19. Influence of Pollen, Chia Seeds and Cranberries Addition on the Physical and Probiotics Characteristics of Yogurt

    Directory of Open Access Journals (Sweden)

    Carmen Pop

    2015-05-01

    Full Text Available Yoghurt is a fermented milk product obtained from fermentation of Lactobacillus bulgaricus and Streptococcus thermophilus strains. The effect of bee pollen, chia seeds and cranberries on the viability of probiotic bacteria in yogurts during storage (21 days at refrigerated temperature (4°C was evaluated. The yogurt supplementation with 1,4 % chia seeds and 7,6% cranberries significantly improves the stability of the lactic acid bacteria, that contained the recommended levels of (106–107 cfu/g probiotic bacteria at the end of 21-day shelf life.

  20. Antibiotic susceptibility of strains in Chinese medical probiotic products

    Institute of Scientific and Technical Information of China (English)

    TANG Huan; YUAN Jing; XIE Cai-hong; WEI Hong

    2007-01-01

    Objective:To investigate the susceptibility of strains separated from probiotic products for medical purpose to 14 antimicrobial agents. Methods:The single aerobic strains were isolated from these products respectively and disc agar diffusion assay was proceeded to determine the susceptibility. Results:Probiotics tested in the study mostly showed multiresistant to the agents. Lactobacillus acidophilus LAP,LAB, Lactobacillus bulgaricus LBJ and Streptococcus thermophilus STJ were resistant to vancomycin. Conclusion: Drug resistance exists in most of commercial probiotics. The evaluation and monitoring of safety of probiotic products for medical purpose should be paid great attention.

  1. Selection of Chinese Cheese Strains%中式干酪生产用菌种的筛选

    Institute of Scientific and Technical Information of China (English)

    鄂卫峰

    2005-01-01

    概述中式干酪的特点及生产用菌种所需具备的特性,对已有的四种菌株进行发酵性能和生化试验,优选出两株优良菌株为嗜热链球菌(Streptococcus thermophilus)和保加利亚德氏乳杆菌(Lactobacillus bulgaricus),并进行两菌株实际的干酪生产,证明表现良好.

  2. The Incidence of Necrotizing Enterocolitis Is Increased Following Probiotic Administration to Preterm Pigs

    DEFF Research Database (Denmark)

    Cilieborg, Malene Skovsted; Thymann, Thomas; Siggers, Richard;

    2011-01-01

    in the immediate postnatal period. Just after birth, caesarean-delivered preterm pigs were inoculated with Lactobacillus paracasei, Bifidobacteria animalis, and Streptococcus thermophilus (total 2.4 x 1010/d) either as live (ProLive, n = 14) or gamma-irradiated dead bacteria (ProDead, n = 12) and compared......Preterm birth and necrotizing enterocolitis (NEC) is associated with inappropriate gut colonization and immunity, which may be improved by probiotic bacteria. Using a preterm pig model of NEC, we investigated the effects of probiotics on intestinal structure, function, microbiology, and immunology...

  3. The influence of heme ruffling on spin densities in ferricytochromes c probed by heme core 13C NMR

    OpenAIRE

    Kleingardner, Jesse G.; Bowman, Sarah E. J.; Bren, Kara L.

    2013-01-01

    The heme in cytochromes c undergoes a conserved out-of-plane distortion known as ruffling. For cytochromes c from the bacteria Hydrogenobacter thermophilus and Pseudomonas aeruginosa, NMR and EPR spectra have been shown to be sensitive to the extent of heme ruffling and to provide insights into the effect of ruffling on electronic structure. Using mutants of each of these cytochromes that differ in the amount of heme ruffling, NMR characterization of the low-spin (S=1/2) ferric proteins has c...

  4. Estudio microbiológico comparativo de yogur fresco y termizado en un modelo animal in vivo Comparative microbiological study of fresh and heat-treated yoghurt in an in vivo animal model

    OpenAIRE

    J. E. García de los Ríos; J. Santos Jiménez Gómez; P. A. Jiménez Gómez; M.ª P. Reche Sainz; F. Álvarez Dávila; A. M.ª Rojas Mendoza

    2003-01-01

    En un ensayo agudo, ratas en ayunas de 24 h fueron sondadas con 2 ml de yogur fresco, yogur termizado o agua destilada, y se sacrificaron a las 2, 4, 8 y 24 h después, siendo comparadas con ratas sin sondar. Se determinó la supervivencia de las bacterias lácticas del yogur administrado en estómago e intestinos de los animales, y si las bacterias aisladas tenían el perfil de las cepas de Lactobacillus delbrueckii subsp. bulgaricus y Streptococcus thermophilus aisladas originalmente del yogur. ...

  5. Utilización de inulina en la formulación de yogur descremado de leche de cabra Using inulin in the formulation of non-fat goats´milk yogurt

    OpenAIRE

    Enzo Goncalvez de Oliveira; Noelia F Paz; Erica N Budde; Andrea P Cravero; Adriana N Ramón

    2012-01-01

    La inulina es una fibra prebiótica que imparte mayor cremosidad y favorece la textura y firmeza en productos lácteos. El objetivo de este trabajo fue utilizarla para reemplazar la grasa en la formulación de un yogur descremado de leche de cabra, de consistencia batida. Para ello se utilizó: leche de cabra biotipo Saanen; cultivos de bacterias lácticas liofilizadas: Streptococcus thermophilus (ST M6) y Lactobacillus delbrueckii subsp. bulgaricus (Lb-12); inulina en polvo; azúcar, gelatina sin ...

  6. Evaluación de la fermentación de bebida de soya con un cultivo láctico

    OpenAIRE

    Quicazán, Martha Cecilia; Sandoval, Angélica; Padilla, Germán

    2011-01-01

    La leche o bebida de soya, extracto acuoso de las proteínas de este grano, elaborada mediante un proceso previamente validado en el Instituto de Ciencia y Tecnología de Alimentos (ICTA), fue fermentada con un cultivo láctico termófilo, constituido por L delbrueckii ss. bulgaricus, L delbrueckii ss. laciis y S.salivarius ss. thermophilus. La composición de la bebida fue estandarizada, antes de la fermentación, en cuanto al contenido de sólidos y de proteína, utilizando como variable de proceso...

  7. Mechanism of CuA assembly

    OpenAIRE

    Luciano A Abriata; Banci, Lucia; Bertini, Ivano; Ciofi-Baffoni, Simone; Gkazonis, Petros; Spyroulias, Georgios A.; Alejandro J. Vila; Wang, Shenlin

    2008-01-01

    Copper is essential for proper functioning of cytochrome c oxidases, and therefore for cellular respiration in eukaryotes and many bacteria. Here we show that a new periplasmic protein (PCuAC) selectively inserts Cu(I) ions into subunit II of Thermus thermophilus ba3 oxidase to generate a native CuA site. The purported metallochaperone Sco1 is unable to deliver copper ions; instead, it works as a thiol-disulfide reductase to maintain the correct oxidation state of the CuA cysteine ligands.

  8. Sensory Evaluation of the Strawberry Flavored Yoghurt with Stevia and Sucrose Addition

    OpenAIRE

    Lisak, Katarina; Lenc, Maja; Jeličić, Irena; Božanić, Rajka

    2012-01-01

    As obesity is a growing problem in the world the target of food industries is to produce low calorie products safe for the human health. According to the non-caloric value of stevia and its beneficial influence on human health, there is a great possibility of its use in the food industry. Plain yoghurts were produced by fermentation with ABT-5 cultures (Lactobacillus acidophilus, Streptococcus thermophilus and Bifi dobacterium spp.) and flavored by addition of 0.2 mL/100 g strawberry aroma. Y...

  9. of intestinal microflora” pursuant to Article 14 of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    of a health claim related to a combination of Lactobacillus delbrueckii subsp. bulgaricus AY/CSL (LMG P-17224) and Streptococcus thermophilus 9Y/CSL (LMG P-17225) and “beneficial modulation of intestinal microflora”. The scope of the application was proposed to fall under a health claim referring to children...... microflora”. The target population, as proposed by the applicant, is children from 3 months to 14 years old. No evidence has been provided by the applicant to establish that the claimed effect, “beneficial modulation of the intestinal microflora”, is a beneficial physiological effect. The Panel concludes...... of the intestinal microflora”. © European Food Safety Authority, 2011...

  10. Fermentation and storage of probiotic yoghurt from goat’s milk

    Directory of Open Access Journals (Sweden)

    Rajka Božanić

    2002-04-01

    Full Text Available Cow’s and goat’s milk supplemented with inulin were fermented withABT4 culture. The population growth of Streptococcus thermophilus,Lactobacillus acidophilus and Bifidobacterium ssp. in plain and inulinsupplemented goat’s milk during fermentation was evaluated. The survival of strains during 28 d of storage was followed in comparison with that of cow’s milk. The time required to reach the desired pH of 4.6 during fermentation was 6 h for both types of milk. At that time the proportion of viable cells of Streptococcus thermophilus, Lactobacillus acidophilus and Bifidobacterium ssp. in all fermented samples was comparable 40 : 33 : 27, respectively. During the storage viable count of streptococci and bifidobacteria have not decreased. In supplemented samples viable counts of bifidobacteria were increased and during 28th day of storage were higher for 0.6 logarithms compared to the non supplemented samples. Surviving of lactobacilli was poorer in fermented goat's milk than in fermented cow's milk during storage. The addition of inulin improved the firmness of fermented goat’s and cow’s milks products. Inulin addition partly masked the goat’s flavour of produced yoghurt. During storage the fermented goat's samples were scored better in comparison with cow's samples. Goat’s milk fermented with probiotic bacteria and fortified with inulin complies with the requirements of functional food.

  11. EFFECTS OF ARTICHOKE (CYNARA SCOLYMUS L. EXTRACT ADDITION ON MICROBIOLOGICAL AND PHYSICO-CHEMICAL PROPERTIES OF PROBIOTIC YOGURT

    Directory of Open Access Journals (Sweden)

    Jalal Ehsani

    2015-06-01

    Full Text Available In this study, the effects of addition of artichoke (Cynara scolymus L. leaf extract into yogurt (0 or 0.5% on biochemical parameters (pH, titrable acidity and the viability of probiotic bacteria (Lactobacillus acidophilus LA-5, Bifidobacterium lactis BB-12 during fermentation and over 28 days of refrigerated storage (4°C were investigated. Moreover, the amounts of syneresis, total phenolic content, antioxidant activity and sensory attributes of yogurts at the end of fermentation were assessed. Yogurts contained the two yogurt bacteria (Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus: ABY or only S. thermophilus (ABT as adjunct culture to probiotics. Yogurts containing Cynara scolymus L. (ABT-C and ABY-C had faster acidity increase, shorter incubation time and greater final titrable acidity than control yogurts (ABT and ABY. Also, yogurts containing Cynara scolymus L. had lower syneresis, higher total phenolic content and greater antioxidant activity. ABT-C yogurt had the ever greatest viability of probiotics. In case of samples sensory evaluation, generally, the highest total score was related to ABT yogurt whereas lowest total score belonged to ABT-C yogurt.

  12. Crystal structure of tRNA m(1)A58 methyltransferase TrmI from Aquifex aeolicus in complex with S-adenosyl-L-methionine.

    Science.gov (United States)

    Kuratani, Mitsuo; Yanagisawa, Tatsuo; Ishii, Ryohei; Matsuno, Michiyo; Si, Shu-Yi; Katsura, Kazushige; Ushikoshi-Nakayama, Ryoko; Shibata, Rie; Shirouzu, Mikako; Bessho, Yoshitaka; Yokoyama, Shigeyuki

    2014-09-01

    The N (1)-methyladenosine residue at position 58 of tRNA is found in the three domains of life, and contributes to the stability of the three-dimensional L-shaped tRNA structure. In thermophilic bacteria, this modification is important for thermal adaptation, and is catalyzed by the tRNA m(1)A58 methyltransferase TrmI, using S-adenosyl-L-methionine (AdoMet) as the methyl donor. We present the 2.2 Å crystal structure of TrmI from the extremely thermophilic bacterium Aquifex aeolicus, in complex with AdoMet. There are four molecules per asymmetric unit, and they form a tetramer. Based on a comparison of the AdoMet binding mode of A. aeolicus TrmI to those of the Thermus thermophilus and Pyrococcus abyssi TrmIs, we discuss their similarities and differences. Although the binding modes to the N6 amino group of the adenine moiety of AdoMet are similar, using the side chains of acidic residues as well as hydrogen bonds, the positions of the amino acid residues involved in binding are diverse among the TrmIs from A. aeolicus, T. thermophilus, and P. abyssi. PMID:24894648

  13. Structural Studies on Cytosolic Domain of Magnesium Transporter MgtE from Enterococcus faecalis

    Energy Technology Data Exchange (ETDEWEB)

    Ragumani, S.; Sauder, J; Burley, S; Swaminathan, S

    2009-01-01

    Magnesium (Mg{sup 2+}) is an essential element for growth and maintenance of living cells. It acts as a cofactor for many enzymes and is also essential for stability of the plasma membrane. There are two distinct classes of magnesium transporters identified in bacteria that convey Mg{sup 2+} from periplasm to cytoplasm [ATPase-dependent (MgtA and MgtB) and constitutively active (CorA and MgtE)]. Previously published work on Mg{sup 2+} transporters yielded structures of full length MgtE from Thermus thermophilus, determined at 3.5 {angstrom} resolution, and its cytoplasmic domain with and without bond Mg{sup 2+} determined at 2.3 and 3.9 {angstrom} resolution, respectively. Here, they report the crystal structure of the Mg{sup 2+} bound form of the cytosolic portion of MgtE (residues 6-262) from Enterococcus faecalis at 2.2 {angstrom} resolution. The present structure and magnesium bound cytosolic domain structure from T. thermophilus (PDB ID: 2YVY) are structurally similar. Three magnesium binding sites are common to both MgtE full length and the present structure. Their work revealed an additional Mg{sup 2+} binding site in the E. faecalis structure. In this report, they discuss the functional significance of Mg{sup 2+} binding sites in the cytosolic domains of MgtE transporters.

  14. Structure of bacterial respiratory complex I.

    Science.gov (United States)

    Berrisford, John M; Baradaran, Rozbeh; Sazanov, Leonid A

    2016-07-01

    Complex I (NADH:ubiquinone oxidoreductase) plays a central role in cellular energy production, coupling electron transfer between NADH and quinone to proton translocation. It is the largest protein assembly of respiratory chains and one of the most elaborate redox membrane proteins known. Bacterial enzyme is about half the size of mitochondrial and thus provides its important "minimal" model. Dysfunction of mitochondrial complex I is implicated in many human neurodegenerative diseases. The L-shaped complex consists of a hydrophilic arm, where electron transfer occurs, and a membrane arm, where proton translocation takes place. We have solved the crystal structures of the hydrophilic domain of complex I from Thermus thermophilus, the membrane domain from Escherichia coli and recently of the intact, entire complex I from T. thermophilus (536 kDa, 16 subunits, 9 iron-sulphur clusters, 64 transmembrane helices). The 95Å long electron transfer pathway through the enzyme proceeds from the primary electron acceptor flavin mononucleotide through seven conserved Fe-S clusters to the unusual elongated quinone-binding site at the interface with the membrane domain. Four putative proton translocation channels are found in the membrane domain, all linked by the central flexible axis containing charged residues. The redox energy of electron transfer is coupled to proton translocation by the as yet undefined mechanism proposed to involve long-range conformational changes. This article is part of a Special Issue entitled Respiratory complex I, edited by Volker Zickermann and Ulrich Brandt. PMID:26807915

  15. Structure of bacterial respiratory complex I.

    Science.gov (United States)

    Berrisford, John M; Baradaran, Rozbeh; Sazanov, Leonid A

    2016-07-01

    Complex I (NADH:ubiquinone oxidoreductase) plays a central role in cellular energy production, coupling electron transfer between NADH and quinone to proton translocation. It is the largest protein assembly of respiratory chains and one of the most elaborate redox membrane proteins known. Bacterial enzyme is about half the size of mitochondrial and thus provides its important "minimal" model. Dysfunction of mitochondrial complex I is implicated in many human neurodegenerative diseases. The L-shaped complex consists of a hydrophilic arm, where electron transfer occurs, and a membrane arm, where proton translocation takes place. We have solved the crystal structures of the hydrophilic domain of complex I from Thermus thermophilus, the membrane domain from Escherichia coli and recently of the intact, entire complex I from T. thermophilus (536 kDa, 16 subunits, 9 iron-sulphur clusters, 64 transmembrane helices). The 95Å long electron transfer pathway through the enzyme proceeds from the primary electron acceptor flavin mononucleotide through seven conserved Fe-S clusters to the unusual elongated quinone-binding site at the interface with the membrane domain. Four putative proton translocation channels are found in the membrane domain, all linked by the central flexible axis containing charged residues. The redox energy of electron transfer is coupled to proton translocation by the as yet undefined mechanism proposed to involve long-range conformational changes. This article is part of a Special Issue entitled Respiratory complex I, edited by Volker Zickermann and Ulrich Brandt.

  16. Identifikace bakterií mléčného kvašení v kysaných mléčných výrobcích s využitím amplifikačních metod

    OpenAIRE

    Tycová, Martina

    2008-01-01

    Polymerázová řetěžová reakce (PCR) je molekulární diagnostická metoda, která umožňuje identifikaci bakterií mléčného kvašení používaných v potravinářském průmyslu. V této práci byla použita PCR pro identifikaci bakterií druhu Streptococcus thermophilus v 10-ti náhodně vybraných komerčně dostupných kysaných mléčných výrobcích a pro identifikaci druhu Streptococcus thermophilus v 25 lyofilizátech kmenů získaných ze Sbírky mlékařských kultur Laktoflóra (CCDM, Tábor, CZ). Druhově specifické PCR p...

  17. 益生菌Lactobacillus helveticus H9在脱脂乳中复配发酵特性研究%Mixed fermentation characteristic of probiotic Lactobacillus helveticus H9 in skim

    Institute of Scientific and Technical Information of China (English)

    陈永福; 李常坤; 王记成

    2015-01-01

    将益生菌Lactobacillus helveticus H9分别与Lactococcus lactis、Streptococcus thermophilus及Lactobacillus delbrueckii subsp.bulgaricus和Streptococcus thermophilus三类不同的菌种复配发酵脱脂乳,对其复配发酵乳的ACE抑制率、Val-Pro-Pro浓度、Ile-Pro-Pro浓度、游离氨基氮浓度、滴定酸度、pH值、活菌数、黏度、脱水收缩性和凝乳时间进行测定.结果表明,L.helveticus H9与Streptococcus thermophilus IMAU40115, Lactococcus lactis subsp.cremoris IMAU60065,Lactococcus lactis subsp.cremoris IMAU60064,L.delbrueckii subsp.bulgaricus IMAU20094和Streptococcus thermophilus IMAU40051的复配发酵乳的ACE抑制率大于60%,而且和IMAU40115、IMAU20094和IMAU40051的复配发酵乳具有较高的Val-Pro-Pro和Ile-Pro-Pro浓度.从复配菌种的种类分析发现,Lactococcus lactis较Streptococcus thermophilus和酸奶发酵剂更适宜与L.helveticus H9复配发酵.本文为进一步研究L helveticus H9发酵乳的开发提供了借鉴.

  18. Combining crystallography and EPR: crystal and solution structures of the multidomain cochaperone DnaJ

    Energy Technology Data Exchange (ETDEWEB)

    Barends, Thomas R. M., E-mail: thomas.barends@mpimf-heidelberg.mpg.de [MPI for Medical Research, Heidelberg (Germany); Brosi, Richard W. W. [Freie Universitat Berlin, Berlin (Germany); Steinmetz, Andrea; Scherer, Anna; Hartmann, Elisabeth; Eschenbach, Jessica; Lorenz, Thorsten [MPI for Medical Research, Heidelberg (Germany); Seidel, Ralf [MPI for Molecular Physiology, Dortmund (Germany); Shoeman, Robert L.; Zimmermann, Sabine [MPI for Medical Research, Heidelberg (Germany); Bittl, Robert [Freie Universitat Berlin, Berlin (Germany); Schlichting, Ilme; Reinstein, Jochen [MPI for Medical Research, Heidelberg (Germany)

    2013-08-01

    The crystal structure of the N-terminal part of T. thermophilus DnaJ unexpectedly showed an ordered GF domain and guided the design of a construct enabling the first structure determination of a complete DnaJ cochaperone molecule. By combining the crystal structures with spin-labelling EPR and cross-linking in solution, a dynamic view of this flexible molecule was developed. Hsp70 chaperones assist in a large variety of protein-folding processes in the cell. Crucial for these activities is the regulation of Hsp70 by Hsp40 cochaperones. DnaJ, the bacterial homologue of Hsp40, stimulates ATP hydrolysis by DnaK (Hsp70) and thus mediates capture of substrate protein, but is also known to possess chaperone activity of its own. The first structure of a complete functional dimeric DnaJ was determined and the mobility of its individual domains in solution was investigated. Crystal structures of the complete molecular cochaperone DnaJ from Thermus thermophilus comprising the J, GF and C-terminal domains and of the J and GF domains alone showed an ordered GF domain interacting with the J domain. Structure-based EPR spin-labelling studies as well as cross-linking results showed the existence of multiple states of DnaJ in solution with different arrangements of the various domains, which has implications for the function of DnaJ.

  19. Role of milk protein-based products in some quality attributes of goat milk yogurt.

    Science.gov (United States)

    Gursel, A; Gursoy, A; Anli, E A K; Budak, S O; Aydemir, S; Durlu-Ozkaya, F

    2016-04-01

    Goat milk yogurts were manufactured with the fortification of 2% (wt/vol) skim goat milk powder (SGMP), sodium caseinate (NaCn), whey protein concentrate (WPC), whey protein isolate (WPI), or yogurt texture improver (YTI). Yogurts were characterized based on compositional, microbiological, and textural properties; volatile flavor components (with gas chromatography); and sensory analyses during storage (21d at 5 °C). Compared with goat milk yogurt made by using SGMP, the other goat milk yogurt variants had higher protein content and lower acidity values. Goat milk yogurts with NaCn and WPC, in particular, had better physical characteristics. Using WPI caused the hardest structure in yogurt, leading to higher syneresis values. Acetaldehyde and ethanol formation increased with the incorporation of WPI, WPC, or YTI to yogurt milk. The tyrosine value especially was higher in the samples with NaCn and YTI than in the samples with WPC and WPI. Counts of Streptococcus thermophilus were higher than the counts of Lactobacillus delbrueckii ssp. bulgaricus, possibly due to a stimulatory effect of milk protein-based ingredients other than SGMP on the growth of S. thermophilus. Yogurt with NaCn was the best accepted among the yogurts. For the parameters used, milk protein-based products such as NaCn or WPC have promising features as suitable ingredients for goat milk yogurt manufacture.

  20. Screening Heat-resistant Strains of Lactic Acid Bacteria in Fermented Goat Milk%发酵羊奶中乳酸菌耐热菌株的筛选

    Institute of Scientific and Technical Information of China (English)

    乌素; 张富新; 乔星; 苏彤

    2012-01-01

    The heat-resistance of 11 Lactobacillus bulgaricus strains and 11 Streptococcus thermophilus strains from the commercial starter cultures was studied in fermented goat milk. The results showed that Lactobacillus bulgaricus L. b-346 strain and L. b-124 strain, Streptococcus thermophilus S. t-883 strain and S. t-124 strain had better heat-resist- ance. When L. b-346 strain and S. t-124 strain were mixed in 1:2 ratio, improved heat resistance was obtained.%对从商业乳酸菌发酵剂分离纯化的11株保加利亚乳杆菌和11株嗜热链球菌在发酵羊奶中的耐热性进行了研究。结果表明,保加利亚乳杆菌Lb-346和L.b-124菌株、嗜热链球菌S.t-883菌株和Js.t-124菌株的耐热性较好;当Lb-346菌株、Lb-124菌株与S.t-883菌株、S.t-124菌株按杆菌和球菌以不同比例配合时,L.b-346菌株与S.£-124菌株以1:2比例配合,可获得较高的耐热性。

  1. Screening low-viscosity strains of lactic acid bacteria in fermented goat milk%发酵羊乳中乳酸菌低产粘菌株的筛选

    Institute of Scientific and Technical Information of China (English)

    乌素; 张富新; 苏彤; 乔星

    2012-01-01

    The viscosity of 11 Lactobacillus bulgaricus strains and 11 Streptococcus thermophilus strains from the commercial starter cultures was studied in fermented goat milk.The results showed that Lactobacillus bulgaricus L.b-124 strain and L.b-346 strain, Streptococcus thermophilus S.t-222 strain and S.t-346 strain had lower viscoproduction capacity and shorter coagulation time.When L.b-346 strain and S.t-346 strain were mixed in 1:1 ratio and fermented in goat milk at 43℃,the lower viscosity was obtained.%对从商业乳酸菌发酵剂分离纯化的11株保加利亚乳杆菌和11株嗜热链球菌在发酵羊乳中的粘度进行了研究。结果表明,保加利亚乳杆菌L.b-124菌株和L.b-346菌株、嗜热链球菌S.t-222菌株和S.t-346菌株具有较低的产粘能力,凝乳时间也较短;将L.b-346菌株与S.t-346菌株按杆菌和球菌1:1比例混合后在43℃下发酵羊乳时,可获得较低的发酵粘度。

  2. The role of resistance to bile salts and acid tolerance of exopolysaccharides (EPSS produced by yogurt starter bacteria

    Directory of Open Access Journals (Sweden)

    Boke Hatice

    2010-01-01

    Full Text Available The aim of this study was to investigate a possible relation between EPS production and resistance to bile salts and tolerance to low pH. Eight strains which produced the highest and lowest amount of EPS (16- 211mg/l were selected among 54 bacteria isolated from yogurt. Additionally, they were tested for resistance to bile salts (0.15, 0.3 % and tolerance to low pH (2.0-3.0. After treatment with bile salts and acid, viable bacteria (log cfu ml-1 were determined by surface plating. The high EPS producing strains (B3, G12, W22 showed a significant (P<0.05 protective effect against low pH (pH 2.0. All Streptococcus thermophilus strains showed a higher tolerance to bile salts than the Lactobacillus delbrueckii subsp. bulgaricus strains. The high EPS-producing S. thermophilus (W22, T12 and L. bulgaricus (B3, G2 strains showed a significant (P<0.01 protective effect against bile salts (0.3 %.

  3. Compositional characteristics of commercial yoghurt based on quantitative determination of viable lactic acid bacteria

    Directory of Open Access Journals (Sweden)

    Niketić Gordana B.

    2009-01-01

    Full Text Available Yoghurt quality is particularly difficult to standardize because of the many forms, varieties, manufacturing methods, ingredients and consumer preferences that exist. Since these factors will always play an important role, it is unlikely that a uniform yoghurt quality concept will ever emerge, such as has been developed for other dairy products. There are a number of common denominators, however that have bearing on yoghurt quality. Since a number of producers are recognized within the broad category entitled yoghurt. This situation makes yoghurt an interesting, challenging, but also a confusing area to work in. The present investigation was undertaken to isolate from commercial yoghurt the strains involved in its manufacture and determine the characteristics of Streptococcus thermophilus and Lactobacillus delbrueckii subsp.bulgaricus. This study is concerned with the lactic acid bacteria (L.delbrueckii subsp. bulgaricus and S. thermophilus growth in yoghurt from involving different procedures and with the determination of the number of lactic acid bacteria in dependence of the temperature and acidity in the period of storage. Predominant samples of yoghurt were with 11-107/ml lactic acid lactococci (44.28%.

  4. Combining crystallography and EPR: crystal and solution structures of the multidomain cochaperone DnaJ

    International Nuclear Information System (INIS)

    The crystal structure of the N-terminal part of T. thermophilus DnaJ unexpectedly showed an ordered GF domain and guided the design of a construct enabling the first structure determination of a complete DnaJ cochaperone molecule. By combining the crystal structures with spin-labelling EPR and cross-linking in solution, a dynamic view of this flexible molecule was developed. Hsp70 chaperones assist in a large variety of protein-folding processes in the cell. Crucial for these activities is the regulation of Hsp70 by Hsp40 cochaperones. DnaJ, the bacterial homologue of Hsp40, stimulates ATP hydrolysis by DnaK (Hsp70) and thus mediates capture of substrate protein, but is also known to possess chaperone activity of its own. The first structure of a complete functional dimeric DnaJ was determined and the mobility of its individual domains in solution was investigated. Crystal structures of the complete molecular cochaperone DnaJ from Thermus thermophilus comprising the J, GF and C-terminal domains and of the J and GF domains alone showed an ordered GF domain interacting with the J domain. Structure-based EPR spin-labelling studies as well as cross-linking results showed the existence of multiple states of DnaJ in solution with different arrangements of the various domains, which has implications for the function of DnaJ

  5. 内蒙古酸马奶中乳酸菌多样性的研究%Biodiversity of Lactic Acid Bacteria in Home-made Koumiss from Inner Mongolia

    Institute of Scientific and Technical Information of China (English)

    刘芳; 都立辉; 杜鹏; 霍贵成

    2008-01-01

    In order to identify and protect home-made Koumiss,the diversity of lactic acid bacteria was investigated using appropriate starter cultures. Analysis of the 16S rRNA gene was used to confirm the presence of the predominant groups indicated by phenotypic analysis and to determine the phylogenetic affiliation of representative strains. The prevalent lactic acid bacteria consisted of Lactobacillus plantarum(10%), Lactobacillus brevis(8%), Lactobacillus casei(7.8%), Enterococcus faecium (17%), Ente rococcus faecalis(14%), Lactococcus lactis(19%), Lactobacillus acidlophilus(5%), Lactobacillus paracasei(2%),Lactobacillus delbrueckii subsp. bulgaricus(4%), Lactobacillus helveticus(4%), Enterococcus durans(4%), Leuconostoc mesenteroides(4%), Leuconostoc garlicum(1%) and Streptococcus thermophilus(1%).%采用16S rRNA基因全序列测定和聚类分析技术,对酸马奶中的乳酸菌进行了准确鉴定并构建了乳酸菌的系统发育树.然后对乳酸菌菌群进行了多样性分析,结果显示,酸马奶中的优势乳酸菌分别为:Lactobacillus plantarum(10%),Lactobacillus brevis(8%),Lactobacillus casei(7.8%),Enterococcus faecium(17%),Enterococcus faecalis(14%),Lactococcuslactis(19%),Lactobacillus acidlophilus(5%),Lactobacillus paracasei(2%),Lactobacillus delbrueckii subsp.bulgaricus(4%),Lactobacillus helveticus(4%),Enterococcus durans(4%),Leuconostoc mesenteroides (4%),Leuconostoc garlicum(1%),Streptococcus thermophilus(1%).

  6. 高加索酸奶中乳酸菌的分离与鉴定%Isolation and identification of Lactic acid bacteria (LAC) from yogurt of Russia's Caucasus region

    Institute of Scientific and Technical Information of China (English)

    杨郁荭; 邵景海; 李亮; 韩迪; 赵霞; 方曙光

    2012-01-01

    从自然发酵的5份酸奶样品中,通过平板划线等方法分离筛选乳酸菌.经形态特征,生理生化特性及糖发酵试验等,筛选到12株乳酸菌,分别为:乳杆菌7株,其中:3株德氏乳杆菌保加利亚亚种(Lactobacillus delbrueckii subsp.bulgaricus),3株瑞士乳杆菌(Lactobacillus helveticus),1株罗伊氏乳杆菌(Lactobacillus reuteri;乳酸球菌5株,包括3株嗜热链球菌(Streptococcus thermophilus),2株乳酸乳球菌乳脂亚种(Lactococcus lactis subsp.Cremoris.%12 strains of Lactic acid bacteria were isolated from 5 traditional fermented yogurt samples of Russia's Caucasus region. According to the morphological, physiological, biochemical characteristics, and the sugar-fermentation testing, among 12 strains, 7 strain were identified as lactobacillus, which include 3 strains of Lactobadllus detbrueckii ssp. Bulgaricus, 3 strains of Lactobacillus helveticus, and 1 strain of Lactobacillus reuteri; 5 strains were identified as lactococcus, which include 3 strains of Streptococcus thermophilus and 2 strains of Lactococcus lactis spp. Cremoris.

  7. Structural and biochemical analysis of nuclease domain of clustered regularly interspaced short palindromic repeat (CRISPR)-associated protein 3 (Cas3).

    Science.gov (United States)

    Mulepati, Sabin; Bailey, Scott

    2011-09-01

    RNA transcribed from clustered regularly interspaced short palindromic repeats (CRISPRs) protects many prokaryotes from invasion by foreign DNA such as viruses, conjugative plasmids, and transposable elements. Cas3 (CRISPR-associated protein 3) is essential for this CRISPR protection and is thought to mediate cleavage of the foreign DNA through its N-terminal histidine-aspartate (HD) domain. We report here the 1.8 Å crystal structure of the HD domain of Cas3 from Thermus thermophilus HB8. Structural and biochemical studies predict that this enzyme binds two metal ions at its active site. We also demonstrate that the single-stranded DNA endonuclease activity of this T. thermophilus domain is activated not by magnesium but by transition metal ions such as manganese and nickel. Structure-guided mutagenesis confirms the importance of the metal-binding residues for the nuclease activity and identifies other active site residues. Overall, these results provide a framework for understanding the role of Cas3 in the CRISPR system.

  8. Thermal adaptation of conformational dynamics in ribonuclease H.

    Directory of Open Access Journals (Sweden)

    Kate A Stafford

    Full Text Available The relationship between inherent internal conformational processes and enzymatic activity or thermodynamic stability of proteins has proven difficult to characterize. The study of homologous proteins with differing thermostabilities offers an especially useful approach for understanding the functional aspects of conformational dynamics. In particular, ribonuclease HI (RNase H, an 18 kD globular protein that hydrolyzes the RNA strand of RNA:DNA hybrid substrates, has been extensively studied by NMR spectroscopy to characterize the differences in dynamics between homologs from the mesophilic organism E. coli and the thermophilic organism T. thermophilus. Herein, molecular dynamics simulations are reported for five homologous RNase H proteins of varying thermostabilities and enzymatic activities from organisms of markedly different preferred growth temperatures. For the E. coli and T. thermophilus proteins, strong agreement is obtained between simulated and experimental values for NMR order parameters and for dynamically averaged chemical shifts, suggesting that these simulations can be a productive platform for predicting the effects of individual amino acid residues on dynamic behavior. Analyses of the simulations reveal that a single residue differentiates between two different and otherwise conserved dynamic processes in a region of the protein known to form part of the substrate-binding interface. Additional key residues within these two categories are identified through the temperature-dependence of these conformational processes.

  9. Phage mutations in response to CRISPR diversification in a bacterial population.

    Science.gov (United States)

    Sun, Christine L; Barrangou, Rodolphe; Thomas, Brian C; Horvath, Philippe; Fremaux, Christophe; Banfield, Jillian F

    2013-02-01

    Interactions between bacteria and their coexisting phage populations impact evolution and can strongly influence biogeochemical processes in natural ecosystems. Periodically, mutation or migration results in exposure of a host to a phage to which it has no immunity; alternatively, a phage may be exposed to a host it cannot infect. To explore the processes by which coexisting, co-evolving hosts and phage populations establish, we cultured Streptococcus thermophilus DGCC7710 with phage 2972 and tracked CRISPR (clustered regularly interspaced short palindromic repeats) diversification and host-phage co-evolution in a population derived from a colony that acquired initial CRISPR-encoded immunity. After 1 week of co-culturing, the coexisting host-phage populations were metagenomically characterized using 454 FLX Titanium sequencing. The evolved genomes were compared with reference genomes to identify newly incorporated spacers in S. thermophilus DGCC7710 and recently acquired single-nucleotide polymorphisms (SNPs) in phage 2972. Following phage exposure, acquisition of immune elements (spacers) led to a genetically diverse population with multiple subdominant strain lineages. Phage mutations that circumvented three early immunization events were localized in the proto-spacer adjacent motif (PAM) or near the PAM end of the proto-spacer, suggesting a strong selective advantage for the phage that mutated in this region. The sequential fixation or near fixation of these single mutations indicates selection events so severe that single phage genotypes ultimately gave rise to all surviving lineages and potentially carried traits unrelated to immunity to fixation.

