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Sample records for natto production strain

  1. Whole genome assembly of a natto production strain Bacillus subtilis natto from very short read data.

    Science.gov (United States)

    Nishito, Yukari; Osana, Yasunori; Hachiya, Tsuyoshi; Popendorf, Kris; Toyoda, Atsushi; Fujiyama, Asao; Itaya, Mitsuhiro; Sakakibara, Yasubumi

    2010-04-16

    Bacillus subtilis natto is closely related to the laboratory standard strain B. subtilis Marburg 168, and functions as a starter for the production of the traditional Japanese food "natto" made from soybeans. Although re-sequencing whole genomes of several laboratory domesticated B. subtilis 168 derivatives has already been attempted using short read sequencing data, the assembly of the whole genome sequence of a closely related strain, B. subtilis natto, from very short read data is more challenging, particularly with our aim to assemble one fully connected scaffold from short reads around 35 bp in length. We applied a comparative genome assembly method, which combines de novo assembly and reference guided assembly, to one of the B. subtilis natto strains. We successfully assembled 28 scaffolds and managed to avoid substantial fragmentation. Completion of the assembly through long PCR experiments resulted in one connected scaffold for B. subtilis natto. Based on the assembled genome sequence, our orthologous gene analysis between natto BEST195 and Marburg 168 revealed that 82.4% of 4375 predicted genes in BEST195 are one-to-one orthologous to genes in 168, with two genes in-paralog, 3.2% are deleted in 168, 14.3% are inserted in BEST195, and 5.9% of genes present in 168 are deleted in BEST195. The natto genome contains the same alleles in the promoter region of degQ and the coding region of swrAA as the wild strain, RO-FF-1. These are specific for gamma-PGA production ability, which is related to natto production. Further, the B. subtilis natto strain completely lacked a polyketide synthesis operon, disrupted the plipastatin production operon, and possesses previously unidentified transposases. The determination of the whole genome sequence of Bacillus subtilis natto provided detailed analyses of a set of genes related to natto production, demonstrating the number and locations of insertion sequences that B. subtilis natto harbors but B. subtilis 168 lacks

  2. Whole genome assembly of a natto production strain Bacillus subtilis natto from very short read data

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    Fujiyama Asao

    2010-04-01

    Full Text Available Abstract Background Bacillus subtilis natto is closely related to the laboratory standard strain B. subtilis Marburg 168, and functions as a starter for the production of the traditional Japanese food "natto" made from soybeans. Although re-sequencing whole genomes of several laboratory domesticated B. subtilis 168 derivatives has already been attempted using short read sequencing data, the assembly of the whole genome sequence of a closely related strain, B. subtilis natto, from very short read data is more challenging, particularly with our aim to assemble one fully connected scaffold from short reads around 35 bp in length. Results We applied a comparative genome assembly method, which combines de novo assembly and reference guided assembly, to one of the B. subtilis natto strains. We successfully assembled 28 scaffolds and managed to avoid substantial fragmentation. Completion of the assembly through long PCR experiments resulted in one connected scaffold for B. subtilis natto. Based on the assembled genome sequence, our orthologous gene analysis between natto BEST195 and Marburg 168 revealed that 82.4% of 4375 predicted genes in BEST195 are one-to-one orthologous to genes in 168, with two genes in-paralog, 3.2% are deleted in 168, 14.3% are inserted in BEST195, and 5.9% of genes present in 168 are deleted in BEST195. The natto genome contains the same alleles in the promoter region of degQ and the coding region of swrAA as the wild strain, RO-FF-1. These are specific for γ-PGA production ability, which is related to natto production. Further, the B. subtilis natto strain completely lacked a polyketide synthesis operon, disrupted the plipastatin production operon, and possesses previously unidentified transposases. Conclusions The determination of the whole genome sequence of Bacillus subtilis natto provided detailed analyses of a set of genes related to natto production, demonstrating the number and locations of insertion sequences that B

  3. Phylogenetic analysis of Bacillus subtilis strains applicable to natto (fermented soybean) production.

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    Kubo, Yuji; Rooney, Alejandro P; Tsukakoshi, Yoshiki; Nakagawa, Rikio; Hasegawa, Hiromasa; Kimura, Keitarou

    2011-09-01

    Spore-forming Bacillus strains that produce extracellular poly-γ-glutamic acid were screened for their application to natto (fermented soybean food) fermentation. Among the 424 strains, including Bacillus subtilis and B. amyloliquefaciens, which we isolated from rice straw, 59 were capable of fermenting natto. Biotin auxotrophism was tightly linked to natto fermentation. A multilocus nucleotide sequence of six genes (rpoB, purH, gyrA, groEL, polC, and 16S rRNA) was used for phylogenetic analysis, and amplified fragment length polymorphism (AFLP) analysis was also conducted on the natto-fermenting strains. The ability to ferment natto was inferred from the two principal components of the AFLP banding pattern, and natto-fermenting strains formed a tight cluster within the B. subtilis subsp. subtilis group.

  4. Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

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    Kubo, Yuji; Rooney, Alejandro P.; Tsukakoshi, Yoshiki; Nakagawa, Rikio; Hasegawa, Hiromasa; Kimura, Keitarou

    2011-01-01

    Spore-forming Bacillus strains that produce extracellular poly-γ-glutamic acid were screened for their application to natto (fermented soybean food) fermentation. Among the 424 strains, including Bacillus subtilis and B. amyloliquefaciens, which we isolated from rice straw, 59 were capable of fermenting natto. Biotin auxotrophism was tightly linked to natto fermentation. A multilocus nucleotide sequence of six genes (rpoB, purH, gyrA, groEL, polC, and 16S rRNA) was used for phylogenetic analysis, and amplified fragment length polymorphism (AFLP) analysis was also conducted on the natto-fermenting strains. The ability to ferment natto was inferred from the two principal components of the AFLP banding pattern, and natto-fermenting strains formed a tight cluster within the B. subtilis subsp. subtilis group. PMID:21764950

  5. Surfactins in Natto: The Surfactin Production Capacity of the Starter Strains and the Actual Surfactin Contents in the Products

    DEFF Research Database (Denmark)

    Juola, Mari; Kinnunen, Kristiina; Nielsen, Kristian Fog

    2014-01-01

    product, natto, were isolated in order to clarify their potential to produce the suspect lipopeptides. The isolated bacilli were characterized as Bacillus subtilis. They were b-hemolytic and gave a positive signal in the PCR screen for genes associated with surfactin production, and their culture extracts...... were cytotoxic to boar sperm cells. Organic extracts of both Bacillus cultures and the natto samples were analyzed for their surfactin content using ultrahigh-performance liquid chromatography with high-resolution mass spectrometry. All the strains proved to be surfactin producers (15 to 39 mg....../ml culture medium); the natto samples contained as much as 2.2 mg g21 of surfactins. This means that consumers can ingest at least approximately 80 to 100 mg of surfactins per single 50-g natto serving apparently without suffering any ill effects, indicating a very low human toxicity....

  6. Phylogenetic Analysis of Bacillus subtilis Strains Applicable to Natto (Fermented Soybean) Production

    OpenAIRE

    Kubo, Yuji; Rooney, Alejandro P.; Tsukakoshi, Yoshiki; Nakagawa, Rikio; Hasegawa, Hiromasa; Kimura, Keitarou

    2011-01-01

    Spore-forming Bacillus strains that produce extracellular poly-γ-glutamic acid were screened for their application to natto (fermented soybean food) fermentation. Among the 424 strains, including Bacillus subtilis and B. amyloliquefaciens, which we isolated from rice straw, 59 were capable of fermenting natto. Biotin auxotrophism was tightly linked to natto fermentation. A multilocus nucleotide sequence of six genes (rpoB, purH, gyrA, groEL, polC, and 16S rRNA) was used for phylogenetic analy...

  7. Surfactins in natto: the surfactin production capacity of the starter strains and the actual surfactin contents in the products.

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    Juola, Mari; Kinnunen, Kristiina; Nielsen, Kristian Fog; von Wright, Atte

    2014-12-01

    Surfactin-type lipopeptides are suspected of being implicated in the rare food poisonings caused by Bacillus species outside the Bacillus cereus cluster. In order to get information on surfactin levels in actual human foods, bacilli from three commercial samples of a Japanese traditional bean product, natto, were isolated in order to clarify their potential to produce the suspect lipopeptides. The isolated bacilli were characterized as Bacillus subtilis. They were β-hemolytic and gave a positive signal in the PCR screen for genes associated with surfactin production, and their culture extracts were cytotoxic to boar sperm cells. Organic extracts of both Bacillus cultures and the natto samples were analyzed for their surfactin content using ultrahigh-performance liquid chromatography with high-resolution mass spectrometry. All the strains proved to be surfactin producers (15 to 39 μg/ml culture medium); the natto samples contained as much as 2.2 mg g(-1) of surfactins. This means that consumers can ingest at least approximately 80 to 100 mg of surfactins per single 50-g natto serving apparently without suffering any ill effects, indicating a very low human toxicity.

  8. Phylogenetic analysis of Bacillus subtilis strains applicable to natto (fermented soybean) production

    Science.gov (United States)

    Spore-forming Bacillus strains that produce extracellular poly-'-glutamic acid were screened for their application to natto (fermented soybean food) fermentation. Among the 365 strains, including B. subtilis and B. amyloliquefaciens, which we isolated from rice straw, 59 were capable of fermenting n...

  9. Analisa Asam Organik Dan Asam Amino Pada Natto Yang Difermentasi Oleh 7 Strain Bacillus Natto

    OpenAIRE

    SULISTYO, JOKO

    1986-01-01

    JOKO SUUSTYO. 1986. Analysis of organic acid and amino acid in natto fermented by 7 strains of Bacillus natto. Berita Biologi 3 (6): 277 - 282. The ability of seven strains of Bacillus natto produced a number of organic and amino acid components in natto were investigated. Raw, submerced and autoclaved soybeans for natto contained 1.15% 0.46% and 0.77% in organic acid and 0.19%, 0.30% and 0.30% in amino acid, while after fermentation those range were 0.36 - 0.90% in organic acid and 2.86 -6.0...

  10. Biogenic amine formation and bacterial contribution in Natto products.

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    Kim, Bitna; Byun, Bo Young; Mah, Jae-Hyung

    2012-12-01

    Twenty-one Natto products currently distributed in Korea were analysed for biogenic amine contents and tested to determine physicochemical and bacterial contributions to biogenic amine formation. Among them, nine products (about 43%) had β-phenylethylamine or tyramine contents greater than the toxic dose (30mg/kg and 100mg/kg, respectively) of each amine, although no products showed total amounts of biogenic amines above the harmful level (1000mg/kg), which indicates that the amounts of biogenic amines in some Natto products are not within the safe level for human health. From four different Natto products, that contained noticeable levels of β-phenylethylamine and tyramine, 80 bacterial strains were isolated. All the strains were identified to be Bacillus subtilis and highly capable of producing β-phenylethylamine and tyramine. Therefore, it seems likely that the remarkable contents of β-phenylethylamine and tyramine in Natto predominantly resulted from the strains highly capable of producing those amines present in the food. Copyright © 2012 Elsevier Ltd. All rights reserved.

  11. Effect of corona electric field on the production of gamma-poly glutamic acid based on bacillus natto

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    Qi, Hong; Na, Ri; Xin, Jiletu; Jie Xie, Ya; Guo, Jiu Feng

    2013-03-01

    Bacillus Natto is an important strain for gamma-poly glutamic acid (γ-PGA) production. The mutagenesis of Bacillus Natto 20646 under corona electric field and the screening of high γ-PGA producing mutant were investigated. A new mutant bacillus natto Ndlz01 was isolated from Bacillus Natto 20646 after mutation in corona electric field at 9kV for 2min. The Ndlz01 exhibited genetic stability of high γ-PGA producing ability even after five generation cultures. When the bacterium was mutated in streamer discharge state at 9kV for 2min, its death rate was more than 90%. Compared with the yield of γ-PGA based on the original Bacillus Natto 20646, the γ-PGA yield of mutant bacillus natto Ndlz01 increased from 2.6 to 5.94 g/L, with an increase rate of 129.78%.

  12. Effect of corona electric field on the production of gamma-poly glutamic acid based on bacillus natto

    International Nuclear Information System (INIS)

    Qi, Hong; Na, Ri; Xin, Jiletu; Xie, Ya Jie; Guo, Jiu Feng

    2013-01-01

    Bacillus Natto is an important strain for gamma-poly glutamic acid (γ-PGA) production. The mutagenesis of Bacillus Natto 20646 under corona electric field and the screening of high γ-PGA producing mutant were investigated. A new mutant bacillus natto Ndlz01 was isolated from Bacillus Natto 20646 after mutation in corona electric field at 9kV for 2min. The Ndlz01 exhibited genetic stability of high γ-PGA producing ability even after five generation cultures. When the bacterium was mutated in streamer discharge state at 9kV for 2min, its death rate was more than 90%. Compared with the yield of γ-PGA based on the original Bacillus Natto 20646, the γ-PGA yield of mutant bacillus natto Ndlz01 increased from 2.6 to 5.94 g/L, with an increase rate of 129.78%.

  13. Complete nucleotide sequence of Bacillus subtilis (natto) bacteriophage PM1, a phage associated with disruption of food production.

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    Umene, Kenichi; Shiraishi, Atsushi

    2013-06-01

    "Natto", considered a traditional food, is made by fermenting boiled soybeans with Bacillus subtilis (natto), which is a natto-producing strain related to B. subtilis. The production of natto is disrupted by phage infections of B. subtilis (natto); hence, it is necessary to control phage infections. PM1, a phage of B. subtilis (natto), was isolated during interrupted natto production in a factory. In a previous study, PM1 was classified morphologically into the family Siphoviridae, and its genome, comprising approximately 50 kbp of linear double-stranded DNA, was assumed to be circularly permuted. In the present study, the complete nucleotide sequence of the PM1 genomic DNA of 50,861 bp (41.3 %G+C) was determined, and 86 open reading frames (ORFs) were deduced. Forty-one ORFs of PM1 shared similarities with proteins deduced from the genome of phages reported so far. Twenty-three ORFs of PM1 were associated with functions related to the phage multiplication process of gene control, DNA replication/modification, DNA packaging, morphogenesis, and cell lysis. Bacillus subtilis (natto) produces a capsular polypeptide of glutamate with a γ-linkage (called poly-γ-glutamate), which appears to serve as a physical barrier to phage adsorption. One ORF of PM1 had similarity with a poly-γ-glutamate hydrolase, which is assumed to degrade the capsular barrier to allow phage progenies to infect encapsulated host cells. The genome analysis of PM1 revealed the characteristics of the phage that are consistent as Bacillus subtilis (natto)-infecting phage.

  14. Chemical changes associated with lotus and water lily natto production

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    Lestari, S. D.; Fatimah, N.; Nopianti, R.

    2017-04-01

    Natto is a traditional Japanese food made by fermenting whole soybean seeds with pure culture of Bacillus subtilis subsp. natto. The purpose of this study was to investigate the suitability of lotus (Nelumbo nucifera) and water lily (Nymphaea stellata) seeds as the raw materials for natto production. Chemical (proximate, amino acids and minerals) changes were observed on raw, steamed and fermented seeds. Proximate compositions of all samples were calculated in both wet basis and dry basis. In wet basis calculation, steaming and fermentation tended to lower the carbohydrates, ashes, fats and protein content which were attributed to the increase of moisture. The total amino acid, iron and magnesium contents of raw lotus seeds were 24.29%, 5.08 mg 100g-1 and 174.23 mg 100g-1 dry matter, respectively. After a 24h-fermentation at 40°C, the total amino acids decreased while iron and magnesium contents increased significantly reaching, in respective order, 9.9 mg 100g-1 and 411.36 mg 100g-1 dry matter. Changes in chemical composition after fermentation were more pronounced in lotus seeds than water lily seeds indicating that their nutrient composition were more suitable to support Bacillus subtilis growth.

  15. Changes in biotin levels during production of natto, Japanese fermented soybean

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    Makoto Muratsugu

    2017-09-01

    Full Text Available The change of biotin level during production of natto (Japanese fermented soybean was investigated in this study.  The total biotin level was measured by an agar plate bioassay using Lactobacillus plantarum ATCC 8014.  The total biotin level decreased during water soaking, but increased after the fermentation of soybeans using Bacillus subtilis var. natto (B. natto and reached a maximum level.  The increase of total biotin was not affected by Asp, Arg, and Ile which promoted the growth of L. plantarum in high concentrations.  The peak level of biotin in the fermented soybeans was significantly higher than that of dry soybeans.  The fermented soybeans at the biotin peak level were adequate for food.  In addition, we detected 9 and 4 biotinylated polypeptides in the soybeans and B. natto used in this study, respectively.  We speculated that the increase of biotin level may depend on the increase of the 4 biotinylated polypeptides and free biotin in B. natto.

  16. Extract from the fermented soybean product Natto inhibits Vibrio biofilm formation and reduces shrimp mortality from Vibrio harveyi infection.

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    Yatip, Pattanan; Nitin Chandra Teja, D; Flegel, Timothy W; Soowannayan, Chumporn

    2018-01-01

    Many bacteria, including Vibrio pathogens of shrimp, need to colonize and/or form biofilms in hosts or the environment to cause disease. Thus, one possible control strategy for shrimp Vibriosis is biofilm inhibition. With this objective, an extract from the Japanese fermented soybean product, Natto was tested with the luminescent shrimp pathogen Vibrio harveyi (VH) for its ability to inhibit or degrade biofilm and to interfere with cell growth in broth. Natto is a traditional fermentation product of Bacillus subtilis var Natto (BSN1). Using 96 well microtiter plates coated with 0.4% chitosan, we found that biofilm formation by VH was inhibited, while growth in parallel broth cultures was not. When an extract from Natto prepared using BSN1 was mixed with feed for the whiteleg shrimp Penaeus vannamei before immersion challenge with V. harveyi at 10 6  cfu/ml, survival was significantly higher (p≤0.05) than for control shrimp given feed without these additives. Further work done to test whether d-amino acids were involved in biofilm formation as previously reported for B. subtilis, Staphylococus aureus and Pseudomonas aeruginosa gave negative results. In conclusion, we discovered that Natto extract can inhibit Vibrio biofilm formation and that it or BSN1 alone added to shrimp feed can significantly reduce shrimp mortality in immersion challenges with pathogenic VH. This shows some promise for possible application against Vibriosis in shrimp since Natto is generally regarded as safe (GRAS) for human consumption. Copyright © 2017 Elsevier Ltd. All rights reserved.

  17. Utilization of low-cost substrates for the production of nystose by Bacillus subtilis natto cct 7712

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    Dieyssi Alves dos Santos

    2016-08-01

    Full Text Available Current analysis describes the capacity of Bacillus subtilis natto CCT 7712 to produce high amounts of nystose by low-cost substrates available in Brazil, such as commercial sucrose, sugarcane molasses and sugarcane juice. Optimization resulted in a maximum production of 179.77 g L-1 of nystose, averaging 7.49 g L-1 hour-1 of productivity and a 71.73% yield in a medium with 400 g L-1of commercial sucrose and 0.8 g L-1 of MnSO4. Fermentations with sugarcane molasses and sugarcane juice also resulted in a satisfactory production reaching 97.93 and 42.58 g L-1 nystose, respectively. High nystose production in a medium with sugarcane derivatives suggests submerged fermentation with Bacillus subtillis natto CT 7712 as a promising strategy to produce nystose at industrial level.

  18. Conjoint analysis on the purchase intent for traditional fermented soy product (natto) among Japanese housewives.

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    Kimura, Atsushi; Kuwazawa, Shigetaka; Wada, Yuji; Kyutoku, Yasushi; Okamoto, Masako; Yamaguchi, Yui; Masuda, Tomohiro; Dan, Ippeita

    2011-04-01

    The effect of sensory and extrinsic attributes on consumer intentions to purchase the Japanese traditional fermented soybean product natto was evaluated using conjoint analysis. Six attributes with 2 levels each were chosen and manipulated: price (high compared with low), the country of origin of the soybeans (domestic compared with imported), stickiness (strong compared with moderate), smell (rich compared with moderate), attached seasonings (attached compared with no attached seasonings), and the environmental friendliness of the packaging (high compared with low). A fractional factorial design was applied and 8 hypothetical product labels were produced. A sample of 479 Japanese housewives ranked these product labels based on their purchase intentions. Overall purchase intention was affected by country of origin, attached seasonings, and price; those attributes accounted for 81.0%, while the sensory attributes of the product accounted for 19.0% of purchase intents. In order to estimate market segments for the natto products based on consumer preference, a cluster analysis was performed. It identified 4 segments of consumers: 1 oriented to attached seasonings, another conscious of the price, and the other 2 oriented to origins. The behavioral and demographic characteristics of the respondents had a limited influence on segment membership.   This research was conducted to understand how consumers valuate various sensory and nonsensory product attributes based on their assessment of the overall product in the case of Japanese fermented soy product (natto). The data of this research would be of great importance both in understanding consumer behavior and in designing strategies for product development.

  19. Effects of Bacillus subtilis var. natto products on symptoms caused by blood flow disturbance in female patients with lifestyle diseases

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    Hitosugi, Masahito; Hamada, Katsuo; Misaka, Kazutaka

    2015-01-01

    The fermented soybean product natto is a popular traditional food in Japan and is considered a health supplement. NKCP®, a natto-derived dietary food supplement whose main component is bacillopeptidase F, has antithrombotic, fibrinolytic, and blood pressure-lowering effects. We examined whether daily intake of NKCP® effectively improves subjective symptoms in patients with lifestyle diseases in this cross-over, double-blind study. Fermented soya extract with subtilisin NAT (nattokinase) as the main component was used as an active placebo. A 4-week course of NKCP® significantly decreased the visual analog scale (VAS) score for shoulder stiffness from 42.3 to 32.4 (P=0.009), the VAS score for low back pain from 25.5 to 18.8 (P=0.02), and the VAS score for coldness of the extremities from 33.1 to 25.7 (P=0.002). However, no significant difference was found in the VAS score for headache. After a 4-week course of active placebo, no significant changes in the VAS score were found for any symptoms. The significant improvement in the symptoms secondary to blood flow disturbance was caused by the improvement in blood flow by NKCP®. The use of dietary supplements based on the Japanese traditional food natto helps to relieve subjective symptoms for patients with lifestyle diseases receiving medical care. PMID:25653551

  20. A simple and cost-saving approach to optimize the production of subtilisin NAT by submerged cultivation of Bacillus subtilis natto.

    Science.gov (United States)

    Ku, Ting-Wei; Tsai, Ruei-Lan; Pan, Tzu-Ming

    2009-01-14

    Subtilisin NAT, formerly designated nattokinase or subtilisin BSP, is a potent cardiovascular drug because of its strong fibrinolytic activity and safety. In this study, one Bacillus subtilis natto strain with high fibrinolytic activity was isolated. We further studied the optimal conditions for subtilisin NAT production by submerged cultivation and three variables/three levels of response surface methodology (RSM) using various inoculum densities, glucose concentrations, and defatted soybean concentrations as the three variables. According to the RSM analysis, while culturing by 2.93% defatted soybean, 1.75% glucose, and 4.00% inoculum density, we obtained an activity of 13.78 SU/mL. Processing the batch fermentation with this optimal condition, the activity reached 13.69 SU/mL, which is equal to 99.3% of the predicted value.

  1. Effects of Bacillus subtilis var. natto products on symptoms caused by blood flow disturbance in female patients with lifestyle diseases

    Directory of Open Access Journals (Sweden)

    Hitosugi M

    2015-01-01

    Full Text Available Masahito Hitosugi,1,2 Katsuo Hamada,2 Kazutaka Misaka2 1Department of Legal Medicine, Shiga University of Medical Science, Otsu, Shiga, Japan; 2Department of Internal Medicine, Nagareyama Central Hospital, Nagareyama, Chiba, Japan Abstract: The fermented soybean product natto is a popular traditional food in Japan and is considered a health supplement. NKCP®, a natto-derived dietary food supplement whose main component is bacillopeptidase F, has antithrombotic, fibrinolytic, and blood pressure-lowering effects. We examined whether daily intake of NKCP® effectively improves subjective symptoms in patients with lifestyle diseases in this cross-over, double-blind study. Fermented soya extract with subtilisin NAT (nattokinase as the main component was used as an active placebo. A 4-week course of NKCP® significantly decreased the visual analog scale (VAS score for shoulder stiffness from 42.3 to 32.4 (P=0.009, the VAS score for low back pain from 25.5 to 18.8 (P=0.02, and the VAS score for coldness of the extremities from 33.1 to 25.7 (P=0.002. However, no significant difference was found in the VAS score for headache. After a 4-week course of active placebo, no significant changes in the VAS score were found for any symptoms. The significant improvement in the symptoms secondary to blood flow disturbance was caused by the improvement in blood flow by NKCP®. The use of dietary supplements based on the Japanese traditional food natto helps to relieve subjective symptoms for patients with lifestyle diseases receiving medical care. Keywords: bacillopeptidase F, Japanese traditional food, lifestyle disease, subjective symptom, supplement

  2. Co-production of Fructooligosaccharides and Levan by Levansucrase from Bacillus subtilis natto with Potential Application in the Food Industry.

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    Bersaneti, Gabrielly Terassi; Pan, Nicole Caldas; Baldo, Cristiani; Celligoi, Maria Antonia Pedrine Colabone

    2018-03-01

    Fructooligosaccharides and levan have a wide range of applications in the food industry due to their physiological and functional properties. The enzymatic synthesis of these molecules exhibits great advantages when compared with microbial fermentation. In this study, the production of levansucrase from Bacillus subtilis natto and its utilization in fructooligosaccharides and levan syntheses using different reaction conditions were described. The best condition for levansucrase production was 420.7 g L -1 of sucrose at pH 7.0, which reached 23.9 U ml -1 of transfructosylation activity. In a bioreactor, the highest production of fructooligosaccharides was 41.3 g L -1 using a medium containing 350 g L -1 sucrose at 35 °C for 36 h. The enzymatic synthesis of levan resulted in 86.9 g L -1 when conditions similar to those used for fructooligosaccharides synthesis were applied. These results indicate that the levansucrase from B. subtilis natto could be applied for the co-production of fructooligosaccharides and levan, which are biomolecules that have health benefits and are used successfully in the food industry.

  3. Enhanced secretion of natto phytase by Bacillus subtilis.

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    Tsuji, Shogo; Tanaka, Kosei; Takenaka, Shinji; Yoshida, Ken-ichi

    2015-01-01

    Phytases comprise a group of phosphatases that trim inorganic phosphates from phytic acid (IP6). In this study, we aimed to achieve the efficient secretion of phytase by Bacillus subtilis. B. subtilis laboratory standard strain 168 and its derivatives exhibit no phytase activity, whereas a natto starter secretes phytase actively. The natto phytase gene was cloned into strain RIK1285, a protease-defective derivative of 168, to construct a random library of its N-terminal fusions with 173 different signal peptides (SPs) identified in the 168 genome. The library was screened to assess the efficiency of phytase secretion based on clear zones around colonies on plates, which appeared when IP6 was hydrolyzed. The pbp SP enhanced the secretion of the natto phytase most efficiently, i.e. twice that of the original SP. Thus, the secreted natto phytase was purified and found to remove up to 3 phosphates from IP6.

  4. Toxicological assessment of nattokinase derived from Bacillus subtilis var. natto.

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    Lampe, Bradley J; English, J Caroline

    2016-02-01

    Subtilisin NAT, commonly known as "nattokinase," is a fibrinolytic enzyme produced by the bacterial strain B. subtilis var. natto, which plays a central role in the fermentation of soybeans into the popular Japanese food natto. Recent studies have reported on the potential anticoagulatory and antihypertensive effects of nattokinase administration in humans, with no indication of adverse effects. To evaluate the safety of nattokinase in a more comprehensive manner, several GLP-compliant studies in rodents and human volunteers have been conducted with the enzyme product, NSK-SD (Japan Bio Science Laboratory Co., Ltd., Japan). Nattokinase was non-mutagenic and non-clastogenic in vitro, and no adverse effects were observed in 28-day and 90-day subchronic toxicity studies conducted in Sprague-Dawley rats at doses up to 167 mg/kg-day and 1000 mg/kg-day, respectively. Mice inoculated with 7.55 × 10(8) CFU of the enzyme-producing bacterial strain showed no signs of toxicity or residual tissue concentrations of viable bacteria. Additionally consumption of 10 mg/kg-day nattokinase for 4 weeks was well tolerated in healthy human volunteers. These findings suggest that the oral consumption of nattokinase is of low toxicological concern. The 90-day oral subchronic NOAEL for nattokinase in male and female Sprague-Dawley rats is 1000 mg/kg-day, the highest dose tested. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  5. Evaluation of seed chemical quality traits and sensory properties of natto soybean.

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    Yoshikawa, Yoko; Chen, Pengyin; Zhang, Bo; Scaboo, Andrew; Orazaly, Moldir

    2014-06-15

    Natto is a popular soyfood in Japan, and the U.S. is the largest supplier of natto soybeans. However, information on natto seed chemical and sensory properties is very limited. The objectives of this study were to evaluate differences of seed chemical and sensory properties among natto types and determine heritability and correlation. A total of 15 small-seeded natto genotypes (three superior, nine moderate and three inferior) were evaluated for protein, oil, calcium, manganese, boron and sugar content and processed into a natto product to evaluate appearance, stickiness, flavor, texture and shelf-life. The superior natto group had a higher sugar content but lower protein plus oil, calcium, manganese and boron content than other two groups. Most seed quality traits exhibited high heritability. The natto sensory preference was positively correlated with sucrose and oil content, but negatively correlated with seed hardness, protein, protein plus oil, calcium, manganese, and boron contents. Selecting soybean lines with low protein, protein plus oil, calcium, manganese, and boron content while with high sucrose will be an effective approach for soybean breeding for natto production. Published by Elsevier Ltd.

  6. Identification of a Key Gene Involved in Branched-Chain Short Fatty Acids Formation in Natto by Transcriptional Analysis and Enzymatic Characterization in Bacillus subtilis.

    Science.gov (United States)

    Hong, Chenlu; Chen, Yangyang; Li, Lu; Chen, Shouwen; Wei, Xuetuan

    2017-03-01

    Natto as a fermented soybean product has many health benefits for human due to its rich nutritional and functional components. However, the unpleasant odor of natto, caused by the formation of branched-chain short fatty acids (BCFAs), prohibits the wide acceptance of natto products. This work is to identify the key gene of BCFAs formation and develop the guidance to reduce natto odor. Transcriptional analysis of BCFAs synthesis pathway genes was conducted in two Bacillus subtilis strains with obvious different BCFAs synthesis abilities. The transcriptional levels of bcd, bkdAA, and ptb in B. subtilis H-9 were 2.7-fold, 0.7-fold, and 8.9-fold higher than that of B. subtilis H-4, respectively. Therefore, the ptb gene with the highest transcriptional change was considered as the key gene in BCFAs synthesis. The ptb encoded enzyme Ptb was further characterized by inducible expression in Escherichia coli. The recombinant Ptb protein (about 32 kDa) was verified by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis analysis. The catalysis functions of Ptb were confirmed on substrates of isovaleryl-CoA and isobutyryl-CoA, and the higher catalysis efficiency of Ptb on isovaleryl-CoA explained the higher level of isovaleric acid in natto. The optimal activities of Ptb were observed at 50 °C and pH 8.0, and the enzymatic activity was inhibited by Ca 2+ , Zn 2+ , Ba 2+ , Mn 2+ , Cu 2+ , SDS, and EDTA. Collectively, this study reports a key gene responsible for BCFAs formation in natto fermentation and provides potential strategies to solve the odor problem.

  7. Dietary supplement with fermented soybeans, natto, improved the neurobehavioral deficits after sciatic nerve injury in rats.