  10. Phage mutations in response to CRISPR diversification in a bacterial population.

    Science.gov (United States)

    Sun, Christine L; Barrangou, Rodolphe; Thomas, Brian C; Horvath, Philippe; Fremaux, Christophe; Banfield, Jillian F

    2013-02-01

    Interactions between bacteria and their coexisting phage populations impact evolution and can strongly influence biogeochemical processes in natural ecosystems. Periodically, mutation or migration results in exposure of a host to a phage to which it has no immunity; alternatively, a phage may be exposed to a host it cannot infect. To explore the processes by which coexisting, co-evolving hosts and phage populations establish, we cultured Streptococcus thermophilus DGCC7710 with phage 2972 and tracked CRISPR (clustered regularly interspaced short palindromic repeats) diversification and host-phage co-evolution in a population derived from a colony that acquired initial CRISPR-encoded immunity. After 1 week of co-culturing, the coexisting host-phage populations were metagenomically characterized using 454 FLX Titanium sequencing. The evolved genomes were compared with reference genomes to identify newly incorporated spacers in S. thermophilus DGCC7710 and recently acquired single-nucleotide polymorphisms (SNPs) in phage 2972. Following phage exposure, acquisition of immune elements (spacers) led to a genetically diverse population with multiple subdominant strain lineages. Phage mutations that circumvented three early immunization events were localized in the proto-spacer adjacent motif (PAM) or near the PAM end of the proto-spacer, suggesting a strong selective advantage for the phage that mutated in this region. The sequential fixation or near fixation of these single mutations indicates selection events so severe that single phage genotypes ultimately gave rise to all surviving lineages and potentially carried traits unrelated to immunity to fixation. PMID:23057534

  11. Comparison of culture-dependent and -independent methods for bacterial community monitoring during Montasio cheese manufacturing.

    Science.gov (United States)

    Carraro, Lisa; Maifreni, Michela; Bartolomeoli, Ingrid; Martino, Maria Elena; Novelli, Enrico; Frigo, Francesca; Marino, Marilena; Cardazzo, Barbara

    2011-04-01

    The microbial community in milk is of great importance in the manufacture of traditional cheeses produced using raw milk and natural cultures. During milk curdling and cheese ripening, complex interactions occur in the microbial community, and accurate identification of the microorganisms involved provides essential information for understanding their role in these processes and in flavor production. Recent improvements in molecular biological methods have led to their application to food matrices, and thereby opened new perspectives for the study of microbial communities in fermented foods. In this study, a description of microbial community composition during the manufacture and ripening of Montasio cheese was provided. A combined approach using culture-dependent and -independent methods was applied. Culture-dependent identification was compared with 16S clone libraries sequencing data obtained from both DNA and reverse-transcribed RNA (cDNA) amplification and real-time quantitative PCR (qPCR) assays developed to detect and quantify specific bacterial species/genera (Streptococcus thermophilus, Lactobacillus casei, Pediococcus pentosaceus, Enterococcus spp., Pseudomonas spp.). S. thermophilus was the predominant LAB species throughout the entire ripening period of Montasio cheese. The culture-independent method demonstrates the relevant presence of Pseudomonas spp. and Lactococcus piscium at the beginning of ripening. The culture-dependent approach and the two culture-independent approaches produced complementary information, together generating a general view of cheese microbial ecology.

  12. Study on Mixture Starter of Lactic Bacteria and Yeast to Prepare Stirring Fermentation Bean-Milk%发酵型含醇酸豆奶的研制

    Institute of Scientific and Technical Information of China (English)

    蒋立文; 付海平

    2001-01-01

    为了开发酸豆奶新产品,对乳酸菌、酵母菌混和发酵生产酸豆奶进行了研究。正交试验结果表明,生产品质优良产品的最佳工艺参数为:豆水比 1∶ 10;菌种:保加利亚乳杆菌 (Lactobacillus bulgaricus)+嗜热链球菌 (Streptococcus thermophilus),啤酒酵母 (Saccharomyces cerevisiae),菌种配比 1∶ 2;蔗糖、明胶、羧甲基纤维素纳、黄原胶、乙基麦芽酚等适量,发酵时间为 6h,发酵温度 35℃,发酵剂接种量 3%。%In order to develop the new product of fermentation bean-milk.The co-fermentation of soymilk by Lactobacillus bulgaricus Streptococcus thermophilus and Saccharomyces cerevisiae has been studied.The results of orthogonal experiments indicates that the best process factors are:bean-water proportion 1∶ 10;the proportion of specices lactic bacteria and Saccbaromyces cerevisiae is a ration of 1∶ 2,sugar、 gelation、 sodium carboxymethyl cellulose、 ethyl maltol,fermentation time 6h,temperature 35℃ ,inoculum size 3% .The quality of the product is good.

  13. Selection of dairy culture and changes of Podravec cheese acidity during production

    Directory of Open Access Journals (Sweden)

    Slavko Kirin

    2002-06-01

    Full Text Available The selection and characteristics of dairy culture play a basic role in all types of cheese production process. The most important characteristic is acidification ability i.e. lactic acid formation, which regulates manufacturing and maturing conditions of cheese, thus affecting its organoleptic characteristics as well. In this work the results on control of acidity increase in Podravec cheeseproduction are presented. In the production process, a technical culture as well as identical frozen and concentrated culture, with and without auxiliary Streptococcus thermophilus for direct milk inoculation, were used. It was established that the acidity, expressed as pH value, is more intensively developed in cheeses produced with culture for direct inoculation. This was especially evident in the first phases of production i.e. before cheese salting. During salting the acidity of cheeses, in both cases, was almoust identical. Cheeses produced with identical frozen culture and auxiliary Streptococcus thermophilus culture had more pronounced acidity before salting and lower after salting in comparison with cheeses with the mentioned two cultures. Organoleptic and other characteristics of mature cheeses were identical.

  14. An intermolecular binding mechanism involving multiple LysM domains mediates carbohydrate recognition by an endopeptidase

    Energy Technology Data Exchange (ETDEWEB)

    Wong, Jaslyn E. M. M. [Aarhus University, Gustav Wieds Vej 10C, 8000 Aarhus (Denmark); Midtgaard, Søren Roi [University of Copenhagen, Universitetsparken 5, 2100 Copenhagen (Denmark); Gysel, Kira [Aarhus University, Gustav Wieds Vej 10C, 8000 Aarhus (Denmark); Thygesen, Mikkel B.; Sørensen, Kasper K.; Jensen, Knud J. [University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg C (Denmark); Stougaard, Jens; Thirup, Søren; Blaise, Mickaël, E-mail: mickael.blaise@cpbs.cnrs.fr [Aarhus University, Gustav Wieds Vej 10C, 8000 Aarhus (Denmark)

    2015-03-01

    The crystal and solution structures of the T. thermophilus NlpC/P60 d, l-endopeptidase as well as the co-crystal structure of its N-terminal LysM domains bound to chitohexaose allow a proposal to be made regarding how the enzyme recognizes peptidoglycan. LysM domains, which are frequently present as repetitive entities in both bacterial and plant proteins, are known to interact with carbohydrates containing N-acetylglucosamine (GlcNAc) moieties, such as chitin and peptidoglycan. In bacteria, the functional significance of the involvement of multiple LysM domains in substrate binding has so far lacked support from high-resolution structures of ligand-bound complexes. Here, a structural study of the Thermus thermophilus NlpC/P60 endopeptidase containing two LysM domains is presented. The crystal structure and small-angle X-ray scattering solution studies of this endopeptidase revealed the presence of a homodimer. The structure of the two LysM domains co-crystallized with N-acetyl-chitohexaose revealed a new intermolecular binding mode that may explain the differential interaction between LysM domains and short or long chitin oligomers. By combining the structural information with the three-dimensional model of peptidoglycan, a model suggesting how protein dimerization enhances the recognition of peptidoglycan is proposed.

  15. Estudio microbiológico comparativo de yogur fresco y termizado en un modelo animal in vivo Comparative microbiological study of fresh and heat-treated yoghurt in an in vivo animal model

    Directory of Open Access Journals (Sweden)

    J. E. García de los Ríos

    2003-08-01

    Full Text Available En un ensayo agudo, ratas en ayunas de 24 h fueron sondadas con 2 ml de yogur fresco, yogur termizado o agua destilada, y se sacrificaron a las 2, 4, 8 y 24 h después, siendo comparadas con ratas sin sondar. Se determinó la supervivencia de las bacterias lácticas del yogur administrado en estómago e intestinos de los animales, y si las bacterias aisladas tenían el perfil de las cepas de Lactobacillus delbrueckii subsp. bulgaricus y Streptococcus thermophilus aisladas originalmente del yogur. Tambien se hizo el recuento del número de microorganismos totales que crecían en el medios para lactobacilos y estreptococos lácticos. Se hicieron análisis de los diferentes tipos de colonias resultantes en los medios para lactobacilos y estreptococos lácticos. Ninguna de las colonias caracterizadas en este ensayo agudo resultó ser L. delbrueckii subsp. bulgaricus o S. thermophilus. El análisis estadístico de los resultados no mostró diferencias significativas en el log UFC · g-1, de la microbiota intestinal de los cuatro grupos de animales en ninguno de los tiempos. En un ensayo crónico, se sacrificaron tres grupos de animales, alojados en células individuales de metabolismo, tras 30 días de alimentación ad libitum con dieta semisintética con 10% de yogur fresco, con 10% de yogur termizado o solo con dieta semisintética. Los resultados no mostraron diferencias significativas en el log UFC · g-1 de los microorganismos de la microbiota intestinal entre los tres grupos de animales en ninguno de los tiempos. A su vez, se caracterizaron los diferentes tipos de colonias resultantes en los medios para lactobacilos y estreptococos lácticos, y ninguna de las colonias caracterizadas en este ensayo crónico resultó ser L. delbrueckii subsp. bulgaricus o S. thermophilus.In an acute assay, rats fasted for 24 h were sounded with 2 ml of fresh yogurt, sterilised yogurt or distilled water, and sacrificed at 2, 4, 8 and 24 h. They were compared with

  16. Probiotic viability and storage stability of yogurts and fermented milks prepared with several mixtures of lactic acid bacteria.

    Science.gov (United States)

    Mani-López, E; Palou, E; López-Malo, A

    2014-05-01

    Currently, the food industry wants to expand the range of probiotic yogurts but each probiotic bacteria offers different and specific health benefits. Little information exists on the influence of probiotic strains on physicochemical properties and sensory characteristics of yogurts and fermented milks. Six probiotic yogurts or fermented milks and 1 control yogurt were prepared, and we evaluated several physicochemical properties (pH, titratable acidity, texture, color, and syneresis), microbial viability of starter cultures (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and probiotics (Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus reuteri) during fermentation and storage (35 d at 5°C), as well as sensory preference among them. Decreases in pH (0.17 to 0.50 units) and increases in titratable acidity (0.09 to 0.29%) were observed during storage. Only the yogurt with S. thermophilus, L. delbrueckii ssp. bulgaricus, and L. reuteri differed in firmness. No differences in adhesiveness were determined among the tested yogurts, fermented milks, and the control. Syneresis was in the range of 45 to 58%. No changes in color during storage were observed and no color differences were detected among the evaluated fermented milk products. Counts of S. thermophilus decreased from 1.8 to 3.5 log during storage. Counts of L. delbrueckii ssp. bulgaricus also decreased in probiotic yogurts and varied from 30 to 50% of initial population. Probiotic bacteria also lost viability throughout storage, although the 3 probiotic fermented milks maintained counts ≥ 10(7)cfu/mL for 3 wk. Probiotic bacteria had variable viability in yogurts, maintaining counts of L. acidophilus ≥ 10(7) cfu/mL for 35 d, of L. casei for 7d, and of L. reuteri for 14 d. We found no significant sensory preference among the 6 probiotic yogurts and fermented milks or the control. However, the yogurt and fermented milk made with L. casei were better accepted. This

  17. Short communication: survival of the characteristic microbiota in probiotic fermented camel, cow, goat, and sheep milks during refrigerated storage.

    Science.gov (United States)

    Varga, L; Süle, J; Nagy, P

    2014-01-01

    The objective of this study was to monitor the viability during storage of Lactobacillus acidophilus LA-5 (A), Bifidobacterium animalis ssp. lactis BB-12 (B), and Streptococcus thermophilus CHCC 742/2130 (T) in probiotic cultured dairy foods made from pasteurized camel, cow, goat, and sheep milks fermented by an ABT-type culture. The products manufactured were stored at 4°C for 42d. Microbiological analyses were performed at weekly intervals. Streptococcus thermophilus CHCC 742/2130 was the most numerous culture component in all 4 products both at the beginning and at the end of storage. The viable counts of streptococci showed no significant decline in fermented camel milk throughout the entire storage period. The initial numbers of Lb. acidophilus LA-5 were over 2 orders of magnitude lower than those of Strep. thermophilus CHCC 742/2130. With the progress of time, a slow and constant decrease was observed in lactobacilli counts; however, the final viability percentages of this organism did not differ significantly in the probiotic fermented milks tested. The cultured dairy foods made from cow, sheep, and goat milks had comparable B. animalis ssp. lactis BB-12 counts on d 0, exceeding by approximately 0.5 log10 cycle those in the camel milk-based product. No significant losses occurred in viability of bifidobacteria in fermented camel, cow, and sheep milks during 6wk of refrigerated storage. In conclusion, all 4 varieties of milk proved to be suitable raw materials for the manufacture of ABT-type fermented dairy products that were microbiologically safe and beneficial for human consumption. It was suggested that milk from small ruminants be increasingly used to produce probiotic fermented dairy foods. The development of camel milk-based probiotic cultured milks appears to be even more promising because new markets could thus be conquered. It must be emphasized, however, that further microbiological and sensory studies, technology development activities, and

  18. Effects of synbiotic fermented milk containing Lactobacillus acidophilus La-5 and Bifidobacterium animalis ssp. lactis BB-12 on the fecal microbiota of adults with irritable bowel syndrome: A randomized double-blind, placebo-controlled trial.

    Science.gov (United States)

    Bogovič Matijašić, Bojana; Obermajer, Tanja; Lipoglavšek, Luka; Sernel, Tjaša; Locatelli, Igor; Kos, Mitja; Šmid, Alenka; Rogelj, Irena

    2016-07-01

    We conducted a randomized double-blind, placebo-controlled multicentric study to investigate the influence of a synbiotic fermented milk on the fecal microbiota composition of 30 adults with irritable bowel syndrome (IBS). The synbiotic product contained Lactobacillus acidophilus La-5, Bifidobacterium animalis ssp. lactis BB-12, Streptococcus thermophilus, and dietary fiber (90% inulin, 10% oligofructose), and a heat-treated fermented milk without probiotic bacteria or dietary fiber served as placebo. Stool samples were collected after a run-in period, a 4-wk consumption period, and a 1-wk follow-up period, and were subjected to real-time PCR and 16S rDNA profiling by next-generation sequencing. After 4wk of synbiotic (11 subjects) or placebo (19 subjects) consumption, a greater increase in DNA specific for L. acidophilus La-5 and Bifidobacterium animalis ssp. lactis was detected in the feces of the synbiotic group compared with the placebo group by quantitative real-time PCR. After 1wk of follow-up, the content of L. acidophilus La-5 and B. animalis ssp. lactis decreased to levels close to initial levels. No significant changes with time or differences between the groups were observed for Lactobacillus, Enterobacteriaceae, Bifidobacterium, or all bacteria. The presence of viable BB-12- and La-5-like bacteria in the feces resulting from the intake of synbiotic product was confirmed by random amplification of polymorphic DNA (RAPD)-PCR. At the end of consumption period, the feces of all subjects assigned to the synbiotic group contained viable bacteria with a BB-12-like RAPD profile, and after 1wk of follow-up, BB-12-like bacteria remained in the feces of 87.5% of these subjects. The presence of La-5-like colonies was observed less frequently (37.5 and 25% of subjects, respectively). Next-generation sequencing of 16S rDNA amplicons revealed that only the percentage of sequences assigned to Strep. thermophilus was temporarily increased in both groups, whereas the

  19. Screening of yogurt starter strains with low post-acidification capacity by ultraviolet mutagenesis%弱后酸化酸奶发酵菌株的紫外诱变选育

    Institute of Scientific and Technical Information of China (English)

    韩雪; 张兰威; 张爽; 易华西; 杜明

    2011-01-01

    To obtain yogurt starter strains with low post-acidification capacity, Lactobacillus delbrueckii subsp. bulgaricus 3 4.5 and Streptococcus thermophilus sp. 1.1 were treated by ultraviolet, and then were inoculated into low pH MRS medium and skimmed milk, respectively. Being cultivated at 37℃ for 24h, the mutants, which could grow in skimmed milk but not in low pH MRS medium, were identified to have low post-acidification capacity. The mutants were preserved and their genetic stabilities were tested. The results showed that the optimal ultraviolet exposure time was 50s for both S. thermophilus sp1.1 and L. delbtueckii 3 4.5. The post-acidification capacity of S. thermophilus mutant S1 was 8.0% lower than original strain. The post-acidification capacities of L. delbrueckii mutant L2 and L34 were 10.3% and 9.5% lower than original strain,respectively. After generation, the mutants were mixed-inoculated for milk fermentation test. The results showed the mutants were genetically stable.%为获得弱后酸化酸奶发酵菌株,采用紫外线对保加利亚乳杆菌3 4.5和嗜热链球菌sp1.1进行诱变处理,将诱变菌株同时接到低pH液体培养基和脱脂乳中,经37℃培养24h,选出低pH液体培养基不发生混浊的菌所对应的脱脂乳管,即为弱后酸化菌株,并对诱变菌株进行贮藏及传代稳定性实验.结果表明,球菌sp1.1与杆菌3 4.5紫外最佳诱变时间50S.经诱变后得到一株弱后酸化球菌S1,与原始菌株比后酸化程度降低了8.0%;两株杆菌L2、L34,与原始菌比后酸化分别降低10.3%、9.5%,经传代组合发酵乳实验,突变菌可稳定遗传.

  20. Study on improving the fermentation performance of Lactobacillus acidophilus in soybean milk%提高嗜酸乳杆菌在豆乳中发酵性能的研究

    Institute of Scientific and Technical Information of China (English)

    吕慧超; 熊宇生; 刘伦伦; 夏延斌

    2011-01-01

    测定了嗜酸乳杆菌、保加利亚乳杆菌和嗜热链球菌发酵豆乳的单菌产酸曲线与混菌产酸曲线,并采用正交实验分析了它们对发酵豆乳的品质影响及与单菌发酵豆乳贮存期质量变化的比较.结果表明,嗜酸乳杆菌、保加利亚乳杆菌是影响酸豆乳质量的主要因素,嗜热链球菌次之.按体积分数为2.2%嗜酸乳杆菌、体积分数为3.0%保加利亚乳杆菌、体积分数为1.8%嗜热链球菌配比共同发酵,可得到组织状态、口感风味较好的酸豆乳,克服了单一嗜酸乳杆菌发酵豆乳产酸慢、凝乳时间长、活菌数不高且贮存期活菌数下降快的缺陷.%In this study,Single curves and the mixed lactic acid bacteria acid production curves of Lactobacillus acidophilus, Lactobacillus bulgaricus and Streptococcus thermophilus bacteria fermented soybean milk were determined. By using the cross-experimental analysis method,their impact on the quality of fermented soybean milk were studied .Also,the quality changes of mixed bacteria and single-strain fermented soybean milk were compared during storage period. The results shows that Lactobacillus acidophilus, Lactobacillus bulgaricus are the main factots in fermented soybean milk, followed by Streptococcus thermophilus; In accordance with the proportion of 2.2%Lactobacillus acidophilus,3.0%Lactoba cillusbulgaricusandl.8%Streptococc-us thermophilus to co-ferment soybean milk, we can get organized state and good taste ,which overcomes the defects of single Lactobacillus acidophilus in fermented soybean milk that produce acid slowly, curd for a long time, not high number of viable cells and fast -decreasing viable cells during storage period.

  1. Screening low-goaty flavor fermentation strains for goat's yogurt%羊奶发酵中低膻昧菌株的筛选

    Institute of Scientific and Technical Information of China (English)

    苏伟丽; 张富新; 艾对; 葛萍; 于玲玲; 云丹; 侯院林; 杜远华

    2015-01-01

    The lipase activity,FFA content and goaty flavor of goat milks fermented with 6 Lactobacillus bulgaricus strains and 6 Streptococcus thermophilus strains from the commercial starter cultures were evaluated.The results showed that when a single strain was used to ferment,Lactobacillus bulgaricus L.b-883 strain and L.b-211 strain,Streptococcus thermophilus S.t-187 strain and S.t-300 strain had lower lipase activity,FFA content and goaty flavor in fermentation goat milk compared to other strains.When Lactobacillus bulgaricus was used with Streptococcus thermophilus in fermentation,L.b-211 strain and S.t-187 strain with 1∶2 ratio had lowest lipase activity (11.62 mol/mL · min)and FFA content (3.28 μequ/mL),and goat yogurt almost had not goaty flavor.%从商业发酵剂分离纯化出6株保加利亚乳杆菌和6株嗜热链球菌,研究其在羊奶酸奶发酵中脂肪酶活性、FFA以及膻味的变化.结果表明,用单一菌株发酵羊奶时,保加利亚乳杆菌L.b-883和L.b-211菌株、嗜热链球菌S.t-187和S.t-300菌株制作的羊奶酸奶中脂肪酶活性和FFA含量较低,羊奶酸奶膻味较轻;当杆菌和球菌配合应用时,L.b-211菌株与S.t-187菌株以2∶1比例配合,羊奶酸奶中脂肪酶活性和FFA含量最低,分别是11.62 μmol/mL·min和3.28 μequ/mL,感官评定几乎无膻味.

  2. Recognition of tRNAs with a long variable arm by aminoacyl-tRNA synthetases

    Directory of Open Access Journals (Sweden)

    Tukalo M. A.

    2013-07-01

    Full Text Available In prokaryotic cells three tRNA species, tRNASer, tRNALeu and tRNATyr, possess a long variable arm of 11–20 nucleotides (type 2 tRNA rather than usual 4 or 5 nucleotides (type 1 tRNA. In this review we have summarized the results of our research on the structural basis for recognition and discrimination of type 2 tRNAs by Thermus thermophilus seryl-, tyrosyl- and leucyl-tRNA synthetases (SerRS, TyrRS and LeuRS obtained by X-ray crystallography and chemical probing tRNA in solution. Crystal structures are now known of all three aminoacyl-tRNA synthetases complexed with type 2 tRNAs and the different modes of tRNA recognition represented by these structures will be discussed. In particular, emphasis will be given to the results on recognition of characteristic shape of type 2 tRNAs by cognate synthetases. In tRNASer, tRNATyr and tRNALeu the orientation of the long variable arm with respect to the body of the tRNA is different and is controlled by different packing of the core. In the case of SerRS the N-terminal domain and in the case of TyrRS, the C-terminal domain, bind to the characteristic long variable arm of the cognate RNA, thus recognizing the unique shape of the tRNA. The core of T. thermophilus tRNALeu has several layers of unusual base-pairs, which are revealed by the crystal structure of tRNALeu complexed with T. thermophilus LeuRS and by probing a ligand-free tRNA by specific chemical reagents in solution. In the crystal structure of the LeuRS-tRNALeu complex the unique D-stem structure is recognized by the C-terminal domain of LeuRS and these data are in good agreement with those obtained in solution. LeuRS has canonical class I mode of tRNA recognition, approaching the tRNA acceptor stem from the D-stem and minor groove of the acceptor stem side. SerRS also has canonical class II mode of tRNA recognition and approaches tRNASer from opposite, variable stem and major groove of acceptor stem site. And finally, TyrRS in strong

  3. Phenotypic Suppression of Streptomycin Resistance by Mutations in Multiple Components of the Translation Apparatus

    Science.gov (United States)

    Carr, Jennifer F.; Lee, Hannah J.; Jaspers, Joshua B.; Dahlberg, Albert E.; Jogl, Gerwald

    2015-01-01

    ABSTRACT The bacterial ribosome and its associated translation factors are frequent targets of antibiotics, and antibiotic resistance mutations have been found in a number of these components. Such mutations can potentially interact with one another in unpredictable ways, including the phenotypic suppression of one mutation by another. These phenotypic interactions can provide evidence of long-range functional interactions throughout the ribosome and its functional complexes and potentially give insights into antibiotic resistance mechanisms. In this study, we used genetics and experimental evolution of the thermophilic bacterium Thermus thermophilus to examine the ability of mutations in various components of the protein synthesis apparatus to suppress the streptomycin resistance phenotypes of mutations in ribosomal protein S12, specifically those located distant from the streptomycin binding site. With genetic selections and strain constructions, we identified suppressor mutations in EF-Tu or in ribosomal protein L11. Using experimental evolution, we identified amino acid substitutions in EF-Tu or in ribosomal proteins S4, S5, L14, or L19, some of which were found to also relieve streptomycin resistance. The wide dispersal of these mutations is consistent with long-range functional interactions among components of the translational machinery and indicates that streptomycin resistance can result from the modulation of long-range conformational signals. IMPORTANCE The thermophilic bacterium Thermus thermophilus has become a model system for high-resolution structural studies of macromolecular complexes, such as the ribosome, while its natural competence for transformation facilitates genetic approaches. Genetic studies of T. thermophilus ribosomes can take advantage of existing high-resolution crystallographic information to allow a structural interpretation of phenotypic interactions among mutations. Using a combination of genetic selections, strain constructions

  4. Ionic liquids for the concomitant use in extremophiles lysis and extremozymes extraction.

    Science.gov (United States)

    Gutiérrez-Arnillas, E; Deive, F J; Sanromán, M A; Rodríguez, A

    2015-06-01

    Ionic liquids have been successfully proposed to modify membrane permeability in cultures of a model extremophilic bacterium Thermus thermophilus HB27, which makes up the first time that aqueous solutions of these molten salts are applied in downstream stages of this kind of microorganisms. The presence of 1g/L of C10MIMCl entails a great solubilisation of cell biomass, thus allowing the release of intracellular and membrane-bound enzyme. The influence on the enzyme activity of two inorganic salts such as Na2CO3 and (NH4)2SO4, selected on the basis of their high salting out potential and biocompatibility with enzymes, respectively, was investigated. In parallel, their ability to trigger phase segregation was confirmed in the presence of the enzyme crude, leading to very high levels of enzyme extraction (96%). The validity of the strategy was confirmed by operating at bioreactor scale, and the main bioprocess parameters were obtained by modelling the experimental data.

  5. Mozzarella干酪生产用菌种的筛选%Selection of Mozzarella cheese strains

    Institute of Scientific and Technical Information of China (English)

    刘会平; 南庆贤; 马长伟

    2003-01-01

    概述了Mozzarella干酪的特点以及生产用菌株所需具备的特性,从现有的菌种和市售优质酸乳中分离纯化得到25株乳酸菌,经初步的生产性能试验,筛选出12株性能较好的菌株.经进一步的发酵性能和生化试验,优选出两株优良的菌株,并初步鉴定为唾液链球菌嗜热亚种(Streptococcus salivarius subsp.Thermophilus)CH9和保加利亚乳杆菌德氏亚种(Lactobacillus delbrueckil ssp.Bulgaricus)LB,经过实际的干酪生产,证明表现良好.

  6. Improved immobilization of laccase on a glassy carbon electrode by oriented covalent attachment

    Directory of Open Access Journals (Sweden)

    Liu Xin

    2014-01-01

    Full Text Available A laccase from Thermus thermophilus HB27 was reported to be potentially useful in the design of a temperature controlled biofuel cell. For enhancing its application in different thermal conditions, we engineered a laccase-oriented immobilized electrode. A site-directed mutant N323C of the laccase was constructed. A photometric assay was employed in order to compare the catalytic properties of wild-type laccase and mutant. The mutant was attached to a glass carbon electrode by covalent cross-linking. The electrochemical properties of the immobilized laccase were investigated by cyclic voltammetry. This immobilization allowed the active electrode to function at temperatures up to 95°C. The thermal and pH dependence profiles were similar to those of the soluble enzyme investigated by spectrophotometry.

  7. Concentration dependent effects of dextran on the physical properties of acid milk gels.

    Science.gov (United States)

    Mende, Susann; Peter, Michaela; Bartels, Karin; Dong, Tingting; Rohm, Harald; Jaros, Doris

    2013-11-01

    The effect of dextran from Leuconostoc mesenteroides (DEX500), added to milk prior to acidification with glucono-δ-lactone (GDL) or Streptococcus thermophilus DSM20259, was studied with respect to polysaccharide concentration. The incorporation of 5-30 g/kg DEX500 significantly affected gelation behavior. Increasing DEX500 concentrations resulted in a linear increase of gel stiffness (GDL gels: R(2)=0.96; microbial acidification: R(2)=0.94; Psyneresis, which decreased from 30.4% (0 g/kg DEX500) to 22.0% (30 g/kg DEX500) for chemically acidified gels after 1 d of storage. Physical characteristics of DEX500 in aqueous solution were helpful to explain its behavior in the complex system milk.

  8. Crystal structure of ribosomal protein L1 from the bacterium Aquifex aeolicus

    Science.gov (United States)

    Nikonova, E. Yu.; Tishchenko, S. V.; Gabdulkhakov, A. G.; Shklyaeva, A. A.; Garber, M. B.; Nikonov, S. V.; Nevskaya, N. A.

    2011-07-01

    The crystal structure of ribosomal protein L1 from the bacterium Aquifex aeolicus was solved by the molecular-replacement method and refined to R cryst = 19.4% and R free = 25.1% at 2.1 Å protein consists of two domains linked together by a flexible hinge region. In the structure under consideration, the domains are in close proximity and adopt a closed conformation. Earlier, this conformation has been found in the structure of protein L1 from the bacterium Thermus thermophilus, whereas the structures of archaeal L1 proteins and the structures of all L1 proteins in the RNA-bound form have an open conformation. The fact that a closed conformation was found in the structures of two L1 proteins which crystallize in different space groups and belong to different bacteria suggests that this conformation is a characteristic feature of L1 bacterial proteins in the free form.

  9. Food environments select microorganisms based on selfish energetic behavior

    Directory of Open Access Journals (Sweden)

    Diego eMora

    2013-11-01

    Full Text Available Nutrient richness, and specifically the abundance of mono- and disaccharides that characterize several food matrixes, such as milk and grape juice, has allowed the speciation of lactic acid bacteria and yeasts with a high fermentation capacity instead of energetically favorable respiratory metabolism. In these environmental contexts, rapid sugar consumption and lactic acid or ethanol production, accumulation and tolerance, together with the ability to propagate in the absence of oxygen, are several of the ‘winning’ traits that have apparently evolved and become specialized to perfection in these fermenting microorganisms. Here, we summarize and discuss the evolutionary context that has driven energetic metabolism in food-associated microorganisms, using the dairy species Lactococcus lactis and Streptococcus thermophilus among prokaryotes and the bakers’ yeast Saccharomyces cerevisiae among eukaryotes as model organisms.

  10. Yeast surface display of dehydrogenases in microbial fuel-cells.

    Science.gov (United States)

    Gal, Idan; Schlesinger, Orr; Amir, Liron; Alfonta, Lital

    2016-12-01

    Two dehydrogenases, cellobiose dehydrogenase from Corynascus thermophilus and pyranose dehydrogenase from Agaricus meleagris, were displayed for the first time on the surface of Saccharomyces cerevisiae using the yeast surface display system. Surface displayed dehydrogenases were used in a microbial fuel cell and generated high power outputs. Surface displayed cellobiose dehydrogenase has demonstrated a midpoint potential of -28mV (vs. Ag/AgCl) at pH=6.5 and was used in a mediator-less anode compartment of a microbial fuel cell producing a power output of 3.3μWcm(-2) using lactose as fuel. Surface-displayed pyranose dehydrogenase was used in a microbial fuel cell and generated high power outputs using different substrates, the highest power output that was achieved was 3.9μWcm(-2) using d-xylose. These results demonstrate that surface displayed cellobiose dehydrogenase and pyranose dehydrogenase may successfully be used in microbial bioelectrochemical systems.

  11. Exploring assembly energetics of the 30S ribosomal subunit using an implicit solvent approach.

    Science.gov (United States)

    Trylska, Joanna; McCammon, J Andrew; Brooks Iii, Charles L

    2005-08-10

    To explore the relationship between the assembly of the 30S ribosomal subunit and interactions among the constituent components, 16S RNA and proteins, relative binding free energies of the T. thermophilus 30S proteins to the 16S RNA were studied based on an implicit solvent model of electrostatic, nonpolar, and entropic contributions. The late binding proteins in our assembly map were found not to bind to the naked 16S RNA. The 5' domain early kinetic class proteins, on average, carry the highest positive charge, get buried the most upon binding to 16S RNA, and show the most favorable binding. Some proteins (S10/S14, S6/S18, S13/S19) have more stabilizing interactions while binding as dimers. Our computed assembly map resembles that of E. coli; however, the central domain path is more similar to that of A. aeolicus, a hyperthermophilic bacteria.

  12. A study of the thermophilic ribosomal protein S7 binding to the truncated S12-S7 intercistronic region provides more insight into the mechanism of regulation of the str operon of E. coli(1).

    Science.gov (United States)

    Spiridonova, V A; Rozhdestvensky, T S; Kopylov, A M

    1999-10-29

    A study of the ability of His6-tagged ribosomal protein S7 of Thermus thermophilus to interact with the truncated S12-S7 intercistronic region of str mRNA of Escherichia coli has been described. A minimal S7 binding mRNA fragment is a part of the composite hairpin, with the termination codon of the S12 cistron on one side and the initiation codon of the next S7 cistron on the other. It has a length in the range of 63-103 nucleotides. The 63 nucleotide mRNA fragment, which corresponds to a putative S7 binding site, binds very poorly with S7. Tight RNA structure models, which behave as integral systems and link the S7 binding site with the translational regulation region of the hairpin, are suggested. This observation provides more insight into the mechanism of S7-directed autogenous control of translational coupling of str mRNA.

  13. Affinity of ribosomal protein S8 from mesophilic and (hyper)thermophilic archaea and bacteria for 16S rRNA correlates with the growth temperatures of the organisms.

    Science.gov (United States)

    Gruber, Thomas; Köhrer, Caroline; Lung, Birgit; Shcherbakov, Dmitri; Piendl, Wolfgang

    2003-08-14

    The ribosomal protein S8 plays a pivotal role in the assembly of the 30S ribosomal subunit. Using filter binding assays, S8 proteins from mesophilic, and (hyper)thermophilic species of the archaeal genus Methanococcus and from the bacteria Escherichia coli and Thermus thermophilus were tested for their affinity to their specific 16S rRNA target site. S8 proteins from hyperthermophiles exhibit a 100-fold and S8 from thermophiles exhibit a 10-fold higher affinity than their mesophilic counterparts. Thus, there is a striking correlation of affinity of S8 proteins for their specific RNA binding site and the optimal growth temperatures of the respective organisms. The stability of individual rRNA-protein complexes might modulate the stability of the ribosome, providing a maximum of thermostability and flexibility at the growth temperature of the organism.

  14. 酸乳饮料中杂菌的变化及作用%Changing Principles of Infectious Microbe and Its Reaction in Fermented Milk Drinks

    Institute of Scientific and Technical Information of China (English)

    刘绍军; 李军; 周丽艳; 刘卫华; 赵希艳

    2004-01-01

    对酸乳饮料中的微生物进行分离鉴定,除两种发酵剂乳酸菌--嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus) 以外,其中酵母主要是酿酒酵母(Saccharomyces cerevisive);细菌主要有植物乳杆菌(Lactobacillus plantarum)、发酵乳杆菌(Lactobacillus fermentum)和大肠埃希氏菌(E.coli);霉菌主要有黑曲霉(Aspergillus niger)和桔青霉(Penicillum citrinum).在酸乳饮料腐败变质过程中,其中的微生物数量、酸碱度及含糖量存在一定的变化规律.