    Science.gov (United States)

    Pan, Hung-Chuan; Cheng, Fu-Chou; Chen, Chun-Jung; Lai, Shu-Zhen; Liu, Mu-Jung; Chang, Ming-Hong; Wang, Yeou-Chih; Yang, Dar-Yu; Ho, Shu-Peng

    2009-06-01

    Clearance of fibrin and associated inflammatory cytokines by tissue-type plasminogen activator (t-PA) is related to improved regeneration in neurological disorder. The biological activity of fermented soybean (natto) is very similar to that of t-PA. We investigated the effect of the dietary supplement of natto on peripheral nerve regeneration. The peripheral nerve injury was produced by crushing the left sciatic nerve with a vessel clamp in Sprague-Dawley rats. The injured animals were fed orally either with saline or natto (16 mg/day) for seven consecutive days after injury. Increased functional outcome such as sciatic nerve functional index, angle of ankle, compound muscle action potential and conduction latency were observed in natto-treated group. Histological examination demonstrated that natto treatment improved injury-induced vacuole formation, S-100 and vessel immunoreactivities and axon loss. Oral intake of natto prolonged prothrombin time and reduced fibrinogen but did not change activated partial thromboplastin time and bleeding time. Furthermore, natto decreased injury-induced fibrin deposition, indicating a tolerant fibrinolytic activity. The treatment of natto significantly improved injury-induced disruption of blood-nerve barrier and loss of matrix component such as laminin and fibronectin. Sciatic nerve crush injury induced elevation of tumor necrosis factor alpha (TNF-alpha) production and caused apoptosis. The increased production of TNF-alpha and apoptosis were attenuated by natto treatment. These findings indicate that oral intake of natto has the potential to augment regeneration in peripheral nerve injury, possibly mediated by the clearance of fibrin and decreased production of TNF-alpha.

  8. Whole genome complete resequencing of Bacillus subtilis natto by combining long reads with high-quality short reads.

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    Mayumi Kamada

    Full Text Available De novo microbial genome sequencing reached a turning point with third-generation sequencing (TGS platforms, and several microbial genomes have been improved by TGS long reads. Bacillus subtilis natto is closely related to the laboratory standard strain B. subtilis Marburg 168, and it has a function in the production of the traditional Japanese fermented food "natto." The B. subtilis natto BEST195 genome was previously sequenced with short reads, but it included some incomplete regions. We resequenced the BEST195 genome using a PacBio RS sequencer, and we successfully obtained a complete genome sequence from one scaffold without any gaps, and we also applied Illumina MiSeq short reads to enhance quality. Compared with the previous BEST195 draft genome and Marburg 168 genome, we found that incomplete regions in the previous genome sequence were attributed to GC-bias and repetitive sequences, and we also identified some novel genes that are found only in the new genome.

  9. Effects of Bacillus subtilis natto and Different Components in Culture on Rumen Fermentation and Rumen Functional Bacteria In Vitro.

    Science.gov (United States)

    Sun, Peng; Li, Jinan; Bu, Dengpan; Nan, Xuemei; Du, Hong

    2016-05-01

    This study was to investigate the effects of live or autoclaved Bacillus subtilis natto, their fermented products and media on rumen fermentation and rumen functional bacteria in vitro. Rumen fluid from three multiparous lactating Holstein cows was combined and transferred into serum bottles after diluted. Fifteen serum bottles were divided into five treatments, which were designed as following: CTR (the fermentation of 0.5 g TMR and ruminal fluids from dairy cows), LBS (CTR plus a minimum of 10(11) cfu live Bacillus subtilis natto), ABS (CTR plus a minimum of 10(11) cfu autoclaved Bacillus subtilis natto), BSC (CTR plus 1 ml Bacillus subtilis natto fermentation products without bacteria), and BSM (CTR plus 1 ml liquid fermentation medium). When separated from the culture, live Bacillus subtilis natto individually increased the concentrations of ammonia-N (P Bacillus subtilis natto has the similar function with the live bacteria except for the ratio of acetate and propionate. Except B. fibrisolvens, live or autoclaved Bacillus subtilis natto did not influence or decreased the 16S rRNA gene quantification of the detected bacteria. BSC and BSM altered the relative expression of certain functional bacteria in the rumen. These results indicated that it was Bacillus subtilis natto thalli that played the important role in promoting rumen fermentation when applied as a probiotic in dairy ration.

  10. Comparison of soybean cultivars for enhancement of the polyamine contents in the fermented soybean natto using Bacillus subtilis (natto).

    Science.gov (United States)

    Kobayashi, Kazuya; Horii, Yuichiro; Watanabe, Satoshi; Kubo, Yuji; Koguchi, Kumiko; Hoshi, Yoshihiro; Matsumoto, Ken-Ichi; Soda, Kuniyasu

    2017-03-01

    Polyamines have beneficial properties to prevent aging-associated diseases. Raw soybean has relatively high polyamine contents; and the fermented soybean natto is a good source of polyamines. However, detailed information of diversity of polyamine content in raw soybean is lacking. The objectives of this study were to evaluate differences of polyamines among raw soybeans and select the high polyamine-containing cultivar for natto production. Polyamine contents were measured chromatographically in 16 samples of soybean, which showed high variation among soybeans as follows: 93-861 nmol/g putrescine, 1055-2306 nmol/g spermidine, and 177-578 nmol/g spermine. We then confirmed the high correlations of polyamine contents between raw soybean and natto (r = 0.96, 0.95, and 0.94 for putrescine, spermidine, and spermine, respectively). Furthermore, comparison of the polyamine contents among 9 Japanese cultivars showed that 'Nakasen-nari' has the highest polyamine contents, suggesting its suitability for enhancement of polyamine contents of natto.

  11. Which characteristic of Natto: appearance, odor, or taste most affects preference for Natto

    OpenAIRE

    Tsumura, Yuki; Ohyane, Aki; Yamashita, Kuniko; Sone, Yoshiaki

    2012-01-01

    Abstract Background In Japan, consumption of Natto, a fermented bean dish, is recommended because of its high quality protein, digestibility in the gut and its preventive effect on blood clot formation due to high vitamin K content. However, consumption of Natto in Kansai and the Chugoku area (the western part of Honshu) is less than that in the other areas of Japan probably because of a “food related cultural inhibition”. In this study, we determined which characteristic of Natto (appearance...

  12. Isolation of antifungal bacteria from Japanese fermented soybeans, natto.

    Science.gov (United States)

    Murata, Daichi; Sawano, Sayaka; Ohike, Tatsuya; Okanami, Masahiro; Ano, Takashi

    2013-12-01

    An inhibitory effect of a traditional Japanese fermented food, natto, was found against plant pathogens such as Rhizoctonia solani and Fusarium oxysporum, and the bacteria which showed inhibition were isolated from the natto. Among isolated bacteria, BC-1 and GAc exhibited a strong antagonistic effect in vitro against plant pathogens on an agar medium. The supernatant of bacterial culture also showed strong activity against R. solani, which meant the antimicrobial substances were produced and secreted into the medium. Both of the bacteria were estimated as Bacillus amyloliquefaciens from a partial sequence of the 16s rRNA gene. High performance liquid chromatography analysis clearly showed the production of the lipopeptide antibiotic iturin A by BC-1 and GAc. Copyright © 2013 The Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences. Published by Elsevier B.V. All rights reserved.

  13. Characterization of Bacillus subtilis strains in Thua nao, a traditional fermented soybean food in northern Thailand.

    Science.gov (United States)

    Inatsu, Y; Nakamura, N; Yuriko, Y; Fushimi, T; Watanasiritum, L; Kawamoto, S

    2006-09-01

    To clarify the diversity of Bacillus subtilis strains in Thua nao that produce high concentrations of products useful in food manufacturing and in health-promoting compounds. Production of amylase, protease, subtilisin NAT (nattokinase), and gamma-polyglutamic acid (PGA) by the Bacillus subtilis strains in Thua nao was measured. Productivity of protease NAT by these strains tended to be higher than by Japanese commercial natto-producing strains. Molecular diversity of isolated strains was analysed via randomly amplified polymorphic DNA-PCR fingerprinting. The strains were divided into 19 types, including a type with the same pattern as a Japanese natto-producing strain. B. subtilis strains that could be a resource for effective production of protease, amylase, subtilisin NAT, or PGA were evident in Thua nao produced in various regions in northern Thailand. This study clearly demonstrated the value of Thua nao as a potential resource of food-processing enzymes and health-promoting compounds.

  14. The Antithrombotic and Fibrinolytic Effect of Natto in Hypercholesterolemia Rats

    OpenAIRE

    Park, Kum-Ju; Kang, Jung Il; Kim, Tae-Seok; Yeo, Ik-Hyun

    2012-01-01

    Antithrombotic and fibrinolytic activity of natto was evaluated on platelet aggregation in vitro and in vivo. Natto showed inhibitory effects on platelet aggregation induced by adenosine 5′diphosphate (ADP) and collagen. Orally administered natto also showed fibrinolytic activity in hypercholesterolemia rats. Normal levels of natto, when administered for four weeks, shortened euglobulin clot lysis time (ECLT) and prolonged partial thromboplastin time (PATT) significantly compared to non-treat...

  15. Which characteristic of Natto: appearance, odor, or taste most affects preference for Natto

    Directory of Open Access Journals (Sweden)

    Tsumura Yuki

    2012-05-01

    Full Text Available Abstract Background In Japan, consumption of Natto, a fermented bean dish, is recommended because of its high quality protein, digestibility in the gut and its preventive effect on blood clot formation due to high vitamin K content. However, consumption of Natto in Kansai and the Chugoku area (the western part of Honshu is less than that in the other areas of Japan probably because of a “food related cultural inhibition”. In this study, we determined which characteristic of Natto (appearance, odor or taste most affect subjects’ perception of sensory attributes by observation of brain hemodynamics in relation to subjects’ preference for Natto. Findings In this experiment, we defined each subject’s changes in brain hemodynamics as (+ or (− corresponding to an increase or a decrease in total hemoglobin concentration after stimuli compared to that before stimuli. As a result, there was no relation between preference for Natto and change in brain hemodynamics by the stimuli of “looking at” or “smelling”, while there was a significant relationship between preference and stimulus of “ingestion”; (+ : (− = 21:15 in the subjects of the “favorite” group and (+:(− = 30:7 in the subjects of the “non-favorite” group (P = 0.034. Conclusion This result indicated that characteristic “taste” of Natto most affects preference for Natto.

  16. Which characteristic of Natto: appearance, odor, or taste most affects preference for Natto.

    Science.gov (United States)

    Tsumura, Yuki; Ohyane, Aki; Yamashita, Kuniko; Sone, Yoshiaki

    2012-05-28

    In Japan, consumption of Natto, a fermented bean dish, is recommended because of its high quality protein, digestibility in the gut and its preventive effect on blood clot formation due to high vitamin K content. However, consumption of Natto in Kansai and the Chugoku area (the western part of Honshu) is less than that in the other areas of Japan probably because of a "food related cultural inhibition". In this study, we determined which characteristic of Natto (appearance, odor or taste) most affect subjects' perception of sensory attributes by observation of brain hemodynamics in relation to subjects' preference for Natto. In this experiment, we defined each subject's changes in brain hemodynamics as (+) or (-) corresponding to an increase or a decrease in total hemoglobin concentration after stimuli compared to that before stimuli. As a result, there was no relation between preference for Natto and change in brain hemodynamics by the stimuli of "looking at" or "smelling", while there was a significant relationship between preference and stimulus of "ingestion"; (+) : (-) = 21:15 in the subjects of the "favorite" group and (+):(-) = 30:7 in the subjects of the "non-favorite" group (P = 0.034). This result indicated that characteristic "taste" of Natto most affects preference for Natto.

  17. Characterization of fermented black soybean natto inoculated with Bacillus natto during fermentation.

    Science.gov (United States)

    Hu, Yongjin; Ge, Changrong; Yuan, Wei; Zhu, Renjun; Zhang, Wujiu; Du, Lijuan; Xue, Jie

    2010-05-01

    To make nutrients more accessible and further increase biological activity, cooked black soybeans were inoculated with Bacillus natto and fermented at 37 degrees C for 48 h. The changes in physiochemical properties of fermented black soybean natto were investigated. The inoculation procedure significantly increased moisture, viscosity, color, polyphenol compounds and anthocyanin, and significantly decreased hardness after 48 h fermentation. Fibrinolytic and caseinolytic protease, beta-glucosidase activities, TCA-soluble nitrogen, and ammonia nitrogen contents in the inoculated samples significantly increased as fermentation time increased. Genistin and daidzin concentrations gradually decreased with increased fermentation time. However, genistein and daidzein increased with fermentation time, which reached 316.8 and 305.2 microg g(-1) during 48 h fermentation, respectively. DPPH radical scavenging activities of the fermented black soybeans increased linearly with fermentation time and concentration. Compared with the soaked black soybeans and cooked black soybeans, the fermented black soybeans with B. natto resulted in higher scavenging activity towards DPPH radicals, which correlated well with the content of total phenols (r = 0.9254, P natto fermented by B. natto has the potential to become a functional food because of its high antioxidant activity.

  18. Whole-Genome Sequencing and Comparative Genome Analysis of Bacillus subtilis Strains Isolated from Non-Salted Fermented Soybean Foods.

    Directory of Open Access Journals (Sweden)

    Mayumi Kamada

    Full Text Available Bacillus subtilis is the main component in the fermentation of soybeans. To investigate the genetics of the soybean-fermenting B. subtilis strains and its relationship with the productivity of extracellular poly-γ-glutamic acid (γPGA, we sequenced the whole genome of eight B. subtilis stains isolated from non-salted fermented soybean foods in Southeast Asia. Assembled nucleotide sequences were compared with those of a natto (fermented soybean food starter strain B. subtilis BEST195 and the laboratory standard strain B. subtilis 168 that is incapable of γPGA production. Detected variants were investigated in terms of insertion sequences, biotin synthesis, production of subtilisin NAT, and regulatory genes for γPGA synthesis, which were related to fermentation process. Comparing genome sequences, we found that the strains that produce γPGA have a deletion in a protein that constitutes the flagellar basal body, and this deletion was not found in the non-producing strains. We further identified diversity in variants of the bio operon, which is responsible for the biotin auxotrophism of the natto starter strains. Phylogenetic analysis using multilocus sequencing typing revealed that the B. subtilis strains isolated from the non-salted fermented soybeans were not clustered together, while the natto-fermenting strains were tightly clustered; this analysis also suggested that the strain isolated from "Tua Nao" of Thailand traces a different evolutionary process from other strains.

  19. THE ENDOGENOUS BACILLUS-SUBTILIS (NATTO) PLASMIDS PTA1015 AND PTA1040 CONTAIN SIGNAL PEPTIDASE-ENCODING GENES - IDENTIFICATION OF A NEW STRUCTURAL MODULE ON CRYPTIC PLASMIDS

    NARCIS (Netherlands)

    MEIJER, WJJ; DEJONG, A; BEA, G; WISMAN, A; TJALSMA, H; VENEMA, G; BRON, S; MAARTEN, J; VANDIJL, JM

    Various strains of Bacillus subtilis (natto) contain small cryptic plasmids that replicate via the rolling-circle mechanism. Like plasmids from other Gram-positive bacteria, these plasmids are composed of several distinct structural modules. A new structural module was identified on the B. subtilis

  20. Hydrogen production from microbial strains

    Science.gov (United States)

    Harwood, Caroline S; Rey, Federico E

    2012-09-18

    The present invention is directed to a method of screening microbe strains capable of generating hydrogen. This method involves inoculating one or more microbes in a sample containing cell culture medium to form an inoculated culture medium. The inoculated culture medium is then incubated under hydrogen producing conditions. Once incubating causes the inoculated culture medium to produce hydrogen, microbes in the culture medium are identified as candidate microbe strains capable of generating hydrogen. Methods of producing hydrogen using one or more of the microbial strains identified as well as the hydrogen producing strains themselves are also disclosed.

  1. Fiscal 1998 regional consortium R and D project. Report of research results on venture business promoting type regional consortium for medium and small business innovative foundation (Development and comercialization of ecology-oriented new biodegradable packing materials with natto resin used - 2nd year); 1998 nendo nattoo jushi wo oyoshita eko taio shinki seibunkaisei hoso shizai no kaihatsu jigyoka seika hokokusho. 2

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    2000-03-01

    This paper describes fiscal 1998 results of development of a new biodegradable packing material. Using natto resin (synthesized by irradiating fermented soybeans with {gamma} rays) for compound materials, existing biodegradable plastics resin was improved. Analysis was made on a correlation for example between honeycomb structure and water absorption rate of natto resin and between crosslinked density and mechanical strength. The production process of natto resin by electron-beam irradiation was developed, with the patent applied. The optimization of natto resin synthesis was established by using chemical regent (epoxy resin). After a compound sheet was formed by varying the mixing ratio of natto resin into poly(lactic acid), conditions were set such as formability, plasticity, etc. The production capacity of natto resin gel was improved over one thousand times more than before by electron-beam irradiation technology, with a production process established for one ton/day. The production cost of 40,000 yen/kg was achieved. No abnormality was recognized in a safety test using male rats nor in the mucosa-stimulative test using rabbits. Variability was negative. Using a microbial oxidative degradation analyzer, a completeness degradability test was carried out by analyzing the generating CO2 quantitatively, which revealed about 80% of natto resin was decomposed in two weeks. (NEDO)

  2. Levan from Bacillus subtilis Natto: its effects in normal and in streptozotocin-diabetic rats

    Directory of Open Access Journals (Sweden)

    Fernando Cesar Bazani Cabral de Melo

    2012-12-01

    Full Text Available Levan is an exopolysaccharide of fructose primarily linked by β-(2→6 glycosidic bonds with some β-(2→1 branched chains. Due to its chemical properties, levan has possible applications in both the food and pharmaceutical industries. Bacillus subtilis is a promising industrial levan producer, as it ferments sucrose and has a high levan-formation capacity. A new strain of B. subtilis was recently isolated from Japanese food natto, and it has produced levan in large quantities. For future pharmaceutical applications, this study aimed to investigate the effects of levan produced by B. subtilis Natto, mainly as potential hypoglycemic agent, (previously optimized with a molecular weight equal to 72.37 and 4,146 kDa in Wistar male rats with diabetes induced by streptozotocin and non-diabetic rats and to monitor their plasma cholesterol and triacylglycerol levels. After 15 days of experimentation, the animals were sacrificed, and their blood samples were analyzed. The results, compared using analysis of variance, demonstrated that for this type of levan, a hypoglycemic effect was not observed, as there was no improvement of diabetes symptoms during the experiment. However, levan did not affect any studied parameters in normal rats, indicating that the exopolysaccharide can be used for other purposes.

  3. Natto (fermented soybean) extract extends the adult lifespan of Caenorhabditis elegans.

    Science.gov (United States)

    Ibe, Sachie; Kumada, Kaoru; Yoshida, Keiko; Otobe, Kazunori

    2013-01-01

    We investigated the effects of a water extract of natto on the aging of the nematode Caenorhabditis elegans. The water extract significantly prolonged the adult lifespan of the wild-type worms and rendered them resistant to oxidative and thermal stress. In addition, treatment with natto extract significantly delayed the accumulation of lipofuscin, a characteristic of aging cells. Our findings suggest that components of natto have a beneficial anti-aging effect in vivo.

  4. [Transformation of baicalin and wogonoside through liquid fermentation with Bacillus natto].

    Science.gov (United States)

    Long, Hou-ning; Zhang, Shuo; Yao, Lei; Zhang, Min; Wang, Peng-jiao; Meng, Xiao-xia; Gao, Xiu; Zhang, Rong-ping

    2015-12-01

    This experiment aimed to explore and research the process of preparing baicalein and wogonin through liquid fermentation with Bacillus natto. Active enzymes of produced by B. natto was used for the biological transformation of baclin and wogonoside, in order to increase the content of the haicalein and wogonin in the scutellaria. With the content of the baicalein and wogonin as evaluating indexes, the effects of carbon source, nitrogen source, the types and suitable concentration of inorganic salt, medium pH, granularities of medical materials, liquid volume in flask, shaking speed, liquid-to-solid ratio, fermentation time on the fermentation process were studied. The optimal process conditions for liquid fermentation of scutellaria were 1.0% of peptone, 0.05% of NaCl, pH at 6, the granularities of medical materials of the scutellaria screened through 40-mesh sifter, 33% of liquid, shaker incubator speed at 200 r x min(-1), liquid-to-solid ratio of 5:1, temperature at 37 degrees C, fermentation for 6 days, baclin's conversion rate at 97.6% and wogonoside's conversion rate at 97% in the scutellaria. According to the verification test, the process was stable and feasible, and could provide data reference for the industrial production.

  5. Effects of natto extract on endothelial injury in a rat model.

    Science.gov (United States)

    Chang, Chin-Hsien; Chen, Kuo-Ti; Lee, Tsong-Hai; Wang, Chao-Hung; Kuo, Yi-Wen; Chiu, Ya-Huang; Hsieh, Ching-Liang; Wu, Chang-Jer; Chang, Yen-Lin

    2010-12-01

    Vascular endothelial damage has been found to be associated with thrombus formation, which is considered to be a risk factor for cardiovascular disease. A diet of natto leads to a low prevalence of cardiovascular disease. The aim of the present study was to investigate the effects of natto extract on vascular endothelia damage with exposure to laser irradiation. Endothelial damage both in vitro and in vivo was induced by irradiation of rose bengal using a DPSS green laser. Cell viability was determined by MTS assay, and the intimal thickening was verified by a histological approach. The antioxidant content of natto extract was determined for the free radical scavenging activity. Endothelial cells were injured in the presence of rose bengal irradiated in a dose-dependent manner. Natto extract exhibits high levels of antioxidant activity compared with purified natto kinase. Apoptosis of laser-injured endothelial cells was significantly reduced in the presence of natto extract. Both the natto extract and natto kinase suppressed intimal thickening in rats with endothelial injury. The present findings suggest that natto extract suppresses vessel thickening as a synergic effect attributed to its antioxidant and anti-apoptosis properties.

  6. Hydrogen production by recombinant Escherichia coli strains

    Science.gov (United States)

    Maeda, Toshinari; Sanchez‐Torres, Viviana; Wood, Thomas K.

    2012-01-01

    Summary The production of hydrogen via microbial biotechnology is an active field of research. Given its ease of manipulation, the best‐studied bacterium Escherichia coli has become a workhorse for enhanced hydrogen production through metabolic engineering, heterologous gene expression, adaptive evolution, and protein engineering. Herein, the utility of E. coli strains to produce hydrogen, via native hydrogenases or heterologous ones, is reviewed. In addition, potential strategies for increasing hydrogen production are outlined and whole‐cell systems and cell‐free systems are compared. PMID:21895995

  7. Effect of Bacillus subtilis natto on growth performance in Muscovy ducks

    Directory of Open Access Journals (Sweden)

    T Sheng-Qiu

    2013-09-01

    Full Text Available The aim of the present study was to determine whether dietary Bacillus subtilis natto could affect growth performance of Muscovy ducks. A total of 120 hundred Muscovy ducks at the age of 1 day were randomly assigned to four groups (30 Muscovy ducks/group, and fed with diets supplemented with 0% (control group, 0.1%, 0.2%, and 0.4% Bacillus subtilis natto, respectively during the 6-week feeding period. Weight gain, feed intake and feed conversion efficiency of Muscovy ducks were significantly improved by the dietary addition of Bacillus subtilis natto, and the results were more significant in 0.4% dietary Bacillus subtilis natto treatment group; Also, Bacillus subtilis natto reduced Escherichia coli and Salmonella colonies, and increased lactobacilli population in the ileum and the cecum. Biochemical parameters, including total protein, GOT (glutamic oxaloacetic transaminase, GPT (glutamic pyruvic transaminase, AKP (alkaline phosphatase, triiodothyronine (T3 and tetraiodothyronine (T4 contents (pBacillus subtilis natto was added to the diets (p0.05. The results of the present study indicate that diets with 0.4% Bacillus subtilis natto improved the growth performance of Muscovy ducks by increasing the absorption of protein, simulating hormone secretion, suppressing harmful microflora, and improving the duodenal structure and immune functions of Muscovy ducks. It is suggested that Bacillus subtilis natto is a potential candidate to be used use as a probiotic to improve the growth performance of Muscovy ducks.

  8. Distribution of radioactive cesium ((134)Cs Plus(137)Cs) in a contaminated Japanese soybean cultivar during the preparation of tofu, natto, and nimame (Boiled Soybean).

    Science.gov (United States)

    Hachinohe, Mayumi; Kimura, Keitarou; Kubo, Yuji; Tanji, Katsuo; Hamamatsu, Shioka; Hagiwara, Shoji; Nei, Daisuke; Kameya, Hiromi; Nakagawa, Rikio; Matsukura, Ushio; Todoriki, Setsuko; Kawamoto, Shinichi

    2013-06-01

    We investigated the fate of radioactive cesium ((134)Cs plus (137)Cs) during the production of tofu, natto, and nimame (boiled soybean) from a contaminated Japanese soybean cultivar harvested in FY2011. Tofu, natto, and nimame were made from soybean grains containing radioactive cesium (240 to 340 Bq/kg [dry weight]), and the radioactive cesium in the processed soybean foods and in by-product fractions such as okara, broth, and waste water was measured with a germanium semiconductor detector. The processing factor is the ratio of radioactive cesium concentration of a product before and after processing. For tofu, natto, nimame, and for the by-product okara, processing factors were 0.12, 0.40, 0.20, and 0.18, respectively; this suggested that these three soybean foods and okara, used mainly as an animal feed, can be considered safe for human and animal consumption according to the standard limit for radioactive cesium of soybean grains. Furthermore, the ratio of radioactive cesium concentrations in the cotyledon, hypocotyl, and seed coat portions of the soybean grain was found to be approximately 1:1:0.4.

  9. Evaluation of a lipopeptide biosurfactant from Bacillus natto TK-1 as a potential source of anti-adhesive, antimicrobial and antitumor activities Avaliação de um lipopeptídio biosurfactante de Bacillus natto TK-1 com fonte potencial de atividade antiadesiva, antimicrobiana e antitumoral

    Directory of Open Access Journals (Sweden)

    Xiao-Hong Cao

    2009-06-01

    Full Text Available A lipopeptide biosurfactant produced by Bacillus natto TK-1 has a strong surface activity. The biosurfactant was found to be an anti-adhesive agent against several bacterial strains, and also showed a broad spectrum of antimicrobial activity. The biosurfactant induced a significant reduction in tumor cells viability in a dose- dependent manner.Um lipopeptídio biosurfactante produzido por Bacillus natto TK-1 apresenta intensa atividade de superfície. Verificou-se que o biosurfactante apresentou atividade antiadesiva contra várias cepas bacterianas, e também atividade antimicrobiana de amplo espectro. O biosurfactante causou uma redução significativa na viabilidade de células tumorais, de forma dose-dependente.

  10. Fumonisin and Ochratoxin Production in Industrial Aspergillus niger Strains

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Larsen, Thomas Ostenfeld; Thrane, Ulf

    2011-01-01

    as safe). However, A. niger has the potential to produce two groups of potentially carcinogenic mycotoxins: fumonisins and ochratoxins. In this study all available industrial and many non-industrial strains of A. niger (180 strains) as well as 228 strains from 17 related black Aspergillus species were...... examined for mycotoxin production. None of the related 17 species of black Aspergilli produced fumonisins. Fumonisins (B(2), B(4), and B(6)) were detected in 81% of A. niger, and ochratoxin A in 17%, while 10% of the strains produced both mycotoxins. Among the industrial strains the same ratios were 83......%, 33% and 26% respectively. Some of the most frequently used strains in industry NRRL 337, 3112 and 3122 produced both toxins and several strains used for citric acid production were among the best producers of fumonisins in pure agar culture. Most strains used for other biotechnological processes also...

  11. Agar Plates Made from Common Supermarket Substances and Bacillus subtilis Natto as an Inexpensive Approach to Microbiology Education

    Directory of Open Access Journals (Sweden)

    Franz-Josef Scharfenberg

    2015-08-01

    Full Text Available To address the possible limitations that financial restrictions may have on students’ independent experimentation at school, we developed and implemented an inexpensive approach for basic microbiology education. We describe four nutrient agars consisting only of everyday substances available from the supermarket or online that we developed to replace standard agars and specific agars. Additionally, we selected Bacillus subtilis natto as an example of a pure-culture species. Our tip first reports the four supermarket-substance agar variants; second, it suggests utilizing them to introduce basic microbiological techniques; and third, it introduces B. subtilis natto in the context of the antibacterial effects of antibiotics as well as of supermarket products which students can bring to class from home. We implemented our approach in microbiology education at school as well as in pre-service teacher education and in in-service teacher professional development courses at our university. Finally, our paper provides worksheets for all the experiments. Editor's Note:The ASM advocates that students must successfully demonstrate the ability to explain and practice safe laboratory techniques. For more information, read the laboratory safety section of the ASM Curriculum Recommendations: Introductory Course in Microbiology and the Guidelines for Biosafety in Teaching Laboratories, available at www.asm.org. The Editors of JMBE recommend that adopters of the protocols included in this article follow a minimum of Biosafety Level 1 practices. If the soil plates described in the activity are opened, a minimum of Biosafety Level 2 is required.

  12. Mead production: selection and characterization assays of Saccharomyces cerevisiae strains.

    Science.gov (United States)

    Pereira, Ana Paula; Dias, Teresa; Andrade, João; Ramalhosa, Elsa; Estevinho, Letícia M

    2009-08-01

    Mead is a traditional drink, which results from the alcoholic fermentation of diluted honey carried out by yeasts. However, when it is produced in a homemade way, mead producers find several problems, namely, the lack of uniformity in the final product, delayed and arrested fermentations, and the production of "off-flavours" by the yeasts. These problems are usually associated with the inability of yeast strains to respond and adapt to unfavourable and stressful growth conditions. The main objectives of this work were to evaluate the capacity of Saccharomyces cerevisiae strains, isolated from honey of the Trás-os-Montes (Northeast Portugal), to produce mead. Five strains from honey, as well as one laboratory strain and one commercial wine strain, were evaluated in terms of their fermentation performance under ethanol, sulphur dioxide and osmotic stress. All the strains showed similar behaviour in these conditions. Two yeasts strains isolated from honey and the commercial wine strain were further tested for mead production, using two different honey (a dark and a light honey), enriched with two supplements (one commercial and one developed by the research team), as fermentation media. The results obtained in this work show that S. cerevisiae strains isolated from honey, are appropriate for mead production. However it is of extreme importance to take into account the characteristics of the honey, and supplements used in the fermentation medium formulation, in order to achieve the best results in mead production.

  13. Dietary soy and natto intake and cardiovascular disease mortality in Japanese adults: the Takayama study.

    Science.gov (United States)

    Nagata, Chisato; Wada, Keiko; Tamura, Takashi; Konishi, Kie; Goto, Yuko; Koda, Sachi; Kawachi, Toshiyuki; Tsuji, Michiko; Nakamura, Kozue

    2017-02-01

    Whether soy intake is associated with a decreased risk of cardiovascular disease (CVD) remains unclear. A traditional Japanese soy food, natto, contains a potent fibrinolytic enzyme. However, its relation to CVD has not been studied. We aimed to examine the association of CVD mortality with the intake of natto, soy protein, and soy isoflavones in a population-based cohort study in Japan. The study included 13,355 male and 15,724 female Takayama Study participants aged ≥35 y. At recruitment in 1992, each subject was administered a validated semiquantitative food-frequency questionnaire. Deaths from CVD were ascertained over 16 y. A total of 1678 deaths from CVD including 677 stroke and 308 ischemic heart disease occurred during follow-up. The highest quartile of natto intake compared with the lowest intake was significantly associated with a decreased risk of mortality from total CVD after control for covariates: the HR was 0.75 (95% CI: 0.64, 0.88, P-trend = 0.0004). There were no significant associations between the risk of mortality from total CVD and intakes of total soy protein, total soy isoflavone, and soy protein or soy isoflavone from soy foods other than natto. The highest quartiles of total soy protein and natto intakes were significantly associated with a decreased risk of mortality from total stroke (HR = 0.75, 95% CI: 0.57, 0.99, P-trend = 0.03 and HR = 0.68, 95% CI: 0.52, 0.88, P-trend = 0.0004, respectively). The highest quartile of natto intake was also significantly associated with a decreased risk of mortality from ischemic stroke (HR = 0.67, 95% CI:0.47, 0.95, P-trend = 0.03). Data suggest that natto intake may contribute to the reduction of CVD mortality. © 2017 American Society for Nutrition.