  15. Examining the possibilities of applying high pressure to preserve yoghurt supplemented with probiotic bacteria

    Science.gov (United States)

    Jankowska, Agnieszka; Grześkiewicz, Aleksandra; Wiśniewska, Krystyna; Reps, Arnold

    2012-09-01

    Natural yoghurt was subject to pressures of 200 and 250 MPa/4 and 18°C/15 min, after which milk-activated inocula of Lactobacillus acidophilus and Bifidobacterium sp. were added. The yoghurts were stored for 4 weeks at refrigeration temperature. After preparation and each week of storage, the count of bacteria, acidity, antibacterial property and an organoleptic assessment was determined. The highest survival rate was demonstrated by the bacteria of Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus and Bifidobacterium sp. in the yoghurt pressurised 200 MPa/15min at 4°C. Acidity increases in the control yoghurts were higher than in the pressurised ones. Pressurised yoghurts demonstrated weaker antibacterial effect in comparison to control yoghurts. Slight changes in the smell and taste were observed after pressurisation. Yoghurts pressurised at 18°C were characterised by more favourable organoleptic properties. Better conciseness of the curd and lower whey seep out were observed in pressurised yoghurt.

  16. Cloning vectors based on cryptic plasmids isolated from lactic acid bacteria: their characteristics and potential applications in biotechnology.

    Science.gov (United States)

    Shareck, Julie; Choi, Young; Lee, Byong; Miguez, Carlos B

    2004-01-01

    Lactic acid bacteria (LAB) are Gram positive bacteria, widely distributed in nature, and industrially important as they are used in a variety of industrial food fermentations. The use of genetic engineering techniques is an effective means of enhancing the industrial applicability of LAB. However, when using genetic engineering technology, safety becomes an essential factor for the application of improved LAB to the food industry. Cloning and expression systems should be derived preferably from LAB cryptic plasmids that generally encode genes for which functions can be proposed, but no phenotypes can be observed. However, some plasmid-encoded functions have been discovered in cryptic plasmids originating from Lactobacillus, Streptococcus thermophilus, and Pediococcus spp. and can be used as selective marker systems in vector construction. This article presents information concerning LAB cryptic plasmids, and their structures, functions, and applications. A total of 134 cryptic plasmids collated are discussed.

  17. Dicty_cDB: Contig-U16217-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available .. 38 0.78 3 ( BX119979 ) Zebrafish DNA sequence from clone CH211-201G8 in ... 44 0.81 7 ( EV866087 ) rpbfe0...T677259 ) Danio rerio EST, clone ZF_mu_162m07 5'. 48 1.1 2 ( EV868478 ) rpbfe0111... sequence. 38 1.6 2 ( CO953317 ) UMC-pfeto1-017-f08 Fetal ovary pfeto Sus scrofa c... 38 1.6 2 ( EV870684 ) rpbf... 1 ( AF316886 ) Streptococcus thermophilus oligopeptide binding p... 48 1.7 1 ( EV867579 ) rpbfe0102_d3.y1 b...fe Sus scrofa cDNA 5', mRNA sequ... 38 1.8 2 ( EV873666 ) rpbfe0136_c13.y1 bfe Sus scrofa cDNA 5', mRNA seq.

  18. Determination of reactive oxygen generated from natural medicines and their antibacterial activity$

    Institute of Scientific and Technical Information of China (English)

    Noriko Tajima; Makiko Takasaki; Haruka Fukamachi; Takeshi Igarashi; Yoshijiro Nakajima; Hidetoshi Arakawa

    2016-01-01

    Extracts of 16 natural medicine powders (Galla chinensis, Malloti cortex, Cassiae semen, Sophorae radix, Myricae cortex, Crataegi fructus, Gambir, Mume fructus, Geranii herba, Phellodendri cortex, Coptidis rhizoma, Swertiae herba, and Cinnamomi cortex) were assayed for reactive oxygen concentrations using the per-oxyoxalate chemiluminescent detection system. High luminescence intensity was observed in Galla chinensis, Geranii herba, Malloti cortex, Myricae cortex, and Cinnamomi cortex. Additional experiments identified the reactive oxygen species as hydrogen peroxide. Galla chinensis generated 2.4 ? 10 ? 4 mol/L hydrogen peroxide from a 1 mg/mL solution. In bacterial growth tests, Galla chinensis extract had antibacterial activity against Escherichia coli, Staphylococcus aureus, Bacteroides thetaiotaomicron, Campylobacter sputorum biovar sputorum, Streptococcus salivarius thermophilus, Lactobacillus casei, and Bifidobacterium longum infantis. This antibacterial activity was de-creased by the addition of catalase. It revealed that hydrogen peroxide which Galla chinensis produced participated in antibacterial activity.

  19. STUDY OF GUSTATORY AND STRUCTURAL PROPERTIES OF KEFIR PRODUCT

    Directory of Open Access Journals (Sweden)

    MILENA H. MOMCHILOVA

    2012-06-01

    Full Text Available The subject of the study is the improvement of Kefir culture. For this investigation it was used Kefir culture and two series of experiments were carried out. Yeasts from probiotic strain Saccharomyces boulardii were added in the first series and the lactic acid bacteria (LAB of Streptococcus thermophilus strain, with a polysaccharide activity were added to Kefir culture in the second series. The fermentation conditions were 30°C, duration 16 hours and cooling 4 hours up to 4°C. The characteristics of Kefir were analyzed by determination of: pH, acidity, qualitative reaction for existence of diacetyl, cell number of LAB and yeasts. The structural properties of Kefir were evaluated by microscopic study.

  20. Aminotransferase and glutamate dehydrogenase activities in lactobacilli and streptococci.

    Science.gov (United States)

    Peralta, Guillermo Hugo; Bergamini, Carina Viviana; Hynes, Erica Rut

    2016-01-01

    Aminotransferases and glutamate dehydrogenase are two main types of enzymes involved in the initial steps of amino acid catabolism, which plays a key role in the cheese flavor development. In the present work, glutamate dehydrogenase and aminotransferase activities were screened in twenty one strains of lactic acid bacteria of dairy interest, either cheese-isolated or commercial starters, including fifteen mesophilic lactobacilli, four thermophilic lactobacilli, and two streptococci. The strains of Streptococcus thermophilus showed the highest glutamate dehydrogenase activity, which was significantly elevated compared with the lactobacilli. Aspartate aminotransferase prevailed in most strains tested, while the levels and specificity of other aminotransferases were highly strain- and species-dependent. The knowledge of enzymatic profiles of these starter and cheese-isolated cultures is helpful in proposing appropriate combinations of strains for improved or increased cheese flavor. PMID:27266631

  1. Using PacBio Long-Read High-Throughput Microbial Gene Amplicon Sequencing To Evaluate Infant Formula Safety.

    Science.gov (United States)

    Zheng, Yi; Xi, Xiaoxia; Xu, Haiyan; Hou, Qiangchuan; Bian, Yanfei; Yu, Zhongjie; Kwok, Lai-Yu; Zhang, Wenyi; Sun, Zhihong; Zhang, Heping

    2016-09-21

    Infant formula (IF) requires a strict microbiological standard because of the high vulnerability of infants to foodborne diseases. The current study used the PacBio single-molecule real-time (SMRT) sequencing platform to generate full-length 16S rRNA-based bacterial microbiota profiles of 30 Chinese domestic and imported IF samples. A total of 600 species were identified, dominated by Streptococcus thermophilus, Lactococcus lactis, and Lactococcus piscium. Distinctive bacterial profiles were observed between the two sample groups, as confirmed with both principal coordinate analysis and multivariate analysis of variance. Moreover, the product whey protein nitrogen index (WPNI), representing the degree of preheating, negatively correlated with the relative abundances of the Bacillus genus. This study has demonstrated the application of the PacBio SMRT sequencing platform in assessing the bacterial contamination of IF products, which is of interest to the dairy industry for effective monitoring of microbial quality and safety during production. PMID:27500310

  2. Site-specific Incorporation of 3-Iodo-L-tyrosine into Proteins and Single-wavelength Anomalous Dispersion Phasing with Soft X-ray in Protein Crystallography

    Science.gov (United States)

    Murayama, Kazutaka; Sakamoto, Kensaku

    Iodine is a good anomalous scatter for radiations from in-house X-ray generators (Cu/CrKα). Non-natural amino acid, 3-iodo-L-tyrosine, is able to be site-specifically incorporated into proteins with amber suppresser tRNA and mutated TyrRS from M. jannaschii in the E. coli expression system. To determine the crystal structure of acetyl transferase from T. thermophilus, iodotyrosine-containing proteins were prepared and crystallized. Structure determination was successfully conducted with the protein variant with iodotyrosine at position 111. Anomalous signals from iodotyrosine with Cu/CrKα radiations were both sufficient to calculate clear electron density map. In the crystal structure, iodotyrosine did not significantly disturb the native structure.

  3. Efficiency of new fungal cellulase systems in boosting enzymatic degradation of barley straw lignocellulose

    DEFF Research Database (Denmark)

    Rosgaard, L.; Pedersen, S.; Meyer, Anne Boye Strunge

    2006-01-01

    This study examined the cellulytic effects on steam-pretreated barley straw of cellulose-degrading enzyme systems from the five thermophilic fungi Chaetomium thermophilum, Thielavia terrestris, Thermoascus aurantiacus, Corynascus thermophilus, and Myceliophthora thermophila and from the mesophile...... Penicillum funiculosum. The catalytic glucose release was compared after treatments with each of the crude enzyme systems when added to a benchmark blend of a commercial cellulase product, Celluclast, derived from Trichoderma reesei and a P-glucosidase, Novozym 188, from Aspergillus niger. The enzymatic...... treatments were evaluated in an experimental design template comprising a span of pH (3.5-6.5) and temperature (35-65 degrees C) reaction combinations. The addition to Celluclast + Novozym 188 of low dosages of the crude enzyme systems, corresponding to 10 wt % of the total enzyme protein load, increased...

  4. 不同乳酸菌对羊酸奶的感官和质构特性的影响%The Effect of Different Lactobacillus on the Sensory and Textural Properties of Goat Yoghurt

    Institute of Scientific and Technical Information of China (English)

    祝静; 丁武

    2011-01-01

    研究分析不同乳酸菌对羊酸奶感官与质构特性的影响作用,为筛选优良的羊酸奶发酵剂提供依据.使用双歧杆菌、保加利亚乳杆菌、嗜热链球菌、丁二酮乳链球菌发酵羊奶,对羊酸奶的感官品质、酸度及质构特性进行评定.结果表明,单一菌种发酵,四种菌发酵的羊酸奶的感官评分排序是双歧杆菌>嗜热链球菌>保加利亚乳杆菌>丁二酮乳链球菌,不同乳酸菌发酵羊酸奶的质构特性差异显著,使用双歧杆菌发酵可以获得最佳的质构特性,其发酵的羊酸奶的硬度为34.96g、黏度为802.28g·s、凝聚性为44.65g、粘性指数为137.17g·s、保水力为85.00%;混合发酵剂发酵羊酸奶具有各自不同的感官和质构特性优势,双歧杆菌和嗜热链球菌发酵的羊酸奶的硬度和黏度最好,分别达到29.93g和715.09g·s,但风味欠佳;双歧杆菌和丁二酮乳链球菌发酵的羊酸奶的凝聚性、粘性指数和保水力最好,分别达到45.86g、143.49g·s、91.00%,且风味良好.综合感官评价与质构特性,复合菌种双歧杆菌和丁二酮乳链球菌适宜作为羊酸奶的发酵剂.%The effect of different lactobacillus on the sensory and textural properties of goat yoghurt was investigated, and a basis for selectiving better starters of goat yoghurt was provided. Goat milk was fermented by Bifidobacterium, Streptococcus thermophilus,Lactobacillus bulgaricus and Streptococcus diacetilucs, and their sensory evaluation, acidity and texture were determind. The result showed that fermentation by singlelactobocillus, the sensory order of the four kind oflactobocillus was Bifidobacterium>Streptococcus thermophilus >Lactobocillus bulgaricus >Streptococcus diacetilucs. Different lactobacillus was significantly different on textural properties, and the firmness, consistency, cohesiveness, index of viscosity and water holding capacity of goat yogurt fermented by Bifidobactedum were the best, the were 34

  5. 自然发酵面团中乳酸菌菌株的分离与鉴定%Isolation and Identification of Lactic Acid Bacteria from Nature Fermented Dough

    Institute of Scientific and Technical Information of China (English)

    2013-01-01

    We obtained two lactic acid bacteria strains from fermented dough by MRS medium and M17 medium. Through the observation of colony and mycelia morphologies and physiological and biochemical experiment, the strain 1 was Lactobacillus plantarum, the strain 2 was Streptococcus thermophilus.%  利用MRS和M17培养基,从自然发酵面团中分离纯化出2株乳酸菌菌株,通过菌落形态、菌体形态观察和生理生化试验鉴定,菌株1为植物乳杆菌(Lactobacillus plantarum),菌株2为嗜热链球菌(Streptococcus thermophilus)。

  6. Characterization of non-starter lactic acid bacteria in traditionally produced home-made Radan cheese during ripening

    Directory of Open Access Journals (Sweden)

    Jokovic Natasa

    2011-01-01

    Full Text Available Two hundred thirteen non-starter lactic acid bacteria isolated from Radan cheese during ripening were identified with both a classical biochemical test and rep-PCR with (GTG5 primer. For most isolates, which belong to the Lactococcus lactis subsp. lactis, Leuconostoc mesenteroides, Lactobacillus plantarum, Lactobacillus paraplantarum and Enterococcus faecium, a phenotypic identification was in good agreement with rep-PCR identification. Lactococeus lactis subsp. lactis, Enterococcus faecium and subspecies from the Lenconostoc mesenteroides group were the dominant population of lactic acid bacteria in cheese until 10 days of ripening and only one Streptococcus thermophilus strain was isolated from the 5-day-old cheese sample. As ripening progressed, Lactobacillus plantarum became the predominant species together with the group of heterofermentative species of lactobacilli that could not be precisely identified with rep-PCR.

  7. Isolation,identificiation and and its probiotic characteristics of Lactic acid bacteria from market yoghurts in Zhengzhou%郑州部分市售酸奶中乳酸菌的分离鉴定与益生特性的研究

    Institute of Scientific and Technical Information of China (English)

    杨霞; 陈陆; 杜魏红; 邓瑞坡; 刘红英; 姚惠霞; 王川庆

    2011-01-01

    目的 调查郑州部分市售的酸奶中嗜热乳酸链球菌与保加利亚乳酸杆菌存在情况与益生特性.方法 无菌采取样品进行乳酸菌分离,并对分离菌进行形态学、生化特性、产酸性、耐酸性、耐胆汁性、高温耐受性和抑菌性等生物学特性研究.结果 从8种品牌中分离到4株保加利亚乳酸杆菌、7株嗜热链球菌,且分离菌株均能产酸、对高温具有一定的耐受性,在pH 3.0以上均能生长良好,而且分离菌的抑菌物质具有一定的热稳定性,保加利亚乳酸杆菌经70、100 ℃热处理抑菌活性基本不变.嗜热链球菌经70 ℃热处理后保留90%以上的抑菌活性,但经100 ℃高温处理后抑菌活性有所降低.分离菌产生的抑菌物质对胰蛋白酶敏感,保加利亚乳酸杆菌在偏中性条件下抑菌活性较高,而嗜热链球菌在偏中性条件下丧失抑菌活性.结论 本研究为河南市售酸奶质量以及为消费者提供有利信息奠定基础.%Objective To separate Streptococcus thermophilus and Lactobacillus bulgaricus from various yojurt in Zhengzhou and discuss the probiotic characteristics of them. Method The gathered yoghurt samples were used to isolate the strain, and a series tests were carried out, such ss morphology, biochemical characteristics, acidity-producing ability,acidity-enduring ability, tolerance to bile salts and high temperature tolerance, and so on. Result 4 Lactobacillus bulgaricus and 7 Streptococcus thermophilus were separated from 8 kinds of brand yojurt, and isolated strains could produce acid,also had certain high temperature tolerance. It could grow well above pH 3.0, and the isolated antibacterial material had certain thermal stability. The antibacterial activity of Lactobacillus bulgaricus was almost no change under 70 ℃ and 100 ℃ treatment; The antibacterial activity of Streptococcus thermophilus was retain 90% under 70 ℃ treatment, but lower after 100 ℃ heated treatment. The

  8. Functional Starters for Functional Yogurt

    Directory of Open Access Journals (Sweden)

    Mattia P. Arena

    2015-02-01

    Full Text Available In this study, we investigated the multifunctionality (microbial starters and probiotics of Lactobacillus plantarum WCFS1 and Lactobacillus plantarum CECT 8328 strains used as microbial starters for the production of yogurt in combination with Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus. The ability of the probiotic strains to survive oro-gastrointestinal stresses was monitored by an in vitro assay simulating the human digestive tract. The transcriptional level of several genes involved in the immune response suggested that the probiotic strains may have a favorable influence on immunomodulation. Overall, this study revealed that the tested Lactobacilli exhibited suitable technological features for yogurt production and might be used to formulate novel food with immunomodulating effects.

  9. Lysogeny detection of lactic acid bacteria and analysis on protein expression of lysogenic strain after UV mutagenesis%乳酸菌溶源性的检测及紫外诱变后溶源菌蛋白表达分析

    Institute of Scientific and Technical Information of China (English)

    张鹭; 郭承宇; 王庆忠; 路福平

    2006-01-01

    用紫外诱变的方法对本实验室保藏的嗜热链球菌(Streptococcus thermophilus)和保加利亚乳杆菌(Lactobacillus bulgaricus)进行溶源性的检测;并采用单向聚丙稀酰氨凝胶电泳,对不同诱变剂量下溶源菌菌体蛋白质组分的变化做了进一步分析,发现诱变前后菌体的蛋白组成发生了明显的变化,即随着诱变剂量的增加,菌体内蛋白种类,尤其是小分子蛋白有上升的趋势.

  10. CARACTERÍSTICAS FÍSICAS E QUÍMICAS DE BEBIDAS LÁCTEAS FERMENTADAS E PREPARADAS COM SORO DE QUEIJO MINAS FRESCAL PHYSICAL AND CHEMICAL CHARACTERISTICS OF FERMENTED DAIRY BEVERAGES USING MINAS CHEESE WHEY

    Directory of Open Access Journals (Sweden)

    Keila Emílio de ALMEIDA

    2001-08-01

    Full Text Available Nesta pesquisa procurou-se verificar as características físicas e químicas de bebidas lácteas preparadas com três concentrações de soro de queijo Minas Frescal (30, 40 e 50%, empregando-se dois tipos de culturas lácticas: uma tradicional para iogurte (YC-180 contendo cepas mistas de Streptococcus salivarus subsp. thermophilus, Lactobacillus delbrueckii subsp. lactis e Lactobacillus delbrueckii subsp. bulgaricus e outra (ABY-1 contendo cepas mistas de Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus acidophillus, Bifidobacteria e Streptococcus salivarius subsp. thermophilus. Constatou-se que as bebidas lácteas apresentaram diferença estatística no tempo zero para os teores de gordura e de extrato seco. À medida em que se elevou a proporção de soro em relação ao leite, os teores de gordura e de extrato seco diminuíram. O teor de proteína também diminuiu à medida em que se aumentou o teor de soro nas bebidas lácteas, embora a diferença não tenha sido tão acentuada quanto as observadas para os teores de gordura e de extrato seco. Em relação à lactose, não se constatou diferença entre os tratamentos. Os teores de soro não influenciaram o índice de proteólise das bebidas lácteas. Verificou-se todavia que as bebidas elaboradas com a cultura probiótica ABY-1 apresentaram valores superiores para proteólise quando comparadas às bebidas elaboradas com as culturas YC-180. As bebidas lácteas elaboradas com 30% de soro apresentaram maiores valores para viscosidade. As bebidas elaboradas com a cultura YC-180 apresentaram valores superiores para viscosidade durante o período de armazenamento.This research studies the physical and chemical characteristics of dairy beverages formulated with three different concentrations (30, 40 and 50 % of Minas cheese whey, using two kinds of dairy cultures: the traditional yogurt (YC--180 culture, holding mixed strains of Streptococcus salivarus subsp. thermophilus, Lactobacillus

  11. Evaluation the Sensory and Probiotics Properties of the Yogurt Supplemented with Carrot Juice

    Directory of Open Access Journals (Sweden)

    Carmen R. Pop

    2015-11-01

    Full Text Available This study aimed to develop a new type of yogurt through addition of carrot juice. Yogurt is a fermented milk product obtained from fermentation of Lactobacillus bulgaricus and Streptococcus thermophilus strains. The effect of carrot juice at different levels (0%, 8%, 16%, 24% on sensory properties and the viability of probiotic bacteria in yogurts during storage (21 days at refrigerated temperature (4°C was evaluated. The yogurt supplementation with 24 % carrot juice significantly improves the stability of the lactic acid bacteria, that contained the recommended levels of 107 cfu/g probiotic bacteria at the end of 21-days shelf life. The yogurt with 24% carrot juice was the most appreciated (7.07 points, followed by the classic yogurt (6 points, yogurt with 8% (5.28 points and yogurt with16% carrot juice (5.5 points.

  12. Bathymodiolus growth dynamics in relation to environmental fluctuations in vent habitats

    Science.gov (United States)

    Nedoncelle, K.; Lartaud, F.; Contreira Pereira, L.; Yücel, M.; Thurnherr, A. M.; Mullineaux, L.; Le Bris, N.

    2015-12-01

    The deep-sea mussel Bathymodiolus thermophilus is a dominant species in the East Pacific Rise (EPR) hydrothermal vent fields. On the EPR volcanically unstable area, this late colonizer reaches high biomass within 4-5 years on new habitats created by lava flows. The environmental conditions and growth rates characterizing the reestablishment of B. thermophilus populations are however largely unknown, leaving unconstrained the role of this foundation species in the ecosystem dynamics. A typical example from the vent field at 9°50'N that was affected by the last massive eruption was the Bio-9 hydrothermal vent site. Here, six years later, a large mussel population had reestablished. The von Bertalanffy growth model estimates the oldest B. thermophilus specimens to be 1.3 year-old in March 2012, consistent with the observation of scarce juveniles among tubeworms in 2010. Younger cohorts were also observed in 2012 but the low number of individuals, relatively to older cohorts, suggests limited survival or growth of new recruits at this site, that could reflect unsuitable habitat conditions. To further explore this asumption, we investigated the relationships between mussel growth dynamics and habitat properties. The approach combined sclerochronology analyses of daily shell growth with continuous habitat monitoring for two mussel assemblages; one from the Bio-9 new settlement and a second from the V-vent site unreached by the lava flow. At both vent sites, semi-diurnal fluctuations of abiotic conditions were recorded using sensors deployed in the mussel bed over 5 to 10 days. These data depict steep transitions from well oxygenated to oxygen-depleted conditions and from alkaline to acidic pH, combined with intermittent sulfide exposure. These semi-diurnal fluctuations exhibited marked changes in amplitude over time, exposing mussels to distinct regimes of abiotic constraints. The V-vent samples allowed growth patterns to be examined at the scale of individual life and

  13. Current taxonomy of phages infecting lactic acid bacteria

    Directory of Open Access Journals (Sweden)

    Jennifer eMahony

    2014-01-01

    Full Text Available Phages infecting lactic acid bacteria have been the focus of significant research attention over the past three decades. Through the isolation and characterization of hundreds of phage isolates, it has been possible to classify phages of the dairy starter and adjunct bacteria Lactococus lactis, Streptococcus thermophilus, Leuconostoc spp. and Lactobacillus spp. Among these, phages of L. lactis have been most thoroughly scrutinized and serve as an excellent model system to address issues that arise when attempting taxonomic classification of phages infecting other LAB species. Here, we present an overview of the current taxonomy of phages infecting LAB genera of industrial significance, the methods employed in these taxonomic efforts and how these may be employed for the taxonomy of phages of currently underrepresented and emerging phage species.

  14. Aminotransferase and glutamate dehydrogenase activities in lactobacilli and streptococci.

    Science.gov (United States)

    Peralta, Guillermo Hugo; Bergamini, Carina Viviana; Hynes, Erica Rut

    2016-01-01

    Aminotransferases and glutamate dehydrogenase are two main types of enzymes involved in the initial steps of amino acid catabolism, which plays a key role in the cheese flavor development. In the present work, glutamate dehydrogenase and aminotransferase activities were screened in twenty one strains of lactic acid bacteria of dairy interest, either cheese-isolated or commercial starters, including fifteen mesophilic lactobacilli, four thermophilic lactobacilli, and two streptococci. The strains of Streptococcus thermophilus showed the highest glutamate dehydrogenase activity, which was significantly elevated compared with the lactobacilli. Aspartate aminotransferase prevailed in most strains tested, while the levels and specificity of other aminotransferases were highly strain- and species-dependent. The knowledge of enzymatic profiles of these starter and cheese-isolated cultures is helpful in proposing appropriate combinations of strains for improved or increased cheese flavor.

  15. Yeast surface display of dehydrogenases in microbial fuel-cells.

    Science.gov (United States)

    Gal, Idan; Schlesinger, Orr; Amir, Liron; Alfonta, Lital

    2016-12-01

    Two dehydrogenases, cellobiose dehydrogenase from Corynascus thermophilus and pyranose dehydrogenase from Agaricus meleagris, were displayed for the first time on the surface of Saccharomyces cerevisiae using the yeast surface display system. Surface displayed dehydrogenases were used in a microbial fuel cell and generated high power outputs. Surface displayed cellobiose dehydrogenase has demonstrated a midpoint potential of -28mV (vs. Ag/AgCl) at pH=6.5 and was used in a mediator-less anode compartment of a microbial fuel cell producing a power output of 3.3μWcm(-2) using lactose as fuel. Surface-displayed pyranose dehydrogenase was used in a microbial fuel cell and generated high power outputs using different substrates, the highest power output that was achieved was 3.9μWcm(-2) using d-xylose. These results demonstrate that surface displayed cellobiose dehydrogenase and pyranose dehydrogenase may successfully be used in microbial bioelectrochemical systems. PMID:27459246

  16. A direct electron transfer-based glucose/oxygen biofuel cell operating in human serum

    Energy Technology Data Exchange (ETDEWEB)

    Coman, V.; Gorton, L. [Department of Analytical Chemistry/Biochemistry, Lund University, 22100 Lund (Sweden); Ludwig, R. [Research Centre Applied Biocatalysis, 8010 Graz (Austria); Department of Food Sciences and Technology, BOKU-University of Natural Resources and Applied Life Sciences, 1190 Wien (Austria); Harreither, W.; Haltrich, D. [Department of Food Sciences and Technology, BOKU-University of Natural Resources and Applied Life Sciences, 1190 Wien (Austria); Ruzgas, T. [Biomedical Laboratory Science, Health and Society, Malmoe University, 20506 Malmoe (Sweden); Laboratory of Chemical Enzymology, A.N. Bach Institute of Biochemistry, 119071 Moscow (Russian Federation); Shleev, S.

    2010-02-15

    We report on the fabrication and characterisation of the very first direct electron transfer-based glucose/oxygen biofuel cell (BFC) operating in neutral glucose-containing buffer and human serum. Corynascus thermophilus cellobiose dehydrogenase and Myrothecium verrucaria bilirubin oxidase were used as anodic and cathodic bioelements, respectively. The following characteristics of the mediator-, separator- and membrane-less, a priori, non-toxic and simple miniature BFC, was obtained: an open-circuit voltage of 0.62 and 0.58 V, a maximum power density of ca. 3 and 4 {mu}W cm{sup -2} at 0.37 and 0.19 V of cell voltage, in phosphate buffer and human serum, respectively. (Abstract Copyright [2010], Wiley Periodicals, Inc.)

  17. Study on the removal of fish odour and bitter from protein hydrolysates of tilapia by-products by microorganism fermentation%微生物发酵对罗非鱼下脚料蛋白酶解液脱腥去苦效果比较

    Institute of Scientific and Technical Information of China (English)

    曾少葵; 杨萍; 陈秀红

    2009-01-01

    以嗜热链球菌(Streptococus thermophilus)、保加利亚乳杆菌(Lactobacillus bulgaricus)和瑞士乳杆菌(L.helveticus)及2种酿酒酵母菌(Saccharomyces cerevisiae)为菌种,对罗非鱼(Oreochromis niloticus)下脚料蛋白酶解液进行发酵,利用感官评定法,比较不同微生物发酵对酶解液脱腥去苦效果.试验结果表明,乳酸菌脱腥效果优于酵母菌,其中以嗜热链球菌脱腥效果最好,其适宜的发酵条件为接种量10%,发酵温度42℃,发酵时间9 h.微生物发酵去苦效果不明显.

  18. Production optimization of probiotic soft cheese made from goat's and cow's milk

    Directory of Open Access Journals (Sweden)

    Ida Drgalić

    2006-07-01

    Full Text Available The aim of this study was to determine optimal rennet share and fat content in milk for probiotic soft cheeseproduction made from goat's and cow's milk using DVS mixed probiotic culture ABT-4 (Lactobacillus acidophilus, Bifidobacterium spp. and Streptococcus thermophilus, with desirable sensory properties, which will be acceptable by consumers. The best sensory scores had samples of probiotic soft cheese produced in laboratory conditions from milk with 1% of milk fat, pasteurized at 65°C/30 min., fermented at 38°C with 2% culture. To achieve characteristic consistency of traditional soft cheese, 0.01% of rennet was added to goat's milk. Probiotic soft cheese made from cow's and goat's milk, produced under optimal conditions, were 100% acceptable by the tested consumers.

  19. Structure of branching enzyme- and amylomaltase modified starch produced from well-defined amylose to amylopectin substrates

    DEFF Research Database (Denmark)

    Sorndecha, Waraporn; Sagnelli, Domenico; Meier, Sebastian;

    2016-01-01

    Thermostable branching enzyme (BE, EC 2.4.1.18) from Rhodothermus obamensis in combination with amylomaltase (AM, EC 2.4.1.25) from Thermus thermophilus was used to modify starch structure exploring potentials to extensively increase the number of branch points in starch. Amylose is an important...... constituent in starch and the effect of amylose on enzyme catalysis was investigated using amylose-only barley starch (AO) and waxy maize starch (WX) in well-defined ratios. All products were analysed for amylopectin chain length distribution, α-1,6 glucosidic linkages content, molar mass distribution...... and digestibility by using rat intestinal α-glucosidases. For each enzyme treatment series, increased AO content resulted in a higher rate of α-1,6 glucosidic linkage formation but as an effect of the very low initial branching of the AO, the final content of α-1,6 glucosidic linkages was slightly lower as compared...

  20. Microbiological characterization and functionality of set-type yogurt fermented with potential prebiotic substrates Cudrania tricuspidata and Morus alba L. leaf extracts.

    Science.gov (United States)

    Oh, Nam Su; Lee, Ji Young; Joung, Jae Yeon; Kim, Kyung Su; Shin, Yong Kook; Lee, Kwang-Won; Kim, Sae Hun; Oh, Sangnam; Kim, Younghoon

    2016-08-01

    The objective of this study was to investigate the effect of 2 plant leaf extracts on fermentation mechanisms and health-promoting activities and their potential as a nutraceutical prebiotics ingredient for application in dairy products. The individual active phenolic compounds in the plant extract-supplemented milk and yogurts were also identified. Compared with control fermentation, the plant extracts significantly increased the growth and acidification rate of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. In particular, plant extract components, including monosaccharides, formic acid, and hydroxycinnamic acid, such as neo-chlorogenic, chlorogenic, and caffeic acid, together play a stimulatory role and cause this beneficial effect on the growth of yogurt culture bacteria through fermentation. In addition, supplementation with the plant extracts enhanced antioxidant activities with increased total phenolic contents, especially the highest antioxidant activity was observed in yogurt supplemented with Cudrania tricuspidata leaf extract. PMID:27236762

  1. THE SEARCH AND PROPERTIES OF LACTIC ACID BACTERIA PERSPECTIVE FOR BIOTECHNOLOGY

    Directory of Open Access Journals (Sweden)

    Naumenko О. V.

    2014-10-01

    Full Text Available Search of biologically active Lactobacillus strains prospective for functional milk food production was the aim of the research. The study involved the lactic acid bacteria isolated from biological material of healthy humen and non- dairy lactic products. Using modern methodological approaches, the strains of lactic acid bacteria such as Lactobacillus casei 302, Lactobacillus acidophilus 35 and Streptococcus thermophilus 21 having high level of biological activity were selected. High biological potential of selected cultures of lactic acid bacteria, which could provide stability for the technological process of production and essential characteristics of bacterial preparations and fermented their products, was set. In vitro the experiments demonstrated that selected strains had valuable production properties, namely the ability to reduce level of cholesterol and lactose during development in milk, were resistant to virulent bacteriophages and aggressive compounds of the gastrointestinal tract, and high adhesive and antagonistic activities as well.

  2. of gastro-intestinal discomfort (ID 2946, 2951, 2974), decreasing potentially pathogenic gastro-intestinal microorganisms (ID 2946, 2951, 2974), improved lactose digestion (ID 2946, 2951, 2974), and reduction in numbers of circulating CD34+ cells (ID 2947, 2952, 2975) (further assessment) pursuant

    DEFF Research Database (Denmark)

    Tetens, Inge

    , might be a beneficial physiological effect for the general population. The claimed effect, improved lactose digestion, is a beneficial physiological effect for individuals with lactose maldigestion. No human intervention studies were provided from which conclusions could be drawn for the scientific...... of Lactobacillus acidophilus BCCM/LMG P-18806, Lactobacillus delbrueckii BCCM/LMG P-18805 and Streptococcus thermophilus BCCM/LMG P-18807 and reduction of gastro-intestinal discomfort, decreasing potentially pathogenic gastro-intestinal microorganisms, improved lactose digestion, and reduction in numbers......, reduction in numbers of circulating CD34+ cells, is a beneficial physiological effect. The claimed effect, reduction of gastro-intestinal discomfort, is a beneficial physiological effect for the general population. The claimed effect, a decrease in potentially pathogenic gastro-intestinal microorganisms...

  3. Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate oxidizing bacteria

    DEFF Research Database (Denmark)

    Wang, Han; Fotidis, Ioannis; Angelidaki, Irini

    2015-01-01

    Ammonia-rich substrates can cause inhibition on anaerobic digestion process. Syntrophic acetate oxidizing bacteria (SAOB) and hydrogenotrophic methanogens are important for the ammonia inhibitory mechanism on anaerobic digestion. The roles and interactions of SAOB and hydrogenotrophic methanogens...... to ammonia inhibition effect are still unclear. The aim of the current study was to determine the ammonia toxicity levels of various pure strains of SAOB and hydrogenotrophic methanogens. Moreover, ammonia toxicity on the syntrophic cultivated strains of SAOB and hydrogenotrophic methanogens was tested. Thus......, four hydrogenotrophic methanogens (i.e. Methanoculleus bourgensis, Methanobacterium congolense, Methanoculleus thermophilus and Methanothermobacter thermautotrophicus), two SAOB (i.e. Tepidanaerobacter acetatoxydans and Thermacetogenium phaeum) and their syntrophic cultivation, were assessed under 0...

  4. Microbiological characterization and functionality of set-type yogurt fermented with potential prebiotic substrates Cudrania tricuspidata and Morus alba L. leaf extracts.

    Science.gov (United States)

    Oh, Nam Su; Lee, Ji Young; Joung, Jae Yeon; Kim, Kyung Su; Shin, Yong Kook; Lee, Kwang-Won; Kim, Sae Hun; Oh, Sangnam; Kim, Younghoon

    2016-08-01

    The objective of this study was to investigate the effect of 2 plant leaf extracts on fermentation mechanisms and health-promoting activities and their potential as a nutraceutical prebiotics ingredient for application in dairy products. The individual active phenolic compounds in the plant extract-supplemented milk and yogurts were also identified. Compared with control fermentation, the plant extracts significantly increased the growth and acidification rate of Streptococcus thermophilus and Lactobacillus delbrueckii ssp. bulgaricus. In particular, plant extract components, including monosaccharides, formic acid, and hydroxycinnamic acid, such as neo-chlorogenic, chlorogenic, and caffeic acid, together play a stimulatory role and cause this beneficial effect on the growth of yogurt culture bacteria through fermentation. In addition, supplementation with the plant extracts enhanced antioxidant activities with increased total phenolic contents, especially the highest antioxidant activity was observed in yogurt supplemented with Cudrania tricuspidata leaf extract.