  14. Canthaxanthin production with modified Mucor circinelloides strains.

    Science.gov (United States)

    Papp, Tamás; Csernetics, Arpád; Nagy, Gábor; Bencsik, Ottó; Iturriaga, Enrique A; Eslava, Arturo P; Vágvölgyi, Csaba

    2013-06-01

    Canthaxanthin is a natural diketo derivative of β-carotene primarily used by the food and feed industries. Mucor circinelloides is a β-carotene-accumulating zygomycete fungus and one of the model organisms to study the carotenoid biosynthesis in fungi. In this study, the β-carotene ketolase gene (crtW) of the marine bacterium Paracoccus sp. N81106 fused with fungal promoter and terminator regions was integrated into the M. circinelloides genome to construct stable canthaxanthin-producing strains. Different transformation methods including polyethylene glycol-mediated transformation with linear DNA fragments, restriction enzyme-mediated integration and Agrobacterium tumefaciens-mediated transformation were tested to integrate the crtW gene into the Mucor genome. Mitotic stability, site of integration and copy number of the transferred genes were analysed in the transformants, and several stable strains containing the crtW gene in high copy number were isolated. Carotenoid composition of selected transformants and effect of culturing conditions, such as temperature, carbon sources and application of certain additives in the culturing media, on their carotenoid content were analysed. Canthaxanthin-producing transformants were able to survive at higher growth temperature than the untransformed strain, maybe due to the effect of canthaxanthin on the membrane fluidity and integrity. With the application of glucose, trehalose, dihydroxyacetone and L-aspartic acid as sole carbon sources in minimal medium, the crtW-expressing M. circinelloides strain, MS12+pCA8lf/1, produced more than 200 μg/g (dry mass) of canthaxanthin.

  15. Fumonisin and Ochratoxin Production in Industrial Aspergillus niger Strains

    Science.gov (United States)

    Frisvad, Jens C.; Larsen, Thomas O.; Thrane, Ulf; Meijer, Martin; Varga, Janos; Samson, Robert A.; Nielsen, Kristian F.

    2011-01-01

    Aspergillus niger is perhaps the most important fungus used in biotechnology, and is also one of the most commonly encountered fungi contaminating foods and feedstuffs, and occurring in soil and indoor environments. Many of its industrial applications have been given GRAS status (generally regarded as safe). However, A. niger has the potential to produce two groups of potentially carcinogenic mycotoxins: fumonisins and ochratoxins. In this study all available industrial and many non-industrial strains of A. niger (180 strains) as well as 228 strains from 17 related black Aspergillus species were examined for mycotoxin production. None of the related 17 species of black Aspergilli produced fumonisins. Fumonisins (B2, B4, and B6) were detected in 81% of A. niger, and ochratoxin A in 17%, while 10% of the strains produced both mycotoxins. Among the industrial strains the same ratios were 83%, 33% and 26% respectively. Some of the most frequently used strains in industry NRRL 337, 3112 and 3122 produced both toxins and several strains used for citric acid production were among the best producers of fumonisins in pure agar culture. Most strains used for other biotechnological processes also produced fumonisins. Strains optimized through random mutagenesis usually maintained their mycotoxin production capability. Toxigenic strains were also able to produce the toxins on media suggested for citric acid production with most of the toxins found in the biomass, thereby questioning the use of the remaining biomass as animal feed. In conclusion it is recommended to use strains of A. niger with inactive or inactivated gene clusters for fumonisins and ochratoxins, or to choose isolates for biotechnological uses in related non-toxigenic species such as A. tubingensis, A. brasiliensis, A vadensis or A. acidus, which neither produce fumonisins nor ochratoxins. PMID:21853139

  16. Effect of Natto on Meat Quality and Skatole Content in TOPIGS Pigs

    Directory of Open Access Journals (Sweden)

    Q. K. Sheng

    2016-05-01

    Full Text Available This study investigated the effect of Bacillus subtilis (B. subtilis natto on meat quality and skatole in TOPIGS pigs. Sixty TOPIGS pigs were randomly assigned to 3 groups (including 5 pens per group, with 4 pigs in each pen and fed with basic diet (control group, basic diet plus 0.1% B. subtilis natto (B group, and basic diet plus 0.1% B. subtilis natto plus 0.1% B. coagulans (BB group, respectively. All pigs were sacrificed at 100 kg. Growth performance, meat quality, serum parameters and oxidation status in the three groups were assessed and compared. Most parameters regarding growth performance and meat quality were not significantly different among the three groups. However, compared with the control group, meat pH24, fat and feces skatole and the content of Escherichia coli (E. Coli, Clostridium, NH3-N were significantly reduced in the B and BB groups, while serum total cholesterol, high density lipoprotein, the levels of liver P450, CYP2A6, and CYP2E1, total antioxidant capability (T-AOC and glutathione peroxidase and Lactobacilli in feces were significantly increased in the B and BB groups. Further, the combined supplementation of B. subtilis natto and B. coagulans showed more significant effects on the parameters above compared with B. subtilis, and Clostridium, and NH3-N. Our results indicate that the supplementation of pig feed with B. subtilis natto significantly improves meat quality and flavor, while its combination with B. coagulans enhanced these effects.

  17. Ethanol production potential of local yeast strains isolated from ripe ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-05-16

    May 16, 2008 ... ... of these studies, the preferred candidate for industrial production of ethanol ... The yeast strains were isolated using the method of Ameh et al. (1989), on ... gas in the Durham tube during the incubation period. Fermentation ...

  18. Antimicrobial Resistance of Staphylococcal Strains Isolated from Various Pathological Products

    Directory of Open Access Journals (Sweden)

    Laura-Mihaela SIMON

    2010-12-01

    Full Text Available Background: The optimal choice of antimicrobial therapy is an important problem in hospital environment in which the selection of resistant and virulent strains easy occurs. S. aureus and especially MRSA(methicillin-resistant S. aureus creates difficulties in both treatment and prevention of nosocomial infections. Aim: The purpose of this study is to determine the sensitivity and the resistance to chemotherapy of staphylococci strains isolated from various pathological products. Material and Method: We identified Staphylococccus species after morphological appearance, culture properties, the production of coagulase, hemolisines and the enzyme activity. The susceptibility tests were performed on Mueller-Hinton medium according to CLSI (Clinical and Laboratory Standards Institute. Results: The strains were: MSSA (methicillin-susceptible S. aureus (74%, MRSA (8%, MLS B (macrolides, lincosamides and type B streptogramines resistance (12% and MRSA and MLS B (6%. MRSA strains were more frequently isolated from sputum. MRSA associated with the MLS B strains were more frequently isolated from pus. MLS B strains were more frequently isolated from sputum and throat secretions. All S. aureus strains were susceptible to vancomycin and teicoplanin. Conclusions: All staphylococcal infections require resistance testing before treatment. MLS B shows a high prevalence among strains of S. aureus. The association between MLS B and MRSA remains a major problem in Romania.

  19. Xylitol production from colombian native yeast strains

    Directory of Open Access Journals (Sweden)

    Isleny Andrea Vanegas Córdoba

    2004-07-01

    Full Text Available Xylitol is an alternative sweetener with similar characteristics to sucrose that has become of great interest, due mainly to its safe use in diabetic patients and those deficient in glucose-6-phosphate-dehydrogenase. Its chemical production is expensive and generates undesirable by-products, whereas biotechnological process, which uses different yeasts genera, is a viable production alternative because it is safer and specific. Colombia has a privilege geographic location and offers a great microbial variety, this can be taken advantage of with academic and commercial goals. Because of this, some native microorganisms with potential to produce xylitol were screened in this work. It were isolated 25 yeasts species, from which was possible to identify 84% by the kit API 20C-AUX. Three yeasts: Candida kefyr, C. tropicalis y C. parapsilosis presented greater capacity to degrade xylose compared to the others, therefore they were selected for the later evaluation of its productive capacity. Discontinuous cellular cultures were developed in shaken flasks at 200 rpm and 35°C by 30 hours, using synthetic media with xylose as carbon source. Xylose consumption and xylitol production were evaluated by thin layer chromatography and high performance liquid chromatography. The maximal efficiency were obtained with Candida kefyr and C. tropicalis (Yp/s 0.5 y 0.43 g/g, respectively, using an initial xylose concentration of 20 g/L. Key words: Xylitol, xylose, yeasts, Candida kefyr, C. tropicalis, C. parapsilosis.

  20. Penicillin production by mutant strains of penicillium chrysogenum

    International Nuclear Information System (INIS)

    Tawfik, Z.S.; Ashour, M.S.; Shihab, A.

    1986-01-01

    The mutagenic agent 8-rays was used to initiate the penicillium chrysogenum isolated from local spices. After irradiation, colonies invariably differing from the parent strain in their morphological and cultural characteristics were tested for antibiotic production on fermentation agar medium. Twenty two isolates were found to be penicillin producing mutant strains. Mutant strain M 24 forming small colonies with white conidia was found to be a high yielding penicillin producer (9550 i.u/ml). All of the 22 isolates obtained lost their ability to produce the antibiotic after 11 months storage at 4 0 with subsequent subculturing

  1. Growth and α-amylase production by strains of Lactobacillus ...

    African Journals Online (AJOL)

    A cassava starch medium was used to analyse the dynamics of batch growth and α-amylase production of strains of Lactobacillus plantarum and Rhizopus oryzae isolated from cassava dried chips. The strains displayed a growth of 0.5h-1 and 0.55 h-1, a biomass yield on cassava starch of 0.49g/g and 0.5g/g, a maximum ...

  2. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto

    OpenAIRE

    Yanagisawa, Yasuhide; Chatake, Toshiyuki; Chiba-Kamoshida, Kaori; Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki; Yasuda, Ichiro; Morimoto, Yukio

    2010-01-01

    Nattokinase, a protein found in high levels in the traditional Japanese food natto, has been reported to have high thrombolytic activity. In the present study, the crystallization of native nattokinase and the collection of X-ray diffraction date from a nattokinase crystal to a resolution of 1.74 Å are reported.

  3. Purification and characterization of nattokinase from Bacillus subtilis natto B-12.

    Science.gov (United States)

    Wang, Cong; Du, Ming; Zheng, Dongmei; Kong, Fandong; Zu, Guoren; Feng, Yibing

    2009-10-28

    Bacillus subtilis natto B-12 was isolated from natto, a traditional fermented soybean food in Japan. A fibrinolytic enzyme (B-12 nattokinase) was purified from the supernatant of B. subtilis natto B-12 culture broth and showed strong fibrinolytic activity. The enzyme was homogenously purified to 56.1-fold, with a recovery of 43.2% of the initial activity. B-12 nattokinase was demonstrated to be homogeneous by SDS-PAGE and was identified as a monomer of 29000 +/- 300 Da in its native state by SDS-PAGE and size exclusion methods. The optimal pH value and temperature were 8.0 and 40 degrees C, respectively. Purified nattokinase showed high thermostability at temperatures from 30 to 50 degrees C and alkaline stability within the range of pH 6.0-9.0. The enzyme activity was activated by Zn(2+) and obviously inhibited by Fe(3+) and Al(3+). This study provides some important information for the effect factors of fibrinolytic activity, the purification methods, and characterization of nattokinase from B. subtilis natto B-12, which enriches the theoretical information of nattokinase for the research and development of nattokinase as a functional additive of food.

  4. Yeast strains designed for 2. generation bioethanol production. Final report

    Energy Technology Data Exchange (ETDEWEB)

    Roennow, B.

    2013-04-15

    The aim of the project was to develop a suitable fermentation organism for 2G bioethanol production that would efficiently ferment all of the sugars in lignocellulosic biomass into ethanol at a commercially viable rate (comparable to yeast based 1G ethanol production). More specifically, a yeast strain would be developed with the ability to ferment also the pentoses in lignocellulosic biomass and thereby increase the ethanol yield of the process by 30-45% with a profound positive effect on the total process economy. The project has succeeded in developing a new industrial yeast strain V1. The yeast strain can transform the difficult C5 sugars to ethanol from waste products such as straw and the like from the agricultural sector. The classic issues relating to industrial uses such as inhibitor and ethanol tolerance and high ethanol production is resolved satisfactorily. The potential of the use of the new strain for 2nd generation bioethanol production is that the ethanol yields increase by 30-45%. With the increased ethanol yield follows a marked improvement in the overall process economics. (LN)

  5. Ochratoxin A production by strains of Aspergillus niger var. niger.

    Science.gov (United States)

    Abarca, M L; Bragulat, M R; Castellá, G; Cabañes, F J

    1994-01-01

    In a survey of the occurrence of ochratoxin A (OA)-positive strains isolated from feedstuffs, two of the 19 isolates of Aspergillus niger var. niger that were studied produced OA in 2% yeast extract-15% sucrose broth and in corn cultures. This is the first report of production of OA by this species. PMID:8074536

  6. LYOPHILIZATION EFFECT ON PRODUCTIVITY OF BUTANOL-PRODUCING STRAINS

    Directory of Open Access Journals (Sweden)

    O. O. Tigunova

    2016-10-01

    Full Text Available Investigation of lyophilization effect on the productivity of butanol-producing strains was the aim of our research. For this purpose we used butanol-producing strains; technical glycerol; biomass of switchgrass Panicum virgatum L. Lyophilization was performed using a lyophilization-drying. The effect of the protective medium on residual moisture of freezedrying cultures suspensions depending on the concentration of glucose and sucrose was studed. It was shown that the lowest residual moisture was attained by using glucose and sucrose in amount of 10% and if the samples of freeze-drying bacteria had been saved for one month at 4 οC the productivity did not decrease. As temperature preservation was increased the productivity of the cultures was gradually decreased and it was greatly reduced at 30 οC. So the protective medium composition was optimized for lyophilization of butanol-producing strains as follows: sucrose 10.0%; gelatin 10.0%; agar 0.02%. It was shown that the preservation of samples of freeze-drying bacteria for six months at a temperature of 4 οC did not affect the productivity of strains. It was found that cultures could use glycerol as a carbon source for butanol accumulation before lyophilization.

  7. Elevated temperature stress strain behavior of beryllium powder product

    International Nuclear Information System (INIS)

    Abeln, S.P.; Field, R.; Mataya, M.C.

    1995-01-01

    Several grades of beryllium powder product were tested under isothermal conditions in compression over a temperature range of room temperature to 1000 C and a strain rate range from 0.001 s -1 to 1 s -1 . Samples were compressed to a total strain of 1 (64% reduction in height). It is shown that all the grades are strain rate sensitive and that strain rate sensitivity increases with temperature. Yield points were exhibited by some grades up to a temperature of 500 C, and appeared to be primarily dependent on prior thermal history which determined the availability of mobile dislocations. Serrated flow in the form of stress drops was seen in all the materials tested and was most pronounced at 500 C. The appearance and magnitude of the stress drops were dependent on accumulated strain, strain rate, sample orientation, and composition. The flow stress and shape of the flow curves differed significantly from grade to grade due to variations in alloy content, the size and distribution of BeO particles, aging precipitates, and grain size. The ductile-brittle transition temperature (DBTT) was determined for each grade of material and shown to be dependent on composition and thermal treatment. Structure/property relationships are discussed using processing history, microscopy (light and transmission), and property data

  8. Primary Screening of 10 - Hydroxy - 2 - Decenoic Acid Productive Strains

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    In this paper, eleven strains, which vere screened strictly from raw royal.jelly, soil and honeycomb etc. by means of dilution plate and spread plate methods, were cultured at 28°C for60 h with shaking. To determine whether they could yield 10-Hydroxy-2-decenoic acid during fermentation, gas chromatography and gas chromatography-mass spectrometry methods were used. The results showed that the strains BH002 and BH004. were both identified as Crvtococcaceae. where BH002 was primarily classified into Candida for possessing the abilities. The 10-HDA productivity of Candida BH002 and that of BH004 were 0.327% and 0.2648% respectively.

  9. OPTIMIZATION OF ENNIATINS PRODUCTION BY AN ENDOPHYTIC STRAIN FUSARIUM DIMERUM

    Directory of Open Access Journals (Sweden)

    Eva Buchtová

    2014-10-01

    Full Text Available The goal of this study was to find suitable composition of cultivation media for enniatin production by isolated endophytic strain Fusarium dimerum. In order to find optimal cultivation media, mono- and di- saccharides, complex nitrogen sources and L-amino acids directed biosynthesis of enniatins were tested. Submerged cultivation experiments were carried out in cultivation flasks. Most promising medium for enniatin accumulation contained fructose, malt extract and peptone for bacteriology. Finally, quite expensive carbon source fructose was replaced by more available syrups. Optimization resulted in 4-times elevated enniatin biosynthesis by metabolites production microorganism. Moreover, this is the strain obtained from Magnolia soulangeana, which has similar metabolites spectrum as the isolated Fusarium dimerum. Comparison of these results with published ones revealed that this endophyte is a potential strain for enniatins biosynthesis in submerged cultivation in which the maximum accumulation 1.27 g.L-1 of enniatin in culture medium was reached in a short period (96 h. The results proved that the endophytic strain F. dimerum may potentially be applied for efficient production of bioactive enniatins.

  10. Rapid purification and plasticization of D-glutamate-containing poly-γ-glutamate from Japanese fermented soybean food natto.

    Science.gov (United States)

    Ashiuchi, Makoto; Oike, Shota; Hakuba, Hirofumi; Shibatani, Shigeo; Oka, Nogiho; Wakamatsu, Taisuke

    2015-12-10

    Poly-γ-glutamate (PGA) is a major component of mucilage derived from natto, a Japanese fermented food made from soybeans, and PGAs obtained under laboratory's conditions contain numerous d-glutamyl residues. Natto foods are thus promising as a source for nutritionally safe d-amino acids present in intact and digested polymers, although there is little information on the stereochemistry of PGA isolated directly from natto. Here, we describe the development of a new process for rapid purification of PGA using alum and determined the D-glutamate content of natto PGA by chiral high-performance liquid chromatographic analysis. Further, using hexadecylpyridinium cation (HDP(+)), which is a compound of toothpaste, we chemically transformed natto PGA into a new thermoplastic material, called DL-PGAIC. (1)H nuclear magnetic resonance and calorimetric measurements indicate that DL-PGAIC is a stoichiometric complex of natto PGA and HDP(+) with glass transition points of -16.8 °C and -3.1 °C. Then, DL-PGAIC began decomposing at 210°C, suggesting thermal stability suitable for use as a supramolecular soft plastic. Copyright © 2015 Elsevier B.V. All rights reserved.

  11. Bioactivity characterization of Lactobacillus strains isolated from dairy products

    Science.gov (United States)

    Haghshenas, Babak; Nami, Yousef; Haghshenas, Minoo; Abdullah, Norhafizah; Rosli, Rozita; Radiah, Dayang; Yari Khosroushahi, Ahmad

    2015-01-01

    This study aimed to find candidate strains of Lactobacillus isolated from sheep dairy products (yogurt and ewe colostrum) with probiotic and anticancer activity. A total of 100 samples were randomly collected from yogurt and colostrum and 125 lactic acid bacteria were isolated. Of these, 17 Lactobacillus strains belonging to five species (L. delbrueckii, L. plantarum, L. rhamnosus, L. paracasei, and L. casei) were identified. L. plantarum 17C and 13C, which isolated from colostrums, demonstrated remarkable results such as resistant to low pH and high concentrations of bile salts, susceptible to some antibiotics and good antimicrobial activity that candidate them as potential probiotics. Seven strains (1C, 5C, 12C, 13C, 17C, 7M, and 40M), the most resistant to simulated digestion, were further investigated to evaluate their capability to adhere to human intestinal Caco-2 cells. L. plantarum 17C was the most adherent strain. The bioactivity assessment of L. plantarum 17C showed anticancer effects via the induction of apoptosis on HT-29 human cancer cells and negligible side effects on one human epithelial normal cell line (FHs 74). The metabolites produced by this strain can be used as alternative pharmaceutical compounds with promising therapeutic indices because they are not cytotoxic to normal mammalian cells. PMID:26219634

  12. Strain-related acid production by oral streptococci

    DEFF Research Database (Denmark)

    de Soet, JJ; Nyvad, Bente; Kilian, Mogens

    2000-01-01

    Acid production, in particular at low pH, is thought to be an important ecological determinant in dental caries. The aim of the present study was to determine the acid producing capability at different pH levels of 47 streptococcal strains, representing 9 species, isolated from human dental plaque....... The bacteria were grown until mid log-phase under anaerobic conditions and acid production was measured in a pH-stat system at pH 7.0, 6.0, 5.5 and 5.0. At all pH values, the mean velocity of acid production (V(ap)) by Streptococcus mutans and S. sobrinus was significantly higher (p... that of the other oral streptococci, including S. mitis, S. oralis, S. gordonii, S. sanguis, S. intermedius, S. anginosus, S. constellatus, and S. vestibularis. However, the V(ap) of some strains of S. mitis biovar 1 and S. oralis, particularly at pH values of 7.0 and 6.0, exceeded that of some strains of S. mutans...

  13. Radioprotective action of milk product fermented by strain LBL 4

    International Nuclear Information System (INIS)

    Minkova, M.; G'osheva, L.; Brankova, R.

    1992-01-01

    A food product containing L. Bulgaricus LBL 4 strain and lysozyme was studied for influence upon resistance of experimental animals to nonlethal radiation exposure. The effect was assessed by recording the response of the most radiosensitive body system, that of blood formation. Male Wistar rats were used. The milk product was given by mouth daily for 15 days (3x5 days) prior to 3-Gy gamma irradiation. On day 3 and day 10 in the postradiation period, measurements were made of spleen weight, spleen and bone-marrow cellularity, and peripheral leukocyte counts. The evidence obtained indicated that pretreatment by dietary intake of LBL--4-containing milk product increased the resistance of the blood forming system to nonlethal gamma irradiation, which could be explained by strengthening of the immune activity of the body.

  14. Impact of exopolysaccharide production on functional properties of some Lactobacillus salivarius strains.

    Science.gov (United States)

    Mercan, Emin; İspirli, Hümeyra; Sert, Durmuş; Yılmaz, Mustafa Tahsin; Dertli, Enes

    2015-11-01

    The aim of this work was to characterize functional properties of Lactobacillus salivarius strains isolated from chicken feces. Detection of genes responsible for exopolysaccharide (EPS) production revealed that all strains harbored a dextransucrase gene, but p-gtf gene was only detected in strain E4. Analysis of EPS production levels showed significant alterations among strains tested. Biofilm formation was found to be medium composition dependant, and there was a negative correlation with biofilm formation and EPS production. Autoaggregation properties and coaggregation of L. salivarius strains with chicken pathogens were appeared to be specific at strain level. An increment in bacterial adhesion to chicken gut explants was observed in L. salivarius strains with the reduction in EPS production levels. This study showed that strain-specific properties can determine the functional properties of L. salivarius strains, and the interference of these properties might be crucial for final selection of these strains for technological purposes.

  15. Growth characteristics and enzyme production optimization of lipase Producing Strain

    Science.gov (United States)

    Zheng, Chaocheng

    2018-01-01

    55 samples from different regions were selected and screened by Rhodamine B flat transparent circle method to observe lipase producing effect, among which, LHY-1, identified as Serratia sp. has the characteristics of fast growth, high enzyme production and stable ability. The colony of this strain is white, the edge is smooth and tidy, the surface is moist, the cell is straight, rod-shaped, gram negative, 0.1-0.2 μm in diameter and, length 0.3-0.5 μm in length.

  16. Solving ethanol production problems with genetically modified yeast strains

    Directory of Open Access Journals (Sweden)

    A. Abreu-Cavalheiro

    2013-09-01

    Full Text Available The current world demand for bioethanol is increasing as a consequence of low fossil fuel availability and a growing number of ethanol/gasoline flex-fuel cars. In addition, countries in several parts of the world have agreed to reduce carbon dioxide emissions, and the use of ethanol as a fuel (which produces fewer pollutants than petroleum products has been considered to be a good alternative to petroleum products. The ethanol that is produced in Brazil from the first-generation process is optimized and can be accomplished at low cost. However, because of the large volume of ethanol that is produced and traded each year, any small improvement in the process could represent a savings of billions dollars. Several Brazilian research programs are investing in sugarcane improvement, but little attention has been given to the improvement of yeast strains that participate in the first-generation process at present. The Brazilian ethanol production process uses sugarcane as a carbon source for the yeast Saccharomyces cerevisiae. Yeast is then grown at a high cellular density and high temperatures in large-capacity open tanks with cells recycle. All of these culture conditions compel the yeast to cope with several types of stress. Among the main stressors are high temperatures and high ethanol concentrations inside the fermentation tanks during alcohol production. Moreover, the competition between the desired yeast strains, which are inoculated at the beginning of the process, with contaminants such as wild type yeasts and bacteria, requires acid treatment to successfully recycle the cells. This review is focused on describing the problems and stressors within the Brazilian ethanol production system. It also highlights some genetic modifications that can help to circumvent these difficulties in yeast.

  17. Solving ethanol production problems with genetically modified yeast strains.

    Science.gov (United States)

    Abreu-Cavalheiro, A; Monteiro, G

    2013-01-01

    The current world demand for bioethanol is increasing as a consequence of low fossil fuel availability and a growing number of ethanol/gasoline flex-fuel cars. In addition, countries in several parts of the world have agreed to reduce carbon dioxide emissions, and the use of ethanol as a fuel (which produces fewer pollutants than petroleum products) has been considered to be a good alternative to petroleum products. The ethanol that is produced in Brazil from the first-generation process is optimized and can be accomplished at low cost. However, because of the large volume of ethanol that is produced and traded each year, any small improvement in the process could represent a savings of billions dollars. Several Brazilian research programs are investing in sugarcane improvement, but little attention has been given to the improvement of yeast strains that participate in the first-generation process at present. The Brazilian ethanol production process uses sugarcane as a carbon source for the yeast Saccharomyces cerevisiae. Yeast is then grown at a high cellular density and high temperatures in large-capacity open tanks with cells recycle. All of these culture conditions compel the yeast to cope with several types of stress. Among the main stressors are high temperatures and high ethanol concentrations inside the fermentation tanks during alcohol production. Moreover, the competition between the desired yeast strains, which are inoculated at the beginning of the process, with contaminants such as wild type yeasts and bacteria, requires acid treatment to successfully recycle the cells. This review is focused on describing the problems and stressors within the Brazilian ethanol production system. It also highlights some genetic modifications that can help to circumvent these difficulties in yeast.

  18. Production of functionalized polyhydroxyalkanoates by genetically modified Methylobacterium extorquens strains

    Directory of Open Access Journals (Sweden)

    Miguez Carlos B

    2010-09-01

    Full Text Available Abstract Background Methylotrophic (methanol-utilizing bacteria offer great potential as cell factories in the production of numerous products from biomass-derived methanol. Bio-methanol is essentially a non-food substrate, an advantage over sugar-utilizing cell factories. Low-value products as well as fine chemicals and advanced materials are envisageable from methanol. For example, several methylotrophic bacteria, including Methylobacterium extorquens, can produce large quantities of the biodegradable polyester polyhydroxybutyric acid (PHB, the best known polyhydroxyalkanoate (PHA. With the purpose of producing second-generation PHAs with increased value, we have explored the feasibility of using M. extorquens for producing functionalized PHAs containing C-C double bonds, thus, making them amenable to future chemical/biochemical modifications for high value applications. Results Our proprietary M. extorquens ATCC 55366 was found unable to yield functionalized PHAs when fed methanol and selected unsaturated carboxylic acids as secondary substrates. However, cloning of either the phaC1 or the phaC2 gene from P. fluorescens GK13, using an inducible and regulated expression system based on cumate as inducer (the cumate switch, yielded recombinant M. extorquens strains capable of incorporating modest quantities of C-C double bonds into PHA, starting from either C6= and/or C8=. The two recombinant strains gave poor results with C11=. The strain containing the phaC2 gene was better at using C8= and at incorporating C-C double bonds into PHA. Solvent fractioning indicated that the produced polymers were PHA blends that consequently originated from independent actions of the native and the recombinant PHA synthases. Conclusions This work constitutes an example of metabolic engineering applied to the construction of a methanol-utilizing bacterium capable of producing functionalized PHAs containing C-C double bonds. In this regard, the PhaC2 synthase

  19. Production, optimization and characterization of fibrinolytic enzyme by Bacillus subtilis RJAS19.

    Science.gov (United States)

    Kumar, D J Mukesh; Rakshitha, R; Vidhya, M Annu; Jennifer, P Sharon; Prasad, Sandip; Kumar, M Ravi; Kalaichelvan, P T

    2014-04-01

    The present study aimed at the production, purification and characterization of fibrinolytic nattokinase enzyme from the bacteria isolated from natto food. For the purpose, a fibrinolytic bacterium was isolated and identified as Bacillus subtilis based on 16S rDNA sequence analysis. The strain was employed for the production and optimization of fibrinolytic enzyme. The strain showed better enzyme production during 72nd h of incubation time with 50 degrees C at the pH 9. The lactose and peptone were found to be increasing the enzyme production rate. The enzyme produced was purified and also characterized with the help of SDS-PAGE analysis. The activity and stability profile of the purified enzyme was tested against different temperature and pH. The observations suggesting that the potential of fibrinolytic enzyme produced by Bacillus subtilis RJAS 19 for its applications in preventive medicines.

  20. Lactic acid production from xylose by Geobacillus stearothermophilus strain 15

    Science.gov (United States)

    Kunasundari, B.; Naresh, S.; Chu, J. E.

    2017-09-01

    Lactic acid is an important compound with a wide range of industrial applications. The present study tested the efficiency of xylose, as a sole carbon source to be converted to lactic acid by Geobacillus stearothermophilus strain 15. To the best of our knowledge, limited information is available on the directed fermentation of xylose to lactic acid by this bacterium. The effects of different parameters such as temperature, pH, incubation time, agitation speed, concentrations of nitrogen and carbon sources on the lactic acid production were investigated statistically. It was found that the bacterium exhibited poor assimilation of xylose to lactic acid. Temperature, agitation rate and incubation time were determined to improve the lactic acid production slightly. The highest lactic acid yield obtained was 8.9% at 45°C, 300 RPM, 96 h, pH of 6.0 with carbon and nitrogen source concentrations were fixed at 5% w/v.

  1. Cellulase production by a strain of Myrothecium sp

    Energy Technology Data Exchange (ETDEWEB)

    Kassim, E A

    1982-01-01

    A selected strain of Myrothecium sp. was grown on various carbon sources. Cellulose was found to be the highest inducer of cellulase. CMC resulted in a moderate yield. Cellobiose resulted in a low yield. Glucose, lactose, maltose and soluble starch resulted in negligible amounts. Sucrose, glycerol and salicin were extremely unsuitable. Continuous addition of glucose or cellobiose during fermentation to cellulosic culture media reduced cellulase production, whereas addition of the entire amount of glucose or cellobiose at the beginning did not affect the enzyme production. The enzyme was precipitated from the culture filtrate with ammonium sulfate giving crude cellulase, 3854 units/g. The culture filtrate was concentrated to a one-tenth volume, 97 units/ml. The purified cellulase was prepared by dialysis 6700 units/g of enzyme precipitate.

  2. Late-onset anaphylaxis due to poly (γ-glutamic acid) in the soup of commercial cold Chinese noodles in a patient with allergy to fermented soybeans (natto).