  5. Evolution of the lactic acid bacterial community during malt whisky fermentation: a polyphasic study.

    Science.gov (United States)

    van Beek, Sylvie; Priest, Fergus G

    2002-01-01

    The development of the lactic acid bacterial community in a commercial malt whisky fermentation occurred in three broad phases. Initially, bacteria were inhibited by strong yeast growth. Fluorescence microscopy and environmental scanning electron microscopy revealed, in this early stage, both cocci and rods that were at least partly derived from the wort and yeast but also stemmed from the distillery plant. Denaturing gradient gel electrophoresis (DGGE) of partial 16S rRNA genes and sequence analysis revealed cocci related to Streptococcus thermophilus or Saccharococcus thermophilus, Lactobacillus brevis, and Lactobacillus fermentum. The middle phase began 35 to 40 h after yeast inoculation and was characterized by exponential growth of lactobacilli and residual yeast metabolism. Lactobacillus casei or Lactobacillus paracasei, L. fermentum, and Lactobacillus ferintoshensis were detected in samples of fermenting wort examined by DGGE during this stage. Bacterial growth was accompanied by the accumulation of acetic and lactic acids and the metabolism of residual maltooligosaccharides. By 70 h, two new PCR bands were detected on DGGE gels, and the associated bacteria were largely responsible for the final phase of the fermentation. The bacteria were phylogenetically related to Lactobacillus acidophilus and Lactobacillus delbrueckii, and strains similar to the former had previously been recovered from malt whisky fermentations in Japan. These were probably obligately homofermentative bacteria, required malt wort for growth, and could not be cultured on normal laboratory media, such as MRS. Their metabolism during the last 20 to 30 h of fermentation was associated with yeast death and autolysis and further accumulation of lactate but no additional acetate.

  6. Widespread distribution of archaeal reverse gyrase in thermophilic bacteria suggests a complex history of vertical inheritance and lateral gene transfers

    Directory of Open Access Journals (Sweden)

    Céline Brochier-Armanet

    2006-01-01

    Full Text Available Reverse gyrase, an enzyme of uncertain funtion, is present in all hyperthermophilic archaea and bacteria. Previous phylogenetic studies have suggested that the gene for reverse gyrase has an archaeal origin and was transferred laterally (LGT to the ancestors of the two bacterial hyperthermophilic phyla, Thermotogales and Aquificales. Here, we performed an in-depth analysis of the evolutionary history of reverse gyrase in light of genomic progress. We found genes coding for reverse gyrase in the genomes of several thermophilic bacteria that belong to phyla other than Aquificales and Thermotogales. Several of these bacteria are not, strictly speaking, hyperthermophiles because their reported optimal growth temperatures are below 80 °C. Furthermore, we detected a reverse gyrase gene in the sequence of the large plasmid of Thermus thermophilus strain HB8, suggesting a possible mechanism of transfer to the T. thermophilus strain HB8 involving plasmids and transposases. The archaeal part of the reverse gyrase tree is congruent with recent phylogenies of the archaeal domain based on ribosomal proteins or RNA polymerase subunits. Although poorly resolved, the complete reverse gyrase phylogeny suggests an ancient acquisition of the gene by bacteria via one or two LGT events, followed by its secondary distribution by LGT within bacteria. Finally, several genes of archaeal origin located in proximity to the reverse gyrase gene in bacterial genomes have bacterial homologues mostly in thermophiles or hyperthermophiles, raising the possibility that they were co-transferred with the reverse gyrase gene. Our new analysis of the reverse gyrase history strengthens the hypothesis that the acquisition of reverse gyrase may have been a crucial evolutionary step in the adaptation of bacteria to high-temperature environments. However, it also questions the role of this enzyme in thermophilic bacteria and the selective advantage its presence could provide.

  7. Role of antioxidant enzymes in bacterial resistance to organic acids.

    Science.gov (United States)

    Bruno-Bárcena, Jose M; Azcárate-Peril, M Andrea; Hassan, Hosni M

    2010-05-01

    Growth in aerobic environments has been shown to generate reactive oxygen species (ROS) and to cause oxidative stress in most organisms. Antioxidant enzymes (i.e., superoxide dismutases and hydroperoxidases) and DNA repair mechanisms provide protection against ROS. Acid stress has been shown to be associated with the induction of Mn superoxide dismutase (MnSOD) in Lactococcus lactis and Staphylococcus aureus. However, the relationship between acid stress and oxidative stress is not well understood. In the present study, we showed that mutations in the gene coding for MnSOD (sodA) increased the toxicity of lactic acid at pH 3.5 in Streptococcus thermophilus. The inclusion of the iron chelators 2,2'-dipyridyl (DIP), diethienetriamine-pentaacetic acid (DTPA), and O-phenanthroline (O-Phe) provided partial protection against 330 mM lactic acid at pH 3.5. The results suggested that acid stress triggers an iron-mediated oxidative stress that can be ameliorated by MnSOD and iron chelators. These findings were further validated in Escherichia coli strains lacking both MnSOD and iron SOD (FeSOD) but expressing a heterologous MnSOD from S. thermophilus. We also found that, in E. coli, FeSOD did not provide the same protection afforded by MnSOD and that hydroperoxidases are equally important in protecting the cells against acid stress. These findings may explain the ability of some microorganisms to survive better in acidified environments, as in acid foods, during fermentation and accumulation of lactic acid or during passage through the low pH of the stomach. PMID:20305033

  8. Identification and Characterization of the Nickel Uptake System for Urease Biogenesis in Streptococcus salivarius 57.I

    Science.gov (United States)

    Chen, Yi-Ywan M.; Burne, Robert A.

    2003-01-01

    Ureases are multisubunit enzymes requiring Ni2+ for activity. The low pH-inducible urease gene cluster in Streptococcus salivarius 57.I is organized as an operon, beginning with ureI, followed by ureABC (structural genes), and ureEFGD (accessory genes). Urease biogenesis also requires a high-affinity Ni2+ uptake system. By searching the partial genome sequence of a closely related organism, Streptococcus thermophilus LMG18311, three open reading frame (ORFs) homologous to those encoding proteins involved in cobalamin biosynthesis and cobalt transport (cbiMQO) were identified immediately 3′ to the ure operon. To determine whether these genes were involved in urease biogenesis by catalyzing Ni2+ uptake in S. salivarius, regions 3′ to ureD were amplified by PCRs from S. salivarius by using primers identical to the S. thermophilus sequences. Sequence analysis of the products revealed three ORFs. Reverse transcriptase PCR was used to demonstrate that the ORFs are transcribed as part of the ure operon. Insertional inactivation of ORF1 with a polar kanamycin marker completely abolished urease activity and the ability to accumulate 63Ni2+ during growth. Supplementation of the growth medium with NiCl2 at concentrations as low as 2.5 μM partially restored urease activity in the mutant. Both wild-type and mutant strains showed enhanced urease activity when exogenous Ni2+ was provided at neutral pH. Enhancement of urease activity by adding nickel was regulated at the posttranslational level. Thus, ORF1, ORF2, and ORF3 are part of the ure operon, and these genes, designated ureM, ureQ, and ureO, respectively, likely encode a Ni2+-specific ATP-binding cassette transporter. PMID:14617641

  9. Changes in oxidation-reduction potential during milk fermentation by wild lactic acid bacteria.

    Science.gov (United States)

    Morandi, Stefano; Silvetti, Tiziana; Tamburini, Alberto; Brasca, Milena

    2016-08-01

    Oxidation-reduction potential (E h) is a fundamental physicochemical property of lactic acid bacteria that determines the microenvironment during the cheese manufacture and ripening. For this reason the E h is of growing interest in dairy research and the dairy industry. The objective of the study was to perform a comprehensive study on the reduction activity of wild lactic acid bacteria strains collected in different periods (from 1960 to 2012) from Italian dairy products. A total of 709 strains belonging to Lactococcus lactis, Enterococcus durans, E. faecium, E. faecalis and Streptococcus thermophilus species were studied for their reduction activity in milk. Kinetics of milk reduction were characterised by the minimum redox potential (E h7) and time of reaching E h7 (t min), the maximum difference between two measures (Δmax) and the time at which these maximum differences occurred (t*). Broad diversity in kinetic parameters was observed at both species and strain levels. E. faecalis and L. lactis resulted to be the most reducing species, while S. thermophilus was characterised by the lowest reducing power while the greatest heterogeneity was pointed out among E. durans and E. faecium strains. Considering the period of collection (1960-2012) we observed that the more recently isolated strains generally showed less reducing activity. This trend was particularly evident for the species E. durans, E. faecium and L. lactis while an opposite trend was observed in E. faecalis species. Data reported in this research provide new information for a deeper understanding of redox potential changes during milk fermentation due to bacterial growth. Gain knowledge of the redox potential of the LAB cultures could allow a better control and standardisation of cheesemaking process. PMID:27600976

  10. The influence of the cell free solution of lactic acid bacteria on tyramine production by food borne-pathogens in tyrosine decarboxylase broth.

    Science.gov (United States)

    Toy, Nurten; Özogul, Fatih; Özogul, Yesim

    2015-04-15

    The function of cell-free solutions (CFSs) of lactic acid bacteria (LAB) on tyramine and other biogenic amine production by different food borne-pathogens (FBPs) was investigated in tyrosine decarboxylase broth (TDB) using HPLC. Cell free solutions were prepared from four LAB strains. Two different concentrations which were 50% (5 ml CFS+5 ml medium/1:1) and 25% (2.5 ml CFS+7.5 ml medium/1:3) CFS and the control without CFS were prepared. Both concentration of CFS of Streptococcus thermophilus and 50% CFS of Pediococcus acidophilus inhibited tyramine production up to 98% by Salmonella paratyphi A. Tyramine production by Escherichia coli was also inhibited by 50% CFS of Lactococcus lactis subsp. lactis and 25% CFS of Leuconostoc lactis. subsp. cremoris. The inhibitor effect of 50% CFS of P. acidophilus was the highest on tyramine production (55%) by Listeria monocytogenes, following Lc. lactis subsp. lactis and Leuconostoc mesenteroides subsp. cremoris (20%) whilst 25% CFS of Leu. mes. subsp. cremoris and Lc. lactis subsp. lactis showed stimulator effects (160%). The stimulation effects of 50% CFS of S. thermophilus and Lc. lactis subsp. lactis were more than 70% by Staphylococcus aureus comparing to the control. CFS of LAB strains showed statistically inhibitor effect since lactic acid inhibited microbial growth, decreased pH quickly and reduced the formation of AMN and BAs. Consequently, in order to avoid the formation of high concentrations of biogenic amines in fermented food by bacteria, it is advisable to use CFS for food and food products. PMID:25465993

  11. Effect of green tea supplementation on the microbiological, antioxidant, and sensory properties of probiotic milks.

    Science.gov (United States)

    Najgebauer-Lejko, Dorota

    2014-01-01

    Green tea and its constituents are known for a wide range of health-promoting properties. They may exert antimicrobial action but without altering lactic acid bacteria. The aim of the present study was to estimate the effect of green tea addition on the selected properties of probiotic milks. Bioyogurts (fermented with ABT-1 coculture of Streptococcus thermophilus, Lactobacillus acidophilus LA-5, Bifidobacterium animalis subsp. lactis BB-12) and acidophilus milks (fermented with pure L. acidophilus LA-5 culture) with addition of 0, 5, 10, or 15% (v/v) green tea infusion (GTI) were produced and analyzed for the antioxidant capacity by the "diphenyl picrylhydrazyl" (DPPH) and "ferric-reducing antioxidant power" (FRAP) methods, acidity, the count of starter bacteria, and sensory properties at the 1st, 7th, 14th, and 21st day of cold storage. The 15% addition of GTI to the acidophilus milk significantly reduced the lactic acid production during the whole study. The GTI had no impact on the level of S. thermophilus and B. lactis BB-12 in bioyogurts, and its effect on the count of L. acidophilus LA-5 depended on the concentration and probiotic milk type. GTI similarly and in a dose-dependent manner enhanced the antioxidant capacity of both milk types. There were no significant differences between the sensory notes received for bioyogurts, whereas acidophilus milks with tea were less appreciated by the panelists. In conclusion, green tea could be successfully used as a functional additive for selected probiotic milks enhancing their health benefits, but the proper selection of tea additive and starter culture is recommended. PMID:24883178

  12. Drink fermented to the base of serum of milk and isolated soy protein / Bebida fermentada à base de soro de leite e isolado protéico de soja

    Directory of Open Access Journals (Sweden)

    Ivone Yurika Mizubuti

    2000-04-01

    Full Text Available The whey and soy protein were used as substrate to obtain a basic beverage, fermented by yogurt culture (Lactobacillus delbrueckii subsp. buigaricus / Streptococcus salivarius subsp. thermophilus YC-180 and by probiotic culture (Lactobacillus acidophilus La-5. The formulation ( whey 10g/100mL; soy protein 2,4 g/100mL; fiber 0,5 g/100mL and sucrose 15 g/100mL was fermented separately, and mixed (1:1 to obtain the final fermented product with 0.72% (lactic acid acidity, pH 4.3 and lactic acid bacteria count of 2,0 x 108 cfu/mL, adequate for production of the proposed beverage.O soro de leite e o isolado protéico de soja foram utilizados para obtenção de uma bebida base, fermentada com cultura de iogurte (Lactobacillus delbrueckii subsp. bulgaricus / Streptococcus salivarius subsp. thermophilus YC - 180 e cultura probiótica (Lactobacillus acidophilus La-5. A formulação (soro de leite 10 g/100mL; isolado protéico de soja 2,4 g/100mL; fibras 0,5 g/100mL e sacarose 15 g/100mL indicou que procedendo-se a fermentação seguida de mistura na proporção de 1:1, produziu-se um produto final com acidez de 0,72% de ácido láctico; pH 4,3 e contagem de bactérias lácticas de 2,0 x 108 ufc/mL adequado para a obtenção da bebida proposta.

  13. Proteolytic specificity of Lactobacillus delbrueckli subsp. bulgaricus influences functional properties of mozzarella cheese.

    Science.gov (United States)

    Oommen, B S; McMahon, D J; Oberg, C J; Broadbent, J R; Strickland, M

    2002-11-01

    Low-moisture part-skim Mozzarella cheeses were manufactured from 2% fat milk and aged for 21 d. Treatments included cheeses made with one of three different strains of Lactobacillus delbrueckii ssp. bulgaricus in combination with a single strain of Streptococcus thermophilus. A fourth, control treatment consisted of cheeses made with only S. thermophilus. Although total proteolytic ability of these strains, as indicated by the o-phthaldialdehyde analysis, was similar in each of the three strains of L. bulgaricus, these strains exhibited different proteolytic specificities toward the peptide, alpha(s1)-CN (f 1-23). On the basis of their alpha(s1)-CN (f 1-23) cleavage patterns and a previously described classification, these strains were assigned to the groups I, III, and V. The objective of this study was to investigate the influence of lactobacilli proteolytic systems, based on specificity toward alpha(s1)-CN (f 1-23), on functionality of part-skim Mozzarella cheese. Moisture, fat, protein, salt-in-moisture, and moisture in nonfat substances content of cheeses made with groups I, III, and V strain were similar. Control cheese had a lower moisture content than did other treatments. Significant differences were observed in functional properties between cheeses manufactured using groups III and V strains. Cheeses made with groups I and III strains were similar in their meltability, hardness, cohesiveness, melt strength, and stretch quality. Meltability and cohesiveness increased with age, while melt strength and stretch quality decreased with age for all cheeses. Additionally, HPLC showed that total peak areas of water-soluble peptides derived from cleavage of alpha(s1)-CN (f 1-23) by different strains of lactobacilli could be highly correlated to meltability and stretch characteristics of cheeses made with those strains.

  14. The choice of fermentation strains of lactic acid bacteria in conserving process%保藏过程中乳酸菌发酵菌种的选择

    Institute of Scientific and Technical Information of China (English)

    杜磊; 袁超; 杜杨

    2011-01-01

    Lactobacillus bulgaricus and Streptococcus thermophilus are very primary in yogurt production. They play an important role in yogurt production. But they are easily being subject to degenerate activity in refrigeration so that the activity of preserved zymogen will consumedly debase. And the degenerating activity affected the zymogen's property. The Lactobacillus bulgaricus, Streptococcus thermophilus and their mixture were subcultured. Contrast the experimental data,it was found that mixing the two kinds of lactobacillus was the best way to conserve without exceptional case. It can greatly reduce the change of the lactic acid bacteria activity during the conserving time.%保加利亚乳杆菌和嗜热链球菌是酸奶生产过程的主要发酵菌种,在酸奶制作过程中起着极其重要的作用。但是由于菌种在其保藏过程中极易出现活性退化的现象,使得保藏后的菌种活性大大降低,影响了菌种的使用特性。分别对单独的保加利亚乳杆菌、嗜热链球菌及这两种菌的混合菌种进行菌种在保藏期间的传代实验,通过对实验数据的分析比较可得出,在没有特殊要求的情况下,乳酸菌菌种的保藏最好是使用保加刺亚乳杆菌和嗜热链球菌的混合菌种.这样可以大大降低乳酸菌菌种在保藏期内的特性变化。

  15. Characterization of the cysK2-ctl1-cysE2 gene cluster involved in sulfur metabolism in Lactobacillus casei.

    Science.gov (United States)

    Bogicevic, Biljana; Irmler, Stefan; Portmann, Reto; Meile, Leo; Berthoud, Hélène

    2012-01-16

    The up- and downstream regions of ctl1 and ctl2 that encode a cystathionine lyase were analyzed in various Lactobacillus casei strains. ctl1 and ctl2 were found to be part of a gene cluster encoding two other open reading frames. One of the two open reading frames precedes ctl1 and encodes a putative cysteine synthase. The other open reading frame lies downstream of ctl1 and encodes a putative serine acetyltransferase. The gene cluster is not present in the publicly available genome sequences of L. casei ATCC 334, BL23 and Zhang. Apparently, the gene cluster was acquired by a horizontal gene transfer event and can also be found in other lactic acid bacteria such as Lactobacillus helveticus, Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. RT-PCR was used to analyze the expression of the gene cluster. Additionally, an mass spectrometry-based selected reaction monitoring method was developed for quantifying Ctl1 in a cell-free extract of lactic acid bacteria. The gene cluster cysK2-ctl1-cysE2 was expressed as single transcript, and expression was down-regulated by cysteine. In addition, cystathionine lyase activity present in cell-free extracts disappeared when L. casei was grown in the presence of cysteine. Whereas the transcript and the gene product of ctl1 protein were found in all studied ctl1(+)L. casei strains, only the transcript but not the protein or cystathionine lyase activity was detected in L. helveticus FAM2888, L. delbrueckii subsp. bulgaricus ATCC 11842 and S. thermophilus FAM17014, which actually possess a homolog of the cysK2-ctl1-cysE2 gene cluster.

  16. Identification and characterisation of an iron-responsive candidate probiotic.

    Directory of Open Access Journals (Sweden)

    Jennifer R Bailey

    Full Text Available BACKGROUND: Iron is an essential cofactor in almost all biological systems. The lactic acid bacteria (LAB, frequently employed as probiotics, are unusual in having little or no requirement for iron. Iron in the human body is sequestered by transferrins and lactoferrin, limiting bacterial growth. An increase in the availability of iron in the intestine by bleeding, surgery, or under stress leads to an increase in the growth and virulence of many pathogens. Under these high iron conditions, LAB are rapidly out-competed; for the levels of probiotic bacteria to be maintained under high iron conditions they must be able to respond by increasing growth rate to compete with the normal flora. Despite this, iron-responsive genera are poorly characterised as probiotics. METHODOLOGY/PRINCIPAL FINDINGS: Here, we show that a panel of probiotics are not able to respond to increased iron availability, and identify an isolate of Streptococcus thermophilus that can increase growth rate in response to increased iron availability. The isolate of S. thermophilus selected was able to reduce epithelial cell death as well as NF-κB signalling and IL-8 production triggered by pathogens. It was capable of crossing an epithelial cell barrier in conjunction with E. coli and downregulating Th1 and Th17 responses in primary human intestinal leukocytes. CONCLUSIONS/SIGNIFICANCE: We propose that an inability to compete with potential pathogens under conditions of high iron availability such as stress and trauma may contribute to the lack of efficacy of many LAB-based probiotics in treating disease. Therefore, we offer an alternative paradigm which considers that probiotics should be able to be competitive during periods of intestinal bleeding, trauma or stress.

  17. The microbiota of high-moisture mozzarella cheese produced with different acidification methods.

    Science.gov (United States)

    Guidone, Angela; Zotta, Teresa; Matera, Attilio; Ricciardi, Annamaria; De Filippis, Francesca; Ercolini, Danilo; Parente, Eugenio

    2016-01-01

    The microbiota of high-moisture Mozzarella cheese made from cow's milk and produced with different acidification methods was evaluated at the end of refrigerated storage by pyrosequencing of the 16S rRNA gene. The cheeses were clearly separated on the basis of the acidification methods. Cheeses produced with the addition of starters were dominated by Streptococcus thermophilus, but a variety of lactic acid bacteria and spoilage microorganisms appeared at low levels (0.01-1%). Cheeses produced by direct addition of citric acid were dominated by a diverse microbiota, including both lactic acid bacteria and psychrotrophic γ-proteobacteria. For five brands the acidification system was not declared on the label: the microbiota was dominated by thermophilic lactic acid bacteria (S. thermophilus, Lactobacillus delbrueckii, Lactobacillus helveticus) but a variety of other subdominant lactic acid bacteria, psychrotrophs and Enterobacteriaceae were present, with a diversity comparable or higher to cheeses produced by direct acid addition. This led to the conclusion that undefined starters were used for acidification. Both ordination methods and network analysis were used for the representation of beta-diversity: matrix cluster analysis, principal coordinate analysis and OTU networks uncovered different aspects of the microbial community structure. For three cheese brands both biological replicates (cheeses from different lots) and technical replicates (replicate cheeses from the same lot) were analyzed. Repeatability was acceptable for OTUs appearing at frequencies >1%, but was low otherwise. A linear mixed model showed that the starter system was responsible for most differences related to dairies, while difference due to psychrotrophic contaminants was more related to lot-to-lot variability.

  18. 化妆品用微生物源天然防腐剂的抑菌效能研究%Antimicrobial effectiveness evaluation of natural microorganism source preservatives used for cosmetic formulation

    Institute of Scientific and Technical Information of China (English)

    李馨恩; 何秋星; 区梓聪; 方电力; 关建云; 赵红

    2015-01-01

    以微生物源的嗜热链球菌发酵产物和曲霉发酵产物作为研究对象,根据其抑菌作用进行复配研究,参考美国个人护理品委员会(PCPC)的抑菌效能测试方法对乳液配方进行微生物挑战试验。结果表明,在乳液配方中,质量分数为0.5%的嗜热链球菌发酵产物和0.3%曲霉发酵产物可以使配方中的微生物菌落数在第14天降到100 CFU/g以下,可通过微生物挑战试验。%Two kinds of new natural microorganism source preservatives nominated Streptococus thermophilus Ferment and Aspergillus Ferment were taken as the object for research work. Formulation study based upon the bacterial inhibition action of the aforesaid preservatives was carried out. With reference of the bacterial inhibition efficacy test method of the American Personal Care Product Committee (PCPC),microbial challenge tests for emulsion formulations were processed. The test results demonstrated that an emulsion consisting of Streptococus thermophilus Ferment at 0.5% of mass fraction and Aspergillus Ferment at 0.3% of mass fraction can reduce the amount of microbial to be lower than 100 CFU/g on the 14th day and can pass the microbial challenge test meanwhile.

  19. Antimicrobial effectiveness evaluation of natural microorganism source preservatives used for cosmetic formulation%化妆品用微生物源天然防腐剂的抑菌效能研究

    Institute of Scientific and Technical Information of China (English)

    李馨恩; 何秋星; 区梓聪; 方电力; 关建云; 赵红

    2015-01-01

    以微生物源的嗜热链球菌发酵产物和曲霉发酵产物作为研究对象,根据其抑菌作用进行复配研究,参考美国个人护理品委员会(PCPC)的抑菌效能测试方法对乳液配方进行微生物挑战试验。结果表明,在乳液配方中,质量分数为0.5%的嗜热链球菌发酵产物和0.3%曲霉发酵产物可以使配方中的微生物菌落数在第14天降到100 CFU/g以下,可通过微生物挑战试验。%Two kinds of new natural microorganism source preservatives nominated Streptococus thermophilus Ferment and Aspergillus Ferment were taken as the object for research work. Formulation study based upon the bacterial inhibition action of the aforesaid preservatives was carried out. With reference of the bacterial inhibition efficacy test method of the American Personal Care Product Committee (PCPC),microbial challenge tests for emulsion formulations were processed. The test results demonstrated that an emulsion consisting of Streptococus thermophilus Ferment at 0.5% of mass fraction and Aspergillus Ferment at 0.3% of mass fraction can reduce the amount of microbial to be lower than 100 CFU/g on the 14th day and can pass the microbial challenge test meanwhile.

  20. Diversity of lactic acid bacteria associated with traditional fermented dairy products in Mongolia.

    Science.gov (United States)

    Yu, J; Wang, W H; Menghe, B L G; Jiri, M T; Wang, H M; Liu, W J; Bao, Q H; Lu, Q; Zhang, J C; Wang, F; Xu, H Y; Sun, T S; Zhang, H P

    2011-07-01

    Spontaneous milk fermentation has a long history in Mongolia, and beneficial microorganisms have been handed down from one generation to the next for use in fermented dairy products. The objective of this study was to investigate the diversity of lactic acid bacteria (LAB) communities in fermented yak, mare, goat, and cow milk products by analyzing 189 samples collected from 13 different regions in Mongolia. The LAB counts in these samples varied from 3.41 to 9.03 log cfu/mL. Fermented yak and mare milks had almost identical mean numbers of LAB, which were significantly higher than those in fermented goat milk but slightly lower than those in fermented cow milk. In total, 668 isolates were obtained from these samples using de Man, Rogosa, and Sharpe agar and M17 agar. Each isolate was considered to be presumptive LAB based on gram-positive and catalase-negative properties, and was identified at the species level by 16S rRNA gene sequencing, multiplex PCR assay, and restriction fragment length polymorphism analysis. All isolates from Mongolian dairy products were accurately identified as Enterococcus faecalis (1 strain), Enterococcus durans (3 strains), Lactobacillus brevis (3 strains), Lactobacillus buchneri (2 strains), Lactobacillus casei (16 strains), Lactobacillus delbrueckii ssp. bulgaricus (142 strains), Lactobacillus diolivorans (17 strains), Lactobacillus fermentum (42 strains), Lactobacillus helveticus (183 strains), Lactobacillus kefiri (6 strains), Lactobacillus plantarum ssp. plantarum (7 strains), Lactococcus lactis ssp. lactis (7 strains), Leuconostoc lactis (22 strains), Leuconostoc mesenteroides (21 strains), Streptococcus thermophilus (195 strains), and Weissella cibaria (1 strain). The predominant LAB were Strep. thermophilus and Lb. helveticus, which were isolated from all sampling sites. The results demonstrate that traditional fermented dairy products from different regions of Mongolia have complex compositions of LAB species. Such diversity of

  1. Lactose fermentation at Camembert, made by classic and stabilised

    Directory of Open Access Journals (Sweden)

    Bogdan Perko

    2002-01-01

    Full Text Available In our experiments the fermentation of lactose at Camembert type cheese by classic and stabilised technology was monitored. In each of the two technologies two experiments were made. The difference between these two technologies is in pH level, which drops below 5 by classic technology and remains above 5 by stabilised technology at all times. To achieve the criteria of stabilised technology the fermentation was stopped at a desired level of pH, by dropping the cheese in brine at 14 °C. After salting and moulding cheesewas transferred from the first three experiments into a ripening chamber at 11 °C. With the last experiment (stabilised technology the cheese ripened for 3 days at 5 °C. During ripening process pH dropped below 5 in all experiments. The process of fermentation was performed by the following lactic acid bacteria: Streptococcus thermophilus, Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris. As these lactic acid bacteria ferment differently D-galactose during manufacture and ripening process, the contentof lactose and D-galactose was measured. Based on the results of our research the following conclusion can be made: the action of lactic acid bacteria can not be stropped even at 5 °C. The native microflora, which remains in the milk after thermisation, might be responsible for the fermentation of Dgalactose. It could be possible that the Streptococcus thermophilus enzymes were not inactivated, causing continuation of lactose fermentation even at low temperature. Mesophilic lactococci were inhibited and for that reason Dgalactose accumulated in cheese. Only after cheese were transferred into a ripening chamber at 11 °C and with low lactose concentration in medium left, mesophilic lactococci started to ferment D-galactose.

  2. Characterization of bifunctional L-glutathione synthetases from Actinobacillus pleuropneumoniae and Actinobacillus succinogenes for efficient glutathione biosynthesis.

    Science.gov (United States)

    Yang, Jianhua; Li, Wei; Wang, Dezheng; Wu, Hui; Li, Zhimin; Ye, Qin

    2016-07-01

    Glutathione (GSH), an important bioactive substance, is widely applied in pharmaceutical and food industries. In this work, two bifunctional L-glutathione synthetases (GshF) from Actinobacillus pleuropneumoniae (GshFAp) and Actinobacillus succinogenes (GshFAs) were successfully expressed in Escherichia coli BL-21(DE3). Similar to the GshF from Streptococcus thermophilus (GshFSt), GshFAp and GshFAs can be applied for high titer GSH production because they are less sensitive to end-product inhibition (Ki values 33 and 43 mM, respectively). The active catalytic forms of GshFAs and GshFAp are dimers, consistent with those of GshFPm (GshF from Pasteurella multocida) and GshFSa (GshF from Streptococcus agalactiae), but are different from GshFSt (GshF from S. thermophilus) which is an active monomer. The analysis of the protein sequences and three dimensional structures of GshFs suggested that the binding sites of GshFs for substrates, L-cysteine, L-glutamate, γ-glutamylcysteine, adenosine-triphosphate, and glycine are highly conserved with only very few differences. With sufficient supply of the precursors, the recombinant strains BL-21(DE3)/pET28a-gshFas and BL-21(DE3)/pET28a-gshFap were able to produce 36.6 and 34.1 mM GSH, with the molar yield of 0.92 and 0.85 mol/mol, respectively, based on the added L-cysteine. The results showed that GshFAp and GshFAs are potentially good candidates for industrial GSH production. PMID:26996628

  3. Optimization of fermentation process in skim milk with ST-ⅢLactobacillus plantarum%植物乳杆菌ST-Ⅲ脱脂乳的发酵工艺优化

    Institute of Scientific and Technical Information of China (English)

    华宝珍; 李莎; 徐爱才; 徐志平; 马成杰

    2014-01-01

    Fermented milk is increasingly used as a carrier of probiotics for their potential health functions. Because the concentration of viable probiotics is the key factor to health functions, it should be higher than the recommended concentration for probiotics (106 CFU/g). However, there are many products with low viability of probiotics in the market. It is very important and necessary for the milk industry to increase the count of viable probiotics in yogurt. In addition, survival during the passage through the gastrointestinal tract is generally considered a key feature for probiotics to preserve their expected health functions. However, the traditional yogurt starters (Streptococcus thermophilus and Lactobacillus bulgaricus) have weak tolerance to acid and bile salt and hence limit therapeutic effects. Lactobacillus plantarum has been demonstrated that it can survive in the human intestine and tolerate acid and bile salt. Moreover, it has a lot of precious therapeutic effects, such as precipitating and assimilating cholesterols, lowering blood sugar, diminishing inflammation and improving immunity. Hence, L.plantarumhas become one of research hotspots in recent years. Lactobacillus plantarum ST-Ⅲ strain (CGMCC No.0847) is a probiotics and has ability to tolerate acid and bile salts as well as grow in the lower intestinal tract. It also be proved to have ability to precipitate and assimilate cholesterols in vitro andin vivo. However L.plantarumST-Ⅲ strain is auxotrophic and has weak ability to grow in skim milk and clot milk by acidification. In this study, to increase the concentration of viableL.plantarumST-Ⅲ and elucidate the factors restricting growth ofL.plantarumST-Ⅲ in skim milk, the fermentation conditions were researched and optimized. The effects of soybean polypeptide concentration, manganese gluconate concentration, inoculum size of S. thermophilus and fermentation temperature on the pH and living cell count ofL.plantarumST-Ⅲ of fermented milk

  4. Effects of Xylo-oligosaccharide on Growth and Acid-producing of Yogurt Strains%低聚木糖对酸奶菌株生长及产酸的影响

    Institute of Scientific and Technical Information of China (English)

    曹吉利; 谭源; 舒国伟; 陈合

    2011-01-01

    研究了益生元——低聚木糖及酸奶菌株比例对酸奶发酵过程中总菌数、pH值及酸度的影响。低聚木糖质量分数为0,0.5%,0.6%,0.7%,0.8%,0.9%,1.0%,保加利亚乳杆菌和嗜热乳链球菌的比例(Lb∶St)分别为2∶1,1∶1和1∶2。结果表明:Lb∶St的最佳比例为1∶1,低聚木糖的最适添加量为0.8%,42℃发酵3.5h,总菌数达到最大值为7.14×108CFU/mL(对照为5.92×108CFU/mL),pH值为4.51(对照为5.09),酸度达到72°T(对照为64°T),乳酸量为6.45mg/mL(对照为5.77mg/mL)。低聚木糖的添加可促进酸奶发酵菌株的生长,促进产酸和缩短发酵时间。%Effects of xylo-oligosaccharide on growth and acid-producing of yogurt fermentation strains were researched.The concentration range of xylo-oligosaccharide was 0%,0.5%,0.6%,0.7%,0.8%,0.9% and 1.0%,the ratio of Lactobacillus bulgaricus to Streptococcus thermophilus was 2:1,1:1 and 1:2.The result shows as follows:the optimal ratio L.bulgaricus to S.thermophilus was 1:1,the optimal concentration of xylo-oligosaccharide was 0.8%,42 ℃ for 3.5 h,and the total bacteria number reached 7.14×108 CFU/mL(the control 5.92×108 CFU/mL),pH value,acidity and lactic acid were 4.51(the control 5.09),72°T(the control 64°T) and 6.45mg/mL(the control 5.77mg/mL) respectively.Adding xylo-oligosaccharide could promote growth and acid-producing of L.bulgaricus and S.thermophilus,which could shorten the fermentation time.

  5. Prevalence and comparison of Streptococcus infantarius subsp. infantarius and Streptococcus gallolyticus subsp. macedonicus in raw and fermented dairy products from East and West Africa.

    Science.gov (United States)

    Jans, Christoph; Kaindi, Dasel Wambua Mulwa; Böck, Désirée; Njage, Patrick Murigu Kamau; Kouamé-Sina, Sylvie Mireille; Bonfoh, Bassirou; Lacroix, Christophe; Meile, Leo

    2013-10-15

    Streptococcus infantarius subsp. infantarius (Sii) and Streptococcus gallolyticus subsp. macedonicus are members of the Streptococcus bovis/Streptococcus equinus complex (SBSEC) associated with human infections. SBSEC-related endocarditis was furthermore associated with rural residency in Southern Europe. SBSEC members are increasingly isolated as predominant species from fermented dairy products in Europe, Asia and Africa. African variants of Sii displayed dairy adaptations to lactose metabolism paralleling those of Streptococcus thermophilus including genome decay. In this study, the aim was to assess the prevalence of Sii and possibly other SBSEC members in dairy products of East and West Africa in order to identify their habitat, estimate their importance in dairy fermentation processes and determine geographic areas affected by this potential health risk. Presumptive SBSEC members were isolated on semi-selective M17 and SM agar media. Subsequent genotypic identification of isolates was based on rep-PCR fingerprinting and SBSEC-specific16S rRNA gene PCR assay. Detailed identification was achieved through application of novel primers enhancing the binding stringency in partial groES/groEL gene amplification and subsequent DNA sequencing. The presence of S. thermophilus-like lacS and lacZ genes in the SBSEC isolates was determined to elucidate the prevalence of this dairy adaptation. Isolates (n = 754) were obtained from 72 raw and 95 fermented milk samples from Côte d'Ivoire and Kenya on semi-selective agar media. Colonies of Sii were not detected from raw milk despite high microbial titers of approximately 10(6)CFU/mL on M17 agar medium. However, after spontaneous milk fermentation Sii was genotypically identified in 94.1% of Kenyan samples and 60.8% of Kenyan isolates. Sii prevalence in Côte d'Ivoire displayed seasonal variations in samples from 32.3% (June) to 40.0% (Dec/Jan) and isolates from 20.5% (June) to 27.7% (Dec/Jan) present at titers of 10

  6. Lactic Acid and Probiotic Bacteria from Fermented and Probiotic Dairy Products

    Directory of Open Access Journals (Sweden)

    B.K.L. Karna

    2007-12-01

    Full Text Available Lactic acid and probiotic bacteria were enumerated and isolated from commercially available yoghurt andprobiotic milk products. Lactobacillus delbrueckii ssp. bulgaricus were enumerated and isolated usingMRS agar incubated anaerobically at 37oC for 72 hrs. M17 agar was used for the enumeration andisolation of Streptococcus thermophilus incubated aerobically at 37oC for 48 hrs. MRS agar and modifiedMRS agar (MRS + L-cysteine + LiCl + Na propionate were used for the enumeration and isolation ofprobiotic bacteria. Both were incubated anaerobically at 37oC for 72 hrs. Morphological, physiologicaland biochemical reactions were used to characterize the isolates.Str. thermophilus counts ranged from 2.6 x 1011 to 2.9 x 1020 CFU/g with Fruit Yoghurt (FY having thehighest count and Yoghurt Natural (YN with the lowest count. Highest Lactobacillus delbrueckii ssp.bulgaricus count was obtained in Duo Yoghurt (DY, 1.1 x 109 and lowest in Yoghurt Drink (YD, 8.0 x 107CFU/g. The highest probiotic bacterial count of 2.3 x 108 was obtained in Yakult (YK and Neslac (Nesshowed the lowest, 1.6 x 102 CFU/g. The viable counts of all the products examined met the prescribedminimum viable count of 105 to 106CFU/g for the claimed health benefits for the consumer except forChamyto Plain (CP, Nes and Nan-2 (Nan.Morphological, physiological and biochemical characteristics showed that the following genera andspecies were present Pediococcus acidilactici (YN, P. pentosaceus (FY, Lactobacillus delbrueckiidelbrueckii and L. brevis in Non Fat High Calcium Yoghurt (NC, L. acidophilus and L. delbrueckiidelbrueckii (DY, YD, P. damnosus and P. pentosaceus in Chamyto Orange (CO, L. delbrueckii bulgaricus,L. acidophilus, and L. delbrueckii delbrueckii (CP, L. para. paracasei (YK and Bifidobacterium ssp.(Nes and Nan.Of the 28 isolates characterized in this study, 15 were Lactobacillus (5 species, 5 werePediococcus (3 species, 6 were Bifidobacterium (species not identified, and 2 were

  7. 乳酸菌利用罗非鱼下脚料水解液发酵L-乳酸的探讨%Research on L-lactic acid fermentation for hydrolysate of tilapia wastes with Lactobacillus

    Institute of Scientific and Technical Information of China (English)

    邓楚津; 赖丽芬; 曾少葵; 张小红; 李瑞杰

    2012-01-01

    By hot water extraction method, we extracted the protein hydrolysate of tilapia wastes, which is used as a nitrogen source for L-lactic acid fermentation. Compared with yeast extract, we studied the growth of Lactobacillus bulgaricus, Streptococcus thermophilus and L rhamnosus in fermentation medium of hydrolysate of tilapia wastes, and observed the yield of L-lactic acid. Results show that the 3 lactic acid bacteria grow better in 2 fermentation mediums and pH decreases obviously. The production of L-lactic acid varies with different lactic acid bacteria. The net yield of L-lactic acid is 3. 38 g·L-1 after 10 h fermentation with L rhamnosus in the fermentation medium of hydrolysate of tilapia wastes. The yield is considerable to that of the yeast extract Therefore, the hydrolysate of tilapia wastes can be a substitute for yeast extract and is a low-cost nitrogen source for the yield of L-lactic acid.%以罗非鱼(Oreochromis spp.)下脚料为原料,采用高温热水抽提法提取其蛋白质作为乳酸菌发酵L-乳酸的氮源.以酵母膏作为对照,探讨了保加利亚乳杆菌(Lactobacillus bulgaricus)、嗜热链球菌(Streptococcus thermophilus)和鼠李糖乳杆菌(L.rhamnosus)在罗非鱼下脚料水解液培养基中的生长情况及L-乳酸的产量.结果显示,3种乳酸菌在2种培养基中均能良好地生长,pH下降明显;L-乳酸产量随乳酸菌菌种不同而异,其中鼠李糖乳杆菌在罗非鱼下脚料水解液培养基中发酵10 h后,L-乳酸净产量为3.38 g·L-1,与以酵母膏作为氮源的产酸水平相当.可见罗非鱼下脚料水解液可以替代酵母膏作为L-乳酸发酵的一种低成本氮源.

  8. Textura instrumental e avaliação sensorial de queijo fresco cremoso simbiótico: implicações da adição de Lactobacillus paracasei e inulina Influence of Lactobacillus paracasei and inulin on instrumental texture and sensory evaluation of fresh cream cheese

    Directory of Open Access Journals (Sweden)

    Flávia Carolina Alonso Buriti

    2008-03-01

    Full Text Available Avaliou-se o efeito da adição do probiótico Lactobacillus paracasei e da fibra prebiótica inulina sobre o perfil de textura e as características sensoriais de queijo fresco cremoso. Três tratamentos de queijo fresco cremoso foram preparados em quintuplicata, em escala piloto, todos suplementados com uma cultura starter de Streptococcus thermophilus (T1, T2 e T3. L. paracasei subsp. paracasei foi adicionado em T1 e T2. Inulina foi adicionada em T2. O perfil de textura instrumental foi determinado após 1, 7, 14 e 21 dias de armazenamento. A análise sensorial foi conduzida aos 7 dias de armazenamento dos queijos. A presença de Lactobacillus paracasei nos queijos T1 e T2 e de inulina em T2 não alterou significativamente o perfil de textura (p>0,05. Os queijos T1 apresentaram a menor preferência na análise sensorial e diferiram significativamente de T2 e T3 (p0,05. A adição de inulina ao queijo fresco cremoso produzido com a suplementação de uma cepa potencialmente probiótica de Lactobacillus paracasei resultou em um produto com características adequadas e com propriedades funcionais agregadas.The influence of the addition of a potential probiotic culture of Lactobacillus paracasei and of the prebiotic fiber inulin on the texture profile and on the sensory evaluation of probiotic and synbiotic fresh cream-cheeses was monitored. Three cheese-making trials were prepared in quintuplicate, all supplemented with a Streptococcus thermophilus starter culture (T1, T2 and T3. L. paracasei subsp. paracasei was added to T1 and T2, and inulin, to T2. The instrumental texture profile was determined after 1, 7, 14 and 21 days of storage of the cheeses. Sensory evaluation was performed after 7 days of storage. The presence of Lactobacillus paracasei in cheeses T1 and T2 and of inulin in cheeses T2 did not alter the texture profile significantly. Cheeses T1 were the least preferred in the sensory evaluation and differed significantly from T2 and T3

  9. Effects of yogurt starter cultures on the survival of Lactobacillus acidophilus.

    Science.gov (United States)

    Ng, Elizabeth W; Yeung, Marie; Tong, Phillip S

    2011-01-31

    Recognized to confer health benefits to consumers, probiotics such as Lactobacillus acidophilus are commonly incorporated into fermented dairy products worldwide; among which yogurt is a popular delivery vehicle. To materialize most of the putative health benefits associated with probiotics, an adequate amount of viable cells must be delivered at the time of consumption. However, the loss in their viabilities during refrigerated storage has been demonstrated previously. This study focused on the effects of yogurt starter cultures on the survival of five strains of L. acidophilus, with emphases on low pH and acid production. Differential survival behavior between L. acidophilus strains was further analyzed. To this end, viable cell counts of L. acidophilus were determined weekly during 4°C storage in various types of yogurts made with Streptococcus thermophilus alone, L. delbrueckii ssp. bulgaricus alone, both species of the starter cultures, or glucono-delta-lactone (GDL). All yogurt types, except for pasteurized yogurts, were co-fermented with L. acidophilus. Yogurt filtrate was analyzed for the presence of any inhibitory substance and for the amount of hydrogen peroxide. Multiplication of L. acidophilus was not affected by the starter cultures as all strains reached high level on day 0 of the storage period. Throughout the 28-day storage period, cell counts of L. acidophilus PIM703 and SBT2062 remained steady (~6 × 10(7)CFU/g) in yogurts made with both starter cultures, whereas those of ATCC 700396 and NCFM were reduced by a maximum of 3 and 4.6 logs, respectively. When starter cultures were replaced by GDL, all strains survived well, suggesting that a low pH was not a critical factor dictating their survival. In addition, the filtrate collected from yogurts made with starter cultures appeared to have higher inhibitory activities against L. acidophilus than that made with GDL. The presence of viable starter cultures was necessary to adversely affect the

  10. 高产胞外多糖嗜热链球菌混菌培养特性研究%Study on the mixed cultivated properties of high exopolysacchride-producing Streotococcus thermophillus

    Institute of Scientific and Technical Information of China (English)

    马艳; 陈历俊; 王昌禄; 牟光庆; 陈晓璇

    2011-01-01

    探讨高产胞外多糖嗜热链球菌混菌培养的产酸、产香、产黏特性.以发酵酸度、乙醛质量浓度和黏度为综合指标,确定高产胞外多糖的嗜热链球菌(S.t.)、保加利亚乳杆菌(L.b.)、嗜酸乳杆菌(L.a.)混菌最佳培养条件为培养温度42℃,总接菌量体积分数4%,V(S.t.):V(L.b.):V(L.a.)=3:2:1.在此条件下,混菌培养可以显著地提高酸度、乙醛质量浓度、黏度、胞外多糖质量浓度、β-半乳糖苷酶酶活、蛋白酶水解能力(p<0.01).发酵过程中,胞外多糖质量浓度与黏度,β-半乳糖苷酶酶活与酸度.蛋白酶酶活与酸度、乙醛质量浓度的相关性系数很高.%The aim of this research is to study the acidity,acetaldehyde and viscosity of mixed cultivation with high exopolysaccharides-producing Streptococcus thermophilus. The optimal fermentation conditions with mixed cultivation are:42 ℃, inoculation size 4%(v/v), the strains matching ratio of high exopolysaccharides-producing Streptococcus thermophilus, Lactobacillus bulgaricus and Lactobacillus acidophilus is 3:2:1 according to acidity, content of acetaldehyde and viscosity. On this condition, mixed cultivation of Lactic acid bacteria significantly improve acidity,acetaldehyde content,viscosity, exopolysaccharide (EPS), the activity of β-galactosidase and protease (P<0.01).The correlative modulus of EPS and viscosity, the activity of β-galactosidase and acidity, the activity of protease and acidity, acetaldehyde content is high.

  11. Application of a high cholesterol-reducing Lactobacillus acidophilus strain in fermented milk%一株高效降胆固醇嗜酸乳杆菌在发酵乳中的应用

    Institute of Scientific and Technical Information of China (English)

    田建军; 张开屏; 张保军; 靳烨

    2012-01-01

    采用高效降胆固醇嗜酸乳杆菌菌株2-2和嗜热链球菌调制发酵剂B,研究了发酵剂B在发酵乳中的应用。通过与传统保加利亚乳杆菌和嗜热链球菌调制的发酵剂A的对比,结果表明,发酵剂B有较强的胆固醇去除效力和弱的后酸化能力。4℃条件下存放15d后发酵乳的酸度为103.6°T,发酵乳中乳酸菌活菌数为2.6×107cfu/mL,高于标准的最低限制(≥106cfu/mL)。研究表明,在降胆固醇和抑制后酸化意义上,发酵剂B能够取代发酵剂A。%The starter culture B was formulated with strain 2-2 of high cholesterol-reducing Lactobacillus acidophilus and Streptococcus thermophilus.Then the application of starter culture B in fermented milk production was studied.By comparison of starter culture B with A that formulaed traditionally with Lactobacillus bulgaricus and Streptococcus thermophilus,the results indicated that the starter culture B had abilities of strong cholesterol-reducing and weak post-acidification.After 15d storage at 4℃,the acidity of the fermented milk was 103.6°T and the number of living bacteria was 2.6×107cfu/mL,it was much larger than the minimum standard(≥106cfu/mL).The study indicated that the starter culture A could be replaced by B in a sense of reducing cholesterol and inhibiting the extent of post-acidification.

  12. A hybrid zone between Bathymodiolus mussel lineages from eastern Pacific hydrothermal vents

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    Johnson Shannon B

    2013-01-01

    Full Text Available Abstract Background The inhabitants of deep-sea hydrothermal vents occupy ephemeral island-like habitats distributed sporadically along tectonic spreading-centers, back-arc basins, and volcanically active seamounts. The majority of vent taxa undergo a pelagic larval phase, and thus varying degrees of geographical subdivision, ranging from no impedance of dispersal to complete isolation, often exist among taxa that span common geomorphological boundaries. Two lineages of Bathymodiolus mussels segregate on either side of the Easter Microplate, a boundary that separates the East Pacific Rise from spreading centers connected to the Pacific-Antarctic Ridge. Results A recent sample from the northwest flank of the Easter Microplate contained an admixture of northern and southern mitochondrial haplotypes and corresponding alleles at five nuclear gene loci. Genotypic frequencies in this sample did not fit random mating expectation. Significant heterozygote deficiencies at nuclear loci and gametic disequilibria between loci suggested that this transitional region might be a ‘Tension Zone’ maintained by immigration of parental types and possibly hybrid unfitness. An analysis of recombination history in the nuclear genes suggests a prolonged history of parapatric contact between the two mussel lineages. We hereby elevate the southern lineage to species status as Bathymodiolus antarcticus n. sp. and restrict the use of Bathymodiolus thermophilus to the northern lineage. Conclusions Because B. thermophilus s.s. exhibits no evidence for subdivision or isolation-by-distance across its 4000 km range along the EPR axis and Galápagos Rift, partial isolation of B. antarcticus n. sp. requires explanation. The time needed to produce the observed degree of mitochondrial differentiation is consistent with the age of the Easter Microplate (2.5 to 5.3 million years. The complex geomorphology of the Easter Microplate region forces strong cross-axis currents that

  13. REDUÇÃO DE NÍVEIS DE OLIGOSSACÁRIDES EM “IOGURTE” DE SOJA PELO USO DE CULTURAS LÁTICAS

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    ELIZEU ANTôNIO ROSSI

    2007-02-01

    Full Text Available O Streptococcus thermophilus e o Lactobacillus bulgaricus, microorganismos tradicionalmente empregados na elaboração de iogurtes, não são capazes de reduzir satisfatoriamente os níveis de oligossacárides de “iogurtes” preparados à base de soja. Em razão disso, verificou-se o efeito da substituição e/ou associação dessas culturas láticas com culturas de Lactobacillus fermenti e Lactobacillus acidophilus. Nos diversos produtos obtidos foram determinados os níveis de estaquiose, rafinose e sacarose, além da aceitabilidade geral e textura (avaliações sensoriais, viscosidade e consistência. Foi também verificado o tempo de fermentação de cada produto. Com relação à estaquiose, os níveis encontrados nos “iogurtes” de soja representaram de 98 a 115% do nível original do meio de fermentação. A rafinose não foi detectada em nenhum dos produtos, enquanto a sacarose sofreu reduções que variaram de 55,5% a 68,0% do conteúdo inicial. Vários açúcares simples, dentre eles a galactose, foram verificados nos diferentes produtos acabados. Os resultados mostraram que a modificação do inóculo convencional de iogurte (S. thermophilus + L. bulgaricus, através de substituição ou associação com culturas de L. fermenti e L. acidophilus, não se mostrou como alternativa válida no sentido de reduzir oligossacárides. Por outro lado, nos produtos onde o L. bulgaricus foi substituído, verificou-se um significativo aumento do tempo de fermentação.

  14. Effect of Soymilk Heat-processing on the Quality of Soy Yogurt%豆浆保温处理对大豆酸奶品质的影响

    Institute of Scientific and Technical Information of China (English)

    刘爱洁; 李理

    2013-01-01

    以保温不同时间的豆乳为主要原料,经瑞士乳杆菌(Lactobacillus helveticus,LH-B02)、保加利亚乳杆菌(Lactobacillus bulgaricus ASl.1482,L.B 1482)和嗜热链球菌(Streptococcus thermophilus IFFI 6038,S.T6038)组合发酵制备大豆酸奶,应用SDS-PAGE和氨基氮来表征豆浆中蛋白质的降解情况,分析了酸奶产品的pH、可滴定酸度、持水力、流变学性质以及感官风味评价.结果表明,豆浆经过55℃的保温处理后,氨基氮显著提高,其中保温6h的豆浆氨基态氮含量增加了14.04%.经过保温处理后的豆浆在制备成大豆酸奶后,酸度显著提高,粘弹性增加,剪切稀化特性变弱,感官评价表明,保温4h和6h后制备的大豆酸奶在外观、质构以及风味等方面都有明显提高,凝乳细腻柔和,总体可接受性与对照相比差异显著.%Soymilk was incubated at 55 ℃ for 0, 2, 4, 6 hours in a water bath and then fermented with the Lactobacillus helveticus, Lactobacillus bulgaricus and Streptococcus thermophilus combined culture. The hydrolysis of protein in soymilk was determined by SDS-PAGE and amino-nitrigen method, the quality of the soy yogurt was characterized by pH, titratable acidity, water holding capacity, rhelolgy property, and sensery evaluation. The results indicated that the protein was hydrolyzed as the amino nitrigen content in soymilk increasing significantly along with the incubation time, the amino nitrogen content reaching 14.04% after 6 h incubation. The quality of the soy yogurt was also improved significantly through this soymilk heat-processing treatment. Tthe titratable acidity, viscidity and elasticity increased, and the characteristics of shear thinning got weak. Sensory evaluation showed that the texture of the product became exquisite and soft, and the overall acceptance significantly enhanced.

  15. Prevention of heterocyclic amine-induced DNA damage in colon and liver of rats by different lactobacillus strains.

    Science.gov (United States)

    Zsivkovits, Markus; Fekadu, Kassie; Sontag, Gerhard; Nabinger, Ursula; Huber, Wolfgang W; Kundi, Michael; Chakraborty, Asima; Foissy, Helmuth; Knasmüller, Siegfried

    2003-12-01

    The aim of the present study was to investigate the impact of four different lactobacillus (LB) strains, namely Lactobacillus bulgaricus 291, Streptococcus thermophilus F4, S.thermophilus V3 and Bifidobacterium longum BB536, which are used for the production of yogurt, on the DNA-damaging effects of heterocyclic aromatic amines (HCAs). Male F344 rats were treated orally with HCA mixtures containing 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine, 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline, 2-amino-3,4,8-trimethyl-3H-imidazo[4,5-f]quinoxaline, 2-amino-9H-pyrido[2,3-b]indole and 2-amino-3-methyl-3H- imidazo[4,5-f]quinoline, which were representative of the HCA contents found in fried beef ('beef mix') and chicken ('chicken mix'). Suspensions of LB were given by gavage to the animals simultaneously with and at different time periods before administration of the HCAs. Subsequently, the extent of DNA migration was measured in colon and liver cells in single cell gel electrophoresis (SCGE) assays. All four strains caused complete inhibition of DNA damage induced with beef mix after administration of 1 x 1010 LB cells/animal, whereas with chicken mix only marginal (non-significant) effects were seen. The inhibition of beef-induced DNA damage was dose dependent and was still significant when 1 x 107 cells/animal were administered. Kinetics studies showed that the protective effects were still significant when LB was given 12 h before the beef mix. A comparison of the present results with chemical analytical data from in vitro experiments suggests that the strong reduction in DNA migration seen in the animals can be only partly explained by direct binding effects. The results of the present study show that LB are highly protective against the genotoxic effects of HCAs under conditions which are relevant for humans and provide a possible explanation for the reduced colon cancer rates observed in some studies in individuals with either high LB counts in their feces or with

  16. Codon optimization of xylA gene for recombinant glucose isomerase production in Pichia pastoris and fed-batch feeding strategies to fine-tune bioreactor performance.

    Science.gov (United States)

    Ata, Özge; Boy, Erdem; Güneş, Hande; Çalık, Pınar

    2015-05-01

    The objectives of this work are the optimization of the codons of xylA gene from Thermus thermophilus to enhance the production of recombinant glucose isomerase (rGI) in P. pastoris and to investigate the effects of feeding strategies on rGI production. Codons of xylA gene from T. thermophilus were optimized, ca. 30 % of the codons were replaced with those with higher frequencies according to the codon usage bias of P. pastoris, codon optimization resulted in a 2.4-fold higher rGI activity. To fine-tune bioreactor performance, fed-batch bioreactor feeding strategies were designed as continuous exponential methanol feeding with pre-calculated feeding rate based on the pre-determined specific growth rate, and fed-batch methanol-stat feeding. Six feeding strategies were designed, as follows: (S1) continuous exponential methanol- and pulse- sorbitol feeding; (S2) continuous exponential methanol- and peptone- feeding; (S3) continuous exponential methanol- and pulse- mannitol feeding; (S4) continuous exponential methanol- and peptone- feeding and pulse-mannitol feeding; (S5) methanol-stat feeding by keeping methanol concentration at 5 g L(-1); and, (S6) methanol-stat feeding by keeping methanol concentration at 5 g L(-1) and pulse-mannitol feeding. The highest cell and rGI activity was attained as 117 g L(-1) at t = 66 h and 32530 U L(-1) at t = 53 h, in strategy-S5. The use of the co-substrate mannitol does not increase the rGI activity in methanol-stat feeding, where 4.1-fold lower rGI activity was obtained in strategy-S6. The overall cell yield on total substrate was determined at t = 53 h as 0.21 g g(-1) in S5 strategy.

  17. Textura instrumental de queijo petit-suisse potencialmente probiótico: influência de diferentes combinações de gomas Instrumental texture of probiotic petit-suisse cheese: influence of different combinations of gums

    Directory of Open Access Journals (Sweden)

    Larissa Yukie Maruyama

    2006-06-01

    Full Text Available O efeito de diferentes combinações de gomas sobre os parâmetros de textura instrumental de queijo tipo petit-suisse probiótico foi avaliado. Queijos petit-suisse foram produzidos utilizando-se massa-base de queijo quark preparada com o starter Streptococcus thermophilus e os probióticos Lactobacillus acidophilus e Bifidobacterium longum. Três formulações de queijo petit-suisse foram preparadas, a partir de massa-base de queijo quark adicionada de 0,75% do produto final da mistura de gomas - xantana (X, carragena (C, guar (G, pectina (P: F1 = 2,5X:2,5C:5G; F2 = 2X:3C:5P; F3 = 5C:5G. Os parâmetros avaliados após 1, 7, 14 e 21 dias de armazenamento do produto a 4±1ºC incluíram a contagem de probióticos, a textura instrumental, o pH e a umidade. As populações de probióticos mantiveram-se acima de 6,40 log UFC/g para L. acidophilus e de 7,30 log UFC/g para B. longum. Houve diferença significativa entre as formulações (pThe effect of different combinations of gums over texture parameters of probiotic petit-suisse cheese was evaluated. Petit-suisse cheeses were produced using Quark cheese-base prepared with the starter Streptococcus thermophilus and the probiotics Lactobacillus acidophilus and Bifidobacterium longum. Three formulations of petit-suisse were prepared, using the quark cheese-base added of 0.75% of the final product of the mixture of the hydrocolloids xanthan gum (X, carrageenan gum (C, guar gum (G, pectin (P: F1 = 2,5X:2,5C:5G; F2 = 2X:3C:5P; F3 = 5C:5G. Parameters evaluated after 1, 7, 14, and 21 days of storage of the product at 4±1ºC included microbial counts of probiotic microorganisms, instrumental texture parameters, pH and moisture. Probiotic counts were always above 6.40 log CFU/g for L. acidophilus and above 7.30 log CFU/g for B. longum. The formulations were significantly different (p<0.05 for all the texture parameters, except for firmness of F1. The pH and the moisture were similar for the three

  18. 胡萝卜菠萝酸奶的研制%Development of Yoghurt with Carrot and Pineapple

    Institute of Scientific and Technical Information of China (English)

    张继成

    2012-01-01

    以鲜牛乳和胡萝卜、菠萝为主要原料,以保加利亚乳杆菌、嗜热链球菌和双歧杆菌作为发酵剂,采用感官品质评价和正交试验方法,研究胡萝卜菠萝酸奶的加工工艺.结果表明,胡萝卜菠萝酸奶加工制作的最佳配方及工艺条件为:在鲜牛乳中添加10%的胡萝卜和菠萝汁(胡萝卜汁∶菠萝汁=1∶2),8%的白砂糖,菌种添加量为4%(保加利亚乳杆菌∶嗜热链球菌∶双歧杆菌=1∶2∶1),在42 ℃下发酵4h,经冷却后熟后,制得的胡萝卜菠萝酸奶具有均匀的橙红色和独特的胡萝卜菠萝风味,其凝块均匀一致,表面细腻光滑,硬度、黏度适中,酸甜可口,清爽润喉,无异味,无或有少量的乳清析出,符合酸奶标准.%Using fresh milk, carrots and pineapples as the main raw materials, Lactobacillus bulgaricus, Streptococcus thermophilus and Bacillus bifidus as the starter, the processing technology of carrot pineapple yoghurt was studied, through sensory evaluation and orthogonal test method. The results showed that, the optimum formula and processing technology of carrot pineapple yoghurt were as follows: added 10% carrot and pineapple juice (carrot juice :pineapple juice = 1 -2) to fresh milk, 8% white sugar and 4% bacteria (Lactobacillus bulgaricus 'Streptococcus thermophilus: Bacillus bifidus = 1:2:1), fermented for 4 hours at 42 ℃ and then cooled down. The obtained carrot pineapple yoghurt had a uniform orange red color and clot, unique flavor of carrot and pineapple, smooth appearance, moderate hardness and viscosity, suitable acid and sweet flavor, no off-flavor, and no or a little of whey separtion, and was in line with the yogurt standard.

  19. Modern taxonomy of biotechnologically important Aspergillus and Penicillium species.

    Science.gov (United States)

    Houbraken, Jos; de Vries, Ronald P; Samson, Robert A

    2014-01-01

    Taxonomy is a dynamic discipline and name changes of fungi with biotechnological, industrial, or medical importance are often difficult to understand for researchers in the applied field. Species belonging to the genera Aspergillus and Penicillium are commonly used or isolated, and inadequate taxonomy or uncertain nomenclature of these genera can therefore lead to tremendous confusion. Misidentification of strains used in biotechnology can be traced back to (1) recent changes in nomenclature, (2) new taxonomic insights, including description of new species, and/or (3) incorrect identifications. Changes in the recent published International Code of Nomenclature for Algae, Fungi and Plants will lead to numerous name changes of existing Aspergillus and Penicillium species and an overview of the current names of biotechnological important species is given. Furthermore, in (biotechnological) literature old and invalid names are still used, such as Aspergillus awamori, A. foetidus, A. kawachii, Talaromyces emersonii, Acremonium cellulolyticus, and Penicillium funiculosum. An overview of these and other species with their correct names is presented. Furthermore, the biotechnologically important species Talaromyces thermophilus is here combined in Thermomyces as Th. dupontii. The importance of Aspergillus, Penicillium, and related genera is also illustrated by the high number of undertaken genome sequencing projects. A number of these strains are incorrectly identified or atypical strains are selected for these projects. Recommendations for correct strain selection are given here. Phylogenetic analysis shows a close relationship between the genome-sequenced strains of Aspergillus, Penicillium, and Monascus. Talaromyces stipitatus and T. marneffei (syn. Penicillium marneffei) are closely related to Thermomyces lanuginosus and Th. dupontii (syn. Talaromyces thermophilus), and these species appear to be distantly related to Aspergillus and Penicillium. In the last part of

  20. Construction of Co-expressed MntH and Mn-catalase Gene in Escherichia coli and Optimization of Fermentation Conditions%MntH与含锰过氧化氢酶共表达基因工程菌的构建与发酵优化

    Institute of Scientific and Technical Information of China (English)

    王慧; 崔云风; 刘岩; 史吉平; 赵志军; 王绍明

    2015-01-01

    Mn-catalase from Thermus thermophilus HB27 and Mn2+transport protein MntH from Escherichia coli were co-expressed in E. coli BL21(DE3). The optimization of fermentation medium and environment for the production of Mn-catalase was carried out at the shake flask level. The optimal carbon and nitrogen source were 7.0 g/L glycerine, 3.75 g/L yeast extract and 11.25 g/L peptone respectively. The optimum induced concentration of IPTG was 0.05 mmol/L while the Mn2+in media was 1 mmol/L. In addition, the optimal initial pH of the medium and culture temperature were pH 8.0 and 37℃respectively. Under the optimal conditions, the maximal activity of catalase reached 476 U/mL, which was 3-fold of the control. Finally, in a 5 L fermentor the activity of catalase increased to 1 094 U/mL.%构建Mn2+转运蛋白MntH与来源于Thermus thermophilus HB27的含锰过氧化氢酶的共表达基因工程菌,并进行了发酵培养基及培养环境条件的优化,确定培养基中最佳的碳氮源种类及其浓度分别为:甘油7.0 g/L,酵母粉3.75 g/L和蛋白胨11.25 g/L;当培养基中的Mn2+浓度为1 mmol/L时,最佳的IPTG诱导浓度为0.05 mmol/L。此外,最佳的培养基初始pH值及培养温度分别为:pH 8.0和37℃,在最优发酵条件下工程菌摇瓶发酵培养24 h,过氧化氢酶活最高可达476 U/mL是未优化前3倍。在5 L发酵罐的验证实验中,过氧化氢酶的酶活进一步提高至1094 U/mL。

  1. 益生菌混合菌种在发酵豆乳中的优化%Optimizing Fermentation of Soymilk with Combination of Probiotic Bacteria

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    高凤祥; 张艳杰

    2012-01-01

    The objective of this study is to produce a probiotic fermented soymilk with the aim of shortening the fermentation time. For this, soymilk was fermented with probiotic culture AST and yoghurt culture TLB at 42 ℃, during the fermentation and storage of the fermented soymilk (28 days at 4 ℃, the changes in pH —value and viable cells count were observed. Fermentation time significantly reduces to 8 h at 42 ℃ with AST culture (Lactobacillus acidophi-lus, Bifidobacterium spp. , and Streptococcus thermophilus). However, Lactobacillus acidoph-ilus survived poorly during cold storage and the viable cells count was under the probiotic minimum as soon as after the first week of storage. Soymilk was fermented at 42 ℃ with TLB culture (Streptococcus thermophilus, Lactobacillus bulgaricus, Bifidobacterium animalis subsp. lactis Dbl2). Consequently, the fermentation time was shortened to 4 hours whereby the viable cells count of bifidobacteria increased during fermentation for the half of the logarithm scale approximately. During 28 days of cold storage, bacterial count remained over 107 cfu/mL.%采用益生菌混合物AST和TLB分别在42℃进行豆乳的发酵,在发酵豆乳的发酵和贮存过程中(4℃下28 d),观察pH值和活菌数的变化情况.在AST益生茵混合物(嗜酸性乳杆菌,双歧杆菌和嗜热链球菌)的培育下,42℃下发酵时间减少到8h.但嗜酸性乳杆菌在冷藏的过程中生存状态较差,其活菌数在冷藏后一周后未达到标准.将豆乳在42℃下用TLB益生菌(嗜热链球菌,保加利亚乳杆菌和动物双歧杆菌乳酸亚种Bb12)进行发酵,结果发酵时间缩短到4h,双歧杆菌活菌数目的对数增加了约一半,而且经过28d的冷藏,细菌数仍然维持在107 CFU/mL以上.

  2. Goniometer-based femtosecond X-ray diffraction of mutant 30S ribosomal subunit crystals

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    E. Han Dao

    2015-07-01

    Full Text Available In this work, we collected radiation-damage-free data from a set of cryo-cooled crystals for a novel 30S ribosomal subunit mutant using goniometer-based femtosecond crystallography. Crystal quality assessment for these samples was conducted at the X-ray Pump Probe end-station of the Linac Coherent Light Source (LCLS using recently introduced goniometer-based instrumentation. These 30S subunit crystals were genetically engineered to omit a 26-residue protein, Thx, which is present in the wild-type Thermus thermophilus 30S ribosomal subunit. We are primarily interested in elucidating the contribution of this ribosomal protein to the overall 30S subunit structure. To assess the viability of this study, femtosecond X-ray diffraction patterns from these crystals were recorded at the LCLS during a protein crystal screening beam time. During our data collection, we successfully observed diffraction from these difficult-to-grow 30S ribosomal subunit crystals. Most of our crystals were found to diffract to low resolution, while one crystal diffracted to 3.2 Å resolution. These data suggest the feasibility of pursuing high-resolution data collection as well as the need to improve sample preparation and handling in order to collect a complete radiation-damage-free data set using an X-ray Free Electron Laser.

  3. Determination of volatile and non-volatile products of milk fermentation processes using capillary zone electrophoresis and solid phase microextraction coupled to gas chromatography.

    Science.gov (United States)

    Ligor, Magdalena; Jarmalaviciene, Reda; Szumski, Michal; Maruska, Audrius; Buszewski, Boguslaw

    2008-08-01

    The aim of the investigations was to develop analytical methods for the determination of selected volatile and non-volatile organic compounds numbering among the final products of milk fermentation. The analyzed compounds were as follows: biacetyl and carboxylic acids (formic, acetic, citric, and lactic). The model yogurt was prepared under controlled conditions in our laboratory by addition of the selected bacteria (Lactobacillus bulgaricus and Streptococcus thermophilus) to the milk sample. The temperature, time, and stirring were controlled during the fermentation process. Factors considered in SPMPE-GC-FID method development included fiber exposure time, salt addition, temperature of extraction, and temperature of desorption. Various SPME fibers, for example with PDMS, CAR/PDMS, PA, and PDMS/DVB coatings, were tested to obtain the highest recovery of the investigated compounds extracted from yogurt samples. Based on these preliminary experiments, qualitative and quantitative analyses for the determination of biacetyl were performed by SPME-GC-FID. Moreover, a capillary zone electrophoresis method was developed for the determination of carboxylic acids in the yogurt samples. The buffer composition as well as deproteinization by acetonitrile were found to have a crucial effect on the analysis.

  4. Effects of selected factors on rheological and textural properties of probiotic yoghurt

    Directory of Open Access Journals (Sweden)

    Jovana Glušac

    2011-03-01

    Full Text Available The aim of this work was to study the influence of inulin (1 %, combination of inulin (1 % and acacia honey (4 %, heat treatment of milk, and storage time on the rheological and textural properties of probiotic yoghurt. Rheological properties were assessed trough viscosity, syneresis and texture (firmness, consistency, cohesiveness and index of viscosity. Yoghurt was prepared from milk (1,5 % fat with added inulin (1% before heat treatment at 85 °C for 20 min or 95 °C for 10 min. After cooling to 55 °C honey (4 % was added. Samples were inoculated using probiotic starter culture (70 % w/w Streptococcus thermophilus, 10 % w/w Lactobacillus bulgaricus, 10 % w/w Lactobacillus acidophilus, 10 % w/w Bifidobacterium ssp.. Yoghurt samples were held on +5 °C during 21 days. Measurements of pH value, lactic acid, viscosity, syneresis, and textural properties were done after 1, 7, 14 and 21 days of storage. The results of this study show that honey addition significantly decreased fermentation time compared to fermentation time of control samples or samples containing inulin. Furthermore, addition of honey and inulin to milk caused significant lower syneresis (p<0,05 during storage time, while there was no significant influence on viscosity and texture of final product. The applied heat treatment of milk had no significant influence on rheological properties of probiotic yoghurt.