    Science.gov (United States)

    Inomata, Naoko; Chin, Keishi; Nagashima, Mayumi; Ikezawa, Zenro

    2011-09-01

    Fermented soybeans (natto) have been reported to induce IgE-mediated, late-onset anaphylaxis without early-phase responses. However, the relevant allergens of natto allergy have never been identified. A 38-year-old man developed an anaphylactic reaction accompanied by flashing, generalized urticaria, conjunctival redness, and dyspnea 3 hours after ingestion of commercial cold Chinese noodles. He had avoided natto for the past year due to developing several anaphylactic reactions half a day after natto ingestion. The results of skin prick tests (SPTs) were strongly positive for natto and the soup of cold Chinese noodles. Furthermore, SPTs showed positive for poly (γ-glutamic acid) (PGA), which is a major constituent of natto mucilage, alone among all the ingredients of the cold Chinese noodle soup. Therefore, he was diagnosed with late-onset anaphylaxis to PGA contained in natto and the cold Chinese noodle soup. These results indicated that in the present case, the relevant allergen of late-onset anaphylaxis may have been PGA in all episodes and that the patient had been sensitized by PGA through natto ingestion. PGA is produced by Bacillus subtilis during fermentation and is a high-molecular, biodegradable polymer. The late onset is therefore, hypothesized to be due to a delayed absorption of PGA, as PGA biodegrades to peptides sufficiently small to be absorbed in the bowel. PGA has recently been applied to a wide range of fields such as foods, cosmetics, and medicine. Therefore, patients with late-onset anaphylaxis to PGA of natto should avoid not only natto but also other materials containing PGA.

  3. Halotolerance, ligninase production and herbicide degradation ability of basidiomycetes strains.

    Science.gov (United States)

    Arakaki, R L; Monteiro, D A; Boscolo, M; Dasilva, R; Gomes, E

    2013-12-01

    Fungi have been recently recognized as organisms able to grow in presence of high salt concentration with halophilic and halotolerance properties and their ligninolytic enzyme complex have an unspecific action enabling their use to degradation of a number of xenobiotic compounds. In this work, both the effect of salt and polyols on growth of the basidiomycetes strains, on their ability to produce ligninolytic enzyme and diuron degradation were evaluated. Results showed that the presence of NaCl in the culture medium affected fungal specimens in different ways. Seven out of ten tested strains had growth inhibited by salt while Dacryopinax elegans SXS323, Polyporus sp MCA128 and Datronia stereoides MCA167 fungi exhibited higher biomass production in medium containing 0.5 and 0.6 mol.L(-1) of NaCl, suggesting to be halotolerant. Polyols such as glycerol and mannitol added into the culture media improved the biomass and ligninases production by D. elegans but the fungus did not reveal consumption of these polyols from media. This fungus degraded diuron in medium control, in presence of NaCl as well as polyols, produced MnP, LiP and laccase.

  4. Halotolerance, ligninase production and herbicide degradation ability of basidiomycetes strains

    Directory of Open Access Journals (Sweden)

    R.L. Arakaki

    2013-12-01

    Full Text Available Fungi have been recently recognized as organisms able to grow in presence of high salt concentration with halophilic and halotolerance properties and their ligninolytic enzyme complex have an unspecific action enabling their use to degradation of a number of xenobiotic compounds. In this work, both the effect of salt and polyols on growth of the basidiomycetes strains, on their ability to produce ligninolytic enzyme and diuron degradation were evaluated. Results showed that the presence of NaCl in the culture medium affected fungal specimens in different ways. Seven out of ten tested strains had growth inhibited by salt while Dacryopinax elegans SXS323, Polyporus sp MCA128 and Datronia stereoides MCA167 fungi exhibited higher biomass production in medium containing 0.5 and 0.6 mol.L-1 of NaCl, suggesting to be halotolerant. Polyols such as glycerol and mannitol added into the culture media improved the biomass and ligninases production by D. elegans but the fungus did not reveal consumption of these polyols from media. This fungus degraded diuron in medium control, in presence of NaCl as well as polyols, produced MnP, LiP and laccase.

  5. Biodiesel production with microalgae as feedstock: from strains to biodiesel.

    Science.gov (United States)

    Gong, Yangmin; Jiang, Mulan

    2011-07-01

    Due to negative environmental influence and limited availability, petroleum-derived fuels need to be replaced by renewable biofuels. Biodiesel has attracted intensive attention as an important biofuel. Microalgae have numerous advantages for biodiesel production over many terrestrial plants. There are a series of consecutive processes for biodiesel production with microalgae as feedstock, including selection of adequate microalgal strains, mass culture, cell harvesting, oil extraction and transesterification. To reduce the overall production cost, technology development and process optimization are necessary. Genetic engineering also plays an important role in manipulating lipid biosynthesis in microalgae. Many approaches, such as sequestering carbon dioxide from industrial plants for the carbon source, using wastewater for the nutrient supply, and maximizing the values of by-products, have shown a potential for cost reduction. This review provides a brief overview of the process of biodiesel production with microalgae as feedstock. The methods associated with this process (e.g. lipid determination, mass culture, oil extraction) are also compared and discussed.

  6. Acetone production with metabolically engineered strains of Acetobacterium woodii.

    Science.gov (United States)

    Hoffmeister, Sabrina; Gerdom, Marzena; Bengelsdorf, Frank R; Linder, Sonja; Flüchter, Sebastian; Öztürk, Hatice; Blümke, Wilfried; May, Antje; Fischer, Ralf-Jörg; Bahl, Hubert; Dürre, Peter

    2016-07-01

    Expected depletion of oil and fossil resources urges the development of new alternative routes for the production of bulk chemicals and fuels beyond petroleum resources. In this study, the clostridial acetone pathway was used for the formation of acetone in the acetogenic bacterium Acetobacterium woodii. The acetone production operon (APO) containing the genes thlA (encoding thiolase A), ctfA/ctfB (encoding CoA transferase), and adc (encoding acetoacetate decarboxylase) from Clostridium acetobutylicum were cloned under the control of the thlA promoter into four vectors having different replicons for Gram-positives (pIP404, pBP1, pCB102, and pCD6). Stable replication was observed for all constructs. A. woodii [pJIR_actthlA] achieved the maximal acetone concentration under autotrophic conditions (15.2±3.4mM). Promoter sequences of the genes ackA from A. woodii and pta-ack from C. ljungdahlii were determined by primer extension (PEX) and cloned upstream of the APO. The highest acetone production in recombinant A. woodii cells was achieved using the promoters PthlA and Ppta-ack. Batch fermentations using A. woodii [pMTL84151_actthlA] in a bioreactor revealed that acetate concentration had an effect on the acetone production, due to the high Km value of the CoA transferase. In order to establish consistent acetate concentration within the bioreactor and to increase biomass, a continuous fermentation process for A. woodii was developed. Thus, acetone productivity of the strain A. woodii [pMTL84151_actthlA] was increased from 1.2mgL(-1)h(-1) in bottle fermentation to 26.4mgL(-1)h(-1) in continuous gas fermentation. Copyright © 2016 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  7. Escalated regeneration in sciatic nerve crush injury by the combined therapy of human amniotic fluid mesenchymal stem cells and fermented soybean extracts, Natto.

    Science.gov (United States)

    Pan, Hung-Chuan; Yang, Dar-Yu; Ho, Shu-Peng; Sheu, Meei-Ling; Chen, Chung-Jung; Hwang, Shiaw-Min; Chang, Ming-Hong; Cheng, Fu-Chou

    2009-08-23

    Attenuation of inflammatory cell deposits and associated cytokines prevented the apoptosis of transplanted stem cells in a sciatic nerve crush injury model. Suppression of inflammatory cytokines by fermented soybean extracts (Natto) was also beneficial to nerve regeneration. In this study, the effect of Natto on transplanted human amniotic fluid mesenchymal stem cells (AFS) was evaluated. Peripheral nerve injury was induced in SD rats by crushing a sciatic nerve using a vessel clamp. Animals were categorized into four groups: Group I: no treatment; Group II: fed with Natto (16 mg/day for 7 consecutive days); Group III: AFS embedded in fibrin glue; Group IV: Combination of group II and III therapy. Transplanted AFS and Schwann cell apoptosis, inflammatory cell deposits and associated cytokines, motor function, and nerve regeneration were evaluated 7 or 28 days after injury. The deterioration of neurological function was attenuated by AFS, Natto, or the combined therapy. The combined therapy caused the most significantly beneficial effects. Administration of Natto suppressed the inflammatory responses and correlated with decreased AFS and Schwann cell apoptosis. The decreased AFS apoptosis was in line with neurological improvement such as expression of early regeneration marker of neurofilament and late markers of S-100 and decreased vacuole formation. Administration of either AFS, or Natto, or combined therapy augmented the nerve regeneration. In conclusion, administration of Natto may rescue the AFS and Schwann cells from apoptosis by suppressing the macrophage deposits, associated inflammatory cytokines, and fibrin deposits.

  8. Escalated regeneration in sciatic nerve crush injury by the combined therapy of human amniotic fluid mesenchymal stem cells and fermented soybean extracts, Natto

    Directory of Open Access Journals (Sweden)

    Pan Hung-Chuan

    2009-08-01

    Full Text Available Abstract Attenuation of inflammatory cell deposits and associated cytokines prevented the apoptosis of transplanted stem cells in a sciatic nerve crush injury model. Suppression of inflammatory cytokines by fermented soybean extracts (Natto was also beneficial to nerve regeneration. In this study, the effect of Natto on transplanted human amniotic fluid mesenchymal stem cells (AFS was evaluated. Peripheral nerve injury was induced in SD rats by crushing a sciatic nerve using a vessel clamp. Animals were categorized into four groups: Group I: no treatment; Group II: fed with Natto (16 mg/day for 7 consecutive days; Group III: AFS embedded in fibrin glue; Group IV: Combination of group II and III therapy. Transplanted AFS and Schwann cell apoptosis, inflammatory cell deposits and associated cytokines, motor function, and nerve regeneration were evaluated 7 or 28 days after injury. The deterioration of neurological function was attenuated by AFS, Natto, or the combined therapy. The combined therapy caused the most significantly beneficial effects. Administration of Natto suppressed the inflammatory responses and correlated with decreased AFS and Schwann cell apoptosis. The decreased AFS apoptosis was in line with neurological improvement such as expression of early regeneration marker of neurofilament and late markers of S-100 and decreased vacuole formation. Administration of either AFS, or Natto, or combined therapy augmented the nerve regeneration. In conclusion, administration of Natto may rescue the AFS and Schwann cells from apoptosis by suppressing the macrophage deposits, associated inflammatory cytokines, and fibrin deposits.

  9. Genomic analysis of Bacillus subtilis lytic bacteriophage ϕNIT1 capable of obstructing natto fermentation carrying genes for the capsule-lytic soluble enzymes poly-γ-glutamate hydrolase and levanase.

    Science.gov (United States)

    Ozaki, Tatsuro; Abe, Naoki; Kimura, Keitarou; Suzuki, Atsuto; Kaneko, Jun

    2017-01-01

    Bacillus subtilis strains including the fermented soybean (natto) starter produce capsular polymers consisting of poly-γ-glutamate and levan. Capsular polymers may protect the cells from phage infection. However, bacteriophage ϕNIT1 carries a γ-PGA hydrolase gene (pghP) that help it to counteract the host cell's protection strategy. ϕNIT had a linear double stranded DNA genome of 155,631-bp with a terminal redundancy of 5,103-bp, containing a gene encoding an active levan hydrolase. These capsule-lytic enzyme genes were located in the possible foreign gene cluster regions between central core and terminal redundant regions, and were expressed at the late phase of the phage lytic cycle. All tested natto origin Spounavirinae phages carried both genes for capsule degrading enzymes similar to ϕNIT1. A comparative genomic analysis revealed the diversity among ϕNIT1 and Bacillus phages carrying pghP-like and levan-hydrolase genes, and provides novel understanding on the acquisition mechanism of these enzymatic genes.

  10. Inhibition of Lipopolysaccharide-Induced Interleukin 8 in Human Adenocarcinoma Cell Line HT-29 by Spore Probiotics: B. coagulans and B. subtilis (natto).

    Science.gov (United States)

    Azimirad, Masoumeh; Alebouyeh, Masoud; Naji, Tahereh

    2017-03-01

    Probiotics are used as a treatment for different intestinal disorders. They confer health benefits by different ways. This study was aimed to investigate immunomodulatory effect of Bacillus probiotic spores on the production of lipopolysaccharide (LPS)-induced interleukin 8 (IL-8) in HT-29 intestinal epithelial cells. Differentiated intestinal epithelial cell line was used as a model for the study of colonization of purified spores (Bacillus subtilis (natto) and B. coagulans) and their anti-inflammatory effects. MTT assay and trypan blue staining were used for the detection of optimal concentration of the purified spores and LPS. Pre-treatment assay was done by treatment of the cells with the purified spores for 2 h, followed by challenges with LPS for 3 and 18 h. Post-treatment assay was done by initial treatment of the cells with LPS for 18 h, followed by the spores for 3 and 6 h. Levels of IL-8 secretion and its mRNA expression were measured by ELISA and relative Q real-time PCR. Our results showed similar rates of adherence to intestinal epithelial cells by the spore probiotics, while displaying no cytotoxic effect. In the pre-treatment assay, a significant decrease in IL-8, at both protein and mRNA levels, was measured for B. coagulans spores after the addition of LPS, which was higher than those observed for Bacillus subtilis (natto) spores. In the post-treatment assay, while Bacillus subtilis (but not B. coagulans) diminished the LPS-stimulated IL-8 levels after 3 h of incubation, the inhibitory effect was not constant. In conclusion, ability of Bacillus spore probiotics for adherence to intestinal epithelial cell and their anti-inflammatory effects, through interference with LPS/IL-8 signaling, was shown in this study. Further studies are needed to characterize responsible bacterial compounds associated with these effects.

  11. Screening of strains with the high activity and thermostability nattokinase by {sup 60}Co γ-ray irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Shuying, Li; Ying, Nie; Huan, Du; Xuanming, Tang [Institute of Agro-products Processing Science and Technology, Chinese Academy of Agricultural Sciences, Beijing (China); Zhonglin, Zhao [College of Sciences, Henan Agricultural University, Zhengzhou (China); Xin, Ma [Agricultural Information Institute, Chinese Academic of Agricultural Sciences, Beijing (China); Yan, Li [School of Food and Biological Engineering, Zhengzhou University of Light Industry, Zhengzhou (China)

    2013-06-15

    In this study, Bacillus natto was irradiated by {sup 60}Co γ-ray, and activity was determined by Casein plate method in order to get high activity and thermostability strains. 60 strains with high activity were obtained through irradiation by 800 Gy {sup 60}Co γ-ray. In this dose, the positive mutation rate was 45%. Then 60 strains was treated by different tempreture and 11 strains showed thermostability at 65℃. (authors)

  12. Screening of strains with the high activity and thermostability nattokinase by "6"0Co γ-ray irradiation

    International Nuclear Information System (INIS)

    Li Shuying; Nie Ying; Du Huan; Tang Xuanming; Zhao Zhonglin; Ma Xin; Li Yan

    2013-01-01

    In this study, Bacillus natto was irradiated by "6"0Co γ-ray, and activity was determined by Casein plate method in order to get high activity and thermostability strains. 60 strains with high activity were obtained through irradiation by 800 Gy "6"0Co γ-ray. In this dose, the positive mutation rate was 45%. Then 60 strains was treated by different tempreture and 11 strains showed thermostability at 65℃. (authors)

  13. [Production of pertussis toxin by Bordetella pertussis strains isolated from patients with whooping cough].

    Science.gov (United States)

    Zaĭtsev, E M; Mertsalova, N U; Shinkarev, A S; Mazurova, I K; Zakharova, N S

    2011-01-01

    To assess level of pertussin toxin (PT) production by vaccine strains of Bordetella pertussis and strains isolated from patients with whooping cough. Concentration of PT in supernatants of microbial cultures of 3 vaccine strains and 25 strains of B. pertussis isolated from patients with pertussis in 2001 - 2005 was measured with enzyme immunoassay using gamma-globulin fractions of rabbit antiserum to PT as immunosorbent or included in peroxidase conjugates. Level of PT production by strains isolated from infected persons varied from 3 +/- 0.5 to 64.8 +/- 12.2 ng/MFU/ml: in 9 strains--from 3 +/- 0.5 to 9.4 +/- 2.1 ng/MFU/ml, in 7--10.5 +/- 1.8 to 18.4 +/- 2.6 ng/MFU/ml, and in 9--23.6 +/- 4.5 to 64.8 +/- 12.2 ng/MFU/ml. B. pertussis strains isolated from patients were heterogeneous on level of PT production. Difference in expression of PT between strains were as high as 20-fold. Conditionally low, moderate and high levels of PT production had 9 (36%), 7 (28%), and 9 (36%) of 25 studied strains. Three vaccine strains had levels of toxin production similar to recently isolated strains with moderate level of its production.

  14. Segregation of genes from donor strain during the production of recombinant congenic strains.

    Science.gov (United States)

    van Zutphen, L F; Den Bieman, M; Lankhorst, A; Demant, P

    1991-07-01

    Recombinant congenic strains (RCS) constitute a set of inbred strains which are designed to dissect the genetic control of multigenic traits, such as tumour susceptibility or disease resistance. Each RCS contains a small fraction of the genome of a common donor strain, while the majority of genes stem from a common background strain. We tested at two stages of the inbreeding process in 20 RCS, derived from BALB/cHeA and STS/A, to see whether alleles from the STS/A donor strain are distributed over the RCS in a ratio as would theoretically be expected. Four marker genes (Pep-3; Pgm-1; Gpi-1 and Es-3) located at 4 different chromosomes were selected and the allelic distribution was tested after 3-4 and after 12 generations of inbreeding. The data obtained do not significantly deviate from the expected pattern, thus supporting the validity of the concept of RCS.

  15. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto

    International Nuclear Information System (INIS)

    Yanagisawa, Yasuhide; Chatake, Toshiyuki; Chiba-Kamoshida, Kaori; Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki; Yasuda, Ichiro; Morimoto, Yukio

    2010-01-01

    Nattokinase, a protein found in high levels in the traditional Japanese food natto, has been reported to have high thrombolytic activity. In the present study, the crystallization of native nattokinase and the collection of X-ray diffraction date from a nattokinase crystal to a resolution of 1.74 Å are reported. Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27 724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a = 74.3, b = 49.9, c = 56.3 Å, β = 95.2°. Diffraction images were processed to a resolution of 1.74 Å with an R merge of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase

  16. Directed evolution improves the fibrinolytic activity of nattokinase from Bacillus natto.

    Science.gov (United States)

    Yongjun, Cai; Wei, Bao; Shujun, Jiang; Meizhi, Weng; Yan, Jia; Yan, Yin; Zhongliang, Zheng; Goulin, Zou

    2011-12-01

    Nattokinase (subtilisin NAT, NK) is a relatively effective microbial fibrinolytic enzyme that has been identified and characterized from Bacillus natto. In the current report, DNA family shuffling was used to improve the fibrinolytic activity of nattokinase. Three homologous genes from B. natto AS 1.107, Bacillus amyloliquefaciens CICC 20164 and Bacillus licheniformis CICC 10092 were shuffled to generate a mutant library. A plate-based method was used to screen the mutant libraries for improved activity. After three rounds of DNA shuffling, one desirable mutant with 16 amino acid substitutions was obtained. The mutant enzyme was purified and characterized. The kinetic measurements showed that the catalytic efficiency of the mutant NK was approximately 2.3 times higher than that of the wild-type nattokinase. In addition, the molecular modeling analysis suggested that the mutations affect the enzymatic function by changing the surface conformation of the substrate-binding pocket. The current study shows that the evolution of nattokinase with improved fibrinolytic activity by DNA family shuffling is feasible and provides useful references to facilitate the application of nattokinase in thrombolytic therapy. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  17. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto

    Energy Technology Data Exchange (ETDEWEB)

    Yanagisawa, Yasuhide [Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan); Chatake, Toshiyuki [Research Reactor Institute, Kyoto University, Asashironishi 2, Kumatori, Sennan, Osaka 590-0494 (Japan); Chiba-Kamoshida, Kaori [National Institute of Advanced Industrial Science and Technology (AIST), Umezono 1-1-1, Tsukuba, Ibaraki 305-8568 (Japan); Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki [Kurashiki University of Science and Arts, Nishinoura 2640, Tsurajima-cho, Kurashiki, Okayama 712-8505 (Japan); Yasuda, Ichiro [Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan); Morimoto, Yukio [Research Reactor Institute, Kyoto University, Asashironishi 2, Kumatori, Sennan, Osaka 590-0494 (Japan); Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Choshi, Chiba 288-0025 (Japan)

    2010-12-01

    Nattokinase, a protein found in high levels in the traditional Japanese food natto, has been reported to have high thrombolytic activity. In the present study, the crystallization of native nattokinase and the collection of X-ray diffraction date from a nattokinase crystal to a resolution of 1.74 Å are reported. Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27 724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a = 74.3, b = 49.9, c = 56.3 Å, β = 95.2°. Diffraction images were processed to a resolution of 1.74 Å with an R{sub merge} of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase.

  18. Biosurfactant production by Pseudomonas strains isolated from floral nectar.

    Science.gov (United States)

    Ben Belgacem, Z; Bijttebier, S; Verreth, C; Voorspoels, S; Van de Voorde, I; Aerts, G; Willems, K A; Jacquemyn, H; Ruyters, S; Lievens, B

    2015-06-01

    To screen and identify biosurfactant-producing Pseudomonas strains isolated from floral nectar; to characterize the produced biosurfactants; and to investigate the effect of different carbon sources on biosurfactant production. Four of eight nectar Pseudomonas isolates were found to produce biosurfactants. Phylogenetic analysis based on three housekeeping genes (16S rRNA gene, rpoB and gyrB) classified the isolates into two groups, including one group closely related to Pseudomonas fluorescens and another group closely related to Pseudomonas fragi and Pseudomonas jessenii. Although our nectar pseudomonads were able to grow on a variety of water-soluble and water-immiscible carbon sources, surface active agents were only produced when using vegetable oil as sole carbon source, including olive oil, sunflower oil or waste frying sunflower oil. Structural characterization based on thin layer chromatography (TLC) and ultra high performance liquid chromatography-accurate mass mass spectrometry (UHPLC-amMS) revealed that biosurfactant activity was most probably due to the production of fatty acids (C16:0; C18:0; C18:1 and C18:2), and mono- and diglycerides thereof. Four biosurfactant-producing nectar pseudomonads were identified. The active compounds were identified as fatty acids (C16:0; C18:0; C18:1 and C18:2), and mono- and diglycerides thereof, produced by hydrolysis of triglycerides of the feedstock. Studies on biosurfactant-producing micro-organisms have mainly focused on microbes isolated from soils and aquatic environments. Here, for the first time, nectar environments were screened as a novel source for biosurfactant producers. As nectars represent harsh environments with high osmotic pressure and varying pH levels, further screening of nectar habitats for biosurfactant-producing microbes may lead to the discovery of novel biosurfactants with broad tolerance towards different environmental conditions. © 2015 The Society for Applied Microbiology.

  19. Antimutagenic and radioprotective properties of products from Lactobacterium Bulgaricum strains

    International Nuclear Information System (INIS)

    Rupova, I.; Benova, D.; Yagova, A.; Topalova, S.

    1993-01-01

    The antimutagenic properties of the strains LBL-4 and LBL-144 have been studied by the incidence of micronuclei in polychromatic erythrocytes (PE) and endogenous spleen colony forming units (E-CFU) in C57BL mice. The lyophilized lactic acid products have been administered orally in the course of 3 weeks on 5 consecutive days. After the last treatment the mice have been gamma irradiated with 0.5, 3.0, 7.0 and 8.0 Gy. The prophylactic pretreatment of mice with LBL-4 in the schedule 3 x 5 days has shown a reduction of the induced micronuclei in PE after gamma irradiation with 3.0 Gy (37% compared to 50.5%; p < 0.001). The radiation effects caused by a lower dose of 0.5 Gy has not been modified by LBL-4 (36.7% compared to 33.8%). The application of LBL-144 does not reduce the mutagenic activity of both 0.5 and 3.0 Gy gamma rays. Applied independently, both LBL-4 and LBL-144 have shown the same frequency of micro-nucleated PE as the male and female mice controls. The radioprotective properties of LBL-4 have been assessed according to E-CFU and the changes in spleen weights after irradiation with 7.0 and 8.0 Gy gamma rays. The amount of E-CFU have been counted on day 9 after irradiation. A significant increase of spleen colony formation units has been established in mice pretreated with LBL-4. The spleen weights of the protected mice have been increased compared to unprotected animals. The results obtained show a good protection of haemopoietic stem cells by the product LBL-4. (author)

  20. Triacetic acid lactone production in industrial Saccharomyces yeast strains

    Science.gov (United States)

    Triacetic acid lactone (TAL) is a potential platform chemical that can be produced in yeast. To evaluate the potential for industrial yeast strains to produce TAL, the g2ps1 gene encoding 2-pyrone synthase was transformed into thirteen industrial yeast strains of varied genetic background. TAL produ...

  1. Hydrocarbon productivities in different Botryococcus strains: comparative methods in product quantification.

    Science.gov (United States)

    Eroglu, Ela; Okada, Shigeru; Melis, Anastasios

    2011-08-01

    Six different strains of the green microalgae Botryococcus belonging to the A-race or B-race, accumulating alkadiene or botryococcene hydrocarbons, respectively, were compared for biomass and hydrocarbon productivities. Biomass productivity was assessed gravimetrically upon strain growth in the laboratory under defined conditions. Hydrocarbon productivities were measured by three different and independent experimental approaches, including density equilibrium of the intact cells and micro-colonies, spectrophotometric analysis of hydrocarbon extracts, and gravimetric quantitation of eluted hydrocarbons. All three hydrocarbon-quantitation methods yielded similar results for each of the strains examined. The B-race microalgae Botryococcus braunii var. Showa and Kawaguchi-1 constitutively accumulated botryococcene hydrocarbons equivalent to 30% and 20%, respectively, of their overall biomass. The A-race microalgae Botryococcus braunii, varieties Yamanaka, UTEX 2441 and UTEX LB572 constitutively accumulated alkadiene hydrocarbons ranging from 14% to 13% and 10% of their overall biomass, respectively. Botryococcus sudeticus (UTEX 2629), a morphologically different green microalga, had the lowest hydrocarbon accumulation, equal to about 3% of its overall biomass. Results validate the density equilibrium and spectrophotometric analysis methods in the quantitation of botryococcene-type hydrocarbons. These analytical advances will serve in the screening and selection of B. braunii and of other microalgae in efforts to identify those having a high hydrocarbon content for use in commercial applications.

  2. Toxin production ability of Bacillus cereus strains from food product of Ukraine

    Directory of Open Access Journals (Sweden)

    I. Pylypenko

    2017-10-01

    Full Text Available Potential pathogens of foodborne toxic infections – bacterial contaminants Bacillus cereus isolated from plant raw materials and food products from the Ukrainian region were investigated. When determining of the proportion of isolated bacilli from the plant samples, it was established that the epidemiologically significant microorganisms of Bacillus cereus as agents of food poisoning are the second largest. The average value of contaminated samples of Ukrainian plant raw materials and processed products with Bacillus cereus is 36,2 %. The ability of Bacillus cereus strains identified by a complex of morphological, tinctorial, cultural and biochemical properties, to produce specific emetic and enterotoxins was studied. Molecular genetic diagnosis and detection of the toxin-producing ability of isolated 42 Bacillus cereus strains showed both the possibility of their rapid identification and the presence of specific toxicity genes. Multiplex polymerase chain reaction (PCR was carried out with specific primers to detect toxicity determined of various bacilli genes: nheA, hblD, cytK, cesВ. The distribution of toxigenic genes is significantly different among the Bacillus cereus isolates from various sources. The nheA, hblD and cytK enterotoxin genes were detected in 100, 83,3 and 61,9 % of the investigated strains of Bacillus cereus, respectively. The cesB gene encoding emetic toxin was detected in 4,8 % of  strains. Molecular-genetic PCR-method confirmed that all the isolated strains belong to the Bacillus cereus group, and the ability to produce toxins can be attributed to five groups. The main toxins that produce the investigated Bacillus cereus strains were nhe and hbl enterotoxins encoded by the corresponding genes of nheA and hblD. The enterotoxic type of Bacillus cereus was predominant in Ukrainian region.  Studies of domestic plant food raw materials and products have confirmed the need to improve microbiological control of product safety

  3. Bio-succinic acid production: Escherichia coli strains design from genome-scale perspectives

    Directory of Open Access Journals (Sweden)

    Bashir Sajo Mienda

    2017-10-01

    Full Text Available Escherichia coli (E. coli has been established to be a native producer of succinic acid (a platform chemical with different applications via mixed acid fermentation reactions. Genome-scale metabolic models (GEMs of E. coli have been published with capabilities of predicting strain design strategies for the production of bio-based succinic acid. Proof-of-principle strains are fundamentally constructed as a starting point for systems strategies for industrial strains development. Here, we review for the first time, the use of E. coli GEMs for construction of proof-of-principles strains for increasing succinic acid production. Specific case studies, where E. coli proof-of-principle strains were constructed for increasing bio-based succinic acid production from glucose and glycerol carbon sources have been highlighted. In addition, a propose systems strategies for industrial strain development that could be applicable for future microbial succinic acid production guided by GEMs have been presented.

  4. Benchmarking two commonly used Saccharomyces cerevisiae strains for heterologous vanillin-β-glucoside production

    DEFF Research Database (Denmark)

    Strucko, Tomas; Magdenoska, Olivera; Mortensen, Uffe Hasbro

    2015-01-01

    factories for production of specific compounds. To examine this possibility, we have reconstructed a de novo vanillin-β-glucoside pathway in an identical manner in S288c and CEN.PK strains. Characterization of the two resulting strains in two standard conditions revealed that the S288c background strain...... produced up to 10-fold higher amounts of vanillin-β-glucoside compared to CEN.PK. This study demonstrates that yeast strain background may play a major role in the outcome of newly developed cell factories for production of a given product....

  5. Benchmarking two commonly used Saccharomyces cerevisiae strains for heterologous vanillin-β-glucoside production

    Directory of Open Access Journals (Sweden)

    Tomas Strucko

    2015-12-01

    Full Text Available The yeast Saccharomyces cerevisiae is a widely used eukaryotic model organism and a key cell factory for production of biofuels and wide range of chemicals. From the broad palette of available yeast strains, the most popular are those derived from laboratory strain S288c and the industrially relevant CEN.PK strain series. Importantly, in recent years these two strains have been subjected to comparative “-omics” analyzes pointing out significant genotypic and phenotypic differences. It is therefore possible that the two strains differ significantly with respect to their potential as cell factories for production of specific compounds. To examine this possibility, we have reconstructed a de novo vanillin-β-glucoside pathway in an identical manner in S288c and CEN.PK strains. Characterization of the two resulting strains in two standard conditions revealed that the S288c background strain produced up to 10-fold higher amounts of vanillin-β-glucoside compared to CEN.PK. This study demonstrates that yeast strain background may play a major role in the outcome of newly developed cell factories for production of a given product. Keywords: Yeast, Cell factory, Strain choice, Heterologous production, Vanillin-glucoside, Shikimate pathway

  6. Association between vitamin K intake from fermented soybeans, natto, and bone mineral density in elderly Japanese men: the Fujiwara-kyo Osteoporosis Risk in Men (FORMEN) study.