  5. Structures and reaction mechanisms of the two related enzymes, PurN and PurU.

    Science.gov (United States)

    Sampei, Gen-ichi; Kanagawa, Mayumi; Baba, Seiki; Shimasaki, Toshiaki; Taka, Hiroyuki; Mitsui, Shohei; Fujiwara, Shinji; Yanagida, Yuki; Kusano, Mayumi; Suzuki, Sakiko; Terao, Kayoko; Kawai, Hiroya; Fukai, Yoko; Nakagawa, Noriko; Ebihara, Akio; Kuramitsu, Seiki; Yokoyama, Shigeyuki; Kawai, Gota

    2013-12-01

    The crystal structures of glycinamide ribonucleotide transformylases (PurNs) from Aquifex aeolicus (Aa), Geobacillus kaustophilus (Gk) and Symbiobacterium toebii (St), and of formyltetrahydrofolate hydrolase (PurU) from Thermus thermophilus (Tt) were determined. The monomer structures of the determined PurN and PurU were very similar to the known structure of PurN, but oligomeric states were different; AaPurN and StPurN formed dimers, GkPurN formed monomer and PurU formed tetramer in the crystals. PurU had a regulatory ACT domain in its N-terminal side. So far several structures of PurUs have been determined, yet, the mechanisms of the catalysis and the regulation of PurU have not been elucidated. We, therefore, modelled ligand-bound structures of PurN and PurU, and performed molecular dynamics simulations to elucidate the reaction mechanisms. The evolutionary relationship of the two enzymes is discussed based on the comparisons of the structures and the catalytic mechanisms. PMID:24108189

  6. Composition of Hydrothermal Vent Microbial Communities as Revealed by Analyses of Signature Lipids, Stable Carbon Isotopes and Aquificales Cultures

    Science.gov (United States)

    Jahnke, Linda L.; Eder, Wolfgang; Huber, Robert; Hinrichs, Kai-Uwe; Hayes, John M.; Cady, Sherry L.; DesMarais, David J.; Hope, Janet M.; Summons, Roger E.

    2001-01-01

    Extremely thermophilic microbial communities associated with the siliceous vent walls and outflow channel of Octopus Spring, Yellowstone National Park, have been examined for lipid biomarker and carbon isotopic signatures. These data were compared with that obtained from representatives of three Aquificales genera. Thermocrinis ruber, Thermocrinis sp. HI, Hydrogenobacter thermophilus, Aquifex pyrophilus and Aquifex aeolicus all contained phospholipids composed not only of the usual ester-linked fatty acids, but also ether-linked alkyl moieties. The fatty acids of all cultured organisms were dominated by very distinct pattern of n-C-20:1 and cy-C-21 compounds. The alkyl glycerol ethers were present primarily as C-18:0 monoethers with the exception of the Aquifex spp. in which dialkyl glycerol ethers with a boarder carbon-number distribution were also present. These Aquificales biomarker lipids were the major constituents in the lipid extracts of the Octopus Spring microbial samples. Two natural samples, a microbial biofilm growing in association with deposition of amorphous silica on the vent walls at 92 C, and the well-known "pink-streamer community" (PSC), siliceous filaments of a microbial consortia growing in the outflow channel at 87 C were analyzed. Both the biofilm and PSC samples contained mono- and dialkyl glycerol ethers with a prevalence of C-18 and C-20 alkyls. Phospholipid fatty acids were comprised of both the characteristic. Additional information is contained in the original extended abstract.

  7. A differentially conserved residue (Ile42) of GH42 β-galactosidase from Geobacillus stearothermophilus BgaB is involved in both catalysis and thermostability.

    Science.gov (United States)

    Dong, Yi-Ning; Chen, Hai-Qin; Sun, Yan-Hui; Zhang, Hao; Chen, Wei

    2015-04-01

    The glycoside hydrolase family 42 (GH42) of thermophilic microorganisms consists of thermostable β-galactosidases that display significant variations in their temperature optima and stabilities. In this study, we compared the substrate binding modes of 2 GH42 β-galactosidases, BgaB from Geobacillus stearothermophilus and A4-β-Gal from Thermus thermophilus A4. The A4-β-Gal has a catalytic triad (Glu312-Arg32-Glu35) with an extended hydrogen bond network that has not been observed in BgaB. In this study, we performed site-saturation mutagenesis of Ile42 in BgaB (equivalent to Glu312 in A4-β-Gal) to study the effects of different residues on thermostability, catalytic function, and the extended hydrogen bond network. Our experimental results suggest that substitution of Ile42 with polar AA enhanced the thermostability but decreased the catalytic efficiency of BgaB. Polar AA substitution for Ile42 simultaneously affected thermostability, catalytic efficiency, and the hydrogen bond network, suggesting that Ile42 is responsible for functional discrimination between members of the GH42 family. These observations could lead to a novel strategy for investigating the functional evolution of the GH42 β-galactosidases.

  8. A Bio-computing Analysis of the Resting-to-pulsed Conformational Changes in Cytochrome c Oxidase

    Directory of Open Access Journals (Sweden)

    T Alleyne

    2015-09-01

    Full Text Available Cytochrome c oxidase (Cox accepts electrons from its substrate, cytochrome c and passes these to oxygen, which is reduced to water. Kinetic studies show that an active form of the enzyme (pulsed and a slower form (resting exists. More efficient internal electron transfer and the switching of the enzyme’s oxygen/ligand binding site between opened and closed positions are said to account for the different rates of reduction. We employed bio-computing to analyse the structure of the oxygen/ligand binding site of bovine Cox under different redox states; a comparison with Thermus thermophilus Cox was also conducted. The study detected that the ligand binding site of Cox is exposed to the contents of the intermembrane space, and that the side chain of haem a3, located at the enzyme’s oxygen/ligand binding site, approached Pro-69 and Ile-34 in faraway subunit-II. However, no open-to-closed gating structures were detected at the ligand binding site. We concluded that the resting-to-pulse transition in Cox does not involve opening-up of the ligand binding site. We propose that the rates of ligand/oxygen/cyanide binding are partly controlled by “queuing” near the binding site and that the binding of oxygen to haem a3-CuB triggers the resting-to-pulsed transition via long-range conformational changes.

  9. 不同菌种组合对酸奶品质的影响研究%The Effects of Different Groups of Bacteria to Yogurt

    Institute of Scientific and Technical Information of China (English)

    胡豆; 林凤英; 梁钻好; 刘玉冰; 杜冰

    2015-01-01

    This research has performed to study the effect of co-culture of different probiotics bacteria combination on the quality of yogurt. The Bacillus coagulans was inoculated into milk alone or with different lactic acid bacteria such as Lactobacillus bulgaricus and Streptococcus thermophiles, Lactobacillus plantarum and Lactobacillus rhamnosus, respectively. The determination of texture, pH and sensory evaluation of the fermented yogurt has been conducted. We found that co-culture of the combination of Bacillus coagulans with Lactobacillus bulgaricus and Streptococcus thermophilus to be the accepted one, which turned out to be similar to yogurts at present markets.%本文将新型益生菌凝结芽孢杆菌和多种酸奶发酵菌种相组合,通过测定发酵酸奶的质构、酸度,并配以感官评定等三项指标,研究不同菌种组合对酸奶品质的影响。结果表明,在传统型酸奶(保加利亚乳杆菌、嗜热链球菌发酵)基础上加入凝结芽孢杆菌的酸奶在对比组中表现最为出色,与市场所售酸奶品质相仿。

  10. Alkalizing reactions streamline cellular metabolism in acidogenic microorganisms.

    Directory of Open Access Journals (Sweden)

    Stefania Arioli

    Full Text Available An understanding of the integrated relationships among the principal cellular functions that govern the bioenergetic reactions of an organism is necessary to determine how cells remain viable and optimise their fitness in the environment. Urease is a complex enzyme that catalyzes the hydrolysis of urea to ammonia and carbonic acid. While the induction of urease activity by several microorganisms has been predominantly considered a stress-response that is initiated to generate a nitrogen source in response to a low environmental pH, here we demonstrate a new role of urease in the optimisation of cellular bioenergetics. We show that urea hydrolysis increases the catabolic efficiency of Streptococcus thermophilus, a lactic acid bacterium that is widely used in the industrial manufacture of dairy products. By modulating the intracellular pH and thereby increasing the activity of β-galactosidase, glycolytic enzymes and lactate dehydrogenase, urease increases the overall change in enthalpy generated by the bioenergetic reactions. A cooperative altruistic behaviour of urease-positive microorganisms on the urease-negative microorganisms within the same environment was also observed. The physiological role of a single enzymatic activity demonstrates a novel and unexpected view of the non-transcriptional regulatory mechanisms that govern the bioenergetics of a bacterial cell, highlighting a new role for cytosol-alkalizing biochemical pathways in acidogenic microorganisms.

  11. Antibiotic Resistance of Probiotic Strains of Lactic Acid Bacteria Isolated from Marketed Foods and Drugs

    Institute of Scientific and Technical Information of China (English)

    CHANG LIU; ZHUO-YANG ZHANG; KE DONG; JIAN-PING YUAN; XIAO-KUI GUO

    2009-01-01

    Objective To identify the antimicrobial resistance of commercial lactic acid bacteria present in microbial foods and drug additives by analyzing their isolated strains used for fermentation and probioties. Methods Antimicrobial susceptibility of 41 screened isolates was tested with disc diffusion and E-test methods after species-level identification. Resistant strains were selected and examined for the presence of resistance genes by PCR. Results Distribution of resistance was found in different species. All isolates were susceptible to chloramphenicol, tetracycline, ampicillin, amoxicillin/clavulanic acid, cephalothin, and imipenem. In addition, isolates resistant to vancomycin, rifampicin, streptomycin, bacitracin, and erythromycin were detected, although the incidence of resistance to these antibiotics was relatively low. In contrast, most strains were resistant to ciprofloxacin, amikacin, trimethoprim/sulphamethoxazole, and gentamycin. The genes msrC, vanX, and dfrA were detected in strains of Enterococcus faecium, Lactobacillus plantarum, Streptococcus thermophilus, and Lactococcus lactis. Conclusion Antibiotic resistance is present in different species of probiotic strains, which poses a threat to food safety. Evaluation of the safety of lactic acid bacteria for human consumption should be guided by established criteria, guidelines and regulations.

  12. Isolation, crystallization, and investigation of ribosomal protein S8 complexed with specific fragments of rRNA of bacterial or archaeal origin.

    Science.gov (United States)

    Tishchenko, S V; Vassilieva, J M; Platonova, O B; Serganov, A A; Fomenkova, N P; Mudrik, E S; Piendl, W; Ehresmann, C; Ehresmann, B; Garber, M B

    2001-09-01

    The core ribosomal protein S8 binds to the central domain of 16S rRNA independently of other ribosomal proteins and is required for assembling the 30S subunit. It has been shown with E. coli ribosomes that a short rRNA fragment restricted by nucleotides 588-602 and 636-651 is sufficient for strong and specific protein S8 binding. In this work, we studied the complexes formed by ribosomal protein S8 from Thermus thermophilus and Methanococcus jannaschii with short rRNA fragments isolated from the same organisms. The dissociation constants of the complexes of protein S8 with rRNA fragments were determined. Based on the results of binding experiments, rRNA fragments of different length were designed and synthesized in preparative amounts in vitro using T7 RNA-polymerase. Stable S8-RNA complexes were crystallized. Crystals were obtained both for homologous bacterial and archaeal complexes and for hybrid complexes of archaeal protein with bacterial rRNA. Crystals of the complex of protein S8 from M. jannaschii with the 37-nucleotide rRNA fragment from the same organism suitable for X-ray analysis were obtained.

  13. Mapping the ribosomal protein S7 regulatory binding site on mRNA of the E. coli streptomycin operon.

    Science.gov (United States)

    Surdina, A V; Rassokhin, T I; Golovin, A V; Spiridonova, V A; Kopylov, A M

    2010-07-01

    In this work it is shown by deletion analysis that an intercistronic region (ICR) approximately 80 nucleotides in length is necessary for interaction with recombinant E. coli S7 protein (r6hEcoS7). A model is proposed for the interaction of S7 with two ICR sites-region of hairpin bifurcations and Shine-Dalgarno sequence of cistron S7. A de novo RNA binding site for heterologous S7 protein of Thermus thermophilus (r6hTthS7) was constructed by selection of a combinatorial RNA library based on E. coli ICR: it has only a single supposed protein recognition site in the region of bifurcation. The SERW technique was used for selection of two intercistronic RNA libraries in which five nucleotides of a double-stranded region, adjacent to the bifurcation, had the randomized sequence. One library contained an authentic AG (-82/-20) pair, while in the other this pair was replaced by AU. A serwamer capable of specific binding to r6hTthS7 was selected; it appeared to be the RNA68 mutant with eight nucleotide mutations. The serwamer binds to r6hTthS7 with the same affinity as homologous authentic ICR of str mRNA binds to r6hEcoS7; apparent dissociation constants are 89 +/- 43 and 50 +/- 24 nM, respectively.

  14. The location of protein S8 and surrounding elements of 16S rRNA in the 70S ribosome from combined use of directed hydroxyl radical probing and X-ray crystallography.

    Science.gov (United States)

    Lancaster, L; Culver, G M; Yusupova, G Z; Cate, J H; Yusupov, M M; Noller, H F

    2000-05-01

    Ribosomal protein S8, which is essential for the assembly of the central domain of 16S rRNA, is one of the most thoroughly studied RNA-binding proteins. To map its surrounding RNA in the ribosome, we carried out directed hydroxyl radical probing of 16S rRNA using Fe(II) tethered to nine different positions on the surface of protein S8 in 70S ribosomes. Hydroxyl radical-induced cleavage was observed near the classical S8-binding site in the 620 stem, and flanking the other S8-footprinted regions of the central domain at the three-helix junction near position 650 and the 825 and 860 stems. In addition, cleavage near the 5' terminus of 16S rRNA, in the 300 region of its 5' domain, and in the 1070 region of its 3'-major domain provide information about the proximity to S8 of RNA elements not directly involved in its binding. These data, along with previous footprinting and crosslinking results, allowed positioning of protein S8 and its surrounding RNA elements in a 7.8-A map of the Thermus thermophilus 70S ribosome. The resulting model is in close agreement with the extensive body of data from previous studies using protein-protein and protein-RNA crosslinking, chemical and enzymatic footprinting, and genetics.

  15. Transfer, composition and technological characterization of the lactic acid bacterial populations of the wooden vats used to produce traditional stretched cheeses.

    Science.gov (United States)

    Scatassa, Maria Luisa; Gaglio, Raimondo; Macaluso, Giusi; Francesca, Nicola; Randazzo, Walter; Cardamone, Cinzia; Di Grigoli, Antonino; Moschetti, Giancarlo; Settanni, Luca

    2015-12-01

    The biofilms of 12 wooden vats used for the production of the traditional stretched cheeses Caciocavallo Palermitano and PDO Vastedda della valle del Belìce were investigated. Salmonella spp. and Listeria monocytogenes were never detected. Total coliforms were at low numbers with Escherichia coli found only in three vats. Coagulase-positive staphylococci (CPS) were below the enumeration limit, whereas lactic acid bacteria (LAB) dominated the surfaces of all vats. In general, the dominance was showed by coccus LAB. Enterococci were estimated at high numbers, but usually between 1 and 2 Log cycles lower than other LAB. LAB populations were investigated at species and strain level and for their technological properties relevant in cheese production. Eighty-five strains were analysed by a polyphasic genetic approach and allotted into 16 species within the genera Enterococcus, Lactobacillus, Lactococcus, Leuconostoc, Pediococcus and Streptococcus. Enterococcus faecium was found in all wooden vats and the species most frequently isolated were Enterococcus faecalis, Lactococcus lactis, Leuconostoc mesenteroides, Pediococcus acidilactici and Streptococcus thermophilus. The study of the quantitative data on acidification rate, autolysis kinetics, diacetyl production, antibacterial compound generation and proteolysis by cluster and principal component analysis led to the identification of some strains with promising dairy characteristics. Interestingly, a consistent percentage of LAB was bacteriocin-like inhibitory substances (BLIS) producer. Thus, the microbial biofilms of the wooden vats analysed in this study might contribute actively to the stability of the final cheeses.

  16. Characterization of the Deoxynucleotide Triphosphate Triphosphohydrolase (dNTPase) Activity of the EF1143 Protein from Enterococcus faecalis and Crystal Structure of the Activator-Substrate Complex

    Energy Technology Data Exchange (ETDEWEB)

    Vorontsov, Ivan I.; Minasov, George; Kiryukhina, Olga; Brunzelle, Joseph S.; Shuvalova, Ludmilla; Anderson, Wayne F. (NWU)

    2012-06-19

    The EF1143 protein from Enterococcus faecalis is a distant homolog of deoxynucleotide triphosphate triphosphohydrolases (dNTPases) from Escherichia coli and Thermus thermophilus. These dNTPases are important components in the regulation of the dNTP pool in bacteria. Biochemical assays of the EF1143 dNTPase activity demonstrated nonspecific hydrolysis of all canonical dNTPs in the presence of Mn{sup 2+}. In contrast, with Mg{sup 2+} hydrolysis required the presence of dGTP as an effector, activating the degradation of dATP and dCTP with dGTP also being consumed in the reaction with dATP. The crystal structure of EF1143 and dynamic light scattering measurements in solution revealed a tetrameric oligomer as the most probable biologically active unit. The tetramer contains four dGTP specific allosteric regulatory sites and four active sites. Examination of the active site with the dATP substrate suggests an in-line nucleophilic attack on the {alpha}-phosphate center as a possible mechanism of the hydrolysis and two highly conserved residues, His-129 and Glu-122, as an acid-base catalytic dyad. Structural differences between EF1143 apo and holo forms revealed mobility of the {alpha}3 helix that can regulate the size of the active site binding pocket and could be stabilized in the open conformation upon formation of the tetramer and dGTP effector binding.

  17. Effect of Aqueous Extract of the Seaweed Gracilaria domingensis on the Physicochemical, Microbiological, and Textural Features of Fermented Milks.

    Science.gov (United States)

    Tavares Estevam, Adriana Carneiro; Alonso Buriti, Flávia Carolina; de Oliveira, Tiago Almeida; Pereira, Elainy Virginia Dos Santos; Florentino, Eliane Rolim; Porto, Ana Lúcia Figueiredo

    2016-04-01

    The effects of the Gracilaria domingensis seaweed aqueous extract in comparison with gelatin on the physicochemical, microbial, and textural characteristics of fermented milks processed with the mixed culture SAB 440 A, composed of Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium animalis ssp. lactis, were investigated. The addition of G. domingensis aqueous extract did not affect pH, titratable acidity, and microbial viability of fermented milks when compared with the control (with no texture modifier) and the products with added gelatin. Fermented milk with added the seaweed aqueous extract showed firmness, consistency, cohesiveness, and viscosity index at least 10% higher than those observed for the control product (P < 0.05). At 4 h of fermentation, the fermented milks with only G. domingensis extract showed a texture comparable to that observed for products containing only gelatin. At 5 h of fermentation, firmness and consistency increased significantly (P < 0.05) in products with only seaweed extract added, a behavior not observed in products with the full amount of gelatin, probably due to the differences between the interactions of these ingredients with casein during the development of the gel network throughout the acidification of milk. The G. domingensis aqueous extract appears as a promising gelatin alternative to be used as texture modifier in fermented milks and related dairy products.

  18. Effect of bee pollen supplement on antimicrobial, chemical, rheological, sensorial properties and probiotic viability of fermented milk beverages

    Directory of Open Access Journals (Sweden)

    Oktay Yerlikaya

    2014-11-01

    Full Text Available In this research, effect of bee pollen supplement on antimicrobial, chemical, rheological, sensorial properties and probiotic viability of fermented milk beverages was studied. Bee pollens were added in the rate of 2.5 mg•mL-1 (B, 5 mg•mL-1 (C, 7.5 mg•mL-1 (D, 10 mg•mL-1 (E, and 20 mg•mL-1 (F. Control sample (A was not supplemented with bee pollen. Control and supplemented milk samples were fermented by a commercial ABT1 starter culture (Chr. Hansen, Hørsholm, Denmark containing Lactobacillus acidophilus La 5, Bifidobacterium animalis subs. lactis Bb 12, and Streptococcus thermophilus. While no antimicrobial impact was observed against L. monocytogenes, S. aureus, P. fluorescens, P. aeruginosa and A. hydrophilia upto 7.5 mg•mL-1 pollen addition, addition between 10 mg•mL-1 to 20 mg•mL-1 resulted in activity, and positive effect only in inhibition rates against bacteria such as S. thyphimurium and E. coli. Bee pollen supplements has shown a positive effect on probiotic viability and occurred on increase apparent viscosity, but their effect on sensorial properties was negative. Furthermore an improvement with increasing concentration of pollen addition that yielded no negative effect on physicochemical properties was detected.

  19. Sensory profile development of Oaxaca cheese and relationship with physicochemical parameters.

    Science.gov (United States)

    Sandoval-Copado, J; Orozco-Villafuerte, J; Pedrero-Fuehrer, D; Colín-Cruz, M A

    2016-09-01

    A sensory profile of Oaxaca cheese was developed. To represent both industrial and artisanal Oaxaca cheese, 3 types of cheese were manufactured under controlled conditions: 2 with pasteurized milk using a mesophilic lactic acid bacteria culture (Lactococcus lactis ssp. lactis and L. lactis ssp. cremoris) and a thermophilic lactic acid bacteria culture (Streptococcus salivarius ssp. thermophilus), and a third with naturally acidified raw milk. The cheeses were evaluated by a panel of 11 trained judges. Additionally, chemical composition and instrumental color parameters were determined, an instrumental texture profile analysis was performed, and volatile compounds were identified by solid-phase microextraction coupled with gas chromatography/mass spectrometry. The sensory profile developed includes 43 descriptors, of which 13 are discriminant among these 3 cheese types. Descriptors include visual attribute waxy; tactile (manual) attributes moist and oily; orthonasal aroma attributes empyreumatic and cow; basic taste attribute salty; mouth texture attributes chewable, gummy, adherent, fibrous, moist, and fatty; and auditory attribute squeaky. A strong correlation was found between specific sensory data and physicochemical parameters measured by instrumental and chemical methods. PMID:27394949

  20. Effect of Different Storage Period on Lactic Acid Bacterias from Goji Yogurt and Goji Yogurt with Honey

    Directory of Open Access Journals (Sweden)

    Ancuta M. Rotar

    2014-05-01

    Full Text Available Lactic acid bacterias (LAB: Streptococcus thermophilus-ST, Lactobacillus bulgaricus-LB are well known in the food technology area for their ability to produce lactic acid (LA from carbohydrates throught fermentation. In case of goji yogurt the interaction between the two species of LAB has influence on the fermentation period and LA quantity. LAB’s are widely used in the food industry because their growth in the dairy products lowers the carbohydrate content, they can also drop the pH values under 4.0, values were common pathogens are inhibited, and because of all these properties they are capable to prolong the shelf life. The present study aims to observe the evolution of LAB’s – ST and LB, from goji yogurt (7% (A and goji yogurt (7% with honey (B during the shelf life in corelation with other physico-chemical properties such as sugar content, fat content and dry matter. The samples for the evaluation were taken in the first day of storage, at the middle of storage period and in the last day of storage.

  1. Volatile sulphur compounds-forming abilities of lactic acid bacteria: C-S lyase activities.

    Science.gov (United States)

    Bustos, Irene; Martínez-Bartolomé, Miguel A; Achemchem, Fouad; Peláez, Carmen; Requena, Teresa; Martínez-Cuesta, M Carmen

    2011-08-01

    Volatile sulphur compounds (VSCs) are of prime importance in the overall aroma of cheese and make a significant contribution to their typical flavours. Thus, the control of VSCs formation offers considerable potential for industrial applications. Here, lactic acid bacteria (LAB) from different ecological origins were screened for their abilities to produce VSCs from L-methionine. From the data presented, VSC-forming abilities were shown to be strain-specific and were correlated with the C-S lyase enzymatic activities determined using different approaches. High VSCs formation were detected for those strains that were also shown to possess high thiol-producing abilities (determined either by agar plate or spectrophotometry assays). Moreover, differences in C-S lyase activities were shown to correspond with the enzymatic potential of the strains as determined by in situ gel visualization. Therefore, the assessment of the C-S lyase enzymatic potential, by means of either of these techniques, could be used as a valuable approach for the selection of LAB strains with high VSC-producing abilities thus, representing an effective way to enhance cheese sulphur aroma compounds synthesis. In this regard, this study highlights the flavour forming potential of the Streptococcus thermophilus STY-31, that therefore could be used as a starter culture in cheese manufacture. Furthermore, although C-S lyases are involved in both biosynthetic and catabolic pathways, an association between methionine and cysteine auxotrophy of the selected strains and their VSCs-producing abilities could not be found.

  2. PCR-DGGE技术分析混菌发酵乳中马克斯克鲁维酵母与乳酸菌的相互作用%Application of PCR-DGGE to analyze the interaction between Kluyveromyces marxianus and the lactic acid bacteria during fermentation and storage process

    Institute of Scientific and Technical Information of China (English)

    范维; 李路; 姜铁民; 张彧; 闵伟红; 陈历俊

    2015-01-01

    利用变性梯度凝胶电泳(Denaturing Gradient Gel Electrophoresis,DGGE)技术,对混菌发酵及贮藏过程中马克斯克鲁维酵母与乳酸菌之间的相互作用进行分析,进而对整个过程中微生物优势菌群及其稳定性进行跟踪监测.结果表明:混菌发酵及贮藏过程中微生物组成比较稳定,优势菌为嗜热链球菌(Streptococcus thermophilus,ST);发酵过程中,马克斯克鲁维酵母(Kluyveromyces marxianus)的添加对乳酸菌生长起到促进作用,尤其是对保加利亚乳杆菌(Lactobacillus bulgaricus,LB)效果显著,贮藏期间该作用转变为抑制;整个过程乳酸菌的存在对酵母菌的生长具有一定的抑制作用.该研究可为深入探讨乳酸菌与酵母菌共同发酵机理及新型发酵乳制品的开发提供理论基础.

  3. Phages of lactic acid bacteria: the role of genetics in understanding phage-host interactions and their co-evolutionary processes.

    Science.gov (United States)

    Mahony, Jennifer; Ainsworth, Stuart; Stockdale, Stephen; van Sinderen, Douwe

    2012-12-20

    Dairy fermentations are among the oldest food processing applications, aimed at preservation and shelf-life extension through the use of lactic acid bacteria (LAB) starter cultures, in particular strains of Lactococcus lactis, Streptococcus thermophilus, Lactobacillus spp. and Leuconostoc spp. Traditionally this was performed by continuous passaging of undefined cultures from a finished fermentation to initiate the next fermentation. More recently, consumer demands on consistent and desired flavours and textures of dairy products have led to a more defined approach to such processes. Dairy (starter) companies have responded to the need to define the nature and complexity of the starter culture mixes, and dairy fermentations are now frequently based on defined starter cultures of low complexity, where each starter component imparts specific technological properties that are desirable to the product. Both mixed and defined starter culture approaches create the perfect environment for the proliferation of (bacterio)phages capable of infecting these LAB. The repeated use of the same starter cultures in a single plant, coupled to the drive towards higher and consistent production levels, increases the risk and negative impact of phage infection. In this review we will discuss recent advances in tracking the adaptation of phages to the dairy industry, the advances in understanding LAB phage-host interactions, including evolutionary and genomic aspects.

  4. Structure of Vibrio cholerae ribosome hibernation promoting factor

    International Nuclear Information System (INIS)

    The X-ray crystal structure of ribosome hibernation promoting factor from V. cholerae has been determined at 2.0 Å resolution. The crystal was phased by two-wavelength MAD using cocrystallized cobalt. The X-ray crystal structure of ribosome hibernation promoting factor (HPF) from Vibrio cholerae is presented at 2.0 Å resolution. The crystal was phased by two-wavelength MAD using cocrystallized cobalt. The asymmetric unit contained two molecules of HPF linked by four Co atoms. The metal-binding sites observed in the crystal are probably not related to biological function. The structure of HPF has a typical β–α–β–β–β–α fold consistent with previous structures of YfiA and HPF from Escherichia coli. Comparison of the new structure with that of HPF from E. coli bound to the Thermus thermophilus ribosome [Polikanov et al. (2012 ▶), Science, 336, 915–918] shows that no significant structural changes are induced in HPF by binding

  5. Survival and bioactivities of selected probiotic lactobacilli in yogurt fermentation and cold storage: New insights for developing a bi-functional dairy food.

    Science.gov (United States)

    Rutella, Giuseppina Sefora; Tagliazucchi, Davide; Solieri, Lisa

    2016-12-01

    In previous work, we demonstrated that two probiotic strains, namely Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, produce fermented milks with potent angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. Here, we tested these strains for the survivability and the release of antihypertensive and antioxidant peptides in yogurt fermentation and cold storage. For these purposes three yogurt batches were compared: one prepared using yogurt starters alone (Lactobacillus delbrueckii subspecies bulgaricus 1932 and Streptococcus thermophilus 99), and the remaining two containing either PRA205 or PRA331 in addition to yogurt starters. Despite the lower viable counts at the fermentation end compared to PRA331, PRA205 overcame PRA331 in survivability during refrigerated storage for 28 days, leading to viable counts (>10(8) CFU/g) higher than the minimum therapeutic threshold (10(6) CFU/g). Analyses of in vitro ACE-inhibitory and antioxidant activities of peptide fractions revealed that yogurt supplemented with PRA205 displays higher amounts of antihypertensive and antioxidant peptides than that produced with PRA331 at the end of fermentation and over storage. Two ACE-inhibitory peptides, Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP), were identified and quantified. This study demonstrated that L. casei PRA205 could be used as adjunct culture for producing bi-functional yogurt enriched in bioactive peptides and in viable cells, which bring health benefits to the host as probiotics. PMID:27554146

  6. Functional screening of antibiotic resistance genes from a representative metagenomic library of food fermenting microbiota.

    Science.gov (United States)

    Devirgiliis, Chiara; Zinno, Paola; Stirpe, Mariarita; Barile, Simona; Perozzi, Giuditta

    2014-01-01

    Lactic acid bacteria (LAB) represent the predominant microbiota in fermented foods. Foodborne LAB have received increasing attention as potential reservoir of antibiotic resistance (AR) determinants, which may be horizontally transferred to opportunistic pathogens. We have previously reported isolation of AR LAB from the raw ingredients of a fermented cheese, while AR genes could be detected in the final, marketed product only by PCR amplification, thus pointing at the need for more sensitive microbial isolation techniques. We turned therefore to construction of a metagenomic library containing microbial DNA extracted directly from the food matrix. To maximize yield and purity and to ensure that genomic complexity of the library was representative of the original bacterial population, we defined a suitable protocol for total DNA extraction from cheese which can also be applied to other lipid-rich foods. Functional library screening on different antibiotics allowed recovery of ampicillin and kanamycin resistant clones originating from Streptococcus salivarius subsp. thermophilus and Lactobacillus helveticus genomes. We report molecular characterization of the cloned inserts, which were fully sequenced and shown to confer AR phenotype to recipient bacteria. We also show that metagenomics can be applied to food microbiota to identify underrepresented species carrying specific genes of interest. PMID:25243126

  7. Inactivation of the DNA repair genes mutS, mutL or the anti-recombination gene mutS2 leads to activation of vitamin B1 biosynthesis genes.

    Directory of Open Access Journals (Sweden)

    Kenji Fukui

    Full Text Available Oxidative stress generates harmful reactive oxygen species (ROS that attack biomolecules including DNA. In living cells, there are several mechanisms for detoxifying ROS and repairing oxidatively-damaged DNA. In this study, transcriptomic analyses clarified that disruption of DNA repair genes mutS and mutL, or the anti-recombination gene mutS2, in Thermus thermophilus HB8, induces the biosynthesis pathway for vitamin B(1, which can serve as an ROS scavenger. In addition, disruption of mutS, mutL, or mutS2 resulted in an increased rate of oxidative stress-induced mutagenesis. Co-immunoprecipitation and pull-down experiments revealed previously-unknown interactions of MutS2 with MutS and MutL, indicating that these proteins cooperatively participate in the repair of oxidatively damaged DNA. These results suggested that bacterial cells sense the accumulation of oxidative DNA damage or absence of DNA repair activity, and signal the information to the transcriptional regulation machinery for an ROS-detoxifying system.

  8. Novel method based on chromogenic media for discrimination and selective enumeration of lactic acid bacteria in fermented milk products.

    Science.gov (United States)

    Galat, Anna; Dufresne, Jérôme; Combrisson, Jérôme; Thépaut, Jérôme; Boumghar-Bourtchai, Leyla; Boyer, Mickaël; Fourmestraux, Candice

    2016-05-01

    Microbial analyses of fermented milk products require selective methods to discriminate between close species simultaneously present in high amounts. A culture-based method combining novel chromogenic agar media and appropriate incubation conditions was developed to enumerate lactic acid bacteria (LAB) strains in fermented milk. M1 agar, containing two chromogenic substrates, allowed selective enumeration of Lactobacillus rhamnosus, two strains of Lactobacillus paracasei subsp. paracasei and Streptococcus salivarius subsp. thermophilus based on differential β-galactosidase and β-glucosidase activities. Depending on the presence of some or all of the above strains, M1 agar was supplemented with L-rhamnose or vancomycin and incubations were carried out at 37 °C or 44 °C to increase selectivity. A second agar medium, M2, containing one chromogenic substrates was used to selectively enumerate β-galactosidase producing Lactobacillus delbrueckii subsp. bulgaricus at 47 °C. By contrast with the usual culture media, the chromogenic method allowed unambiguous enumeration of each species, including discrimination between the two L. paracasei, up to 10(9) CFU/g of fermented milk. In addition, the relevance of the method was approved by enumerating reference ATCC strains in pure cultures and fermented milk product. The method could also be used for enumerations on non-Danone commercial fermented milk products containing strains different from those used in this study, showing versatility of the method. To our knowledge, this is the first description of a chromogenic culture method applied to selective enumeration of LAB.

  9. Crystal structures of Pseudomonas syringae pv. tomato DC3000 quinone oxidoreductase and its complex with NADPH

    Energy Technology Data Exchange (ETDEWEB)

    Pan, Xiaowei [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China); Graduate University of the Chinese Academy of Sciences, Beijing 100049 (China); Zhang, Hongmei; Gao, Yu; Li, Mei [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China); Chang, Wenrui, E-mail: wrchang@sun5.ibp.ac.cn [National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101 (China)

    2009-12-18

    Zeta-crystallin-like quinone oxidoreductase is NAD(P)H-dependent and catalyzes one-electron reduction of certain quinones to generate semiquinone. Here we present the crystal structures of zeta-crystallin-like quinone oxidoreductase from Pseudomonas syringae pv. tomato DC3000 (PtoQOR) and its complexes with NADPH determined at 2.4 and 2.01 A resolutions, respectively. PtoQOR forms as a homologous dimer, each monomer containing two domains. In the structure of the PtoQOR-NADPH complex, NADPH locates in the groove between the two domains. NADPH binding causes obvious conformational changes in the structure of PtoQOR. The putative substrate-binding site of PtoQOR is wider than that of Escherichia coli and Thermus thermophilus HB8. Activity assays show that PtoQOR has weak 1,4-benzoquinone catalytic activity, and very strong reduction activity towards large substrates such as 9,10-phenanthrenequinone. We propose a model to explain the conformational changes which take place during reduction reactions catalyzed by PtoQOR.

  10. Crystal structure of the 80S yeast ribosome.

    Science.gov (United States)

    Jenner, Lasse; Melnikov, Sergey; Garreau de Loubresse, Nicolas; Ben-Shem, Adam; Iskakova, Madina; Urzhumtsev, Alexandre; Meskauskas, Arturas; Dinman, Jonathan; Yusupova, Gulnara; Yusupov, Marat

    2012-12-01

    The first X-ray structure of the eukaryotic ribosome at 3.0Å resolution was determined using ribosomes isolated and crystallized from the yeast Saccharomyces cerevisiae (Ben-Shem A, Garreau de Loubresse N, Melnikov S, Jenner L, Yusupova G, Yusupov M: The structure of the eukaryotic ribosome at 3.0 A resolution. Science 2011, 334:1524-1529). This accomplishment was possible due to progress in yeast ribosome biochemistry as well as recent advances in crystallographic methods developed for structure determination of prokaryotic ribosomes isolated from Thermus thermophilus and Escherichia coli. In this review we will focus on the development of isolation procedures that allowed structure determination (both cryo-EM and X-ray crystallography) to be successful for the yeast S. cerevisiae. Additionally we will introduce a new nomenclature that facilitates comparison of ribosomes from different species and kingdoms of life. Finally we will discuss the impact of the yeast 80S ribosome crystal structure on perspectives for future investigations.