    Science.gov (United States)

    Fujita, Y; Iki, M; Tamaki, J; Kouda, K; Yura, A; Kadowaki, E; Sato, Y; Moon, J-S; Tomioka, K; Okamoto, N; Kurumatani, N

    2012-02-01

    A cross-sectional analysis of 1,662 community dwelling elderly Japanese men suggested that habitual natto intake was significantly associated with higher bone mineral density (BMD). When adjustment was made for undercarboxylated osteocalcin levels, this association was insignificant, showing the natto-bone association to be primarily mediated by vitamin K. Low vitamin K intake is associated with an increased risk of hip fracture, but reports have been inconsistent on its effect on BMD. Our first aim was to examine the association between BMD and intake of fermented soybeans, natto, which contain vitamin K1 (20 μg/pack) and K2 (380 μg/pack). Our second aim was to examine the association between undercarboxylated osteocalcin (ucOC), a biomarker of vitamin K intake, and BMD to evaluate the role of vitamin K in this association. Of the Japanese men aged ≥65 years who participated in the baseline survey of the Fujiwara-kyo Osteoporosis Risk in Men study, 1,662 men without diseases or medications known to affect bone metabolism were examined for associations between self-reported natto intake or serum ucOC levels with lumbar spine or hip BMD. The subjects with greater intake of natto showed significantly lower level of serum ucOC. Analysis after adjustment for confounding variables showed an association of greater intake of natto with both significantly higher BMD and lower risk of low BMD (T-score natto was associated with a beneficial effect on bone health in elderly men, and this association is primarily due to vitamin K content of natto, although the lack of information on dietary nutrient intake, including vitamin K1 and K2, prevented us from further examining the association.

  7. Lipase production from a wild (LPF-5) and a mutant (HN1) strain of ...

    African Journals Online (AJOL)

    Lipase production from a wild (LPF-5) and a mutant (HN1) strain of Aspergillus niger. ... Several physical parameters (carbon source, nitrogen source, pH, ... for the development of industrial biotechnology for production of extracellular lipase.

  8. Subspecies diversity in bacteriocin production by intestinal Lactobacillus salivarius strains.

    Science.gov (United States)

    O' Shea, Eileen F; O' Connor, Paula M; Raftis, Emma J; O' Toole, Paul W; Stanton, Catherine; Cotter, Paul D; Ross, R Paul; Hill, Colin

    2012-01-01

    A recent comparative genomic hybridization study in our laboratory revealed considerable plasticity within the bacteriocin locus of gastrointestinal strains of Lactobacillus salivarius. Most notably, these analyses led to the identification of two novel unmodified bacteriocins, salivaricin L and salivaricin T, produced by the neonatal isolate L. salivarius DPC6488 with immunity, regulatory and export systems analogous to those of abp118, a two-component bacteriocin produced by the well characterized reference strain L. salivarius UCC118. In this addendum we discuss the intraspecific diversity of our seven bacteriocin-producing L. salivarius isolates on a genome-wide level, and more specifically, with respect to their salivaricin loci.

  9. Purification, crystallization and preliminary X-ray diffraction experiment of nattokinase from Bacillus subtilis natto.

    Science.gov (United States)

    Yanagisawa, Yasuhide; Chatake, Toshiyuki; Chiba-Kamoshida, Kaori; Naito, Sawa; Ohsugi, Tadanori; Sumi, Hiroyuki; Yasuda, Ichiro; Morimoto, Yukio

    2010-12-01

    Nattokinase is a single polypeptide chain composed of 275 amino acids (molecular weight 27,724) which displays strong fibrinolytic activity. Moreover, it can activate other fibrinolytic enzymes such as pro-urokinase and tissue plasminogen activator. In the present study, native nattokinase from Bacillus subtilis natto was purified using gel-filtration chromatography and crystallized to give needle-like crystals which could be used for X-ray diffraction experiments. The crystals belonged to space group C2, with unit-cell parameters a=74.3, b=49.9, c=56.3 Å, β=95.2°. Diffraction images were processed to a resolution of 1.74 Å with an Rmerge of 5.2% (15.3% in the highest resolution shell) and a completeness of 69.8% (30.0% in the highest resolution shell). This study reports the first X-ray diffraction analysis of nattokinase.

  10. Ethanol production potential of local yeast strains isolated from ripe ...

    African Journals Online (AJOL)

    The ability of different yeast strains isolated from ripe banana peels to produce ethanol was investigated. Of the 8 isolates screened for their fermentation ability, 5 showed enhanced performance and were subsequently identified and assessed for important ethanol fermentation attributes such as ethanol producing ability, ...

  11. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin

    OpenAIRE

    Fleige, Christian; Meyer, Florian; Steinbüchel, Alexander

    2016-01-01

    The Gram-positive bacterium Amycolatopsis sp. ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for ...

  12. Engineering Ashbya gossypii strains for de novo lipid production using industrial by-products.

    Science.gov (United States)

    Lozano-Martínez, Patricia; Buey, Rubén M; Ledesma-Amaro, Rodrigo; Jiménez, Alberto; Revuelta, José Luis

    2017-03-01

    Ashbya gossypii is a filamentous fungus that naturally overproduces riboflavin, and it is currently exploited for the industrial production of this vitamin. The utilization of A. gossypii for biotechnological applications presents important advantages such as the utilization of low-cost culture media, inexpensive downstream processing and a wide range of molecular tools for genetic manipulation, thus making A. gossypii a valuable biotechnological chassis for metabolic engineering. A. gossypii has been shown to accumulate high levels of lipids in oil-based culture media; however, the lipid biosynthesis capacity is rather limited when grown in sugar-based culture media. In this study, by altering the fatty acyl-CoA pool and manipulating the regulation of the main ∆9 desaturase gene, we have obtained A. gossypii strains with significantly increased (up to fourfold) de novo lipid biosynthesis using glucose as the only carbon source in the fermentation broth. Moreover, these strains were efficient biocatalysts for the conversion of carbohydrates from sugarcane molasses to biolipids, able to accumulate lipids up to 25% of its cell dry weight. Our results represent a proof of principle showing the promising potential of A. gossypii as a competitive microorganism for industrial biolipid production using cost-effective feed stocks. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  13. A Comparative Study of New Aspergillus Strains for Proteolytic Enzymes Production by Solid State Fermentation

    Directory of Open Access Journals (Sweden)

    Gastón Ezequiel Ortiz

    2016-01-01

    Full Text Available A comparative study of the proteolytic enzymes production using twelve Aspergillus strains previously unused for this purpose was performed by solid state fermentation. A semiquantitative and quantitative evaluation of proteolytic activity were carried out using crude enzymatic extracts obtained from the fermentation cultures, finding seven strains with high and intermediate level of protease activity. Biochemical, thermodynamics, and kinetics features such as optimum pH and temperature values, thermal stability, activation energy (Ea, quotient energy (Q10, Km, and Vmax were studied in four enzymatic extracts from the selected strains that showed the highest productivity. Additionally, these strains were evaluated by zymogram analysis obtaining protease profiles with a wide range of molecular weight for each sample. From these four strains with the highest productivity, the proteolytic extract of A. sojae ATCC 20235 was shown to be an appropriate biocatalyst for hydrolysis of casein and gelatin substrates, increasing its antioxidant activities in 35% and 125%, respectively.

  14. Increased riboflavin production from activated bleaching earth by a mutant strain of Ashbya gossypii.

    Science.gov (United States)

    Tajima, Satoshi; Itoh, Yoko; Sugimoto, Takashi; Kato, Tatsuya; Park, Enoch Y

    2009-10-01

    The production of riboflavin from vegetable oil was increased using a mutant strain of Ashbya gossypii. This mutant was generated by treating the wild-type strain with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). Riboflavin production was 10-fold higher in the mutant compared to the wild-type strain. The specific intracellular catalase activity after 3 d of culture was 6-fold higher in the mutant than in the wild-type strain. For the mutant, riboflavin production in the presence of 40 mM hydrogen peroxide was 16% less than that in the absence of hydrogen peroxide, whereas it was 56% less for the wild-type strain. The isocitrate lyase (ICL) activity of the mutant was 0.26 mU/mg of protein during the active riboflavin production phase, which was 2.6-fold higher than the wild-type strain. These data indicate that the mutant utilizes the carbon flux from the TCA cycle to the glyoxylate cycle more efficiently than the wild-type strain, resulting in enhanced riboflavin production. This novel mutant has the potential to be of use for industrial-scale riboflavin production from waste-activated bleaching earth (ABE), thereby transforming a useless material into a valuable bioproduct.

  15. Fatty acid production by four strains of Mucor hiemalis grown in plant ...

    African Journals Online (AJOL)

    SERVER

    2007-08-06

    Aug 6, 2007 ... Four Mucor hiemalis strains (M1, M2, M3 and M4), isolated from soil at a depth of 0 - 15 ... Plant oils were inhibitors of fatty acid production by these strains. .... The procedure was adapted from Kates (1982) and Kennedy et al.

  16. Cytokine production induced by non-encapsulated and encapsulated Porphyromonas gingivalis strains

    NARCIS (Netherlands)

    Kunnen, A.; Dekker, D.C.; van Pampus, M.G.; Harmsen, H.J.; Aarnoudse, J.G.; Abbas, F.; Faas, M.M.

    Objective: Although the exact reason is not known, encapsulated gram-negative Porphyromonas gingivalis strains are more virulent than non-encapsulated strains. Since difference in virulence properties may be due to difference in cytokine production following recognition of the bacteria or their

  17. [Production of monoclonal antibodies against a wild strain of rabies virus].

    Science.gov (United States)

    Akacem, O; Benmansour, A; Coulon, P; Brahimi, M; Benhassine, M

    1992-01-01

    Production of monoclonal antibodies against a wild strain of rabies virus. Cell fusion of SP 2/O, a murine myeloma against a wild strain of rabies virus has originated five monoclonal antibodies (M.A.) specific for virus nucleocapsid , one M.A. specific for virus glycoprotein and one M.A. specific for a viral membrane protein.

  18. Production of extracellular lipase by a new strain Staphylococcus ...

    African Journals Online (AJOL)

    SAM

    2014-07-09

    Jul 9, 2014 ... mesophilic and solvent tolerant lipase with industrial potential. Key words: ..... amount of enzyme production indicating the inducible nature of the .... biodiesel production, oleochemical industry, polymer syn- thesis and ...

  19. Production of extracellular lipase by a new strain Staphylococcus ...

    African Journals Online (AJOL)

    Based on morphological, biochemical and 16S rRNA sequence analysis, the potent isolate was identified as Staphylococcus aureus. The lipase production of the isolate was increased by improving the conditions of production medium. Maximum lipase production (8.11 U/ml) was achieved when 2% punnakka oil was ...

  20. Inhibition of angiotensin I converting enzyme by subtilisin NAT (nattokinase) in natto, a Japanese traditional fermented food.

    Science.gov (United States)

    Murakami, Keiko; Yamanaka, Naoki; Ohnishi, Katsunori; Fukayama, Minoru; Yoshino, Masataka

    2012-06-01

    Angiotensin I converting enzyme (ACE) was inhibited by the culture medium of Bacillus subtilis subsp. natto, which ferments boiled soy beans to natto, a Japanese traditional food. Subtilisin NAT (nattokinase) produced by B. subtilis also inhibited ACE, and the inhibition was markedly stimulated by heat treatment of subtilisin at 120 °C for 15 min. Inhibition of ACE by subtilisin was of a mixed type: the decrease in V(max) and the increase in K(m) value. SDS-polyacrylamide gel electrophoresis showed that heat treatment of subtilisin caused inactivation with fragmentation of the enzyme protein into small peptides. The inhibitory action of subtilisin was not due to an enzymatic action of protease, but may be ascribed to the potent ACE-inhibitory peptides such as LY and FY, amino acid sequences in subtilisin. HPLC-MS analysis of heat-inactivated subtilisin confirmed that LY and FY were liberated by fragmentation of the enzyme. Inhibition of ACE by subtilisin and its degradation peptides such as LY and FY may participate in the suppression of blood pressure by ingestion of natto.

  1. Maternal intake of Natto, a Japan's traditional fermented soybean food, during pregnancy and the risk of eczema in Japanese babies.

    Science.gov (United States)

    Ozawa, Naoko; Shimojo, Naoki; Suzuki, Yoichi; Ochiai, Shingo; Nakano, Taiji; Morita, Yoshinori; Inoue, Yuzaburo; Arima, Takayasu; Suzuki, Shuichi; Kohno, Yoichi

    2014-06-01

    There are reports that the maternal diet during pregnancy may affect development of babies' eczema. We sought to investigate the association between the maternal diet during pregnancy and the risk of eczema in infancy in Japan. A birth cohort was set up at 2 hospitals in Chiba city. Dietary habits concerning fish, butter, margarine, yogurt and natto during pregnancy was obtained from mothers just after delivery. The intake frequencies of these foods were classified into four groups: 1) daily, 2) 2-3 times a week, 3) once a week and 4) once a month or less. Diagnosis of eczema at 6 months of age was made by the presence of an itchy rash that persisted more than two months. Valid data on 650 mother-baby pairs were obtained. No relationship between frequencies of the maternal intake of fish, margarine and yogurt during pregnancy and the onset rate of the babies' eczema were observed. For butter consumption, the incidence of babies' eczema was significantly higher in the group with daily intake than in those with an intake 2-3 times a week or less (p = 0.044). For natto, incidence of babies' eczema was significantly lower in the group with everyday intake than those eating it 2-3 times a week or less (p = 0.020). High frequency intake of natto during pregnancy possibly reduces the incidence of eczema in children at 6 months of age.

  2. Study on the IAA (Indole acetic acid) Productivity of Soil Yeast Strain Isolats

    International Nuclear Information System (INIS)

    Nwe Nwe Soe Hlaing; Swe Zin Yu; San San Yu

    2011-12-01

    Twelve isolated soil yeast were tested in IAA production in peptone yeast glucose broth (PYG). All strains were screened for the Indole Acetic Acid (IAA) producing activity in PYG broth supplemented with or without L-Tryptophan (L-TRP) as precusor. IAA production was assayed calorimetrically using Salkowski's reagent. The concentration of IAA produced by yeast strains was measured by spectrophotometric method at 530nm. Y6 strain was the highest IAA producer (79ppm) at 9 days incubation period without tryptophan. Y3, Y10 and Y12 strains that were incubated without L-TRP also had the higher ability in the production of IAA than other yeast isolates. The selected yeasts having high IAA production activity were characterized by morphological study and biochemical tests including sugar assimilation and fermentation tests.

  3. Isolation of non-sulphur photosynthetic bacterial strains efficient in hydrogen production at elevated temperatures

    Energy Technology Data Exchange (ETDEWEB)

    Singh, S.P.; Srivastava, S.C. (Banaras Hindu Univ., Varanasi (IN). Centre of Advanced Study in Botany)

    1991-01-01

    Four strains of non-sulphur photosynthetic bacteria were isolated from root zone associations of aquatic plants like Azolla, Salvinia and Eichhornia, as well as the deep-water rice. Based on the gross cell morphology and pigmentation, the isolates resembled Rhodopseudomonas sp. and have been designated as BHU strains 1 to 4, respectively. When subjected to elevated temperature (from 33-45{sup o}C), substantial growth/hydrogen production could be observed only in strains 1 and 4. Strains 2 and 3 on the other hand, showed diminished growth and negligible hydrogen photoproduction. The BHU strains 1 and 4 have been selected as the most active (thermostable) hydrogen producing strains of local origin as far as the Indian tropical climate is concerned. (author).

  4. Screening of Auricularia auricula strains for strong production ability of melanin pigments

    OpenAIRE

    ZOU, Yu; MA, Kun

    2017-01-01

    Abstract Melanin pigments have great application value and development potential in food industry to use as nature functional food colorants. In initial study, twenty-two Auricularia auricula strains were screened for stronger production ability of melanin pigments by solid culture. Three A. auricula strains (RF201, QD2 and QD6) with higher pigment production capacity were selected for further study through submerged culture supplementing 1 g/L l-tyrosine. The maximal pigment yields of A. aur...

  5. Enhanced L-(+)-lactic acid production by an adapted strain of Rhizopus oryzae using corncob hydrolysate

    DEFF Research Database (Denmark)

    Bai, Dongmei; Li, S.Z.; Liu, Z.L.

    2008-01-01

    -added production of a variety of bioproducts. Lactic acid can be used as a precursor for poly-lactic acid production. Although current industrial lactic acid is produced by lactic acid bacteria using enriched medium, production by Rhizopus oryzae is preferred due to its exclusive formation of the......-isomer and a simple nutrition requirement by the fungus. Production of-L-(+)-lactic acid by R. oryzae using xylose has been reported; however, its yield and conversion rate are poor compared with that of using glucose. In this study, we report an adapted R. oryzae strain HZS6 that significantly improved efficiency...... of substrate utilization and enhanced production of L-(+)-lactic acid from corncob hydrolysate. It increased L-(+)-lactic acid final concentration, yield, and volumetric productivity more than twofold compared with its parental strain. The optimized growth and fermentation conditions for Strain HZS6 were...

  6. POTENTIAL PRODUCTION OF CYCLOPIAZONIC ACID BY PENICILLIUM CAMEMBERTI STRAINS ISOLATED FROM CAMEMBERT TYPE CHEESE

    Directory of Open Access Journals (Sweden)

    Miroslava Císarová

    2012-10-01

    Full Text Available The aim of this study was to isolate the strains of fungi from Camembert type cheese, identify them and to test isolated strains of Penicillium camemberti for their ability to produce cyclopiazonic acid. The description of micro- and macromorphological features was used for identification of Penicillium camemberti strains. Strains were subsequently in vitro tested on their potential ability to produce mycotoxin cyclopiazonic acid (CPA. All of the 14 strains of Penicillium camemberti, which were obtained from 20 samples of Camembert type cheese, were cultivated 7, 14, 21, 27 and 30 days on CYA medium at 10±1°C, 15±1°C and 25±1°C in the dark. For determination of CPA production ability by P. camemberti isolates in vitro was TLC used. After 7 days of cultivation cyclopiazonic acid was produced only by 5 from 14 strains cultivated at all cultivation temperatures. After 14 and 21 days of cultivation was CPA produced by 6 strains at all of cultivation temperatures. After 27 and 30 days of cultivation was CPA identified in 7 strains cultivated at all temperatures of cultivation. The other strains also produced mycotoxin, however, not at each temperature. The most productive at all temperatures and after all days were 5 out of 14 tested strains (S9, S10, S13, S18 and S19. Strains S6 and S16 did not produce CPA at any temperature. The lowest production after all days of cultivation was found at 10±1 °C (44% and the highest at 25±1 °C (85%.

  7. Investigation on Strain Development of Azotobacter chroococcum through Chemical Mutagenesis for Indole Acetic Acid Production

    International Nuclear Information System (INIS)

    Ye Lin Win; Myo Myint Han; Myo Myint

    2010-12-01

    Two wild strains of Azotobacter chroococcum were mutagenized by various concentration of NTG (.005% , .0075% and .01%) for 1 hour. The IAA producing activity of 12 effective mutagenized strains were quantitatively measured by UV spectroscopic method. Mutagenized strains M1 and M2 from W1, and M8 and M9 from W2 were selected. Their IAA productivities were 300.498, 216.290, 238.436 and 190.856 ppm, respectively. M1and M9 were higher IAA productivity with greater quality. In germination, M1, M8 and M9 can promote the plant growth compare with commercial IAA.

  8. Improvement of Escherichia coli production strains by modification of the phosphoenolpyruvate:sugar phosphotransferase system

    Directory of Open Access Journals (Sweden)

    Gosset Guillermo

    2005-05-01

    Full Text Available Abstract The application of metabolic engineering in Escherichia coli has resulted in the generation of strains with the capacity to produce metabolites of commercial interest. Biotechnological processes with these engineered strains frequently employ culture media containing glucose as the carbon and energy source. In E. coli, the phosphoenolpyruvate:sugar phosphotransferase system (PTS transports glucose when this sugar is present at concentrations like those used in production fermentations. This protein system is involved in phosphoenolpyruvate-dependent sugar transport, therefore, its activity has an important impact on carbon flux distribution in the phosphoenolpyruvate and pyruvate nodes. Furthermore, PTS has a very important role in carbon catabolite repression. The properties of PTS impose metabolic and regulatory constraints that can hinder strain productivity. For this reason, PTS has been a target for modification with the purpose of strain improvement. In this review, PTS characteristics most relevant to strain performance and the different strategies of PTS modification for strain improvement are discussed. Functional replacement of PTS by alternative phosphoenolpyruvate-independent uptake and phosphorylation activities has resulted in significant improvements in product yield from glucose and productivity for several classes of metabolites. In addition, inactivation of PTS components has been applied successfully as a strategy to abolish carbon catabolite repression, resulting in E. coli strains that use more efficiently sugar mixtures, such as those obtained from lignocellulosic hydrolysates.

  9. Strains for the production of flavonoids from glucose

    Energy Technology Data Exchange (ETDEWEB)

    Stephanopoulos, Gregory; Santos, Christine; Koffas, Mattheos

    2015-11-13

    The invention relates to the production of flavonoids and flavonoid precursors in cells through recombinant expression of tyrosine ammonia lyase (TAL), 4-coumarate:CoA ligase (4CL), chalcone synthase (CHS), and chalcone isomerase (CHI).

  10. RobOKoD: microbial strain design for (over)production of target compounds.

    Science.gov (United States)

    Stanford, Natalie J; Millard, Pierre; Swainston, Neil

    2015-01-01

    Sustainable production of target compounds such as biofuels and high-value chemicals for pharmaceutical, agrochemical, and chemical industries is becoming an increasing priority given their current dependency upon diminishing petrochemical resources. Designing these strains is difficult, with current methods focusing primarily on knocking-out genes, dismissing other vital steps of strain design including the overexpression and dampening of genes. The design predictions from current methods also do not translate well-into successful strains in the laboratory. Here, we introduce RobOKoD (Robust, Overexpression, Knockout and Dampening), a method for predicting strain designs for overproduction of targets. The method uses flux variability analysis to profile each reaction within the system under differing production percentages of target-compound and biomass. Using these profiles, reactions are identified as potential knockout, overexpression, or dampening targets. The identified reactions are ranked according to their suitability, providing flexibility in strain design for users. The software was tested by designing a butanol-producing Escherichia coli strain, and was compared against the popular OptKnock and RobustKnock methods. RobOKoD shows favorable design predictions, when predictions from these methods are compared to a successful butanol-producing experimentally-validated strain. Overall RobOKoD provides users with rankings of predicted beneficial genetic interventions with which to support optimized strain design.

  11. Defect production in Ar irradiated graphene membranes under different initial applied strains

    Energy Technology Data Exchange (ETDEWEB)

    Martinez-Asencio, J., E-mail: jesusmartinez@ua.es [Dept. Física Aplicada, Facultad de Ciencias, Fase II, Universidad de Alicante, Alicante E-036090 (Spain); Ruestes, C.J.; Bringa, E. [CONICET and Facultad de Ciencias Exactas y Naturales, Universidad Nacional de Cuyo, Mendoza 5500 (Argentina); Caturla, M.J. [Dept. Física Aplicada, Facultad de Ciencias, Fase II, Universidad de Alicante, Alicante E-036090 (Spain)

    2017-02-15

    Highlights: • Defects in graphene membranes are formed due to 140 eV Ar ions irradiation using MD. • Different initial strains are applied, which influence the type and number of defects. • Mono-vacancies, di-vacancies and tri-vacancies production behaves linearly with dose. • The total number of defects under compression is slightly higher than under tension. - Abstract: Irradiation with low energy Ar ions of graphene membranes gives rise to changes in the mechanical properties of this material. These changes have been associated to the production of defects, mostly isolated vacancies. However, the initial state of the graphene membrane can also affect its mechanical response. Using molecular dynamics simulations we have studied defect production in graphene membranes irradiated with 140 eV Ar ions up to a dose of 0.075 × 10{sup 14} ions/cm{sup 2} and different initial strains, from −0.25% (compressive strain) to 0.25% (tensile strain). For all strains, the number of defects increases linearly with dose with a defect production of about 80% (80 defects every 100 ions). Defects are mostly single vacancies and di-vacancies, although some higher order clusters are also observed. Two different types of di-vacancies have been identified, the most common one being two vacancies at first nearest neighbours distance. Differences in the total number of defects with the applied strain are observed which is related to the production of a higher number of di-vacancies under compressive strain compared to tensile strain. We attribute this effect to the larger out-of-plane deformations of compressed samples that could favor the production of defects in closer proximity to others.

  12. Comparison of Bacterial Cellulose Production among Different Strains and Fermented Media

    Directory of Open Access Journals (Sweden)

    Maryam Jalili Tabaii

    2015-12-01

    Full Text Available The effect of different carbon sources on bacterial cellulose production by Gluconacetobacter xylinus (PTCC 1734 and two newly isolated strains (from vinegar under static culture conditions was studied. The production of bacterial cellulose was examined in modified Hestrin-Shramm medium by replacing D-glucose with other carbon sources. The results showed that the yield and characteristics of bacterial cellulose were influenced by the type of carbon source. Glycerol gave the highest yield in all of the studied strains (6%, 9.7% and 3.8% for S, A2 strain and Gluconacetobacter xylinus (PTCC 1734, respectively. The maximum dry bacterial cellulose weight in the glycerol containing medium is due to A2 strain (1.9 g l-1 in comparison to Gluconacetobacter xylinus as reference strain (0.76 g l-1. Although all of the studied strains were in Gluconacetobacter family, each used different sugars for maximum production after glycerol (mannitol and fructose for two newly isolated strains and glucose for Gluconacetobacter xylinus. The maximum moisture content was observed when sucrose and food-grade sucrose were used as carbon source. Contrary to expectations, while the maximum thickness of bacterial cellulose membrane was attained when glycerol was used, bacterial cellulose from glycerol had less moisture content than the others. The oxidized cellulose showed antibacterial activities, which makes it as a good candidate for food-preservatives.

  13. Glucoamylase production by a newly isolated strain of Aspergillus niger

    Energy Technology Data Exchange (ETDEWEB)

    Sinkar, V.P.; Lewis, N.F.

    1982-01-01

    Glucoamylase production by Aspergillus niger 57 was studied in complex and synthetic media under stationary vs. submerged conditions. Stationary cultivation resulted in significantly greater yields than did submerged culture. Crude enzyme activity was optimum at 60 degrees and pH 4.0.

  14. Acid production by oral strains of Candida albicans and Lactobacilli

    NARCIS (Netherlands)

    Klinke, T.; Kneist, S.; de Soet, J.J.; Kuhlisch, E.; Mauersberger, S.; Forster, A.; Klimm, W.

    2009-01-01

    Both Candida albicans and lactobacilli are common colonizers of carious lesions in children and adolescents. The purpose of this study is to compare the velocity of acid production between C. albicans and several Lactobacillus species at different pH levels and concentrations of glucose. Washed,

  15. Improvement of halophilic cellulase production from locally isolated fungal strain.

    Science.gov (United States)

    Gunny, Ahmad Anas Nagoor; Arbain, Dachyar; Jamal, Parveen; Gumba, Rizo Edwin

    2015-07-01

    Halophilic cellulases from the newly isolated fungus, Aspergillus terreus UniMAP AA-6 were found to be useful for in situ saccharification of ionic liquids treated lignocelluloses. Efforts have been taken to improve the enzyme production through statistical optimization approach namely Plackett-Burman design and the Face Centered Central Composite Design (FCCCD). Plackett-Burman experimental design was used to screen the medium components and process conditions. It was found that carboxymethylcellulose (CMC), FeSO4·7H2O, NaCl, MgSO4·7H2O, peptone, agitation speed and inoculum size significantly influence the production of halophilic cellulase. On the other hand, KH2PO4, KOH, yeast extract and temperature had a negative effect on enzyme production. Further optimization through FCCCD revealed that the optimization approach improved halophilic cellulase production from 0.029 U/ml to 0.0625 U/ml, which was approximately 2.2-times greater than before optimization.

  16. Isolation and characterization of the E. coli membrane protein production strain Mutant56(DE3)

    DEFF Research Database (Denmark)

    Baumgarten, Thomas; Schlegel, Susan; Wagner, Samuel

    2017-01-01

    Membrane protein production is usually toxic to E. coli. However, using genetic screens strains can be isolated in which the toxicity of membrane protein production is reduced, thereby improving production yields. Best known examples are the C41(DE3) and C43(DE3) strains, which are both derived...... from the T7 RNA polymerase (P)-based BL21(DE3) protein production strain. In C41(DE3) and C43(DE3) mutations lowering t7rnap expression levels result in strongly reduced T7 RNAP accumulation levels. As a consequence membrane protein production stress is alleviated in the C41(DE3) and C43(DE3) strains......, thereby increasing membrane protein yields. Here, we isolated Mutant56(DE3) from BL21(DE3) using a genetic screen designed to isolate BL21(DE3)-derived strains with mutations alleviating membrane protein production stress other than the ones in C41(DE3) and C43(DE3). The defining mutation of Mutant56(DE3...

  17. The mutagenesis and breeding of high productive strains of streptomyces jingyangensis '5406'

    International Nuclear Information System (INIS)

    Qi Hongyan; Yin Xinyun

    1988-03-01

    The purpose of these experiments is to explore the mutagenesis rhythm and breed high productive strains of actinomycete '5406'. The single colony agar pieces of strain F 358 were treated with fast neutron and 60 Co-γ ray irradiation Two mutants have been selected from 20025 treated single colonies. The output of cytokinins from them is higher than from strain F 358 . The original strain 'Mu-Tan-al' rejuvenated by freezing was treated with several physical and chemical mutagens. The mutagenesis rhythm has been summed up tentatively. Eight mutants obtained from 93014 treated single colonies produced more '5406' antibiotics than that of strain 'Mu-Tan-al,. The effect of mutant 'N2-10-Ra3' was the best

  18. Late-Onset Anaphylaxis Due to Poly (γ-glutamic acid in the Soup of Commercial Cold Chinese Noodles in a Patient with Allergy to Fermented Soybeans (Natto

    Directory of Open Access Journals (Sweden)

    Naoko Inomata

    2011-01-01

    Discussion: These results indicated that in the present case, the relevant allergen of late-onset anaphylaxis may have been PGA in all episodes and that the patient had been sensitized by PGA through natto ingestion. PGA is produced by Bacillus subtilis during fermentation and is a high-molecular, biodegradable polymer. The late onset is therefore, hypothesized to be due to a delayed absorption of PGA, as PGA biodegrades to peptides sufficiently small to be absorbed in the bowel. PGA has recently been applied to a wide range of fields such as foods, cosmetics, and medicine. Therefore, patients with late-onset anaphylaxis to PGA of natto should avoid not only natto but also other materials containing PGA.

  19. Characterization and increment of amylase production in mutant strains of Iranian native Bacillus licheniformis

    Directory of Open Access Journals (Sweden)

    Mohsen Mobini-Dehkordi

    2017-03-01

    Results: In this study, two interesting mutant strains were isolated and named B.L.2.M.1 and B.L.2.M.2. Mutations caused many changes in bacteria such as cell growth speed and enzyme production content. Differences in cell growth, production of amylase and other characters were significant at 0.05 level (Pvalue

  20. In Vitro Characterization of Lactobacillus Strains Isolated from Fruit Processing By-Products as Potential Probiotics.

    Science.gov (United States)

    de Albuquerque, Thatyane Mariano Rodrigues; Garcia, Estefânia Fernandes; de Oliveira Araújo, Amanda; Magnani, Marciane; Saarela, Maria; de Souza, Evandro Leite

    2017-08-23

    Nine wild Lactobacillus strains, namely Lactobacillus plantarum 53, Lactobacillus fermentum 56, L. fermentum 60, Lactobacillus paracasei 106, L. fermentum 250, L. fermentum 263, L. fermentum 139, L. fermentum 141, and L. fermentum 296, isolated from fruit processing by-products were evaluated in vitro for a series of safety, physiological functionality, and technological properties that could enable their use as probiotics. Considering the safety aspects, the resistance to antibiotics varied among the examined strains, and none of the strains presented hemolytic and mucinolytic activity. Regarding the physiological functionality properties, none of the strains were able to deconjugate bile salts; all of them presented low to moderate cell hydrophobicity and were able to autoaggregate, coaggregate with Listeria monocytogenes and Escherichia coli, and antagonize pathogenic bacteria. Exposure to pH 2 sharply decreased the survival of the examined strains after 1- or 2-h exposure; variable decreases were noted after 3-h exposure to pH 3. Overall, exposure to pH 5 and to bile salts (0.15, 0.3, and 1%) did not decrease the strains' survival. Examined strains presented better ability to survive from the exposure to simulated gastrointestinal conditions in laboratorial media and milk than in grape juice. Considering the technological properties, all the strains were positive for proteolytic activity and EPS and diacetyl production, and most of them had good tolerance to 1-4% NaCl. These results indicate that wild Lactobacillus strains isolated from fruit processing by-products could present performance compatible with probiotic properties and technological features that enable the development of probiotic foods with distinct characteristics.

  1. Improvement of strain Penicillium sp. EZ-ZH190 for tannase production by induced mutation.

    Science.gov (United States)

    Zakipour-Molkabadi, E; Hamidi-Esfahani, Z; Sahari, M A; Azizi, M H

    2013-11-01

    In the search for an efficient producer of tannase, Penicillium sp. EZ-ZH190 was subjected to mutagenesis using heat treatment and strain EZ-ZH290 was isolated. The maximum tannase in this mutant strain was 4.32 U/mL with an incubation period of 84 h as compared to wild strain EZ-ZH190 where the incubation period was 96 h with a maximum enzyme activity of 4.33 U/mL. Also, the Penicillium sp. EZ-ZH290 tannase had a maximum activity at 40 °C and pH 5.5. Then, the spores of strain EZ-ZH290 were subjected to γ irradiation mutagenesis and strain EZ-ZH390 was isolated. Strain EZ-ZH390 exhibited higher tannase activity (7.66 U/mL) than the parent strain EZ-ZH290. It was also found that Penicillium sp. EZ-ZH390 tannase had an optimum activity at 35 °C and a broad pH profile with an optimum at pH 5.5. The tannase pH stability of Penicillium sp. EZ-ZH390 and its maximum production of tannase followed the same trend for five generations confirming the occurrence of stable mutant. This paper is shown that γ irradiation can mutate the Penicillium sp. leading to increase the tannase production.

  2. Antimicrobial resistance of Enterococcus faecium strains isolated from commercial probiotic products used in cattle and swine.

    Science.gov (United States)

    Amachawadi, Raghavendra G; Giok, Felicia; Shi, Xiaorong; Soto, Jose; Narayanan, Sanjeev K; Tokach, Mike D; Apley, Mike D; Nagaraja, T G

    2018-04-03

    Probiotics, an antibiotic alternative, are widely used as feed additives for performance benefits in cattle and swine production systems. Among bacterial species contained in probiotics, Enterococcus faecium is common. Antimicrobial resistance (AMR), particularly multidrug resistance, is a common trait among enterococci because of their propensity to acquire resistance and horizontally transfer AMR genes. Also, E. faecium is an opportunistic pathogen, and in the United States, it is the second most common nosocomial pathogen. There has been no published study on AMR and virulence potential in E. faecium contained in probiotic products used in cattle and swine in the United States. Therefore, our objectives were to determine phenotypic susceptibilities or resistance to antimicrobials, virulence genes (asa1, gelE, cylA, esp, and hyl) and assess genetic diversity of E. faecium isolated from commercial products. Twenty-two commercially available E. faecium-based probiotic products used in cattle (n = 13) and swine (n = 9) were procured and E. faecium was isolated and species confirmed. Antimicrobial susceptibility testing to determine minimum inhibitory concentrations was done by micro-broth dilution method using National Antimicrobial Resistance Monitoring Systems Gram-positive Sensititre panel plate (CMV3AGPF), and categorization of strains as susceptible or resistant was as per Clinical Laboratory and Standards Institute's guidelines. E. faecium strains from 7 products (3 for swine and 4 for cattle) were pan-susceptible to the 16 antimicrobials tested. Strains from 15 products (6 for swine and 9 for cattle) exhibited resistance to at least one antimicrobial and a high proportion of strains was resistant to lincomycin (10/22), followed by tetracycline (4/22), daptomycin (4/22), ciprofloxacin (4/22), kanamycin (3/22), and penicillin (2/22). Four strains were multidrug resistant, with resistant phenotypes ranging from 3 to 6 antimicrobials or class. None of the E

  3. Warming Affects Growth Rates and Microcystin Production in Tropical Bloom-Forming Microcystis Strains

    Directory of Open Access Journals (Sweden)

    Trung Bui

    2018-03-01

    Full Text Available Warming climate is predicted to promote cyanobacterial blooms but the toxicity of cyanobacteria under global warming is less well studied. We tested the hypothesis that raising temperature may lead to increased growth rates but to decreased microcystin (MC production in tropical Microcystis strains. To this end, six Microcystis strains were isolated from different water bodies in Southern Vietnam. They were grown in triplicate at 27 °C (low, 31 °C (medium, 35 °C (high and 37 °C (extreme. Chlorophyll-a-, particle- and MC concentrations as well as dry-weights were determined. All strains yielded higher biomass in terms of chlorophyll-a concentration and dry-weight at 31 °C compared to 27 °C and then either stabilised, slightly increased or declined with higher temperature. Five strains easily grew at 37 °C but one could not survive at 37 °C. When temperature was increased from 27 °C to 37 °C total MC concentration decreased by 35% in strains with MC-LR as the dominant variant and by 94% in strains with MC-RR. MC quota expressed per particle, per unit chlorophyll-a and per unit dry-weight significantly declined with higher temperatures. This study shows that warming can prompt the growth of some tropical Microcystis strains but that these strains become less toxic.

  4. High-frequency rugose exopolysaccharide production by Vibrio cholerae strains isolated in Haiti.

    Directory of Open Access Journals (Sweden)

    Mustafizur Rahman

    Full Text Available In October, 2010, epidemic cholera was reported for the first time in Haiti in over 100 years. Establishment of cholera endemicity in Haiti will be dependent in large part on the continued presence of toxigenic V. cholerae O1 in aquatic reservoirs. The rugose phenotype of V. cholerae, characterized by exopolysaccharide production that confers resistance to environmental stress, is a potential contributor to environmental persistence. Using a microbiologic medium promoting high-frequency conversion of smooth to rugose (S-R phenotype, 80 (46.5% of 172 V. cholerae strains isolated from clinical and environmental sources in Haiti were able to convert to a rugose phenotype. Toxigenic V. cholerae O1 strains isolated at the beginning of the epidemic (2010 were significantly less likely to shift to a rugose phenotype than clinical strains isolated in 2012/2013, or environmental strains. Frequency of rugose conversion was influenced by incubation temperature and time. Appearance of the biofilm produced by a Haitian clinical rugose strain (altered biotype El Tor HC16R differed from that of a typical El Tor rugose strain (N16961R by confocal microscopy. On whole-genome SNP analysis, there was no phylogenetic clustering of strains showing an ability to shift to a rugose phenotype. Our data confirm the ability of Haitian clinical (and environmental strains to shift to a protective rugose phenotype, and suggest that factors such as temperature influence the frequency of transition to this phenotype.

  5. Production of novel types of antibacterial liamocins by diverse strains of Aureobasidium pullulans grown on different culture media

    Science.gov (United States)

    Objective: The objective was to compare production of antibacterial liamocins by diverse strains of A. pullulans grown on different culture media. Results: Liamocins produced by strains of A. pullulans have potential agricultural and pharmaceutical applications as antibacterials with specificity aga...

  6. Xylitol production by genetically modified industrial strain of Saccharomyces cerevisiae using glycerol as co-substrate.

    Science.gov (United States)

    Kogje, Anushree B; Ghosalkar, Anand

    2017-06-01

    Xylitol is commercially used in chewing gum and dental care products as a low calorie sweetener having medicinal properties. Industrial yeast strain of S. cerevisiae was genetically modified to overexpress an endogenous aldose reductase gene GRE3 and a xylose transporter gene SUT1 for the production of xylitol. The recombinant strain (XP-RTK) carried the expression cassettes of both the genes and the G418 resistance marker cassette KanMX integrated into the genome of S. cerevisiae. Short segments from the 5' and 3' delta regions of the Ty1 retrotransposons were used as homology regions for integration of the cassettes. Xylitol production by the industrial recombinant strain was evaluated using hemicellulosic hydrolysate of the corn cob with glucose as the cosubstrate. The recombinant strain XP-RTK showed significantly higher xylitol productivity (212 mg L -1  h -1 ) over the control strain XP (81 mg L -1  h -1 ). Glucose was successfully replaced by glycerol as a co-substrate for xylitol production by S. cerevisiae. Strain XP-RTK showed the highest xylitol productivity of 318.6 mg L -1  h -1 and titre of 47 g L -1 of xylitol at 12 g L -1 initial DCW using glycerol as cosubstrate. The amount of glycerol consumed per amount of xylitol produced (0.47 mol mol -1 ) was significantly lower than glucose (23.7 mol mol -1 ). Fermentation strategies such as cell recycle and use of the industrial nitrogen sources were demonstrated using hemicellulosic hydrolysate for xylitol production.

  7. CodY Promotes Sporulation and Enterotoxin Production by Clostridium perfringens Type A Strain SM101.

    Science.gov (United States)

    Li, Jihong; Freedman, John C; Evans, Daniel R; McClane, Bruce A

    2017-03-01

    Clostridium perfringens type D strains cause enterotoxemia and enteritis in livestock via epsilon toxin production. In type D strain CN3718, CodY was previously shown to increase the level of epsilon toxin production and repress sporulation. C. perfringens type A strains producing C. perfringens enterotoxin (CPE) cause human food poisoning and antibiotic-associated diarrhea. Sporulation is critical for C. perfringens type A food poisoning since spores contribute to transmission and resistance in the harsh food environment and sporulation is essential for CPE production. Therefore, the current study asked whether CodY also regulates sporulation and CPE production in SM101, a derivative of C. perfringens type A food-poisoning strain NCTC8798. An isogenic codY -null mutant of SM101 showed decreased levels of spore formation, along with lower levels of CPE production. A complemented strain recovered wild-type levels of both sporulation and CPE production. When this result was coupled with the earlier results obtained with CN3718, it became apparent that CodY regulation of sporulation varies among different C. perfringens strains. Results from quantitative reverse transcriptase PCR analysis clearly demonstrated that, during sporulation, codY transcript levels remained high in SM101 but rapidly declined in CN3718. In addition, abrB gene expression patterns varied significantly between codY -null mutants of SM101 and CN3718. Compared to the levels in their wild-type parents, the level of abrB gene expression decreased in the CN3718 codY -null mutant strain but significantly increased in the SM101 codY -null mutant strain, demonstrating CodY-dependent regulation differences in abrB expression between these two strains. This difference appears to be important since overexpression of the abrB gene in SM101 reduced the levels of sporulation and enterotoxin production, supporting the involvement of AbrB repression in regulating C. perfringens sporulation. Copyright © 2017

  8. CHARACTERIZATION OF EXTENDED-SPECTRUM Β-LACTAMASE-PRODUCING ESCHERICHIA COLI STRAINS ISOLATED FROM DAIRY PRODUCTS

    Directory of Open Access Journals (Sweden)

    Rahem Khoshbakht

    2014-02-01

    Full Text Available Extended-spectrum β-lactamases (ESBLs are enzymes that hydrolyze the β-lactam ring, and ESBL-producing E. coli has rapidly spread worldwide with pose a serious hazard for humans. The aim of this study was to determine the prevalence of ESBL producing E. coli and molecular evaluation of four ESBL-associated genes among E. coli strains isolated from milk and cheese in southern Iran. Antibiotic susceptibility test was carried out for a total of 150 isolates of E. coli, previously collected from dairy products. ESBL production was screened using a double-disc synergy test (DDST and presence of four ESBL genes (PER, VEB, TEM and CTX-M was tested using PCR. Among 150 E. coli strains 57 (38% isolates were identified as ESBL-producing strains. All ESBL positive isolates could be typed for one or more genes and the most prevalent ESBL-associated gene was CTX-M (80.7%. The PER gene was not present among isolates. Isolates showed high susceptibility to imipe¬nem and cefoxitin. The results showed the high prevalence of ESBL producing E. coli strains among dairy products and high occurrence of CTX-M-associated ESBL activity among isolates indicating the hazards of increasing the strains with antibiotic resistance which can transfer to human trough the dairy food products.

  9. Isolation and Fatty Acid Profile of Selected Microalgae Strains from the Red Sea for Biofuel Production

    Directory of Open Access Journals (Sweden)

    Khalid M. Abu-Salah

    2013-05-01

    Full Text Available The isolation of lipid-rich autochthonous strains of microalgae is a crucial stage for the development of a microalgae-based biofuel production plant, as these microalgae already have the necessary adaptations to withstand competition, predation and the temperatures observed at each production site. This is particularly important in extreme climates such as in Saudi Arabia. Resorting to fluorescence activated cell sorting (FACS we screened for and isolated several microalgal strains from samples collected from the Red Sea. Relying on the fluorescence of BODIPY 505/515 (4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diazasindacene and growth performance, four promising candidates were identified and the total lipid content and fatty acid profile was assessed for biofuels production. Selected isolates were classified as chlorophytes, belonging to three different genera: Picochlorum, Nannochloris and Desmochloris. The lipid contents were assessed microscopically by means of BODIPY 505/515-associated fluorescence to detect intracellular lipid bodies, which revealed several lipid drops in all selected strains. This result was confirmed by lipid gravimetric determination, which demonstrated that all strains under study presented inner cell lipid contents ranging from 20% to 25% of the biomass dry weight. Furthermore, the fatty acid methyl esters profile of all strains seems ideal for biodiesel production due to a low degree of polyunsaturated fatty acid methyl esters and high amount of palmitic and oleic acids.

  10. Production of 5-ketofructose from fructose or sucrose using genetically modified Gluconobacter oxydans strains.

    Science.gov (United States)

    Siemen, Anna; Kosciow, Konrad; Schweiger, Paul; Deppenmeier, Uwe

    2018-02-01

    The growing consumer demand for low-calorie, sugar-free foodstuff motivated us to search for alternative non-nutritive sweeteners. A promising sweet-tasting compound is 5-keto-D-fructose (5-KF), which is formed by membrane-bound fructose dehydrogenases (Fdh) in some Gluconobacter strains. The plasmid-based expression of the fdh genes in Gluconobacter (G.) oxydans resulted in a much higher Fdh activity in comparison to the native host G. japonicus. Growth experiments with G. oxydans fdh in fructose-containing media indicated that 5-KF was rapidly formed with a conversion efficiency of 90%. 5-KF production from fructose was also observed using resting cells with a yield of about 100%. In addition, a new approach was tested for the production of the sweetener 5-KF by using sucrose as a substrate. To this end, a two-strain system composed of the fdh-expressing strain and a G. oxydans strain that produced the sucrose hydrolyzing SacC was developed. The strains were co-cultured in sucrose medium and converted 92.5% of the available fructose units into 5-KF. The glucose moiety of sucrose was converted to 2-ketogluconate and acetate. With regard to the development of a sustainable and resource-saving process for the production of 5-KF, sugar beet extract was used as substrate for the two-strain system. Fructose as product from sucrose cleavage was mainly oxidized to 5-KF which was detected in a concentration of over 200 mM at the end of the fermentation process. In summary, the two-strain system was able to convert fructose units of sugar beet extract to 5-KF with an efficiency of 82 ± 5%.

  11. Influence of Mode of Fermentation on Production of Polygalacturonase by a Novel Strain of Streptomyces lydicus

    Directory of Open Access Journals (Sweden)

    Nicemol Jacob

    2006-01-01

    Full Text Available Production of different pectinolytic enzymes was attempted using the actinomycete strain Streptomyces lydicus in submerged fermentation. Polygalacturonase and pectin lyase activities were detected in the culture supernatant, but the strain was not able to produce pectin esterase. Polygalacturonase production was studied in submerged, slurry-state and solid-state fermentation systems. All the experiments were carried out under static and shaking conditions. Solid-state fermentation under static condition was found to be promising. Various agroindustrial residues were tried as substrates for solid-state fermentation. Wheat bran was proved to be the best substrate.

  12. Characterization and optimization of antibiotic resistant bacterial strains for polyhydroxyalkanoates (phas) production

    International Nuclear Information System (INIS)

    Rehman, S. U.; Jamil, N.; Hussain, S.

    2005-01-01

    In this investigation, sugarcane soil, sewage water and soil containing long chain hydrocarbons was screened to obtain bacterial strains that were able to synthesize poly-beta-hydroxyalkanoates (PHA). The potential to synthesize PHA was tested qualitatively by Sudan Black staining of colonies growing in glucose and sucrose. Sixteen bacterial strains were isolated, purified and characterized for Gram reaction, biochemical analysis and PHA production. Isolates showed a wide range of tolerance to different commonly used antibiotics. PHA extraction was done by solvent extraction and hypochlorite digestion method. PHA production was optimized for different nitrogen concentrations. (author)

  13. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet.

    Science.gov (United States)

    Sumi, H; Hamada, H; Tsushima, H; Mihara, H; Muraki, H

    1987-10-15

    A strong fibrinolytic activity was demonstrated in the vegetable cheese Natto, which is a typical soybean food eaten in Japan. The average activity was calculated at about 40 CU (plasmin units)/g wet weight. This novel fibrinolytic enzyme, named nattokinase, was easily extracted with saline. The mol. wt and pI were about 20,000 and 8.6, respectively. Nattokinase not only digested fibrin but also the plasmin substrate H-D-Val-Leu-Lys-pNA (S-2251), which was more sensitive to the enzyme than other substrates tried. Diisopropyl fluorophosphate and 2,2,2-trichloro-1-hydroxyethyl-o,o-dimethylphosphate strongly inhibited this fibrinolytic enzyme.

  14. Comparison of some indigenous bacterial strains of pseudomonas ssp. for production of biosurfactants

    International Nuclear Information System (INIS)

    Sahafeeq, M.; Kokub, D.; Khalid, Z.M.; Malik, K.A.

    1991-01-01

    Some indigenous pseudomonas spp. were found to have the ability of emulsification, lowering the surface and interfacial tensions, and formation of high reciprocal CMCs. Six strains of Pseudomonas spp were compared for biosurfactant production grown on hexadecane. Supernatant from whole culture broth of these strains could lower surface tension from 65 mN/m to 28-32 nM/m, interfacial tension from 40 nM/m to 1-3 mN/m and had high reciprocal CMCs. When compared for emulsification ability by the culture broth of these strains, the emulsification index (E24) was found to range between 60-65. Biosurfactant containing culture broth of some strains could retain the property up to 80 C, pH of 13 and sodium chloride concentration for 17% which indicates their possible role in some depleted oil well. (author)

  15. Antimicrobial resistance and production of biofilms in clinical isolates of coagulase-negative Staphylococcus strains.

    Science.gov (United States)

    de Allori, María Cristina Gaudioso; Jure, María Angela; Romero, Cintia; de Castillo, Marta Elena Cecilia

    2006-08-01

    Coagulase-negative Staphylococcus (CNS) strains are frequently associated with bacteremia and hospital-acquired infections. 293 CNS strains were isolated from 744 samples from a dialysis center in S. M. de Tucumán, Argentina, from hemocultures, catheters and urine and identified as S. epidermidis, S. haemolyticus, S. saprophyticus, S. hominis and S. cohnii. 13 antibiotics were tested for antibacterial resistance. 75% of S. saprophyticus, 66% of S. epidermidis and 57% of S. haemolyticus was resistant to erythromycin and 50% of S. haemolyticus was resistant to ciprofloxacin. OXA resistance was found in 43% of S. haemolyticus. Presence of PBP 2a in OXA-R strains was confirmed with the modified agglutination assay (MRSA) and presence of the mecA gene. 15 strains with intermediate halos for vancomycin and teicoplanin showed a MIC in solid and liquid medium resistance to methicillin and biofilm production are decisive for a prompt and appropriate antimicrobial therapy and limited use of inappropriate glycopeptides.

  16. Microbial Fe (III) reduction and hydrogen production by a transposon-mutagenized strain of Pantoea agglomerans BH18

    International Nuclear Information System (INIS)

    Liu, Hongyan; Wang, Guangce

    2015-01-01

    Based on the transposon-mutagenized library of Pantoea agglomerans BH18, mutant screens were conducted to obtain the strain with the highest Fe (III) reduction and hydrogen production. Of these transposon-mutagenized mutants, the mutant strain TB230 was screened for high Fe (III)-reducing efficiency and hydrogen production. The PCR amplification and kanamycin resistance selection results indicated that the transposon insertion of the mutant strain TB230 was stable. Hydrogen production of the mutant strain TB230 was (2.21 ± 0.34) mol H 2 /mol glucose, which increased hydrogen production by over 40% compared with that of the wild type strain. The accumulation concentration of Fe (II) in the medium of the mutant strain TB230 with Fe (OH) 3 as the sole electron acceptor was (7.39 ± 0.49) mmol/l, which was approximately 3-fold greater than that of the wild type strain. The mutant strain TB230 showed high Fe (III)-reducing activity and hydrogen production by adopting glucose and pyruvate as the carbon source. In addition, the mutant strain TB230 was capable of Fe (III) reduction and hydrogen production under fresh or marine conditions. This result indicates that the mutant strain with high microbial Fe (III) reduction and hydrogen production is beneficial for the improvement of anaerobic performance. - Highlights: • The mutant strain TB230 was a transposon-mutagenized strain of Pantoea agglomerans BH18. • Strain TB230 was screened for high Fe (III)-reducing efficiency and hydrogen production. • H 2 yield and Fe (III)-reducing activity were 2.21 ± 0.34 and 7.39 ± 0.49 in marine condition. • Strain TB230 was capable of Fe (III) reduction and hydrogen production in fresh or marine condition

  17. Preliminary evaluation of probiotic potential of Lactobacillus plantarum strains isolated from Italian food products.

    Science.gov (United States)

    Turchi, Barbara; Mancini, Simone; Fratini, Filippo; Pedonese, Francesca; Nuvoloni, Roberta; Bertelloni, Fabrizio; Ebani, Valentina Virginia; Cerri, Domenico

    2013-10-01

    , as well as the ability of the strains to adhere to yeast cells wasn't modified by previous treatments (lysozyme, low pH and bile salts). The present work allows to increase knowledge about non starter lactic acid bacteria from Italian food products. The studied L. plantarum strains showed a good potential for their use as probiotic cultures. However, more in vivo tests are necessary to confirm this potentiality.

  18. Continuous production of ethanol from hexoses and pentoses using immobilized mixed cultures of Escherichia coli strains

    Science.gov (United States)

    Unrean, Pornkamol; Srienc, Friedrich

    2010-01-01

    We have developed highly efficient ethanologenic E. coli strains that selectively consume pentoses and/or hexoses. Mixed cultures of these strains can be used to selectively adjust the sugar utilization kinetics in ethanol fermentations. Based on the kinetics of sugar utilization, we have designed and implemented an immobilized cell system for the optimized continuous conversion of sugars into ethanol. The results confirm that immobilized mixed cultures support a simultaneous conversion of hexoses and pentoses into ethanol at high yield and at a faster rate than immobilized homogenous cells. Continuous ethanol production has been maintained for several weeks at high productivity with near complete sugar utilization. The control of sugar utilization using immobilized mixed cultures can be adapted to any composition of hexoses and pentoses by adjusting the strain distribution of immobilized cells. The approach, therefore, holds promise for ethanol fermentation from lignocellulosic hydrolysates where the feedstock varies in sugar composition. PMID:20699108

  19. Differences in Shiga toxin and phage production among stx2g-positive STEC strains

    Directory of Open Access Journals (Sweden)

    Claudia Viviana Granobles Velandia

    2012-06-01

    Full Text Available Shigatoxigenic E. coli (STEC are characterized by the production of Shiga toxins (Stx encoded by temperate bacteriophages. Stx production is linked to the induction of the phage lytic cycle. Several stx variants have been described and differentially associated with the risk of developing severe illness.The variant named stx2g was first identified in a STEC strain isolated from the faeces of healthy cattle. Analysis of stx2g-positive strains isolated from humans, animals and environmental sources have shown that they have a close relationship. In this study, stx2g-positive STEC isolated from cattle were analyzed for phage and Stx production, with the aim to relate the results to differences observed in cytotoxicity.The presence of inducible phages was assessed by analyzing the bacterial growth/lysis curves and also by plaque assay. Bacterial growth curves in the absence of induction were similar for all isolates, however, notably differed among induced cultures. The two strains that clearly evidenced bacteriolysis under this condition also showed higher phage titers in plaque assays. However, only the phage plaques produced by one of these strains (FB 62 hybridized with a stx2-probe. Furthermore, the production of Stx was evaluated by EIA and Western immunoblotting in overnight supernatants. By EIA, we detected Stx only in supernatants of FB 62, with a higher signal with induced than in uninduced cultures. By immunoblotting, Stx2 could be detected after induction in all stx2g-positive isolates, but with lower amounts of Stx2B subunit in those supernatants where phages could not be detected.Taking into account all the results, several differences could be found among stx2g-positive strains. The strain with the highest cytotoxic titer showed higher levels of stx2-phages and toxin production by EIA, and the opposite was observed for strains that previously showed low cytotoxic titers, confirming that in stx2g-positive strains Stx production is

  20. The suitability of different probiotic strains for the production of fruit-whey beverages.

    Science.gov (United States)

    Sady, Marek; Najgebauer-Lejko, Dorota; Domagała, Jacek

    2017-01-01

    When designing new probiotic products, one of the most important aspects is the selection of bacterial strains with high survival rates in the matrix of the product concerned. The aim of the present research was to evaluate the potential of selected strains of probiotic bacteria for the production of fruit-whey beverages. Orange, apple and blackcurrant whey beverages were produced, and each was inoculated with one of the following probiotic strains: Bifidobacterium lactis HN019TM; Lactobacillus aci- dophilus NCFM®; Lactobacillus paracasei Lpc-37TM; Lactobacillus rhamnosus HN001TM. The count of probiotic bacteria as well as pH and total acidity were evaluated at the 1st, 7th, 14th, 21st and 28th day of storage. Beverages containing L. paracasei Lpc-37TM or L. rhamnosus HN001TM were characterized by a sig- nificantly higher average number of viable cells (7.02 or 7.05 log cfu/g, respectively) than products with lactis HN019TM or L. acidophilus NCFM® (6.43 or 6.37 log cfu/g, respectively). The use of L. paracasei Lpc-37 and L. rhamnosus HN001 strains in orange and apple drinks allows the recommended count for probiotic products, 106 cfu/g for 28 days of storage, to be exceeded. Survival of the B. lactis HN019 strain fulfills the above requirements only in the orange drink. The L. acidophilus NCFM® strain was found to be the least suitable for the production of beverages, as it did not reach 6 log cfu/g in any products after 28 days of stor- age. The highest average number of bacteria was found in the orange beverages (7.14 log cfu/g). In terms of bacteria viability, blackcurrant juice was the least suitable for the production of whey probiotic drinks, due to its high acidity. The results of the present study indicate that careful selection of the fruit juice component, especially in terms of its acidity, is key to designing successful probiotic fruit-whey beverages. Other factors which should be taken into account to ensure a sufficient number of live probiotic

  1. Genotype by production environment interaction in the GIFT strain of Nile tilapia (Oreochromis niloticus)

    NARCIS (Netherlands)

    Khaw, H.L.; Ponzonia, R.W.; Hamzah, A.; Abu-Bakara, K.R.; Bijma, P.

    2012-01-01

    Three discrete generations of GIFT fish (Nile tilapia strain, Oreochromis niloticus; a total of 10,065 fish with pedigree and phenotypic information) were tested in pond and cage culture environments to determine genotype by production environment interaction between both environments in Malaysia.

  2. Development of over-production strain of saccharification enzyme and biomass pretreatment by proton beam irradiation

    International Nuclear Information System (INIS)

    Kim, S. O.; Lee, J. Y.; Song, Y. S.; Shin, H. S.

    2009-04-01

    - The first year : Pre-treatment of biomass by proton beam irradiation and characterization of the pretreated biomass by IR and SEM - The second year : Strain development by proton beam irradiation for the production of cellulase and hemicellulase - The third year : Optimization of Saccharification process by cellulase and hemicellulase

  3. Ethanol production from Jerusalem artichoke by strains of Saccharomyces cheresiensis and Saccharomyces beticus

    Energy Technology Data Exchange (ETDEWEB)

    Pourrat, H.; Barthomeuf, C.; Regerat, F.; Carnat, A.P.; Carnat, A.

    1983-03-01

    Ethanol production from Jerusalem artichoke which is the most interesting autochtonous material has been studied. Two selected and acclimatised strains of Saccharomyces: Saccharomyces cheresiensis and Saccharomyces beticus were retained. The fermentation conditions, exactly definited, makes it possible to obtain in 4 days a theoric yield.

  4. Total synthesis and related studies of large, strained, and bioactive natural products

    Science.gov (United States)

    HIRAMA, Masahiro

    2016-01-01

    Our chemical syntheses and related scientific investigations of natural products with complex architectures and powerful biological activities are described, focusing on the very large 3 nm-long polycyclic ethers called the ciguatoxins, highly strained and labile chromoprotein antitumor antibiotics featuring nine-membered enediyne cores, and extremely potent anthelmintic macrolides called the avermectins. PMID:27725470

  5. Production of xylitol by a Coniochaeta ligniaria strain tolerant of inhibitors and defective in xylose metabolism

    Science.gov (United States)

    In conversion of biomass to fuels or chemicals, inhibitory compounds arising from physical-chemical pretreatment of the feedstock can interfere with fermentation of the sugars to product. Fungal strain Coniochaeta ligniaria NRRL30616, metabolizes the furan aldehydes furfural and 5-hydroxymethylfurfu...

  6. Under pressure: evolutionary engineering of yeast strains for improved performance in fuels and chemicals production

    NARCIS (Netherlands)

    Mans, R.; Daran, J.G.; Pronk, J.T.

    2018-01-01

    Evolutionary engineering, which uses laboratory evolution to select for industrially relevant traits, is a popular strategy in the development of high-performing yeast strains for industrial production of fuels and chemicals. By integrating whole-genome sequencing, bioinformatics, classical

  7. Production of L-lactic acid from metabolically engineered strain of Enterobacter aerogenes ATCC 29007.

    Science.gov (United States)

    Thapa, Laxmi Prasad; Lee, Sang Jun; Park, Chulhwan; Kim, Seung Wook

    2017-07-01

    In this study, L-lactic acid production was investigated from metabolically engineered strain of E. aerogenes ATCC 29007. The engineered strain E. aerogenes SUMI01 (Δpta) was generated by the deletion of phosphate acetyltransferase (pta) gene from the chromosome of E. aerogenes ATCC 29007 and deletion was confirmed by colony PCR. Under the optimized fermentation conditions, at 37°C and pH 6 for 84h, the L-lactic acid produced by engineered strain E. aerogenes SUMI01 (Δpta) in flask fermentation using 100g/L mannitol as the carbon source was 40.05g/L as compared to that of the wild type counterpart 20.70g/L. At the end of the batch fermentation in bioreactor the production of L-lactic acid reached to 46.02g/L and yield was 0.41g/g by utilizing 112.32g/L mannitol. This is the first report regarding the production of L-lactic acid from Enterobacter species. We believe that this result may provide valuable guidelines for further engineering Enterobacter strain for the improvement of L-lactic acid production. Copyright © 2017 Elsevier Inc. All rights reserved.

  8. Moorella Strains for the Production of Biochemicals from Syngas

    DEFF Research Database (Denmark)

    Redl, Stephanie; Jensen, Torbjørn Ølshøj; Nielsen, Alex Toftgaard

    . In the syngasfermentation process even the recalcitrant lignin portion can be fully converted into higher valuecompounds.However, cost-effectiveness of this process requires better understanding of the metabolism andmodification of the acetogenic strains. The present project lays the basis for production of higher...

  9. Enhanced production of dimethyl phthalate-degrading strain Bacillus sp. QD14 by optimizing fermentation medium

    Directory of Open Access Journals (Sweden)

    Jixian Mo

    2015-05-01

    Conclusion: In this work, the key factors affected by the fermentation of DMP-degrading strain Bacillus sp. QD14 were optimized by PBD, SAM and BBD (RSM; the yield was increased by 57,11% in the conditions in our study. We propose that the conditions optimized in the study can be applied to the fermentation for commercialization production.