  11. Identification and quantification of a novel nitrate-reducing community in sediments of Suquia River basin along a nitrate gradient

    Energy Technology Data Exchange (ETDEWEB)

    Reyna, Luciana; Wunderlin, Daniel Alberto [Universidad Nacional de Cordoba-CONICET, Facultad de Ciencias Quimicas, Departamento de Bioquimica Clinica-CIBICI, Haya de la Torre y Medina Allende, Ciudad Universitaria, 5000 Cordoba (Argentina); Genti-Raimondi, Susana, E-mail: sgenti@fcq.unc.edu.a [Universidad Nacional de Cordoba-CONICET, Facultad de Ciencias Quimicas, Departamento de Bioquimica Clinica-CIBICI, Haya de la Torre y Medina Allende, Ciudad Universitaria, 5000 Cordoba (Argentina)

    2010-05-15

    We evaluated the molecular diversity of narG gene from Suquia River sediments to assess the impact of the nitrate concentration and water quality on the composition and structure of the nitrate-reducing bacterial community. To this aim, a library of one of the six monitoring stations corresponding to the highest nitrate concentration was constructed and 118 narG clones were screened. Nucleotide sequences were associated to narG gene from alpha-, beta-, delta-, gammaproteobacteria and Thermus thermophilus. Remarkably, 18% of clones contained narG genes with less than 69% similarity to narG sequences available in databases. Thus, indicating the presence of nitrate-reducing bacteria with novel narG genes, which were quantified by real-time PCR. Results show a variable number of narG copies, ranging from less than 1.0 x 10{sup 2} to 5.0 x 10{sup 4} copies per ng of DNA, which were associated with a decreased water quality index monitored along the basin at different times. - A novel narG community present in Suquia River sediments was quantified; values were in line with the water quality index.

  12. Change of lactose content after milk fermentation using various microbial cultures

    Directory of Open Access Journals (Sweden)

    Ivana Vinko

    2011-06-01

    Full Text Available The purpose of this study was to determine lactose and lactic acid content and acidity changes in typified milk prior to fermentation and in dairy products on 1st and 28th day of their storage at 8 °C in cold environment. In this study 5 different dairy products were observed: yogurt, extra lactose yogurt, bifido milk, sour cream and sour milk. The enzymatic method for determination of lactose has been used. The biggest change in lactose and lactic acid content, according to study results, has happened in the process of fermentation, as expected. About 16-20 % of lactose has been converted by mesophilus, while significantly bigger part (round 30 % of lactose to lactic acid has been converted by thermophilus. The smallest part of lactose conversion was performed by Bifidobacterium therapy culture (just 15 % after the first day and 19 % on 28th day of cold storage which is due to the greater adjustment period of Bifidobacterium in milk for lactose fermentation.

  13. Development of a non-dairy probiotic fermented product based on almond milk and inulin.

    Science.gov (United States)

    Bernat, Neus; Cháfer, Maite; Chiralt, Amparo; González-Martínez, Chelo

    2015-09-01

    A new fermented almond "milk" that combined the properties of both almonds and probiotics was considered to cover the current versatile health-promoting foods' demand. Almond milk fermentation with probiotic Lactobacillus reuteri and Streptococcus thermophilus was studied by using a Central Composite design with response surface methodology, and different factors (glucose, fructose, inulin and starters) were optimised to assure high probiotic survivals in the final product. The optimal formulation was physicochemically characterised throughout cold storage (28 days) and both probiotic survivals to in vitro digestion and proteolysis were quantified. Results showed that a high probiotic population (>10(7) cfu/mL) was obtained in the previously optimised almond milk throughout storage time, which correspond to the addition of 0.75 g of glucose/100 mL, 0.75 g of fructose/100 mL, 2 g/100 mL inulin and 6 mL/100 mL inoculum. Glucose was used as the main nutrient and the production of mannitol by L. reuteri was detected. The fermentation process increased the viscosity values, forming a weak gel structure, whose physical properties hardly changed. Probiotic bacteria notably survived (51%) to the in vitro digestion, surely related to the inulin presence, which would add value to the developed product by enhancing the potential health benefits of its consumption.

  14. Functional screening of antibiotic resistance genes from a representative metagenomic library of food fermenting microbiota.

    Science.gov (United States)

    Devirgiliis, Chiara; Zinno, Paola; Stirpe, Mariarita; Barile, Simona; Perozzi, Giuditta

    2014-01-01

    Lactic acid bacteria (LAB) represent the predominant microbiota in fermented foods. Foodborne LAB have received increasing attention as potential reservoir of antibiotic resistance (AR) determinants, which may be horizontally transferred to opportunistic pathogens. We have previously reported isolation of AR LAB from the raw ingredients of a fermented cheese, while AR genes could be detected in the final, marketed product only by PCR amplification, thus pointing at the need for more sensitive microbial isolation techniques. We turned therefore to construction of a metagenomic library containing microbial DNA extracted directly from the food matrix. To maximize yield and purity and to ensure that genomic complexity of the library was representative of the original bacterial population, we defined a suitable protocol for total DNA extraction from cheese which can also be applied to other lipid-rich foods. Functional library screening on different antibiotics allowed recovery of ampicillin and kanamycin resistant clones originating from Streptococcus salivarius subsp. thermophilus and Lactobacillus helveticus genomes. We report molecular characterization of the cloned inserts, which were fully sequenced and shown to confer AR phenotype to recipient bacteria. We also show that metagenomics can be applied to food microbiota to identify underrepresented species carrying specific genes of interest.

  15. Microflora of Feta cheese from four Greek manufacturers.

    Science.gov (United States)

    Rantsiou, Kalliopi; Urso, Rosalinda; Dolci, Paola; Comi, Giuseppe; Cocolin, Luca

    2008-08-15

    The components of the microflora of four Feta cheeses, produced by different Greek manufacturers, were determined by culture dependent and independent techniques. Isolates from microbiological media were first grouped by Polymerase Chain Reaction-Denaturing Gradient Gel Electrophoresis (PCR-DGGE) and then representatives of each DGGE group were sequenced for identification purposes. DNA and RNA, extracted directly from the cheese, were subjected to PCR-DGGE. Moreover, Feta cheeses were subjected to FISH analysis in order to identify viable bacterial populations. The microbial ecology, as represented by the Lactic Acid Bacteria (LAB) and yeast populations, was different for the four cheeses. The main LAB species isolated were Lactobacillus plantarum, Lactobacillus brevis, Lactobacillus coryniformis and Lactobacillus fermentum. However, some inconsistencies were observed between the results obtained with the culture dependent and the culture independent approach. In the case of the yeasts, the results obtained by PCR-DGGE compared very well with those obtained by the conventional microbiological analysis and the main species found were Kluyveromyces lactis, Pichia fermentans and C. zeylanoides. FISH analysis highlighted viable but not culturable populations of Streptococcus thermophilus and Lactococcus spp. RAPD-PCR performed on the L. plantarum isolates revealed a cheese specific distribution and a temperature dependent clustering. PMID:18555549

  16. SENSORY EVALUATION AND SURVIVAL OF PROBIOTICS IN MODIFIED BANANA FLOUR YOGHURT DURING STORAGE [Evaluasi Sensori dan Sintasan Probiotik dalam Yoghurt Tepung Pisang Modifikasi selama Penyimpanan

    Directory of Open Access Journals (Sweden)

    Widaningrum3

    2013-06-01

    Full Text Available Modified uli banana flour (MUBF rich in resistant starch as prebiotic source was formulated in yoghurt making to substitute skim milk at various concentrations i.e. 40, 50, 60, and 70%. Yoghurts were prepared using Lactobacillus bulgaricus and Streptococcus thermophilus as the starter cultures. The objective of this study was to determine the maximum MUBF concentration to produce yoghurt with good sensory quality and good number of surviving probiotics (Bifidobacterium bifidum and Lactobacillus plantarum BSL in non-pasteurized and pasteurized (90ºC, 30 minutes MUBF yoghurt during 4 week of storage at 10ºC. Hedonic rating and ranking test on yoghurt attributes (aroma, taste, consistency, texture, color and overall preference performed by 35 untrained panelists showed that yoghurt produced with 70% MUBF substitution achieved good preferences in all of the attributes ranging from neutral to like. The pH value of the product decreased with increase in MUBF concentration. In addition, titratable acidity (TA, expressed as % lactic acid, increased. Survival of both probiotics in 70% MUBF yoghurt either in the pasteurized synbiotic or non-pasteurized yoghurts were still relatively high at week 4. The yoghurt still contains 108 CFU/ml of lactic acid bacteria, although this was a log decrease from the initial count. Therefore, the MUBF yoghurt was promising as synbiotic yoghurt based on the probiotic counts throughout 4 week of storage, which was higher than the minimum level recommended (106 CFU/ml to provide the beneficial effect.

  17. Characterization of goat milk and potentially symbiotic non-fat yogurt

    Directory of Open Access Journals (Sweden)

    Noelia Fernanda Paz

    2014-09-01

    Full Text Available Combining prebiotics and probiotic microorganisms improve quality in the formulation of foods. In this paper, the characteristics of goat milk and symbiotic yogurt were studied. Raw goat milk was analyzed and the skimming process was optimized. For the formulation of a potentially non-fat symbiotic yogurt made with skimmed goat milk, inulin, gelatin, sugar, and Streptococcus salivarius subsp. thermophilus, Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus casei subsp. rhamnoshus. Chemical characteristics, acceptability, and viability of lactic acid bacteria and probiotic culture were assessed. The protein and fat content of the raw milk was 2.90 and 3.56 g/100 mL, respectively. The optimum skimming process was obtained at 9,800 rpm and 4 °C for 15 minutes. The product formulated had a protein and fat content of 4.04 to 0.04 g/100 mL, good sensory properties, and acceptability of 95%. The lactic bacteria count was 9 × 10(7 CFU mL- 1, and probiotic culture count was higher than 1 × 10(6 CFU mL- 1, which guarantees their effect and capacity to survive in the digestive tract and spread in the intestine. The yogurt was stable during the 21 days of storage. Therefore, this study shows that goat milk yogurt is an adequate delivery vehicle of the probiotic culture L. casei and inulin.

  18. Development of a non-dairy probiotic fermented product based on almond milk and inulin.

    Science.gov (United States)

    Bernat, Neus; Cháfer, Maite; Chiralt, Amparo; González-Martínez, Chelo

    2015-09-01

    A new fermented almond "milk" that combined the properties of both almonds and probiotics was considered to cover the current versatile health-promoting foods' demand. Almond milk fermentation with probiotic Lactobacillus reuteri and Streptococcus thermophilus was studied by using a Central Composite design with response surface methodology, and different factors (glucose, fructose, inulin and starters) were optimised to assure high probiotic survivals in the final product. The optimal formulation was physicochemically characterised throughout cold storage (28 days) and both probiotic survivals to in vitro digestion and proteolysis were quantified. Results showed that a high probiotic population (>10(7) cfu/mL) was obtained in the previously optimised almond milk throughout storage time, which correspond to the addition of 0.75 g of glucose/100 mL, 0.75 g of fructose/100 mL, 2 g/100 mL inulin and 6 mL/100 mL inoculum. Glucose was used as the main nutrient and the production of mannitol by L. reuteri was detected. The fermentation process increased the viscosity values, forming a weak gel structure, whose physical properties hardly changed. Probiotic bacteria notably survived (51%) to the in vitro digestion, surely related to the inulin presence, which would add value to the developed product by enhancing the potential health benefits of its consumption. PMID:25028153

  19. Optimization Technology of the Production of Lactobacillus casei Yoghurt%干酪乳杆菌发酵酸乳生产工艺优化

    Institute of Scientific and Technical Information of China (English)

    张杰; 郗宏波; 苏贺靖; 王艳萍

    2013-01-01

      以从日本传统酸奶中分离选育出的优质干酪乳杆菌作为试验菌种。通过单因素试验和正交试验确定最佳发酵工艺条件。结果表明:嗜热链球菌、保加利亚乳杆菌、干酪乳杆菌配比为1∶1∶2;接种量为5%;发酵温度37℃;乳固体含量12%为最佳参数组合。%Lactobacillus casei which was separated and identified from the Japanese traditional yoghurt was taken as the test strains. By the single factor experiments and orthogonal tests to determine the best fermentation conditions, the results showed that the best fermentation parameter combination was the ratio of Streptococcus thermophilus:Lactobacillus bulgaricus:Lactobacillus casei of 1∶1∶2, inoculated quantity of 5%, fermentation temperature of 37℃, milk solid content of 12%.

  20. Effect of the vitamin B12-binding protein haptocorrin present in human milk on a panel of commensal and pathogenic bacteria

    Directory of Open Access Journals (Sweden)

    Nexø Ebba

    2011-06-01

    Full Text Available Abstract Background Haptocorrin is a vitamin B12-binding protein present in high amounts in different body fluids including human milk. Haptocorrin has previously been shown to inhibit the growth of specific E. coli strains, and the aim of the present study was to elucidate whether the antibacterial properties of this protein may exert a general defense against pathogens and/or affect the composition of the developing microbiota in the gastrointestinal tracts of breastfed infants. Findings The present work was the first systematic study of the effect of haptocorrin on bacterial growth, and included 34 commensal and pathogenic bacteria to which infants are likely to be exposed. Well-diffusion assays addressing antibacterial effects were performed with human milk, haptocorrin-free human milk, porcine holo-haptocorrin (saturated with B-12 and human apo-haptocorrin (unsaturated. Human milk inhibited the growth of S. thermophilus and the pathogenic strains L. monocytogenes LO28, L. monocytogenes 4446 and L. monocytogenes 7291, but the inhibition could not be ascribed to haptocorrin. Human apo-haptocorrin inhibited the growth of only a single bacterial strain (Bifidobacterium breve, while porcine holo-haptocorrin did not show any inhibitory effect. Conclusions Our results suggest that haptocorrin does not have a general antibacterial activity, and thereby contradict the existing hypothesis implicating such an effect. The study contributes to the knowledge on the potential impact of breastfeeding on the establishment of a healthy microbiota in infants.

  1. Thermodynamic assessment of domain-domain interactions and in vitro activities of mesophilic and thermophilic ribosome recycling factors.

    Science.gov (United States)

    Uchiyama, Susumu; Ohshima, Atsushi; Yoshida, Takuya; Ohkubo, Tadayasu; Kobayashi, Yuji

    2013-07-01

    Ribosome recycling factor (RRF) is essential for bacterial growth. Structural studies revealed that RRF consists of two domains connected by two short polypeptides at the hinge region. Here, we evaluated the intrinsic stabilities (ΔG*s) of the two domains and the free energy of the domain-domain interactions (ΔG(int)) for mesophilic RRF (RRF from Escherichia coli, EcRRF) and thermophilic RRF (RRF from Thermus thermophilus, TtRRF) by using differential scanning calorimetry and circular dichroic spectroscopy. Despite single endothermic peaks, a higher than unity value for the ratio of calorimetric enthalpy to van't Hoff enthalpy of the unfolding indicated the presence of unfolding intermediates for both RRFs. Deconvolution analysis based on statistical thermodynamics employing multiple states of the unfolding process with domain-domain interactions allowed us to determine ΔG*s of each domain and ΔG(int). The results indicated that domain I has a higher unfolding temperature than domain II and that it stabilizes domain II through ΔG(int), giving rise to an apparent single peak of unfolding. Interestingly, the estimated ΔG(int) values of 6.28 kJ/mol for EcRRF and 26.28 kJ/mol for TtRRF reflect the observation that only EcRRF has recycling activity at ambient temperature. Our present study suggests the importance of a moderate ΔG(int) for biological activity of multidomain proteins.

  2. 2',3'-Cyclic nucleotide 3'-phosphodiesterase: a novel RNA-binding protein that inhibits protein synthesis.

    Science.gov (United States)

    Gravel, Michel; Robert, Francis; Kottis, Vicky; Gallouzi, Imed-Eddine; Pelletier, Jerry; Braun, Peter E

    2009-04-01

    2',3'-Cyclic nucleotide 3'-phosphodiesterase (CNP) is one of the earliest myelin-related proteins to be specifically expressed in differentiating oligodendrocytes (ODCs) in the central nervous system (CNS) and is implicated in myelin biogenesis. CNP possesses an in vitro enzymatic activity, whose in vivo relevance remains to be defined, because substrates with 2',3,-cyclic termini have not yet been identified. To characterize CNP function better, we previously determined the structure of the CNP catalytic domain by NMR. Interestingly, the structure is remarkably similar to the plant cyclic nucleotide phosphodiesterase (CPDase) from A. thaliana and the bacterial 2'-5' RNA ligase from T. thermophilus, which are known to play roles in RNA metabolism. Here we show that CNP is an RNA-binding protein. Furthermore, by using precipitation analyses, we demonstrate that CNP associates with poly(A)(+) mRNAs in vivo and suppresses translation in vitro in a dose-dependent manner. With SELEX, we isolated RNA aptamers that can suppress the inhibitory effect of CNP on translation. We also demonstrate that CNP1 can bridge an association between tubulin and RNA. These results suggest that CNP1 may regulate expression of mRNAs in ODCs of the CNS. PMID:19021295

  3. METABOLIC AND BIOCHEMICAL CHARACTERISTICS OF PROBIOTIC CULTURE IN MILK SUPPLEMENTED WITH RYE FLAKES AND MALT EXTRACT

    Directory of Open Access Journals (Sweden)

    Elena Bărăscu

    2010-01-01

    Full Text Available Rye flakes and malt extract were added to milk in order to stimulate growth and fermentative activity of probioticbacteria and to obtain a probiotic product with pleasant sensory attributes. Probiotic culture used in this study containsbifidobacteria, Lb. acidophilus, Lactobacilus lactis and Streptococcus thermophilus.Rye flakes have a stimulating effect more pronounced than malt extract on acidification capacity of the probioticculture, and to achieve an increase of the milk acidity of 7g lactic acid /dm3 (in 6h at 39oC the two ingredients must beadded in concentration of 2% and, respectively, 0.2%..The probiotic culture reach the greatest proteolytic activity when rye flakes are added in the proportion of 3% andmalt extract in the proportion of 0.1% and the amino acids released rate was 764.6 μg%. The lactose bioconversionrate was greater in the milk supplemented with rye flakes 3% and malt extract 0.1% and residual lactose was 3.84%.

  4. Preparation of lactobacillus beverage containing euryale ferox juice%芡实乳酸菌饮料的研制*

    Institute of Scientific and Technical Information of China (English)

    董基; 梁巧荣; 黄志明; 魏义; 刘钊

    2013-01-01

    Euryale ferox was used as main material, mixed strains of lactobacillus bulgaricus and Streptococcus ther-mophilus (1∶1) were used as starter, lactobacillus beverage containing Euryale ferox juice was prepared by fermen-tation. The optimum processing conditions were determined through orthogonal test, that is, fermentation temperature 42 ℃, fermentation time 36 h, inoculation amount of starter 5 g/100g, after fermentation time 24 h. Then 0.1 g/100g CMC and 0.1 g/100g xanthan gum were added to beverage as stabilizer after fermentation.%  以芡实为主要原料,保加利亚乳杆菌和嗜热链球菌混合菌种(1∶1)为发酵剂,经发酵芡实汁制备乳酸菌饮料。通过正交试验确定最优发酵工艺条件:发酵温度42℃,发酵时间36 h,发酵剂接种量5 g/100g,后发酵时间24 h。发酵结束后,在饮料中添加0.1 g/100g羧甲基纤维素钠(CMC)与0.1 g/100g黄原胶作为稳定剂。

  5. Isolation and identification of lactic acid bacteria from fermented milk and selection of excellent strains%发酵乳中乳酸菌的分离鉴定及优良菌株的筛选

    Institute of Scientific and Technical Information of China (English)

    刘芳; 曹新志; 王凤娇; 周龙伟

    2012-01-01

    从市售发酵乳中分离鉴定出保加利亚乳杆菌和嗜热链球菌,对其进行凝乳情况的考察和产酸能力、后酸化能力、产香能力、产黏能力、分解蛋白质能力及保健功能的测试,各筛出一株优良菌株,拟定在后续试验中按一定比例作进一步的应用研究,以期制备高效复合菌株发酵剂。本试验可为实际生产中发酵乳优良菌株的筛选提供理论依据。%Lactobacillus bulgaricus and Streptococcus thermophilus extracted from commercial fermented milk are to be identified. Upon the observation of their renneting and testing of their potentials for acid, post-acidification, aroma-production, mucosity-production and proteolysis and health functions, an excellent strain is to be selected respectively. It is postulated here that application research will be conducted in follow-up experiments in certain proportions for the preparation of efficient compound strain leavens. The experiment is able to lay the theoretical foundation for the selection of excellent strains of fermented milk in physical production.

  6. Antioxidant Activity of Peptides from Fermented Milk with Mix Culture of Lactic Acid Bacteria and Yeast

    Directory of Open Access Journals (Sweden)

    Yun Li

    2015-02-01

    Full Text Available The aim of the present study is to investigate the production of antioxidant peptides during milk fermentation with co-culture of Lactic Acid Bacteria (LAB and yeast. Five LAB strains, previously screened with higher hydrolysis activity and Debaryomyces hansenii H2 which isolated from Tibet kefir were used in the study. The peptides separated from fermented milk were analysed antioxidant activity with DPPH radical scavenging, hydroxyl radical scavenging, chelation of metal ions and reducing power assays. The growth of Streptococcus. thermophilus Lactobacillus. delbrueckii ssp. bulgaricus and Lactococcus. lactis was enhanced with co-cultures and L. acidophilus was inhibited in co-culture with yeast. In co-culture with yeast, a significant decrease of the acidity was observed among all the fermentation and the pH reached higher values than in single LAB cultures. Except for L.delbrueckii ssp. bulgaricus, there was no significant difference of protein hydrolysis with other test LAB strains between co-culture and single culture. The co-incubation of LAB with the yeast developed a stronger antioxidant activity in DPPH radical and hydroxyl radical scavenging and no significant (p>0.05 difference in chelation of metal ions. The reducing power of L.delbrueckii ssp. bulgaricus and L. helveticus in co-culture was significant higher than those of single culture.

  7. The Impact of Proteolytic Pork Hydrolysate on Microbial, Flavor and Free Amino Acids Compounds of Yogurt

    Science.gov (United States)

    Lin, Jinzhong; Hua, Baozhen; Xu, Zhiping; Li, Sha; Ma, Chengjie

    2016-01-01

    The aim of this study was to investigate the influence of proteolytic pork hydrolysate (PPH) on yoghurt production by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Fresh lean pork was cut into pieces and mixed with deionized water and dealt with protease, then the resulting PPH was added to milk to investigate the effects of PPH on yoghurt production. The fermentation time, the viable cell counts, the flavor, free amino acids compounds, and sensory evaluation of yoghurt were evaluated. These results showed that PPH significantly stimulated the growth and acidification of the both bacterial strains. When the content of PPH reached 5% (w/w), the increased acidifying rate occurred, which the fermentation time was one hour less than that of the control, a time saving of up to 20% compared with the control. The viable cell counts, the total free amino acids, and the scores of taste, flavor and overall acceptability in PPH-supplemented yoghurt were higher than the control. Furthermore, the contents of some characteristic flavor compounds including acids, alcohols, aldehydes, ketones and esters were richer than the control. We concluded that the constituents of PPH such as small peptide, vitamins, and minerals together to play the stimulatory roles and result in beneficial effect for the yoghurt starter cultures growth. PMID:27621698

  8. 乳酸发酵澄清猕猴桃汁的研制%Development of Clarifing Yangtao Juice Lactic Acid Beverage

    Institute of Scientific and Technical Information of China (English)

    郜宗茂; 王永斌

    2001-01-01

    Seed culture solution was inoculated with lactobacillus bulgaricus, lactobacillus acidophilus, lactobacillus thermophilus in quantity of 3%,and pH value of solution and the suitable solution for bolting culture were determined. On this base,technological process of lactic acid fermenting yangtao was established through measuring variety of bacterial quantity and selecting optimal production strain.%采用活化的保加利亚乳杆菌、嗜热链球菌、嗜酸乳杆菌按接种量3%接种于不同的种子培养液中,通过测定并分析pH值的变化,筛选合适的培养液。在此基础上,通过测定菌数的变化,选定适宜的复合菌种,确定了乳酸发酵澄清猕猴桃汁的工艺流程。

  9. Efectos beneficiosos en niños sanos del consumo de un producto lácteo que contiene dos cepas probióticas: Lactobacillus coryniformis CECT5711 y Lactobacillus gasseri CECT5714 Beneficial effects of consumption of a dairy product containing two probiotic strains: Lactobacillus coryniformis CECT5711 and Lactobacillus gasseri CECT5714 in healthy children

    Directory of Open Access Journals (Sweden)

    F. Lara-Villoslada

    2007-08-01

    Full Text Available Objetivo o antecedente: En las últimas décadas la manipulación de la microbiota intestinal mediante el uso de probióticos ha adquirido un gran interés en el tratamiento y prevención de determinadas patologías infantiles. Además actualmente existen multitud de estudios que demuestran que las bacterias probióticas podrían tener un importante papel en el desarrollo del sistema inmunitario. Estudios recientes sugieren que dos cepas probióticas, Lactobacillus coryniformis CECT5711 y Lactobacillus gasseri CECT5714 mejoran la función intestinal de adultos sanos y potencian la respuesta inmunitaria. Dado que son muy pocos los estudios que analizan el papel de los probióticos en niños sanos, principales consumidores de estos productos, el objetivo del presente trabajo fue analizar los efectos de la administración conjunta de estas dos cepas probióticas en un producto lácteo fermentado en niños sanos. Intervenciones: Se reclutaron 30 niños de entre 3 y 12 años sin patología gastrointestinal conocida. Además de su dieta habitual, durante las 3 primeras semanas los niños recibieron 200 ml de un yogurt convencional que contenía Lactobacillus bulgaricus y Streptococcus thermophilus. A continuación este yogurt se sustituyó por 80 ml de un producto probiótico (Max Defensas® Puleva FOOD S.L. que contenía la misma cantidad de Streptococcus thermophilus y donde el L. bulgaricus fue substituido por la misma cantidad de una mezcla de las bacterias probióticas objeto del estudio: L. coryniformis CECT5711 y L. gasseri CECT5714. Se tomaron muestras de heces y de saliva al comienzo del estudio, a las 3 semanas y al finalizar el estudio. Durante estas 6 semanas los niños no tomaron otro yogurt o probiótico que no fueran los citados anteriormente. Se analizó la microbiotafecal de los niños, la toxicidad de las aguas fecales y la capacidad de éstas para inhibir la adhesión de Salmonella cholerasusis ssp. cholerasuis a mucinas

  10. 弱后酸化酸奶发酵剂菌株洲的筛选及复配%Screening and studying the strains of weak postacidification in yoghurt

    Institute of Scientific and Technical Information of China (English)

    刘宁宁; 郭红敏; 崔文静; 葛春美

    2012-01-01

    以典型的酸奶发酵剂菌株:德氏乳杆菌保加利亚亚种和嗜热链球菌为实验材料,进行了单菌株产酸特性的测定,并研究了后酸程度不同的球杆菌按照不同比例混合后产酸特性。结果显示:筛选到2株后酸化弱的菌株,当这两株菌的复配比例为100:1时,后酸程度较弱,在42℃放置24h酸度仅上升了10°T,与科汉森的酸奶发酵剂YF—L822相比后酸更弱。%In this paper, the typical yogurt starter strains: Lactobacillus delbrueckii subsp. Bulgaricus and Streptococcus thermophilus were experimental materials. The characteristics of acid producing of single strain and mixing according to different ratio were determined. Results indicated that two trains with weak postacidification were obtained. When the mixing ratio of the two strains was 100:1, its acidity rose only 10 °T after 24 h at 42 ℃. Compared to YF-L822 starter culture from Hansen Corporation, it produced a little postacidification.

  11. Applicability of a Lactobacillus amylovorus strain as co-culture for natural folate bio-enrichment of fermented milk.

    Science.gov (United States)

    Laiño, Jonathan Emiliano; Juarez del Valle, Marianela; Savoy de Giori, Graciela; LeBlanc, Jean Guy Joseph

    2014-11-17

    The ability of 55 strains from different Lactobacillus species to produce folate was investigated. In order to evaluate folic acid productivity, lactobacilli were cultivated in the folate-free culture medium (FACM). Most of the tested strains needed folate for growth. The production and the extent of vitamin accumulation were distinctive features of individual strains. Lactobacillus amylovorus CRL887 was selected for further studies because of its ability to produce significantly higher concentrations of vitamin (81.2 ± 5.4 μg/L). The safety of this newly identified folate producing strain was evaluated through healthy experimental mice. No bacterial translocation was detected in liver and spleen after consumption of CRL887 during 7 days and no undesirable side effects were observed in the animals that received this strain. This strain in co-culture with previously selected folate producing starter cultures (Lactobacillus bulgaricus CRL871, and Streptococcus thermophilus CRL803 and CRL415) yielded a yogurt containing high folate concentrations (263.1 ± 2.4 μg/L); a single portion of which would provide 15% of the recommended dietary allowance. This is the first report where a Lactobacillus amylovorus strain was successfully used as co-culture for natural folate bio-enrichment of fermented milk.

  12. 嗜酸乳杆菌与传统混合菌混合培养条件的优化%Optimizing Conditions for Mixed Culture of Lactobacillus Acidophilus and Traditional Yoghourt Strains

    Institute of Scientific and Technical Information of China (English)

    张树海; 吴子健; 帖航; 蔡平平

    2013-01-01

    In this study Lactobacillus acidophilus (LA-5) is co-cultured with the two traditional yoghourt strains-Streptococcus thermophilus(St) and Lactobacillus bulgaricus(Lb). Through using the orthogonal test, the optimal fermentation condition is obtained:optimal inoculation mixed proportion of La to St&Lb 1∶4;the total adding strain 0.3 g/L, the sugar-adding amount 60 g/L;cultural temperature 41℃. The viscosity, acidity, pH value and LA-5 viablecountoffinalproductsashadbeenjudgedduring fementation,post-fermentationandevenstorageperiod.%  研究嗜酸乳杆菌(LA-5)与嗜热链球菌、德氏乳杆菌保加利亚亚种两种传统菌株混合发酵制备益生菌酸乳,正交试验结果得到最适生长条件为:菌种配比为1∶4、接种量0.3 g/L、加糖量60 g/L、培养温度41℃;同时测定所得益生菌酸乳的黏度、酸度、pH以及酸乳中嗜酸乳杆菌活菌数等在发酵以及后酵乃至贮存期间的变化情况.

  13. Physiological study of Lactobacillus delbrueckii subsp. bulgaricus strains in a novel chemically defined medium.

    Science.gov (United States)

    Chervaux, C; Ehrlich, S D; Maguin, E

    2000-12-01

    We developed a chemically defined medium called milieu proche du lait (MPL), in which 22 Lactobacillus delbrueckii subsp. bulgaricus (L. bulgaricus) strains exhibited growth rates ranging from 0.55 to 1 h(-1). MPL can also be used for cultivation of other lactobacilli and Streptococcus thermophilus. The growth characteristics of L. bulgaricus in MPL containing different carbon sources were determined, including an initial characterization of the phosphotransferase system transporters involved. For the 22 tested strains, growth on lactose was faster than on glucose, mannose, and fructose. Lactose concentrations below 0.4% were limiting for growth. We isolated 2-deoxyglucose-resistant mutants from strains CNRZ397 and ATCC 11842. CNRZ397-derived mutants were all deficient for glucose, fructose, and mannose utilization, indicating that these three sugars are probably transported via a unique mannose-specific-enzyme-II-like transporter. In contrast, mutants of ATCC 11842 exhibited diverse phenotypes, suggesting that multiple transporters may exist in that strain. We also developed a protein labeling method and verified that exopolysaccharide production and phage infection can occur in MPL. The MPL medium should thus be useful in conducting physiological studies of L. bulgaricus and other lactic acid bacteria under well controlled nutritional conditions.

  14. Complete sequencing and pan-genomic analysis of Lactobacillus delbrueckii subsp. bulgaricus reveal its genetic basis for industrial yogurt production.

    Science.gov (United States)

    Hao, Pei; Zheng, Huajun; Yu, Yao; Ding, Guohui; Gu, Wenyi; Chen, Shuting; Yu, Zhonghao; Ren, Shuangxi; Oda, Munehiro; Konno, Tomonobu; Wang, Shengyue; Li, Xuan; Ji, Zai-Si; Zhao, Guoping

    2011-01-17

    Lactobacillus delbrueckii subsp. bulgaricus (Lb. bulgaricus) is an important species of Lactic Acid Bacteria (LAB) used for cheese and yogurt fermentation. The genome of Lb. bulgaricus 2038, an industrial strain mainly used for yogurt production, was completely sequenced and compared against the other two ATCC collection strains of the same subspecies. Specific physiological properties of strain 2038, such as lysine biosynthesis, formate production, aspartate-related carbon-skeleton intermediate metabolism, unique EPS synthesis and efficient DNA restriction/modification systems, are all different from those of the collection strains that might benefit the industrial production of yogurt. Other common features shared by Lb. bulgaricus strains, such as efficient protocooperation with Streptococcus thermophilus and lactate production as well as well-equipped stress tolerance mechanisms may account for it being selected originally for yogurt fermentation industry. Multiple lines of evidence suggested that Lb. bulgaricus 2038 was genetically closer to the common ancestor of the subspecies than the other two sequenced collection strains, probably due to a strict industrial maintenance process for strain 2038 that might have halted its genome decay and sustained a gene network suitable for large scale yogurt production.

  15. Immunostimulatory oligonucleotide, CpG-like motif exists in Lactobacillus delbrueckii ssp. bulgaricus NIAI B6.

    Science.gov (United States)

    Kitazawa, Haruki; Watanabe, Hiroshi; Shimosato, Takeshi; Kawai, Yasushi; Itoh, Takatoshi; Saito, Tadao

    2003-08-15

    The present study was conducted to find an immunostimulatory oligonucleotide derived from yogurt starter cultures. The chromosomal DNA was purified from nine strains of Lactobacillus delbrueckii ssp. bulgaricus and six strains of Streptococcus thermophilus. An immunostimulatory ability of the DNA was examined in a proliferation of peyer's patch and splenic B cells. Only the DNA from L. bulgaricus NIAI B6 induced a significant proliferation of both cells. When the DNA was cloned and amplified using PCR, the mitogenic activities to B cells were significantly increased by 13 of 135 DNA clones. Ten homologous nucleotide sequences were found as possible oligonucleotide sequences of mitogens, and were then chemically synthesized (sOL-LB1 to sOL-LB10). One CpG-like motif (sOL-LB7; 5'-CGGCACGCTCACGATTCTTG-3') was identified as an immunostimulatory oligonucleotide, but it did not contain palindromic CpG structure known as a B cell-specific mitogen. The sOL-LB7 substantially bound to B cells and increased the CD69 positive cells in peyer's patch cells. This study demonstrated that L. bulgaricus NIAI B6 was a good candidate of a starter culture for the production of new functional foods, "Bio-Defense Foods".

  16. CHOLESTEROL ASSIMILATION BY COMMERCIAL YOGHURT STARTER CULTURES

    Directory of Open Access Journals (Sweden)

    Małgorzata Ziarno

    2007-03-01

    Full Text Available The ability to in vitro cholesterol level reduction in laboratory media has been shown for numerous strains of lactic acid bacteria, but not for all strains of lactic bacteria used in the dairy industry. The aim of this work was the determination of the ability of selected thermophilic lactic acid bacteria to cholesterol assimilation during 24 h culture in MRS broth. Commercial starter cultures showed various ability to cholesterol assimilation from laboratory medium. In case of starter cultures used for production of traditional yoghurt, consisting of Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, the quantity of assimilated cholesterol did not exceed 27% of its initial contents (0.7 g in 1 dm3. Starter cultures used for bioyoghurt production, containing also probiotic strains (came from Lactobacillus acidophilus species or Bifidobacterium genus assimilated from almost 18% to over 38% of cholesterol. For one monoculture of Lb. acidophilus, cholesterol assimilation ability of 49-55% was observed, despite that the number of bacterial cells in this culture was not different from number of bacteria in other cultures.