  10. Excessive by-product formation : A key contributor to low isobutanol yields of engineered Saccharomyces cerevisiae strains

    NARCIS (Netherlands)

    Milne, N.S.W.; Wahl, S.A.; Van Maris, A.J.A.; Pronk, J.T.; Daran, J.M.

    2016-01-01

    It is theoretically possible to engineer Saccharomyces cerevisiae strains in which isobutanol is the predominant catabolic product and high-yielding isobutanol-producing strains are already reported by industry. Conversely, isobutanol yields of engineered S. cerevisiae strains reported in the

  11. Study of itaconic acid production by Aspergillus terrus MJL05 strain with different variable

    OpenAIRE

    Juy, M. I.; Orejas, J. A.; Lucca, M. E.

    2010-01-01

    Itaconic acid (IA) production by Aspergillus terreus MJL05 strain was investigated in submerged batch fermentation in a stirred bioreactor to determine the effect of varying the nitrogen, phosphorous and carbon sources concentrations in the production medium. Glycerol, a biodiesel by-product was reported as an efficient substrate to achieve high itaconic acid productivities. This was used as the sole carbon source. The resulting C:N 18, N:P 10.8 and C:P 195 ratios were selected as the best an...

  12. EFFECT OF ESSENTIAL OIL ON BIOFILM PRODUCTION BY DIFFERENT LISTERIA MONOCYTOGENES STRAINS

    Directory of Open Access Journals (Sweden)

    G. Comi

    2008-12-01

    Full Text Available The effects of different essential oil (hexanal, 2-(E-hexenal, carvacrol, citron, red orange, thymol and limonene on biofilm production of some Lmonocytogenes strains are evaluated. The formation of biofilm on certain surfaces or on the food, seems to be related with cross-contamination during processing or with the contamination of the final product, with potential risk for the consumer. Many studies were done on the antimicrobial activity of essential oils and their components, but not too much is known about their capacity to influence and reduce the microbial production of biofilm. Our data showed that essential oils can inhibit or limit the biofilm production.

  13. Cellulase production by two mutant strain of Trichoderma longibrachiatum Qm9414 and Rut C30

    International Nuclear Information System (INIS)

    Blanco, M.J.

    1991-01-01

    Native or pretreated biomass from Onopordum nervosum boiss, has been examined as candidate feedstock for cellulase production by two mutant strain of trichoderma longibrachiatum QM9414 and Rut C30. Batch cultivation methods were evaluated and compared with previous experiments using ball-milled, crystalline cellulose (Solka floc). Batch cultivation of T. longibrachiatum Rut C30 on 55% (W/V) acid pretreated O. nervosum biomass yielded enzyme productivities and activities comparable to those obtained on Solka floc. However, the overall enzyme production performance was lower than on Solka floc at comparable cellulose concentrations. This fact may be due to the accumulation of pretreated by products and lignin in the fermentor.(author)

  14. Strain- and Substrate-Dependent Redox Mediator and Electricity Production by Pseudomonas aeruginosa.

    Science.gov (United States)

    Bosire, Erick M; Blank, Lars M; Rosenbaum, Miriam A

    2016-08-15

    Pseudomonas aeruginosa is an important, thriving member of microbial communities of microbial bioelectrochemical systems (BES) through the production of versatile phenazine redox mediators. Pure culture experiments with a model strain revealed synergistic interactions of P. aeruginosa with fermenting microorganisms whereby the synergism was mediated through the shared fermentation product 2,3-butanediol. Our work here shows that the behavior and efficiency of P. aeruginosa in mediated current production is strongly dependent on the strain of P. aeruginosa We compared levels of phenazine production by the previously investigated model strain P. aeruginosa PA14, the alternative model strain P. aeruginosa PAO1, and the BES isolate Pseudomonas sp. strain KRP1 with glucose and the fermentation products 2,3-butanediol and ethanol as carbon substrates. We found significant differences in substrate-dependent phenazine production and resulting anodic current generation for the three strains, with the BES isolate KRP1 being overall the best current producer and showing the highest electrochemical activity with glucose as a substrate (19 μA cm(-2) with ∼150 μg ml(-1) phenazine carboxylic acid as a redox mediator). Surprisingly, P. aeruginosa PAO1 showed very low phenazine production and electrochemical activity under all tested conditions. Microbial fuel cells and other microbial bioelectrochemical systems hold great promise for environmental technologies such as wastewater treatment and bioremediation. While there is much emphasis on the development of materials and devices to realize such systems, the investigation and a deeper understanding of the underlying microbiology and ecology are lagging behind. Physiological investigations focus on microorganisms exhibiting direct electron transfer in pure culture systems. Meanwhile, mediated electron transfer with natural redox compounds produced by, for example, Pseudomonas aeruginosa might enable an entire microbial

  15. The Impact of Single Amino Acids on Growth and Volatile Aroma Production by Saccharomyces cerevisiae Strains

    OpenAIRE

    Samantha Fairbairn; Alexander McKinnon; Hannibal T. Musarurwa; António C. Ferreira; António C. Ferreira; Florian F. Bauer

    2017-01-01

    Nitrogen availability and utilization by Saccharomyces cerevisiae significantly influence fermentation kinetics and the production of volatile compounds important for wine aroma. Amino acids are the most important nitrogen source and have been classified based on how well they support growth. This study evaluated the effect of single amino acids on growth kinetics and major volatile production of two phenotypically different commercial wine yeast strains in synthetic grape must. Four growth p...

  16. Selection of aggressive pathogenic and solopathogenic strains of Ustilago maydis to improve Huitlacoche production

    Directory of Open Access Journals (Sweden)

    Porfirio Raúl Galicia-García

    Full Text Available ABSTRACT Ustilago maydis is a basidiomycete known as the causative agent of 'common smut', worldwide disease of maize that is recognized by the galls it forms, which have considerable potential as a gourmet food. Results of infection are quite variable, even under optimal greenhouse conditions. In order to find pathogenic strains able to be used as a highly infective and stable inoculum for the successful production of galls either in greenhouses or in the field, ears with gall symptoms containing teliospores were recovered from maize plants. The teliospores were suspended in water and plated on nutrient-rich medium. Twenty-six colonies developed, containing three types of yeast-like colonies: saprotrophic, pathogenic, and solopathogenic. DAPI staining confirmed the presence of solopathogenic strains with diploid sporidia. Groups of different mating types were found when pairs of the 26 strains were arranged resembling partial-diallel combinations. Amplification of the partial b locus revealed that the strains found harbor the alleles b3 and b4, allowing the formation in dikaryotic strains of heterodimeric regulatory proteins associated with fungal development and pathogenicity. In this study, we isolated compatible haploid and solopathogenic diploid strains for their high capacity for inducing smut.

  17. Evaluation of UV-C mutagenized Scheffersomyces stipitis strains for ethanol production.

    Science.gov (United States)

    Geiger, Melanie; Gibbons, Jaimie; West, Thomas; Hughes, Stephen R; Gibbons, William

    2012-12-01

    We evaluated fermentation capabilities of five strains of Scheffersomyces stipitis (WT-2-1, WT-1-11, 14-2-6, 22-1-1, and 22-1-12) that had been produced by UV-C mutagenesis and selection for improved xylose fermentation to ethanol using an integrated automated robotic work cell. They were incubated under both facultative and anaerobic conditions to evaluate ethanol production on glucose, xylose, cellobiose, and a combination of all three sugars. The medium contained 50 g/L total sugar and 5 g/L yeast extract. The strains performed significantly better under facultative compared with anaerobic conditions. As expected, glucose was the most readily fermented sugar with ~100% fermentation efficiency (FE) under facultative conditions but only 5% to 16% FE anaerobically. Xylose utilization was 20% to 40% FE under facultative conditions but 9% to 25% FE anaerobically. Cellobiose was the least fermented sugar, at 18% to 27% FE facultatively and 8% to 11% anaerobically. Similar trends occurred in the sugar mixture. Under facultative conditions, strain 22-1-12 produced 19.6 g/L ethanol on glucose, but strain 14-2-6 performed best on xylose (4.5 g/L ethanol) and the sugar combination (8.0 g/L ethanol). Ethanol titers from glucose under anaerobic conditions were again highest with strain 22-1-12, but none of the strains produced ethanol from xylose. Future trials will evaluate nutrient addition to boost microaerophilic xylose fermentation.

  18. Preliminary Evaluation of Probiotic Properties of Lactobacillus Strains Isolated from Sardinian Dairy Products

    Directory of Open Access Journals (Sweden)

    Maria Barbara Pisano

    2014-01-01

    Full Text Available Twenty-three Lactobacillus strains of dairy origin were evaluated for some functional properties relevant to their use as probiotics. A preliminary subtractive screening based on the abilities to inhibit the growth of microbial pathogens and hydrolyze conjugated bile salts was applied, and six strains were selected for further characterization including survival under gastrointestinal environmental conditions, adhesion to gut epithelial tissue, enzymatic activity, and some safety properties. All selected strains maintained elevated cell numbers under conditions simulating passage through the human gastrointestinal tract, well comparable to the values obtained for the probiotic strain Lactobacillus rhamnosus GG, and were able to adhere to Caco-2 cells to various extents (from 3 to 20%. All strains exhibited high aminopeptidase, and absent or very low proteolytic and strong β-galactosidase activities; none was found to be haemolytic or to produce biogenic amines and all were susceptible to tetracycline, chloramphenicol, erythromycin, ampicillin, and amoxicillin/clavulanic acid. Our results indicate that the Lactobacillus strains analyzed could be considered appropriate probiotic candidates, due to resistance to GIT simulated conditions, antimicrobial activity, adhesion to Caco-2 cell-line, and absence of undesirable properties. They could be used as adjunct cultures for contributing to the quality and health related functional properties of dairy products.

  19. Large-scale bioreactor production of the herbicide-degrading Aminobacter sp. strain MSH1

    DEFF Research Database (Denmark)

    Schultz-Jensen, Nadja; Knudsen, Berith Elkær; Frkova, Zuzana

    2014-01-01

    The Aminobacter sp. strain MSH1 has potential for pesticide bioremediation because it degrades the herbicide metabolite 2,6-dichlorobenzamide (BAM). Production of the BAM-degrading bacterium using aerobic bioreactor fermentation was investigated. A mineral salt medium limited for carbon and with ......The Aminobacter sp. strain MSH1 has potential for pesticide bioremediation because it degrades the herbicide metabolite 2,6-dichlorobenzamide (BAM). Production of the BAM-degrading bacterium using aerobic bioreactor fermentation was investigated. A mineral salt medium limited for carbon...... and with an element composition similar to the strain was generated. The optimal pH and temperature for strain growth were determined using shaker flasks and verified in bioreactors. Glucose, fructose, and glycerol were suitable carbon sources for MSH1 (μ =0.1 h−1); slower growth was observed on succinate and acetic...... acid (μ =0.01 h−1). Standard conditions for growth of theMSH1 strain were defined at pH 7 and 25 °C, with glucose as the carbon source. In bioreactors (1 and 5 L), the specific growth rate of MSH1 increased from μ =0.1 h−1 on traditional mineral salt medium to μ =0.18 h−1 on the optimized mineral salt...

  20. Production of lipids in 10 strains of Chlorella and Parachlorella, and enhanced lipid productivity in Chlorella vulgaris

    Energy Technology Data Exchange (ETDEWEB)

    Pribyl, Pavel; Cepak, Vladislav [Academy of Sciences of the Czech Republic, Trebon (Czech Republic). Algological Centre and Centre for Bioindication and Revitalization; Zachleder, Vilem [Academy of Sciences of the Czech Republic, Trebon (Czech Republic). Lab. of the Cell Cycles of Algae

    2012-04-15

    We tested 10 different Chlorella and Parachlorella strains under lipid induction growth conditions in autotrophic laboratory cultures. Between tested strains, substantial differences in both biomass and lipid productivity as well as in the final content of lipids were found. The most productive strain (Chlorella vulgaris CCALA 256) was subsequently studied in detail. The availability of nitrates and/or phosphates strongly influenced growth and accumulation of lipids in cells by affecting cell division. Nutrient limitation substantially enhanced lipid productivity up to a maximal value of 1.5 g l{sup -1} day{sup -1}. We also demonstrated the production of lipids through large-scale cultivation of C. vulgaris in a thin layer photobioreactor, even under suboptimal conditions. After 8 days of cultivation, maximal lipid productivity was 0.33 g l{sup -1} day{sup -1}, biomass density was 5.7 g l{sup -1} dry weight and total lipid content was more than 30% dry weight. C. vulgaris lipids comprise fatty acids with a relatively high degree of saturation compared with canola oil offering a possible alternative to the use of higher plant oils. (orig.)

  1. Strain improvement and metabolic flux analysis in the wild-type and a mutant Lactobacillus lactis strain for L(+)-lactic acid production.

    Science.gov (United States)

    Bai, Dong-Mei; Zhao, Xue-Ming; Li, Xin-Gang; Xu, Shi-Min

    2004-12-20

    The effects of initial glucose concentration and calcium lactate concentration on the lactic acid production by the parent strain, Lactobacillus lactis BME5-18, were studied. The results of the experiments indicated that glucose and lactate repressed the cell growth and the lactic acid production by Lactobacillus lactis BME5-18. A L(+)-lactic acid overproducing strain, Lactobacillus lactis BME5-18M, was screened by mutagenizing the parent strain with ultraviolet (UV) light irradiation and selecting the high glucose and lactate calcium concentration repression resistant mutant. Starting with a concentration of 100g L(-1) glucose, the mutant produced 98.6 g L(-1) lactic acid after 60 h in flasks, 73.9% higher than that of the parent strain. The L(+)-lactic acid purity was 98.1% by weight based on the amount of total lactic acid. The culture of the parent strain could not be analyzed well by conventional metabolic flux analysis techniques, since some pyruvate were accumulated intracellularly. Therefore, a revised flux analysis method was proposed by introducing intracellular pyruvate pool. Further studies demonstrate that there is a high level of NADH oxidase activity (12.11 mmol mg(-1) min(-1)) in the parent strain. The molecular mechanisms of the strain improvement were proposed, i.e., the high level of NADH oxidase activity was eliminated and the uptake rate of glucose was increased from 82.1 C-mmol (g DW h)(-1) to 98.9 C-mmol (g DW h)(-1) by mutagenizing the parent strain with UV, and therefore the mutant strain converts mostly pyruvate to lactic acid with a higher productivity (1.76 g L(-1) h(-1)) than the parent strain (0.95 g L(-1) h(-1)).

  2. Improving yeast strains using recyclable integration cassettes, for the production of plant terpenoids

    Directory of Open Access Journals (Sweden)

    Johnson Christopher B

    2011-01-01

    Full Text Available Abstract Background Terpenoids constitute a large family of natural products, attracting commercial interest for a variety of uses as flavours, fragrances, drugs and alternative fuels. Saccharomyces cerevisiae offers a versatile cell factory, as the precursors of terpenoid biosynthesis are naturally synthesized by the sterol biosynthetic pathway. Results S. cerevisiae wild type yeast cells, selected for their capacity to produce high sterol levels were targeted for improvement aiming to increase production. Recyclable integration cassettes were developed which enable the unlimited sequential integration of desirable genetic elements (promoters, genes, termination sequence at any desired locus in the yeast genome. The approach was applied on the yeast sterol biosynthetic pathway genes HMG2, ERG20 and IDI1 resulting in several-fold increase in plant monoterpene and sesquiterpene production. The improved strains were robust and could sustain high terpenoid production levels for an extended period. Simultaneous plasmid-driven co-expression of IDI1 and the HMG2 (K6R variant, in the improved strain background, maximized monoterpene production levels. Expression of two terpene synthase enzymes from the sage species Salvia fruticosa and S. pomifera (SfCinS1, SpP330 in the modified yeast cells identified a range of terpenoids which are also present in the plant essential oils. Co-expression of the putative interacting protein HSP90 with cineole synthase 1 (SfCinS1 also improved production levels, pointing to an additional means to improve production. Conclusions Using the developed molecular tools, new yeast strains were generated with increased capacity to produce plant terpenoids. The approach taken and the durability of the strains allow successive rounds of improvement to maximize yields.

  3. Comparative studies on production of cellulases from three strains of aspergillus niger

    International Nuclear Information System (INIS)

    Sohail, M.; Ahmad, A.; Khan, S.

    2014-01-01

    Three strains of Aspergillus niger were retrieved from culture collection of the Department of Microbiology, University of Karachi, Pakistan and were studied for their ability to produce cellulases. Cultivation at different temperatures and in presence of various carbon sources revealed that all the three strains produced more amounts of endoglucanase, glucosidase and filter-paperase activities at 35 degree C; carboxymethyl cellulose promotes the production of filter paperase and endoglucanase activities whereas salicin induced glucosidase activity. Experiments on growth and enzyme production kinetics showed that generation time and hence volumetric rate of biomass production is influenced by the carbon source used in the medium; simple carbon source, such as glucose favored the growth of all the strains. Cellulases from all the strains showed optimum activity at temperature >50 degree C and under acidic range of pH, while melting temperature was 64-65 degree C. These findings affirm that cellulases from A. niger are potential candidates as alternative to Trichoderma cellulases. (author)

  4. Screening and selection of wild strains for L-arabinose isomerase production

    Directory of Open Access Journals (Sweden)

    R. M. Manzo

    2013-12-01

    Full Text Available The majority of L-arabinose isomerases have been isolated by recombinant techniques, but this methodology implies a reduced technological application. For this reason, 29 bacterial strains, some of them previously characterized as L-arabinose isomerase producers, were assayed as L-arabinose fermenting strains by employing conveniently designed culture media with 0.5% (w/v L-arabinose as main carbon source. From all evaluated bacterial strains, Enterococcus faecium DBFIQ ID: E36, Enterococcus faecium DBFIQ ID: ETW4 and Pediococcus acidilactici ATCC ID: 8042 were, in this order, the best L-arabinose fermenting strains. Afterwards, to assay L-arabinose metabolization and L-arabinose isomerase activity, cell-free extract and saline precipitated cell-free extract of the three bacterial cultures were obtained and the production of ketoses was determined by the cysteine carbazole sulfuric acid method. Results showed that the greater the L-arabinose metabolization ability, the higher the enzymatic activity achieved, so Enterococcus faecium DBFIQ ID: E36 was selected to continue with production, purification and characterization studies. This work thus describes a simple microbiological method for the selection of L-arabinose fermenting bacteria for the potential production of the enzyme L-arabinose isomerase.

  5. Effect of oligosaccharides on the growth of Lactobacillus delbrueckii subsp. bulgaricus strains isolated from dairy products.

    Science.gov (United States)

    Ignatova, Tseteslava; Iliev, Ilia; Kirilov, Nikolai; Vassileva, Tonka; Dalgalarrondo, Michèle; Haertlé, Thomas; Chobert, Jean-Marc; Ivanova, Iskra

    2009-10-28

    Eighteen lactic acid bacteria (LAB) strains isolated from dairy products, all identified as Lactobacillus delbrueckii subsp. bulgaricus, were tested for their ability to grow on three different oligosaccharides: fructo-oligosaccharides (FOS), gluco-oligosaccharides (GOS) and galacto-oligosaccharides (GalOS). The growth of LAB on different oligosaccharides was very different. Study of the antimicrobial activities of these LAB indicated that the system of uptake of unusual sugars influenced in a specific way the production of antimicrobial substances (bacteriocins) specific against gram-negative bacteria. The added oligosaccharides induced LAB to form end-products of a typical mixed acid fermentation. The utilization of different types of oligosaccharides may help to explain the ability of Lactobacillus strains to compete with other bacteria in the ecosystem of the human gastro-intestinal tract.

  6. Inhibition of ochratoxigenic moulds by Debaryomyces hansenii strains for biopreservation of dry-cured meat products

    DEFF Research Database (Denmark)

    Andrade, Maria J.; Thorsen, Line; Rodríguez, Alicia

    2014-01-01

    mould growth and OTA accumulation in dry-cured meat products. The inoculation of D. hansenii should be made at the beginning of processing (at the end of post salting) when the a(w) of the product is still high (near 0.94). This action in addition to application of appropriate hygienic actions......The ability of the osmotolerant yeast Debaryomyces hansenii to inhibit Penicillium nordicum, the most common ochratoxigenic mould encountered in dry-cured meat products, was evaluated. The antagonistic effect of ten D. hansenii strains isolated from dry-cured ham was screened in vitro using malt...... extract media and meat extract peptone media with the water activity (a(w)) adjusted to 0.97 and 0.90. A significant inhibition of the two tested P. nordicum strains by D. hansenii cells and cell-free supernatants was observed. At 0.97 a(w), increasing D. hansenii inoculum concentrations significantly...

  7. Biotin-independent strains of Escherichia coli for enhanced streptavidin production.

    Science.gov (United States)

    Jeschek, Markus; Bahls, Maximilian O; Schneider, Veronika; Marlière, Philippe; Ward, Thomas R; Panke, Sven

    2017-03-01

    Biotin is an archetypal vitamin used as cofactor for carboxylation reactions found in all forms of life. However, biotin biosynthesis is an elaborate multi-enzymatic process and metabolically costly. Moreover, many industrially relevant organisms are incapable of biotin synthesis resulting in the requirement to supplement defined media. Here we describe the creation of biotin-independent strains of Escherichia coli and Corynebacterium glutamicum through installation of an optimized malonyl-CoA bypass, which re-routes natural fatty acid synthesis, rendering the previously essential vitamin completely obsolete. We utilize biotin-independent E. coli for the production of the high-value protein streptavidin which was hitherto restricted because of toxic effects due to biotin depletion. The engineered strain revealed significantly improved streptavidin production resulting in the highest titers and productivities reported for this protein to date. Copyright © 2017 International Metabolic Engineering Society. Published by Elsevier Inc. All rights reserved.

  8. Heterologous Production and Yield Improvement of Epothilones in Burkholderiales Strain DSM 7029.

    Science.gov (United States)

    Bian, Xiaoying; Tang, Biao; Yu, Yucong; Tu, Qiang; Gross, Frank; Wang, Hailong; Li, Aiying; Fu, Jun; Shen, Yuemao; Li, Yue-Zhong; Stewart, A Francis; Zhao, Guoping; Ding, Xiaoming; Müller, Rolf; Zhang, Youming

    2017-07-21

    The cloning of microbial natural product biosynthetic gene clusters and their heterologous expression in a suitable host have proven to be a feasible approach to improve the yield of valuable natural products and to begin mining cryptic natural products in microorganisms. Myxobacteria are a prolific source of novel bioactive natural products with only limited choices of heterologous hosts that have been exploited. Here, we describe the use of Burkholderiales strain DSM 7029 as a potential heterologous host for the functional expression of myxobacterial secondary metabolites. Using a newly established electroporation procedure, the 56 kb epothilone biosynthetic gene cluster from the myxobacterium Sorangium cellulosum was introduced into the chromosome of strain DSM 7029 by transposition. Production of epothilones A, B, C, and D was detected despite their yields being low. Optimization of the medium, introduction of the exogenous methylmalonyl-CoA biosynthetic pathway, and overexpression of rare tRNA genes resulted in an approximately 75-fold increase in the total yields of epothilones to 307 μg L -1 . These results show that strain DSM 7029 has the potential to produce epothilones with reasonable titers and might be a broadly applicable host for the heterologous expression of other myxobacterial polyketide synthases and nonribosomal peptide synthetases, expediting the process of genome mining.

  9. Towards efficient bioethanol production from agricultural and forestry residues: Exploration of unique natural microorganisms in combination with advanced strain engineering.

    Science.gov (United States)

    Zhao, Xinqing; Xiong, Liang; Zhang, Mingming; Bai, Fengwu

    2016-09-01

    Production of fuel ethanol from lignocellulosic feedstocks such as agricultural and forestry residues is receiving increasing attention due to the unsustainable supply of fossil fuels. Three key challenges include high cellulase production cost, toxicity of the cellulosic hydrolysate to microbial strains, and poor ability of fermenting microorganisms to utilize certain fermentable sugars in the hydrolysate. In this article, studies on searching of natural microbial strains for production of unique cellulase for biorefinery of agricultural and forestry wastes, as well as development of strains for improved cellulase production were reviewed. In addition, progress in the construction of yeast strains with improved stress tolerance and the capability to fully utilize xylose and glucose in the cellulosic hydrolysate was also summarized. With the superior microbial strains for high titer cellulase production and efficient utilization of all fermentable sugars in the hydrolysate, economic biofuels production from agricultural residues and forestry wastes can be realized. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Improving itaconic acid production through genetic engineering of an industrial Aspergillus terreus strain.

    Science.gov (United States)

    Huang, Xuenian; Lu, Xuefeng; Li, Yueming; Li, Xia; Li, Jian-Jun

    2014-08-11

    Itaconic acid, which has been declared to be one of the most promising and flexible building blocks, is currently used as monomer or co-monomer in the polymer industry, and produced commercially by Aspergillus terreus. However, the production level of itaconic acid hasn't been improved in the past 40 years, and mutagenesis is still the main strategy to improve itaconate productivity. The genetic engineering approach hasn't been applied in industrial A. terreus strains to increase itaconic acid production. In this study, the genes closely related to itaconic acid production, including cadA, mfsA, mttA, ATEG_09969, gpdA, ATEG_01954, acoA, mt-pfkA and citA, were identified and overexpressed in an industrial A. terreus strain respectively. Overexpression of the genes cadA (cis-aconitate decarboxylase) and mfsA (Major Facilitator Superfamily Transporter) enhanced the itaconate production level by 9.4% and 5.1% in shake flasks respectively. Overexpression of other genes showed varied effects on itaconate production. The titers of other organic acids were affected by the introduced genes to different extent. Itaconic acid production could be improved through genetic engineering of the industrially used A. terreus strain. We have identified some important genes such as cadA and mfsA, whose overexpression led to the increased itaconate productivity, and successfully developed a strategy to establish a highly efficient microbial cell factory for itaconate protuction. Our results will provide a guide for further enhancement of the itaconic acid production level through genetic engineering in future.

  11. Production of lipids and docosahexasaenoic acid (DHA) by a native Thraustochytrium strain

    DEFF Research Database (Denmark)

    Shene, Carolina; Leyton, Allison; Rubilar, Mónica

    2013-01-01

    /w. Under this growth condition lipids and DHA productivities were 50 and 23 mg/(L h), respectively. Practical applications: Consumption of long chain‐polyunsaturated fatty acids (LC‐PUFA) belonging to the omega‐3 family such as DHA has several positive effects on human and animal health. However, natural...... sources are restricted to cold‐water fish and their oils. Marine protists are also good candidates for the production of microbial DHA. For evaluating the potential of new strains the effect of the growth medium composition, growth conditions, and cultivation mode on DHA productivity has to be determined...

  12. Study of itaconic acid production by Aspergillus terrus MJL05 strain with different variable

    OpenAIRE

    Mariana Juy; Joaquín Orejas; María Ester Lucca

    2010-01-01

    Título en español: Estudio de la producción de ácido itacónico con Aspergillus terreus de la cepa MJL05 con diferentes variables Abstract Itaconic acid (IA) production by Aspergillus terreus MJL05 strain was investigated in submerged batch fermentation in a stirred bioreactor to determine the effect of varying the nitrogen, phosphorous and carbon sources concentrations in the production medium. Glycerol, a biodiesel by-product was reported as an efficient substrate to achieve high ita...

  13. Study of itaconic acid production by aspergillus terrus mjl05 strain with different variable

    OpenAIRE

    Juy, Mariana; Orejas, Joaquín; Lucca, María Ester

    2010-01-01

    Título en español: Estudio de la producción de ácido itacónico con Aspergillus terreus de la cepa MJL05 con diferentes variables Abstract Itaconic acid (IA) production by Aspergillus terreus MJL05 strain was investigated in submerged batch fermentation in a stirred bioreactor to determine the effect of varying the nitrogen, phosphorous and carbon sources concentrations in the production medium. Glycerol, a biodiesel by-product was reported as an efficient substrate to achieve high ita...

  14. Characterization of cellulose production by a Gluconacetobacter xylinus strain from Kombucha.

    Science.gov (United States)

    Nguyen, Vu Tuan; Flanagan, Bernadine; Gidley, Michael J; Dykes, Gary A

    2008-11-01

    The aims of this work were to characterize and improve cellulose production by a Gluconoacetobacter xylinus strain isolated from Kombucha and determine the purity and some structural features of the cellulose from this strain. Cellulose yield in tea medium with both black tea and green tea and in Hestrin and Schramm (HS) medium under both static and agitated cultures was compared. In the tea medium, the highest cellulose yield was obtained with green tea (approximately 0.20 g/L) rather than black tea (approximately 0.14 g/L). Yield in HS was higher (approximately 0.28 g/L) but did not differ between static and agitated incubation. (1)H-NMR and (13)C-NMR spectroscopy indicated that the cellulose is pure (free of acetan) and has high crystallinity, respectively. Cellulose yield was improved by changing the type and level of carbon and nitrogen source in the HS medium. A high yield of approximately 2.64 g/L was obtained with mannitol at 20 g/L and corn steep liquor at 40 g/L in combination. In the tea medium, tea at a level of 3 g/L gave the highest cellulose yield and the addition of 3 g/L of tea to the HS medium increased cellulose yield to 3.34 g/L. In conclusion, the G. xylinus strain from Kombucha had different cellulose-producing characteristics than previous strains isolated from fruit. Cellulose was produced in a pure form and showed high potential applicability. Our studies extensively characterized cellulose production from a G. xylinus strain from Kombucha for the first time, indicating both similarities and differences to strains from different sources.

  15. Strain improvement of Pichia kudriavzevii TY13 for raised phytase production and reduced phosphate repression.

    Science.gov (United States)

    Qvirist, Linnea; Vorontsov, Egor; Veide Vilg, Jenny; Andlid, Thomas

    2017-03-01

    In this work, we present the development and characterization of a strain of Pichia kudriavzevii (TY1322), with highly improved phytate-degrading capacity. The mutant strain TY1322 shows a biomass-specific phytate degradation of 1.26 mmol g -1  h -1 after 8 h of cultivation in a high-phosphate medium, which is about 8 times higher compared with the wild-type strain. Strain TY1322 was able to grow at low pH (pH 2), at high temperature (46°C) and in the presence of ox bile (2% w/v), indicating this strain's ability to survive passage through the gastrointestinal tract. The purified phytase showed two pH optima, at pH 3.5 and 5.5, and one temperature optimum at 55°C. The lower pH optimum of 3.5 matches the reported pH of the pig stomach, meaning that TY1322 and/or its phytase is highly suitable for use in feed production. Furthermore, P. kudriavzevii TY1322 tolerates ethanol up to 6% (v/v) and shows high osmotic stress tolerance. Owing to the phenotypic characteristics and non-genetically modified organisms nature of TY1322, this strain show great potential for future uses in (i) cereal fermentations for increased mineral bioavailability, and (ii) feed production to increase the phosphate bioavailability for monogastric animals to reduce the need for artificial phosphate fortification. © 2016 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

  16. Screening and Molecular Identification of New Microbial Strains for Production of Enzymes of Biotechnological Interest

    Directory of Open Access Journals (Sweden)

    Imen Ghazala

    Full Text Available ABSTRACT: This research focused on isolation, identification and characterization of new strains of fungi and bacteria, which were able to produce extracellular xylanase, mannanase, pectinase and α-amylase. Fungi isolates were identified on the basis of analyses of 18S gene sequencing and internal transcribed spacer region. The closest phylogenetic neighbors according to 18S gene sequence and ITS region data for the two isolates M1 and SE were Aspergillus fumigatus and Aspergillus sydowii, respectively. I4 was identified as Bacillus mojavensis on the basis of the 16S rRNA gene sequencing and biochemical properties. The enzyme production was evaluated by cultivating the isolated microorganisms in liquid-state bioprocess using wheat bran as carbon source. Two fungi (M1, and SE and one bacterium (I4 strains were found to be xylanase producer, and several were proven to be outstanding producers of microbial xylanase. The strains producing xylanase secreted variable amounts of starch-debranching enzymes and produced low level β-mannan-degrading enzyme systems. The bacterium strain was found to be capable of producing pectinolytic enzymes on wheat bran at high level. Some of the strains have good potential for use as sources of important industrial enzymes.