  17. Complete sequencing and pan-genomic analysis of Lactobacillus delbrueckii subsp. bulgaricus reveal its genetic basis for industrial yogurt production.

    Directory of Open Access Journals (Sweden)

    Pei Hao

    Full Text Available Lactobacillus delbrueckii subsp. bulgaricus (Lb. bulgaricus is an important species of Lactic Acid Bacteria (LAB used for cheese and yogurt fermentation. The genome of Lb. bulgaricus 2038, an industrial strain mainly used for yogurt production, was completely sequenced and compared against the other two ATCC collection strains of the same subspecies. Specific physiological properties of strain 2038, such as lysine biosynthesis, formate production, aspartate-related carbon-skeleton intermediate metabolism, unique EPS synthesis and efficient DNA restriction/modification systems, are all different from those of the collection strains that might benefit the industrial production of yogurt. Other common features shared by Lb. bulgaricus strains, such as efficient protocooperation with Streptococcus thermophilus and lactate production as well as well-equipped stress tolerance mechanisms may account for it being selected originally for yogurt fermentation industry. Multiple lines of evidence suggested that Lb. bulgaricus 2038 was genetically closer to the common ancestor of the subspecies than the other two sequenced collection strains, probably due to a strict industrial maintenance process for strain 2038 that might have halted its genome decay and sustained a gene network suitable for large scale yogurt production.

  18. Evaluation of anti-bacterial and anti-oxidant potential of andrographolide and echiodinin isolated from callus culture of Andrographis paniculata Nees

    Institute of Scientific and Technical Information of China (English)

    Mohammed Arifullah; Nima Dandu Namsa; Manabendra Mandal; Kishore Kumar Chiruvella; Paritala Vikrama; Ghanta Rama Gopal

    2013-01-01

    Objective: To evaluate the anti-bacterial and anti-oxidant activity of andrographolide (AND) and echiodinin (ECH) of Andrographis paniculata. Methods:In this study, an attempt has been made to demonstrate the anti-microbial and anti-oxidant activity of isolated AND and ECH by broth micro-dilution method and 2,2-diphenyl-2-picryl-hydrazyl (DPPH) assay, respectively. Structure elucidation was determined by electro-spray ionization-MSD, NMR (1H and 13C) and IR spectra. Results: AND was effective against most of the strains tested including Mycobacteriumsmegmatis, showing broad spectrum of growth inhibition activity with Minimum inhibitory concentration values against Staphylococcus aureus (100 µg/mL), Streptococcus thermophilus (350 µg/mL) Bacillus subtilis (100 µg/mL), Escherichia coli (50 µg/mL), Mycobacterium smegmatis (200 µg/mL), Klebsiella pneumonia (100 µg/mL), and Pseudomonas aeruginosa (200 µg/mL). ECH showed specific anti-bacterial activity against Staphylococcus aureus, Escherichia coli, Bacillus subtilis and Pseudomonas aeruginosa at a concentration higher than 225 µg/mL. Both AND and ECH were not effective against the two yeast strains, Candida albicans and Saccharomyces cerevisiae tested in this study. Conclusion:This preliminary study showed promising anti-bacterial activity and moderate free radical scavenging activity of AND and ECH, and it may provide the scientific rationale for its popular folklore medicines.

  19. Preparation of Almond Sour Milk%杏仁酸乳的研制

    Institute of Scientific and Technical Information of China (English)

    权美平

    2013-01-01

    Taking bitter almond and fresh milk as raw materials, the optimum parameters were obtained with main technology including peeling off the skin, debittering process, emulsification operation and fermentation. The results showed that, the optimum parameters of almond sour milk were as follows: the ratio of almond milk to fresh milk 30:70, inoculum size 5%, fermentation time 5 h, lactobacillus bulgaricus I streptococcus thermophilus ratio 1:1 and fermentation temperature 42 ℃. The final product under the optimum conditions showed high sensory qulity.%以苦杏仁为原料和鲜牛乳为原料,经脱皮、去苦、乳化和发酵等主要工艺步骤获得杏仁乳生产最佳技术参数.结果表明,杏仁酸乳发酵工艺的条件为:杏仁乳:牛乳比例为30∶70,接种量5%,发酵时间为5h,按照以保加利亚乳杆菌、嗜热链球菌(1∶1)接种,42℃条件下发酵,可得品质优良的杏仁酸乳.

  20. Suitability of a new mixed-strain starter for manufacturing uncooked raw ewe's milk cheeses.

    Science.gov (United States)

    Feutry, Fabienne; Torre, Paloma; Arana, Ines; Garcia, Susana; Pérez Elortondo, Francisco J; Berthier, Françoise

    2016-06-01

    Most raw milk Ossau-Iraty cheeses are currently manufactured on-farm using the same commercial streptococcal-lactococcal starter (S1). One way to enhance the microbial diversity that gives raw milk its advantages for cheese-making is to formulate new starters combining diverse, characterized strains. A new starter (OI) combining 6 raw milk strains of lactococci, recently isolated and characterized, was tested in parallel with the current starter by making 12 Ossau-Iraty raw milk cheeses at 3 farmhouses under the conditions prevailing at each farm. Compliance of the sensory characteristics with those expected by the Ossau-Iraty professionals, physicochemical parameters and coliforms were quantified at key manufacturing steps. The new starter OI gave cheeses having proper compliance but having lower compliance than the S1 cheeses under most manufacturing conditions, while managing coliform levels equally well as starter S1. This lower compliance relied more on the absence of Streptococcus thermophilus in starter OI, than on the nature of the lactoccocal strains present in starter OI. The study also shows that variations in 5 technological parameters during the first day of manufacture, within the range of values applied in the 3 farmhouses, are powerful tools for diversifying the scores for the sensory characteristics investigated.

  1. Reduced-fat Cheddar and Swiss-type cheeses harboring exopolysaccharide-producing probiotic Lactobacillus mucosae DPC 6426.

    Science.gov (United States)

    Ryan, P M; Burdíková, Z; Beresford, T; Auty, M A E; Fitzgerald, G F; Ross, R P; Sheehan, J J; Stanton, C

    2015-12-01

    Exopolysaccharide-producing Lactobacillus mucosae DPC 6426 was previously shown to have promising hypocholesterolemic activity in the atherosclerosis-prone apolipoprotein-E-deficient (apoE(-/-)) murine model. The aim of this study was to investigate the suitability of reduced-fat Cheddar and Swiss-type cheeses as functional (carrier) foods for delivery of this probiotic strain. All cheeses were manufactured at pilot-scale (500-L vats) in triplicate, with standard commercially available starters: for Cheddar, Lactococcus lactis; and for Swiss-type cheese, Streptococcus thermophilus, Lactobacillus helveticus, and Propionibacterium freudenreichii. Lactobacillus mucosae DPC 6426 was used as an adjunct culture during cheese manufacture, at a level of ~10(6) cfu·mL(-1) cheese milk (subsequently present in the cheese curd at>10(7) cfu·g(-1)). The adjunct strain remained viable at >5×10(7) cfu·g(-1) in both Swiss-type and Cheddar cheeses following ripening for 6 mo. Sensory analysis revealed that the presence of the adjunct culture imparted a more appealing appearance in Swiss-type cheese, but had no significant effect on the sensory characteristics of Cheddar cheeses. Moreover, the adjunct culture had no significant effect on cheese composition, proteolysis, pH, or instrumentally quantified textural characteristics of Cheddar cheeses. These data indicate that low-fat Swiss-type and Cheddar cheeses represent suitable food matrices for the delivery of the hypocholesterolemic Lactobacillus mucosae DPC 6426 in an industrial setting.

  2. The effect of high pressures on the yoghurt from milk with the stabilizer

    Energy Technology Data Exchange (ETDEWEB)

    Reps, A; Jankowska, A; Wisniewska, K [Chair of Food Biotechnology, University of Warmia and Mazury, Heweliusza str 1., 10-724 Olsztyn (Poland)], E-mail: arnold.reps@uwm.edu.pl

    2008-07-15

    The effect of high pressures on the microbiological and physio-chemical properties of yoghurt was investigated. The best results were obtained when the yoghurt was manufactured from milk with the addition of MYO 752 stabilizer (starch, gelatin, pectin) selected from 10 stabilizers. Yoghurt manufactured with the addition of 0, 6% MYO 752 stabilizer was processed at the pressure of 400-600 MPa/15 min. in the range of 50 MPa. Pressurization caused a total reduction of number of Lactobacillus delbrueckii ssp. Bulgaricus and reduced the number of Streptococcus thermophilus by 1-2 orders of magnitude. Pressurized and non-pressurized yoghurts characterized of a homogenous consistency and typical plain yoghurt taste. The decrease of the number of living bacteria was observed in yoghurts during the storage. The acidity of pressurized yoghurts remained on the some level at the temperature of 4{sup 0}C and 20{sup 0}C. The more intensive antibacterial activity of microflora was observed in yoghurts storaged at 20{sup 0}C in comparison with yoghurts storaged at 4{sup 0}C. Disadvantageous changes of the pressurized yoghurts consistency were not found. The taste and aroma of yoghurts remained without any changes.

  3. Phages of lactic acid bacteria: The role of genetics in understanding phage-host interactions and their co-evolutionary processes

    Energy Technology Data Exchange (ETDEWEB)

    Mahony, Jennifer, E-mail: j.mahony@ucc.ie [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Ainsworth, Stuart; Stockdale, Stephen [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Sinderen, Douwe van, E-mail: d.vansinderen@ucc.ie [Department of Microbiology, University College Cork, Western Road, Cork (Ireland); Alimentary Pharmabiotic Centre, Biosciences Institute, University College Cork, Western Road, Cork (Ireland)

    2012-12-20

    Dairy fermentations are among the oldest food processing applications, aimed at preservation and shelf-life extension through the use of lactic acid bacteria (LAB) starter cultures, in particular strains of Lactococcus lactis, Streptococcus thermophilus, Lactobacillus spp. and Leuconostoc spp. Traditionally this was performed by continuous passaging of undefined cultures from a finished fermentation to initiate the next fermentation. More recently, consumer demands on consistent and desired flavours and textures of dairy products have led to a more defined approach to such processes. Dairy (starter) companies have responded to the need to define the nature and complexity of the starter culture mixes, and dairy fermentations are now frequently based on defined starter cultures of low complexity, where each starter component imparts specific technological properties that are desirable to the product. Both mixed and defined starter culture approaches create the perfect environment for the proliferation of (bacterio)phages capable of infecting these LAB. The repeated use of the same starter cultures in a single plant, coupled to the drive towards higher and consistent production levels, increases the risk and negative impact of phage infection. In this review we will discuss recent advances in tracking the adaptation of phages to the dairy industry, the advances in understanding LAB phage-host interactions, including evolutionary and genomic aspects.

  4. Survival and bioactivities of selected probiotic lactobacilli in yogurt fermentation and cold storage: New insights for developing a bi-functional dairy food.

    Science.gov (United States)

    Rutella, Giuseppina Sefora; Tagliazucchi, Davide; Solieri, Lisa

    2016-12-01

    In previous work, we demonstrated that two probiotic strains, namely Lactobacillus casei PRA205 and Lactobacillus rhamnosus PRA331, produce fermented milks with potent angiotensin-converting enzyme (ACE)-inhibitory and antioxidant activities. Here, we tested these strains for the survivability and the release of antihypertensive and antioxidant peptides in yogurt fermentation and cold storage. For these purposes three yogurt batches were compared: one prepared using yogurt starters alone (Lactobacillus delbrueckii subspecies bulgaricus 1932 and Streptococcus thermophilus 99), and the remaining two containing either PRA205 or PRA331 in addition to yogurt starters. Despite the lower viable counts at the fermentation end compared to PRA331, PRA205 overcame PRA331 in survivability during refrigerated storage for 28 days, leading to viable counts (>10(8) CFU/g) higher than the minimum therapeutic threshold (10(6) CFU/g). Analyses of in vitro ACE-inhibitory and antioxidant activities of peptide fractions revealed that yogurt supplemented with PRA205 displays higher amounts of antihypertensive and antioxidant peptides than that produced with PRA331 at the end of fermentation and over storage. Two ACE-inhibitory peptides, Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP), were identified and quantified. This study demonstrated that L. casei PRA205 could be used as adjunct culture for producing bi-functional yogurt enriched in bioactive peptides and in viable cells, which bring health benefits to the host as probiotics.

  5. Role of the semi-conserved histidine residue in the light-sensing domain of LitR, a MerR-type photosensory transcriptional regulator.

    Science.gov (United States)

    Takano, Hideaki; Mise, Kou; Maruyama, Takafumi; Hagiwara, Kenta; Ueda, Kenji

    2016-08-01

    The LitR/CarH protein family transcriptional regulator is a new type of photoreceptor based on the function of adenosyl B12 (AdoB12) as a light-sensitive ligand. Here, we studied a semi-conserved histidine residue (His132) in the light-sensing (AdoB12-binding) domain at the C-terminus of LitR from a thermophilic Gram-negative bacterium, Thermus thermophilus HB27. The in vivo mutation of His132 within LitR caused a reduction in the rate of carotenoid production in response to illumination. BIAcore analysis revealed that the illuminated-LitRH132A possesses high DNA-binding activity compared to the wild-type protein. The subunit structure analysis showed that LitRH132A performed an incomplete subunit dissociation. The ability of LitRH132A to associate with AdoB12 was reduced compared with that of the wild-type protein in an equilibration dialysis experiment. Overall, these results suggest that His132 of LitR is involved in the association with AdoB12 as well as the light-sensitive DNA-binding activity based on oligomer dissociation. PMID:27283316

  6. X-ray Crystallographic Structure of Thermophilic Rhodopsin: IMPLICATIONS FOR HIGH THERMAL STABILITY AND OPTOGENETIC FUNCTION.

    Science.gov (United States)

    Tsukamoto, Takashi; Mizutani, Kenji; Hasegawa, Taisuke; Takahashi, Megumi; Honda, Naoya; Hashimoto, Naoki; Shimono, Kazumi; Yamashita, Keitaro; Yamamoto, Masaki; Miyauchi, Seiji; Takagi, Shin; Hayashi, Shigehiko; Murata, Takeshi; Sudo, Yuki

    2016-06-01

    Thermophilic rhodopsin (TR) is a photoreceptor protein with an extremely high thermal stability and the first characterized light-driven electrogenic proton pump derived from the extreme thermophile Thermus thermophilus JL-18. In this study, we confirmed its high thermal stability compared with other microbial rhodopsins and also report the potential availability of TR for optogenetics as a light-induced neural silencer. The x-ray crystal structure of TR revealed that its overall structure is quite similar to that of xanthorhodopsin, including the presence of a putative binding site for a carotenoid antenna; but several distinct structural characteristics of TR, including a decreased surface charge and a larger number of hydrophobic residues and aromatic-aromatic interactions, were also clarified. Based on the crystal structure, the structural changes of TR upon thermal stimulation were investigated by molecular dynamics simulations. The simulations revealed the presence of a thermally induced structural substate in which an increase of hydrophobic interactions in the extracellular domain, the movement of extracellular domains, the formation of a hydrogen bond, and the tilting of transmembrane helices were observed. From the computational and mutational analysis, we propose that an extracellular LPGG motif between helices F and G plays an important role in the thermal stability, acting as a "thermal sensor." These findings will be valuable for understanding retinal proteins with regard to high protein stability and high optogenetic performance. PMID:27129243

  7. Monitoring of the microbial communities involved in the soy sauce manufacturing process by PCR-denaturing gradient gel electrophoresis.

    Science.gov (United States)

    Tanaka, Yasushi; Watanabe, Jun; Mogi, Yoshinobu

    2012-08-01

    Soy sauce is a traditional seasoning produced through the fermentation of soybeans and wheat using microbes. In this study, the microbial communities involved in the soy sauce manufacturing process were analyzed by PCR-Denaturing Gradient Gel Electrophoresis (PCR-DGGE). The bacterial DGGE profile indicated that the bacterial microbes in the koji were Weissella cibaria (Weissella confusa, Weissella kimchii, Weissella salipiscis, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus iners, or Streptococcus thermophilus), Staphylococcus gallinarum (or Staphylococcus xylosus), and Staphylococcus kloosii. In addition to these bacteria, Tetragenococcus halophilus was also detected in the mash during lactic acid fermentation. The fungal DGGE profile indicated that the fungal microbes in the koji were not only Aspergillus oryzae but also several yeasts. In the mash, Zygosaccharomyces rouxii appeared in the early fermentation stage, Candida etchellsii (or Candida nodaensis) and Candida versatilis were detected at the middle fermentation stage, and Candida etchellsii was detected at the mature fermentation stage. These results suggest that the microbial communities present during the soy sauce manufacturing process change drastically throughout its production. This is the first report to reveal the microbial communities involved in the soy sauce manufacturing process using a culture-independent method. PMID:22475947

  8. Ammonia effect on hydrogenotrophic methanogens and syntrophic acetate-oxidizing bacteria.

    Science.gov (United States)

    Wang, Han; Fotidis, Ioannis A; Angelidaki, Irini

    2015-11-01

    Ammonia-rich substrates can cause inhibition on anaerobic digestion process. Syntrophic acetate-oxidizing bacteria (SAOB) and hydrogenotrophic methanogens are important for the ammonia inhibitory mechanism on anaerobic digestion. The roles and interactions of SAOB and hydrogenotrophic methanogens to ammonia inhibition effect are still unclear. The aim of the current study was to determine the ammonia toxicity levels of various pure strains of SAOB and hydrogenotrophic methanogens. Moreover, ammonia toxicity on the syntrophic-cultivated strains of SAOB and hydrogenotrophic methanogens was tested. Thus, four hydrogenotrophic methanogens (i.e. Methanoculleus bourgensis, Methanobacterium congolense, Methanoculleu thermophilus and Methanothermobacter thermautotrophicus), two SAOB (i.e. Tepidanaerobacter acetatoxydans and Thermacetogenium phaeum) and their syntrophic cultivation were assessed under 0.26, 3, 5 and 7 g NH4 (+)-N L(-1). The results showed that some hydrogenotrophic methanogens were equally, or in some cases, more tolerant to high ammonia levels compared to SAOB. Furthermore, a mesophilic hydrogenotrophic methanogen was more sensitive to ammonia toxicity compared to thermophilic methanogens tested in the study, which is contradicting to the general belief that thermophilic methanogens are more vulnerable to high ammonia loads compared to mesophilic. This unexpected finding underlines the fact that the complete knowledge of ammonia inhibition effect on hydrogenotrophic methanogens is still absent.

  9. Evidence for N coordination to Fe in the [2Fe-2S] clusters of Thermus Rieske protein and phthalate dioxygenase from Pseudomonas

    International Nuclear Information System (INIS)

    Rieske-type iron/sulfur proteins and several NADH-dependent oxygenases contain Fe/S clusters with similar spectral and magnetic properties. Purified Rieske iron/sulfur protein from Thermus thermophilus contains two apparently identical [2Fe-2S] clusters in a polypeptide having only four cysteine residues, and it has been proposed that each Fe/S cluster is coordinated to two cysteine S-atoms and to an unknown number of other non-sulfur atoms. The authors have examined the Rieske protein from Thermus and the phthalate dioxygenase from Pseudomonas cepacia with electron nuclear double resonance (ENDOR) and pulsed EPR methods and report here evidence for the direct coordination of nitrogenous ligands to the Fe/S clusters in these proteins. The electron nuclear double resonance signals arising from 14N have been interpreted in terms of a strongly coupled ligand with A/sup N/ = approx.26-28 MHz and a weakly coupled ligand with A/sup N/ = approx.9 MHz. The pulsed EPR spectrum shows a rich pattern of lines in the Fourier transformed data having peaks in the range of 0.8 to 6.7 MHz. The lower frequency resonances are tentatively associated with coupling of the unpaired spin to the remote N-atoms of coordinated imidazole rings. 26 references, 3 figures

  10. Crystal structures of Pseudomonas syringae pv. tomato DC3000 quinone oxidoreductase and its complex with NADPH

    International Nuclear Information System (INIS)

    Zeta-crystallin-like quinone oxidoreductase is NAD(P)H-dependent and catalyzes one-electron reduction of certain quinones to generate semiquinone. Here we present the crystal structures of zeta-crystallin-like quinone oxidoreductase from Pseudomonas syringae pv. tomato DC3000 (PtoQOR) and its complexes with NADPH determined at 2.4 and 2.01 A resolutions, respectively. PtoQOR forms as a homologous dimer, each monomer containing two domains. In the structure of the PtoQOR-NADPH complex, NADPH locates in the groove between the two domains. NADPH binding causes obvious conformational changes in the structure of PtoQOR. The putative substrate-binding site of PtoQOR is wider than that of Escherichia coli and Thermus thermophilus HB8. Activity assays show that PtoQOR has weak 1,4-benzoquinone catalytic activity, and very strong reduction activity towards large substrates such as 9,10-phenanthrenequinone. We propose a model to explain the conformational changes which take place during reduction reactions catalyzed by PtoQOR.

  11. Lactose digestion from yogurt: mechanism and relevance.

    Science.gov (United States)

    Savaiano, Dennis A

    2014-05-01

    Yogurt is traditionally consumed throughout the world among populations who are seemingly unable to digest lactose. This review provides a historical overview of the studies that show lactose digestion and tolerance from yogurt by lactose-intolerant people. The lactose in yogurt is digested more efficiently than other dairy sources of lactose because the bacteria inherent in yogurt assist with its digestion. The bacterial lactase survives the acidic conditions of the stomach, apparently being physically protected within the bacterial cells and facilitated by the buffering capacity of yogurt. The increasing pH as the yogurt enters the small intestine and a slower gastrointestinal transit time allow the bacterial lactase to be active, digesting lactose from yogurt sufficiently to prevent symptoms in lactose-intolerant people. There is little difference in the lactase capability of different commercial yogurts, because they apparently contain Lactobacillus bulgaricus and Streptococcus thermophilus in sufficient quantities (10(8) bacteria/mL). However, Lactobacillus acidophilus appears to require cell membrane disruption to physically release the lactase. Compared with unflavored yogurts, flavored yogurts appear to exhibit somewhat reduced lactase activity but are still well tolerated.

  12. Enhanced Microbial, Functional and Sensory Properties of Herbal Yogurt Fermented with Korean Traditional Plant Extracts.

    Science.gov (United States)

    Joung, Jae Yeon; Lee, Ji Young; Ha, Young Sik; Shin, Yong Kook; Kim, Younghoon; Kim, Sae Hun; Oh, Nam Su

    2016-01-01

    This study evaluated the effects of two Korean traditional plant extracts (Diospyros kaki THUNB. leaf; DK, and Nelumbo nucifera leaf; NN) on the fermentation, functional and sensory properties of herbal yogurts. Compared to control fermentation, all plant extracts increased acidification rate and reduced the time to complete fermentation (pH 4.5). Supplementation of plant extracts and storage time were found to influence the characteristics of the yogurts, contributing to increased viability of starter culture and phenolic compounds. In particular, the increase in the counts of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus was highest (2.95 and 1.14 Log CFU/mL respectively) in DK yogurt. Furthermore, supplementation of the plant extracts significantly influenced to increase the antioxidant activity and water holding capacity and to produce volatile compounds. The higher antioxidant activity and water holding capacity were observed in NN yogurt than DK yogurt. Moreover, all of the sensory characteristics were altered by the addition of plant extracts. Addition of plant extracts increased the scores related to flavor, taste, and texture from plain yogurt without a plant extract, as a result of volatile compounds analysis. Thus, the overall preference was increased by plant extracts. Consequently, supplementation of DK and NN extracts in yogurt enhanced the antioxidant activity and physical property, moreover increased the acceptability of yogurt. These findings demonstrate the possibility of using plant extracts as a functional ingredient in the manufacture of herbal yogurt.

  13. 马铃薯酸奶生产工艺优化%Process Optimization for the Development of Potato Yogurt

    Institute of Scientific and Technical Information of China (English)

    高杨

    2012-01-01

    以马铃薯和脱脂奶粉为主要原料,利用保加利亚乳杆菌和嗜热链球菌组成的混合发酵剂进行乳酸发酵。以感官评价为考察指标,通过正交试验研究确定马铃薯酸奶的最佳发酵工艺参数为:白砂糖加入量8%、发酵时间6h、发酵温度43℃。此工艺制备的马铃薯酸奶,色泽均匀,组织状态较好,口感细腻,符合标准要求。%The water left over from cooking potato and milk powder were used mainly for yogurt production by fermentation with mixed starter cultures of Lactobacillus bulgaria and Streptococcus thermophilus. The optimal fermentation parameters were 8% of white sugar, 6 h of fermentation time and 43 °C of fermentation temperature as determined using orthogonal array design for maximizing sensory evaluation score. Yogurt with a uniform color, a good texture, and a smooth taste was obtained under these parameters. Which could meet the requirements of relevant standards.

  14. Optimization Of Freeze-Dried Starter For Yogurt By Full Factorial Experimental Design

    Directory of Open Access Journals (Sweden)

    Chen He

    2015-12-01

    Full Text Available With the rapidly development of fermented milk product, it is significant for enhancing the performance of starter culture. This paper not only investigated the influence of anti-freeze factors and freeze-drying protective agents on viable count, freeze-drying survival rate and yield of Lactobacillus bulgaricus (LB and Streptococcus thermophilus (ST, but also optimized the bacteria proportion of freeze-dried starter culture for yogurt by full factorial experimental design. The results showed as following: the freeze-drying protective agents or anti-freeze factors could enhanced survival rate of LB and ST; the freeze-dried LB and ST powders containing both of anti-freeze factors and freeze-drying protective agents had higher viable count and freeze-drying survival rate that were 84.7% and 79.7% respectively; In terms of fermentation performance, the best group of freeze-dried starter for yogurt was the compound of LB3 and ST2.

  15. Optimization of Production Technology of Konjac Yogurt by Response Surface Methodology%响应面法优化魔芋酸奶工艺

    Institute of Scientific and Technical Information of China (English)

    葛谦; 袁亚宏; 岳田利

    2013-01-01

      以魔芋精粉、牛奶为主要原料,通过单因素试验及BOX-Behnken设计法对魔芋固体酸奶进行了初步研究。结果表明,嗜热链球菌和保加利亚乳杆菌比例为2∶1,接种量为3.5%(体积分数),魔芋精粉添加量为0.08%,蔗糖添加量为质量分数7.9%,43℃发酵4.5 h,其获得产品组织均匀、口感细腻、风味俱佳。%With milk and konjac as main materials,the processing technology of konjac yogurt was studied by single factor and box-behnken experimental design.The results showed that excellent quality konjac yogurt could be produced under the conditions of 3.5%fermentation agent (2∶1 mixture of streptococcus thermophilus and lactobacillus bulgaricus),and konjac powder 0.08 %,sucrose 7.9 % and fermentation time 4.5 h under 43℃so that the konjac yogurt was homogeneous and smooth with appropriate sweetness andsourness.

  16. Reconstituted yogurt from yogurt cultured milk powder mix has better overall characteristics than reconstituted yogurt from commercial yogurt powder.

    Science.gov (United States)

    Song, Lijie; Aryana, Kayanush J

    2014-10-01

    For manufacture of commercial yogurt powder, yogurt has to go through a drying process, which substantially lowers the yogurt culture counts, so the potential health benefits of the yogurt culture bacteria are reduced. Also, upon reconstitution, commercial yogurt powder does not taste like yogurt and has an off-flavor. The objective was to study the microbial, physicochemical, and sensory characteristics of reconstituted yogurt from yogurt cultured milk powder (YCMP) mix and reconstituted yogurt from commercial yogurt powder (CYP). The CYP reconstituted yogurt was the control and YCMP mix reconstituted yogurt was the treatment. Microbial and physicochemical characteristics of the CYP reconstituted yogurt and YCMP mix reconstituted yogurt were analyzed daily for the first week and then weekly for a period of 8 wk. Sensory consumer testing of CYP reconstituted yogurt and YCMP mix reconstituted yogurt was conducted with 100 consumers. At 56 d, YCMP mix reconstituted yogurt had 5 log cfu/mL higher counts of Streptococcus thermophilus than the control (CYP reconstituted yogurt). Also, Lactobacillus bulgaricus counts of YCMP mix reconstituted yogurt were 6.55 log cfu/mL at 28 d and were 5.35 log cfu/mL at 56 d, whereas the CYP reconstituted yogurt from 28 d onwards had a count of yogurt also had significantly higher apparent viscosity and sensory scores for appearance, color, aroma, taste, thickness, overall liking, consumer acceptability, and purchase intent than CYP reconstituted yogurt. Overall, YCMP mix reconstituted yogurt had more desirable characteristics than CYP reconstituted yogurt.

  17. Coexistence of Lactic Acid Bacteria and Potential Spoilage Microbiota in a Dairy Processing Environment.

    Science.gov (United States)

    Stellato, Giuseppina; De Filippis, Francesca; La Storia, Antonietta; Ercolini, Danilo

    2015-11-01

    Microbial contamination in food processing plants can play a fundamental role in food quality and safety. In this study, the microbiota in a dairy plant was studied by both 16S rRNA- and 26S rRNA-based culture-independent high-throughput amplicon sequencing. Environmental samples from surfaces and tools were studied along with the different types of cheese produced in the same plant. The microbiota of environmental swabs was very complex, including more than 200 operational taxonomic units with extremely variable relative abundances (0.01 to 99%) depending on the species and sample. A core microbiota shared by 70% of the samples indicated a coexistence of lactic acid bacteria with a remarkable level of Streptococcus thermophilus and possible spoilage-associated bacteria, including Pseudomonas, Acinetobacter, and Psychrobacter, with a relative abundance above 50%. The most abundant yeasts were Kluyveromyces marxianus, Yamadazyma triangularis, Trichosporon faecale, and Debaryomyces hansenii. Beta-diversity analyses showed a clear separation of environmental and cheese samples based on both yeast and bacterial community structure. In addition, predicted metagenomes also indicated differential distribution of metabolic pathways between the two categories of samples. Cooccurrence and coexclusion pattern analyses indicated that the occurrence of potential spoilers was excluded by lactic acid bacteria. In addition, their persistence in the environment can be helpful to counter the development of potential spoilers that may contaminate the cheeses, with possible negative effects on their microbiological quality.

  18. Structure of the protein core of translation initiation factor 2 in apo, GTP-bound and GDP-bound forms

    Energy Technology Data Exchange (ETDEWEB)

    Simonetti, Angelita [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Centre National de la Recherche Scientifique (CNRS) UMR 7104/Institut National de la Santé de la Recherche Médicale - INSERM U964/Université de Strasbourg, 1 Rue Laurent Fries, 67404 Illkirch (France); Marzi, Stefano [Architecture et Réactivité de l’ARN, UPR 9002 CNRS, IBMC (Institute of Molecular and Cellular Biology), 15 Rue R. Descartes, 67084 Strasbourg, France, Université de Strasbourg, 67000 Strasbourg (France); Fabbretti, Attilio [University of Camerino, 62032 Camerino (Monaco) (Italy); Hazemann, Isabelle; Jenner, Lasse [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Centre National de la Recherche Scientifique (CNRS) UMR 7104/Institut National de la Santé de la Recherche Médicale -INSERM U964/Université de Strasbourg, 1 Rue Laurent Fries, 67404 Illkirch (France); Urzhumtsev, Alexandre [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Centre National de la Recherche Scientifique (CNRS) UMR 7104/Institut National de la Santé de la Recherche Médicale - INSERM U964/Université de Strasbourg, 1 Rue Laurent Fries, 67404 Illkirch (France); Université de Lorraine, 54506 Vandoeuvre-lès-Nancy (France); Gualerzi, Claudio O. [University of Camerino, 62032 Camerino (Monaco) (Italy); Klaholz, Bruno P., E-mail: klaholz@igbmc.fr [IGBMC (Institute of Genetics and of Molecular and Cellular Biology), Centre National de la Recherche Scientifique (CNRS) UMR 7104/Institut National de la Santé de la Recherche Médicale - INSERM U964/Université de Strasbourg, 1 Rue Laurent Fries, 67404 Illkirch (France)

    2013-06-01

    The crystal structures of the eubacterial translation initiation factor 2 in apo form and with bound GDP and GTP reveal conformational changes upon nucleotide binding and hydrolysis, notably of the catalytically important histidine in the switch II region. Translation initiation factor 2 (IF2) is involved in the early steps of bacterial protein synthesis. It promotes the stabilization of the initiator tRNA on the 30S initiation complex (IC) and triggers GTP hydrolysis upon ribosomal subunit joining. While the structure of an archaeal homologue (a/eIF5B) is known, there are significant sequence and functional differences in eubacterial IF2, while the trimeric eukaryotic IF2 is completely unrelated. Here, the crystal structure of the apo IF2 protein core from Thermus thermophilus has been determined by MAD phasing and the structures of GTP and GDP complexes were also obtained. The IF2–GTP complex was trapped by soaking with GTP in the cryoprotectant. The structures revealed conformational changes of the protein upon nucleotide binding, in particular in the P-loop region, which extend to the functionally relevant switch II region. The latter carries a catalytically important and conserved histidine residue which is observed in different conformations in the GTP and GDP complexes. Overall, this work provides the first crystal structure of a eubacterial IF2 and suggests that activation of GTP hydrolysis may occur by a conformational repositioning of the histidine residue.

  19. Structure and function of CarD, an essential mycobacterial transcription factor

    Science.gov (United States)

    Srivastava, Devendra B.; Leon, Katherine; Osmundson, Joseph; Garner, Ashley L.; Weiss, Leslie A.; Westblade, Lars F.; Glickman, Michael S.; Landick, Robert; Darst, Seth A.; Stallings, Christina L.; Campbell, Elizabeth A.

    2013-01-01

    CarD, an essential transcription regulator in Mycobacterium tuberculosis, directly interacts with the RNA polymerase (RNAP). We used a combination of in vivo and in vitro approaches to establish that CarD is a global regulator that stimulates the formation of RNAP-holoenzyme open promoter (RPo) complexes. We determined the X-ray crystal structure of Thermus thermophilus CarD, allowing us to generate a structural model of the CarD/RPo complex. On the basis of our structural and functional analyses, we propose that CarD functions by forming protein/protein and protein/DNA interactions that bridge the RNAP to the promoter DNA. CarD appears poised to interact with a DNA structure uniquely presented by the RPo: the splayed minor groove at the double-stranded/single-stranded DNA junction at the upstream edge of the transcription bubble. Thus, CarD uses an unusual mechanism for regulating transcription, sensing the DNA conformation where transcription bubble formation initiates. PMID:23858468

  20. Alkalizing Reactions Streamline Cellular Metabolism in Acidogenic Microorganisms

    Science.gov (United States)

    Arioli, Stefania; Ragg, Enzio; Scaglioni, Leonardo; Fessas, Dimitrios; Signorelli, Marco; Karp, Matti; Daffonchio, Daniele; De Noni, Ivano; Mulas, Laura; Oggioni, Marco; Guglielmetti, Simone; Mora, Diego

    2010-01-01

    An understanding of the integrated relationships among the principal cellular functions that govern the bioenergetic reactions of an organism is necessary to determine how cells remain viable and optimise their fitness in the environment. Urease is a complex enzyme that catalyzes the hydrolysis of urea to ammonia and carbonic acid. While the induction of urease activity by several microorganisms has been predominantly considered a stress-response that is initiated to generate a nitrogen source in response to a low environmental pH, here we demonstrate a new role of urease in the optimisation of cellular bioenergetics. We show that urea hydrolysis increases the catabolic efficiency of Streptococcus thermophilus, a lactic acid bacterium that is widely used in the industrial manufacture of dairy products. By modulating the intracellular pH and thereby increasing the activity of β-galactosidase, glycolytic enzymes and lactate dehydrogenase, urease increases the overall change in enthalpy generated by the bioenergetic reactions. A cooperative altruistic behaviour of urease-positive microorganisms on the urease-negative microorganisms within the same environment was also observed. The physiological role of a single enzymatic activity demonstrates a novel and unexpected view of the non-transcriptional regulatory mechanisms that govern the bioenergetics of a bacterial cell, highlighting a new role for cytosol-alkalizing biochemical pathways in acidogenic microorganisms. PMID:21152088