  17. Succinic acid production from xylose mother liquor by recombinant Escherichia coli strain.

    Science.gov (United States)

    Wang, Honghui; Pan, Jiachuan; Wang, Jing; Wang, Nan; Zhang, Jie; Li, Qiang; Wang, Dan; Zhou, Xiaohua

    2014-11-02

    Succinic acid (1,4-butanedioic acid) is identified as one of important building-block chemicals. Xylose mother liquor is an abundant industrial residue in xylitol biorefining industry. In this study, xylose mother liquor was utilized to produce succinic acid by recombinant Escherichia coli strain SD121, and the response surface methodology was used to optimize the fermentation media. The optimal conditions of succinic acid fermentation were as follows: 82.62 g L -1 total initial sugars, 42.27 g L -1 MgCO 3 and 17.84 g L -1 yeast extract. The maximum production of succinic acid was 52.09 ± 0.21 g L -1 after 84 h with a yield of 0.63 ± 0.03 g g -1 total sugar, approaching the predicted value (53.18 g L -1 ). It was 1.78-fold of the production of that obtained with the basic medium. This was the first report on succinic acid production from xylose mother liquor by recombinant E. coli strains with media optimization using response surface methodology. This work suggested that the xylose mother liquor could be an alternative substrate for the economical production of succinic acid by recombinant E. coli strains.

  18. Exopolysaccharide production by a marine Pseudoalteromonas sp. strain isolated from Madeira Archipelago ocean sediments.

    Science.gov (United States)

    Roca, Christophe; Lehmann, Mareen; Torres, Cristiana A V; Baptista, Sílvia; Gaudêncio, Susana P; Freitas, Filomena; Reis, Maria A M

    2016-06-25

    Exopolysaccharides (EPS) are polymers excreted by some microorganisms with interesting properties and used in many industrial applications. A new Pseudoalteromonas sp. strain, MD12-642, was isolated from marine sediments and cultivated in bioreactor in saline culture medium containing glucose as carbon source. Its ability to produce EPS under saline conditions was demonstrated reaching an EPS production of 4.4g/L within 17hours of cultivation, corresponding to a volumetric productivity of 0.25g/Lh, the highest value so far obtained for Pseudoalteromonas sp. strains. The compositional analysis of the EPS revealed the presence of galacturonic acid (41-42mol%), glucuronic acid (25-26mol%), rhamnose (16-22mol%) and glucosamine (12-16mol%) sugar residues. The polymer presents a high molecular weight (above 1000kDa). These results encourage the biotechnological exploitation of strain MD12-642 for the production of valuable EPS with unique composition, using saline by-products/wastes as feedstocks. Copyright © 2016 Elsevier B.V. All rights reserved.

  19. Phylogenetic analysis, fumonisin production and pathogenicity of Fusarium fujikuroi strains isolated from rice in the Philippines.

    Science.gov (United States)

    Cruz, Alejandra; Marín, Patricia; González-Jaén, M Teresa; Aguilar, Kristel Grace I; Cumagun, Christian Joseph R

    2013-09-01

    Fusarium fujikuroi Nirenberg is a maize and rice pathogen causing important agricultural losses and produces fumonisins - mycotoxins which pose health risk to humans and farm animals. However, little information is available about the phylogenetics of this species and its ability to produce fumonisins in rice. We studied 32 strains isolated from rice in the Philippines and performed a phylogenetic analysis using the partial sequence of Elongation Factor 1 alpha (EF-1α) including isolates belonging to closely related species. Fumonisin B1 (FB1 ) production was analyzed in 7-day-old cultures grown in fumonisin-inducing medium by an enzyme-linked immunosorbent assay-based method and by real-time reverse transcriptase-polymerase chain reaction using primers for FUM1 gene, a key gene in fumonisin biosynthesis. Nucleotide diversities per site (π) were 0.00024 ± 0.00022 (standard deviation) for the 32 F. fujikuroi strains from the Philippines and 0.00189 ± 0.00143 for all 34 F. fujikuroi strains, respectively. F. fujikuroi isolates grouped into one cluster separated from the rest of isolates belonging to the closely related F. proliferatum and showed very low variability, irrespective of their geographic origin. The cluster containing strains of F. proliferatum showed higher intraspecific variability than F. fujikuroi. Thirteen of the 32 strains analyzed were FB1 producers (40.62%), with production ranging from 0.386 to 223.83 ppm. All isolates analyzed showed FUM1 gene expression above 1 and higher than the CT value of the non-template control sample. Both seedling stunting and elongation were induced by the isolates in comparison with the control. F. fujikuroi are distinct from F. proliferatum isolates based on phytogenetic analysis and are potential fumonisin producers because all are positive for FUM1 gene expression. No relationship between fumonisin production and pathogenicity could be observed. © 2013 Society of Chemical Industry.

  20. Enhanced production of bacitracin by a mutant strain bacillus licheniformis UV-MN-HN-8 (enhanced bacitracin production by mutagenesis)

    International Nuclear Information System (INIS)

    Aftab, M.N.; Ikram-ul-Haq; Baig, S.

    2010-01-01

    The present study is focused on the improvement of Bacillus licheniformis through random mutagenesis to obtain mutant having enhanced production of bacitracin. Many isolates of Bacillus licheniformis were isolated and the isolate GP-40 produced maximum bacitracin production (16 +- 0.72 IU/mL). Treatment of Bacillus licheniformis GP-40 with ultraviolet (UV) radiations increased bacitracin production to 29 +- 0.69 IU/mL. Similarly, treatment of vegetative cells of GP-40 with chemicals like N-methyl N'-nitro N-nitroso guanidine (MNNG) and Nitrous acid (HNO/sub 2/) increased bacitracin production to 35 +- 1.35 IU/mL and 29 +- 0.89 IU/mL respectively. Studies regarding the combined effect of UV and chemical treatment on parental cells exhibited significantly higher titers of bacitracin with maximum bacitracin production reached to 47.6 +- 0.92 IU/mL. An increase of 2.97 fold production of bacitracin in comparison to wild type was observed. Mutant strain was highly stable and produced consistent yield of bacitracin even after 15 generations. On the basis of kinetic variables, notably mu (h-/sup 1/)max, Yp/x, qp, Qp and Qx mutant strain B. licheniformis UV-MN-HN-8 was found to be a hyper producer of bacitracin. (author)

  1. Bio-hydrogen production from glycerol by a strain of Enterobacter aerogenes

    Energy Technology Data Exchange (ETDEWEB)

    Marques, P.A.S.S; Bartolomeu, M.L.; Tome, M.M.; Rosa, M.F. [INETI, Unit of Biomass/Renewable Energy Department, Estrada do Paco do Lumiar, 22, 1649-038 Lisboa (Portugal)

    2008-07-01

    The goal of this work was to evaluate the H2 production from glycerol-containing byproducts obtained from biodiesel industrial production, using Enterobacter aerogenes ATCC 13048 Sputum. H2 production using as substrate pure glycerol and glycerol-containing biodiesel byproducts was compared. The effect of parameters such as initial substrate concentration and sodium chloride addition on the bio-hydrogen production efficiency was also investigated. The results showed that using 10 g/L of pure glycerol or biodiesel residues, containing the same concentration of glycerol as substrate, lead to similar bio-hydrogen productions (3.46 LH2/L and 3.28 LH2/L fermentation medium, respectively). This indicates that the performance of the E. aerogenes strain used was not influenced by the presence of other components than glycerol in biodiesel residues, at least for the tested waste concentration range. When sodium chloride was added to the fermentation medium with pure 10 g/L glycerol, H2 production was not affected (3.34 LH2/L fermentation medium), showing that metabolism of the E. aerogenes strain was not inhibited by this biodiesel waste component up to 4 g/L chloride concentration. Biodiesel residues used without sterilization provided a higher H2 production (1.03 L) than the ones submitted to previous sterilization in autoclave (0.89 L).

  2. Evaluation of autochthonous micrococcus strains as starter cultures for the production of Kedong sufu.

    Science.gov (United States)

    Feng, Z; Huang, S; Ai, Z W; Zhang, M; Zhai, S; Chen, X

    2016-03-01

    The technological properties of 22 micrococcus strains from traditional fermented Kedong sufu were evaluated in order to develop autochthonous starter cultures. The proteolytic, autolytic and lipolytic activity, salt tolerance, production and degradation of the biogenic amines of six Micrococcus luteus, nine Kocuria kristinae and seven Kocuria rosea were evaluated. The results indicated that these micrococcus strains exhibited a certain technological diversity, and the results also indicated the best properties to be used in mixed starter cultures. Based on the above findings, two sets of autochthonous starters were formulated. Considering the physicochemical properties and sensory characteristics of sufu, the maturation period of sufu was shortened by 30 days. The profiles of free amino acids and peptides partly revealed the mechanism of sensory quality and shorter ripening time of sufu manufactured using autochthonous mixed starters. Compared to back-slopping fermentation, sufu manufactured with selected autochthonous starter cultures exhibited lower levels of total biogenic amines. The selected strains could be used as starter to avoid the accumulation of high concentrations of biogenic amines while also maintaining typical sensory characteristics and preserving the autochthonous strains of the traditional Kedong sufu. The maturation times of Kedong sufu were shortened by 30 days with application of the autochthonous starter. Autochthonous mixed starters can reduce the generation of biogenic amines, speed up the sufu maturation process and preserve typical sensory quality. Furthermore, the rotation of two sets of mixed starter cultures can effectively resist phage attack during the production of sufu. © 2015 The Society for Applied Microbiology.

  3. New Strains Intended for the Production of Inactivated Polio Vaccine at Low-Containment After Eradication.

    Directory of Open Access Journals (Sweden)

    Sarah Knowlson

    2015-12-01

    Full Text Available Poliomyelitis has nearly been eradicated through the efforts of the World Health Organization's Global Eradication Initiative raising questions on containment of the virus after it has been eliminated in the wild. Most manufacture of inactivated polio vaccines currently requires the growth of large amounts of highly virulent poliovirus, and release from a production facility after eradication could be disastrous; WHO have therefore recommended the use of the attenuated Sabin strains for production as a safer option although it is recognised that they can revert to a transmissible paralytic form. We have exploited the understanding of the molecular virology of the Sabin vaccine strains to design viruses that are extremely genetically stable and hyperattenuated. The viruses are based on the type 3 Sabin vaccine strain and have been genetically modified in domain V of the 5' non-coding region by changing base pairs to produce a cassette into which capsid regions of other serotypes have been introduced. The viruses give satisfactory yields of antigenically and immunogenically correct viruses in culture, are without measurable neurovirulence and fail to infect non-human primates under conditions where the Sabin strains will do so.

  4. New Strains Intended for the Production of Inactivated Polio Vaccine at Low-Containment After Eradication.

    Science.gov (United States)

    Knowlson, Sarah; Burlison, John; Giles, Elaine; Fox, Helen; Macadam, Andrew J; Minor, Philip D

    2015-12-01

    Poliomyelitis has nearly been eradicated through the efforts of the World Health Organization's Global Eradication Initiative raising questions on containment of the virus after it has been eliminated in the wild. Most manufacture of inactivated polio vaccines currently requires the growth of large amounts of highly virulent poliovirus, and release from a production facility after eradication could be disastrous; WHO have therefore recommended the use of the attenuated Sabin strains for production as a safer option although it is recognised that they can revert to a transmissible paralytic form. We have exploited the understanding of the molecular virology of the Sabin vaccine strains to design viruses that are extremely genetically stable and hyperattenuated. The viruses are based on the type 3 Sabin vaccine strain and have been genetically modified in domain V of the 5' non-coding region by changing base pairs to produce a cassette into which capsid regions of other serotypes have been introduced. The viruses give satisfactory yields of antigenically and immunogenically correct viruses in culture, are without measurable neurovirulence and fail to infect non-human primates under conditions where the Sabin strains will do so.

  5. Potential of Rhodococcus strains for biotechnological vanillin production from ferulic acid and eugenol.

    Science.gov (United States)

    Plaggenborg, Rainer; Overhage, Jörg; Loos, Andrea; Archer, John A C; Lessard, Philip; Sinskey, Anthony J; Steinbüchel, Alexander; Priefert, Horst

    2006-10-01

    The potential of two Rhodococcus strains for biotechnological vanillin production from ferulic acid and eugenol was investigated. Genome sequence data of Rhodococcus sp. I24 suggested a coenzyme A-dependent, non-beta-oxidative pathway for ferulic acid bioconversion, which involves feruloyl-CoA synthetase (Fcs), enoyl-CoA hydratase/aldolase (Ech), and vanillin dehydrogenase (Vdh). This pathway was proven for Rhodococcus opacus PD630 by physiological characterization of knockout mutants. However, expression and functional characterization of corresponding structural genes from I24 suggested that degradation of ferulic acid in this strain proceeds via a beta-oxidative pathway. The vanillin precursor eugenol facilitated growth of I24 but not of PD630. Coniferyl aldehyde was an intermediate of eugenol degradation by I24. Since the genome sequence of I24 is devoid of eugenol hydroxylase homologous genes (ehyAB), eugenol bioconversion is most probably initiated by a new step in this bacterium. To establish eugenol bioconversion in PD630, the vanillyl alcohol oxidase gene (vaoA) from Penicillium simplicissimum CBS 170.90 was expressed in PD630 together with coniferyl alcohol dehydrogenase (calA) and coniferyl aldehyde dehydrogenase (calB) genes from Pseudomonas sp. HR199. The recombinant strain converted eugenol to ferulic acid. The obtained data suggest that genetically engineered strains of I24 and PD630 are suitable candidates for vanillin production from eugenol.

  6. Saccharomyces cerevisiae strains for second-generation ethanol production: from academic exploration to industrial implementation

    Science.gov (United States)

    Jansen, Mickel L. A.; Bracher, Jasmine M.; Papapetridis, Ioannis; Verhoeven, Maarten D.; de Bruijn, Hans; de Waal, Paul P.; van Maris, Antonius J. A.; Klaassen, Paul

    2017-01-01

    Abstract The recent start-up of several full-scale ‘second generation’ ethanol plants marks a major milestone in the development of Saccharomyces cerevisiae strains for fermentation of lignocellulosic hydrolysates of agricultural residues and energy crops. After a discussion of the challenges that these novel industrial contexts impose on yeast strains, this minireview describes key metabolic engineering strategies that have been developed to address these challenges. Additionally, it outlines how proof-of-concept studies, often developed in academic settings, can be used for the development of robust strain platforms that meet the requirements for industrial application. Fermentation performance of current engineered industrial S. cerevisiae strains is no longer a bottleneck in efforts to achieve the projected outputs of the first large-scale second-generation ethanol plants. Academic and industrial yeast research will continue to strengthen the economic value position of second-generation ethanol production by further improving fermentation kinetics, product yield and cellular robustness under process conditions. PMID:28899031

  7. BIOUTILIZATION OF GRAPE WASTE FOR EXOPOLYSACCHARIDE PRODUCTION USING ALTERNARIA ALTERNATA NON-PIGMENT STRAIN

    International Nuclear Information System (INIS)

    MELEIGY, S.A.

    2009-01-01

    In the present investigation, five mutant strains from A. alternate were isolated after exposure to gamma irradiation at dose level 8 kGy. The mutant isolated strains (MIS) were non-producing dark pigment and producing polysaccharide. The mutant isolated strain (MIS) belong to the Alternaria alternata MIS (1-5). In shake flask experiments, the exopolysaccharide (EPS) production was 2.90-5.24 g/l and biomass 5.8-8.31g/l. The effect of some fermentation conditions on grape wastes by A. alternata were investigated to produce EPS using gamma irradiation and non-pigment strain. The economical optimum fermentation condition for the highest EPS production by MIS4 when grown on grape waste containing 150 g/l sugar, incubation temperature 28 o C, pH 7 with addition of both 2 % yeast extract and 1.5 % surfactant triton x-100 achieved 14.68 g/l EPS and 6.22 g/l biomass

  8. Influence of Growth Medium on Hydrogen Peroxide and Bacteriocin Production of Lactobacillus Strains

    Directory of Open Access Journals (Sweden)

    Edina Németh

    2005-01-01

    Full Text Available This study was conducted to investigate the inhibitory effect of bacteriocin and the production of hydrogen peroxide by four non-starter lactic acid bacteria, Lactobacillus plantarum 2142, Lactobacillus curvatus 2770, Lactobacillus curvatus 2775, Lactobacillus casei subsp. pseudoplantarum 2750 and the probiotic strain Lactobacillus casei Shirota, propagated in de Man Rogosa Sharpe (MRS and tomato juice (TJ broth. The methods were a commonly used agar diffusion technique and a microtiter assay method. The best peroxide-producing Lactobacillus strain was selected for screening the inhibitory activity against Listeria monocytogenes, Bacillus cereus, Escherichia coli and the activity of bacteriocins against Lactobacillus sakei and Candida glabrata. All of the investigated lactic acid bacteria (LAB strains grown in MRS broth produced the highest concentration of hydrogen peroxide ranging from 2–6 g/mL after 72 h of storage. L. plantarum 2142 produced enough hydrogen peroxide already after 24 h at 5 °C in phosphate buffer to inhibit the growth of L. monocytogenes and B. cereus. Crude bacteriocin suspension from the investigated LAB inhibited only slightly the growth of L. sakei, however, the same suspension from MRS completely inhibited the 6-fold diluted yeast suspension. The concentrated bacteriocin suspensions from the both broths inhibited the growth of L. sakei completely. Among the strains, L. plantarum 2142 seemed to be the best peroxide and bacteriocin producer, and the antimicrobial metabolite production was better in MRS than in TJ broth.

  9. Dose effect of the uvsA+ gene product in duplication strains of Aspergillus nidulans

    International Nuclear Information System (INIS)

    Majerfeld, I.H.; Roper, J.A.

    1978-01-01

    Strains of Aspergillus nidulans which carry a particular segment of chromosome I in duplicate - one segment in normal position, the other translocated to chromosome II - are more resistant to uv light than are strains with a balanced haploid genome. A double dose of the uvsA + allele, carried on the duplicate segment, determines this enhanced resistance; this is shown by the descending order of resistance of duplication haploids uvsA + /uvsA + , uvsA1/uvsA + and uvsA1/uvsA1. An unbalanced diploid with three doses of the uvsA + allele also shows greater resistance than a balanced uvsA + //uvsA + diploid. However, in balanced diploids the uvsA1 allele appears to be completely recessive; uvsA + //uvsA + and uvsA + //uvsA1 diploids produce indistinguishable survival curves after uv irradiation. Thus, the uvsA + gene product is not rate-limiting in repair processes in strains with a balanced genome. The rate-limiting effect observed in these unbalanced strains presumably reflects an interaction of the uvsA + product and other functions determined by the rest of the genome. Duplication haploids and normal haploids lose photorepairable lesions at similar rates. This observation may be interpreted to indicate that differences in survival are not due to differences in the efficiency of excision of uv-induced pyrimidime dimers

  10. Progress toward isolation of strains and genetically engineered strains of microalgae for production of biofuel and other value added chemicals: A review

    International Nuclear Information System (INIS)

    Ghosh, Ashmita; Khanra, Saumyakanti; Mondal, Madhumanti; Halder, Gopinath; Tiwari, O.N.; Saini, Supreet; Bhowmick, Tridib Kumar; Gayen, Kalyan

    2016-01-01

    Highlights: • Sample collection, isolation and identification to obtain a pure microalgal species. • Isolation of microalgal strains worldwide based on continent and habitat. • Genetic engineering tools for enhanced production of biodiesel and value added chemicals. • Cultivation systems for genetically modified strain. - Abstract: Microalgae and cyanobacteria are promising sources of biodiesel because of their high oil content (∼10 fold higher) and shorter cultivation time (∼4 fold lesser) than conventional oil producing territorial plants (e.g., soybean, corn and jatropha). These organisms also provide source of several valuable natural chemicals including pigments, food supplements like eicosapentanoic acid [EPA], decosahexaenoic acid [DHA] and vitamins. In addition, many cellular components of these organisms are associated with therapeutic properties like antioxidant, anti-inflammatory, immunostimulating, and antiviral. Isolation and identification of high-yielding strains with the faster growth rate is the key for successful implementation of algal biodiesel (or other products) at a commercial level. A number of research groups in Europe, America, and Australia are thus extensively involved in exploration of novel microalgal strain. Further, genetic engineering provides a tool to engineer the native strain resulting in transgenic strain with higher yields. Despite these efforts, no consensus has yet been reached so far in zeroing on the best microalgal strain for sustainable production of biofuel at reasonable cost. The search for novel microalgal strain and transgenesis of microalgae, are continuing side by side with the hope of commercial scale production of microalgae biofuel in near future. However, no consolidated review report exists which guides to isolate and identify a uncontaminated microalgal strain along with their transgenesis. The present review is focused on: (i) key factors for sample collection, isolation, and identification to

  11. The genetic basis underlying variation in production of the flavour compound diacetyl by Lactobacillus rhamnosus strains in milk.

    Science.gov (United States)

    Lo, Raquel; Ho, Van Thi Thuy; Bansal, Nidhi; Turner, Mark S

    2018-01-16

    Diacetyl and the closely related compound acetoin impart desirable buttery flavour and odour to many foods including cheese and are generated through the metabolism of citrate by lactic acid bacteria (LAB). To increase the levels of these compounds, adjunct cultures capable of producing them can be added to cheese fermentations. In this study, we compared the diacetyl and acetoin producing abilities of 13 Lactobacillus rhamnosus strains from cheese sources. Diacetyl and acetoin production was found to be a common feature of Lb. rhamnosus grown in milk, with 12 strains producing these compounds. Whole genome sequencing of four strains revealed that genes encoding the citrate metabolising pathway present in other LAB are conserved in Lb. rhamnosus. One strain was, however, totally defective in diacetyl and acetoin production. This was likely due to an inability to produce the diacetyl/acetoin precursor compound acetolactate resulting from a frameshift mutation in the acetolactate synthase (als) gene. Complementation of this defective strain with a complete als gene from a diacetyl producing strain restored production of diacetyl and acetoin to levels equivalent to naturally high producing strains. Introduction of the same als-containing plasmid into the probiotic Lb. rhamnosus strain GG also increased diacetyl and acetoin levels. In model cheesemaking experiments, the als-complemented strain produced very high levels of diacetyl and acetoin over 35days of ripening. These findings identify the genetic basis for natural variation in production of a key cheese flavour compound in Lb. rhamnosus strains. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Biotechnological process for obtaining new fermented products from cashew apple fruit by Saccharomyces cerevisiae strains.

    Science.gov (United States)

    Araújo, Suzane Macêdo; Silva, Cristina Ferraz; Moreira, Jane Jesus Silveira; Narain, Narendra; Souza, Roberto Rodrigues

    2011-09-01

    In Brazil, the use of cashew apple (Anacardium occidentale L.) to obtain new products by biotechnological process represents an important alternative to avoid wastage of a large quantity of this fruit, which reaches about 85% of the annual production of 1 million tons. This work focuses on the development of an alcoholic product obtained by the fermentation of cashew apple juice. The inoculation with two different strains of yeast Saccharomyces cerevisiae viz. SCP and SCT, were standardized to a concentration of 10(7 )cells ml(-1). Each inoculum was added to 1,500 ml of cashew must. Fermentation was performed at 28 ± 3°C and aliquots were withdrawn every 24 h to monitor soluble sugar concentrations, pH, and dry matter contents. The volatile compounds in fermented products were analyzed using the gas chromatography/mass spectrometry (GC/MS) system. After 6 days, the fermentation process was completed, cells removed by filtration and centrifugation, and the products were stabilized under refrigeration for a period of 20 days. The stabilized products were stored in glass bottles and pasteurized at 60 ± 5°C/30 min. Both fermented products contained ethanol concentration above 6% (v v(-1)) while methanol was not detected and total acidity was below 90 mEq l(-1), representing a pH of 3.8-3.9. The volatile compounds were characterized by the presence of aldehyde (butyl aldehyde diethyl acetal, 2,4-dimethyl-hepta-2,4-dienal, and 2-methyl-2-pentenal) and ester (ethyl α-methylbutyrate) representing fruity aroma. The strain SCT was found to be better and efficient and this produced 10% more alcohol over that of strain SCP.

  13. Interaction of Wild Strains of Aspergilla with Aspergillus parasiticus ATCC15517 and Aflatoxin Production

    Science.gov (United States)

    Martins, H. Marina; Almeida, Inês; Marques, Marta; Bernardo, Fernando

    2008-01-01

    Aflatoxins are secondary metabolites produced by some competent mould strains of Aspergillus flavus, A. parasiticus and A. nomius. These compounds have been extensively studied with regards to their toxicity for animals and humans; they are able to induce liver cancer and may cause a wide range of adverse effects in living organisms. Aflatoxins are found as natural contaminants of food and feed; the main line of the strategy to control them is based on the prevention of the mould growth in raw vegetable or during its storage and monitoring of each crop batch. Mould growth is conditioned by many ecological factors, including biotic ones. Hazard characterization models for aflatoxins in crops must take into consideration biotic interactions between moulds and their potential effects on growth development. The aim of this work is to study the effect of the biotic interaction of 14 different wild strains of Aspergilla (different species), with a competent strain (Aspergillus parasiticus ATCC 15517) using an in vitro production model. The laboratory model used was a natural matrix (humidified cracked corn), on which each wild strain challenged the aflatoxin production of a producer strain. Cultures were incubated at 28°C for 12 days and sampled at the 8th and 12th. Aflatoxin detection and quantification was performed by HPLC using a procedure with a MRPL = 1 μg/kg. Results of those interactive cultures revealed both synergic and antagonistic effects on aflatoxin biosynthesis. Productivity increases were particularly evident on the 8th day of incubation with wild strains of A. flavipes (+ 70.4 %), A. versicolor (+ 54.9 %) and A. flavus 3 (+ 62.6 %). Antagonistic effects were found with A. niger (− 69.5%), A. fumigatus (− 47.6 %) and A. terreus (− 47.6 %) on the 12th day. The increased effects were more evident on the 8th of incubation and the decreases were more patent on the 12th day. Results show that the development of Aspergilla strains concomitantly with

  14. Production of an extracellular thermohalophilic lipase from a moderately halophilic bacterium, Salinivibrio sp. strain SA-2.

    Science.gov (United States)

    Amoozegar, Mohammad Ali; Salehghamari, Ensieh; Khajeh, Khosro; Kabiri, Mahbube; Naddaf, Saied

    2008-06-01

    Fifty strains of moderately halophilic bacteria were isolated from various salty environments in Iran. A strain designated as SA-2 was shown to be the best producer of extracellular lipase and was selected for further studies. Biochemical and physiological characterization along with 16S rDNA sequence analysis placed SA-2 in the genus Salinivibrio. The optimum salt, pH, temperature and aeration for enzyme production were 0.1 M KCl, pH 8, 35 degrees C and 150 rpm, respectively. The enzyme production was synchronized bacterial growth and reached a maximum level during the early-stationary phase in the basal medium containing 1 M NaCl. Triacylglycerols enhanced lipase production, while carbohydrates had inhibitory effects on it. The maximum lipase activity was obtained at pH 7.5, 50 degrees C and CaCl(2) concentration of 0.01 M. The enzyme was stable at pH range of 7.5-8 and retained 90% of its activity at 80 degrees C for 30 min. Different concentrations of NaNO(3), Na(2)SO(4), KCl and NaCl had no affect on lipase stability for 3 h. These results suggest that the lipase secreted by Salinivibrio sp. strain SA-2 is industrially important from the perspective of its tolerance to a broad temperature range, its moderate thermoactivity and its high tolerance to a wide range of salt concentrations (0-3 M NaCl).

  15. Strain screening, fermentation, separation, and encapsulation for production of nattokinase functional food.

    Science.gov (United States)

    Wei, Xuetuan; Luo, Mingfang; Xie, Yuchun; Yang, Liangrong; Li, Haojian; Xu, Lin; Liu, Huizhou

    2012-12-01

    This study presents a novel and integrated preparation technology for nattokinase functional food, including strain screening, fermentation, separation, and encapsulation. To rapidly screen a nattokinase-productive strain, PCR-based screening method was combined with fibrinolytic activity-based method, and a high productive strain, Bacillus subtilis LSSE-22, was isolated from Chinese soybean paste. Reduction of poly-γ-glutamic acid (γ-PGA) concentration may contribute to separation of nattokinase and reduction of late-onset anaphylaxis risk. Chickpeas were confirmed as the favorable substrate for enhancement of nattokinase production and reduction of γ-PGA yield. Using cracked chickpeas, the nattokinase activity reached 356.25 ± 17.18 FU/g (dry weight), which is much higher than previous reports. To further reduce γ-PGA concentration, ethanol fractional extraction and precipitation were applied for separation of nattokinase. By extraction with 50 % and precipitation with 75 % ethanol solution, 4,000.58 ± 192.98 FU/g of nattokinase powders were obtained, and the activity recovery reached 89 ± 1 %, while γ-PGA recovery was reduced to 21 ± 2 %. To improve the nattokinase stability at acidic pH condition, the nattokinase powders were encapsulated, and then coated with methacrylic acid-ethyl acrylate copolymer. After encapsulation, the nattokinase was protected from being denatured under various acid conditions, and pH-responsible controlled release at simulated intestinal fluid was realized.

  16. Potential Role of Yeast Strains Isolated from Grapes in the Production of Aglianico of Taurasi DOCG

    Directory of Open Access Journals (Sweden)

    Maria eAponte

    2016-05-01

    Full Text Available Twelve samples of Aglianico grapes, collected in different locations of the Taurasi DOCG (Appellation of Controlled and Guaranteed Origin production area were naturally fermented in sterile containers at room temperature. A total of 70 yeast cultures were isolated from countable WL agar plates: 52 in the middle of the fermentation and 18 at the end. On the basis of ITS-RFLP analysis and ITS sequencing, all cultures collected at the end of fermentations were identified as Saccharomyces (S. cerevisiae; while, the 52 isolates, collected after one week, could be referred to the following species: Metschnikowia (M. pulcherrima; Starmerella (Star. bacillaris; Pichia (P. kudriavzevii; Lachancea (L. thermotolerans; Hanseniaspora (H. uvarum; Pseudozyma (Pseud. aphidis; S. cerevisiae. By means of Interdelta analysis, 18 different biotypes of S. cerevisiae were retrieved. All strains were characterized for ethanol production, SO2 resistance, H2S development, β-glucosidasic, esterasic and antagonistic activities. Fermentation abilities of selected strains were evaluated in micro-fermentations on Aglianico must. Within non-Saccharomyces species, some cultures showed features of technological interest. Antagonistic activity was expressed by some strains of M. pulcherrima, L. thermotolerans, P. kudriavzevii and S. cerevisiae. Strains of M. pulcherrima showed the highest β-glucosidase activity and proved to be able to produce high concentrations of succinic acid. L. thermotolerans produced both succinic and lactic acids. The lowest amount of acetic acid was produced by M. pulcherrima and L. thermotolerans; while the highest content was recorded for H. uvarum. The strain of Star. bacillaris produced the highest amount of glycerol and was able to metabolize all fructose and malic acid. Strains of M. pulcherrima and H. uvarum showed a low fermentation power (about 4%, while, L. thermotolerans, Star. bacillaris and P. kudriavzevii of about 10%. Significant

  17. Evaluation of commercial soy sauce koji strains of Aspergillus oryzae for γ-aminobutyric acid (GABA) production.

    Science.gov (United States)

    Ab Kadir, Safuan; Wan-Mohtar, Wan Abd Al Qadr Imad; Mohammad, Rosfarizan; Abdul Halim Lim, Sarina; Sabo Mohammed, Abdulkarim; Saari, Nazamid

    2016-10-01

    In this study, four selected commercial strains of Aspergillus oryzae were collected from soy sauce koji. These A. oryzae strains designated as NSK, NSZ, NSJ and NST shared similar morphological characteristics with the reference strain (A. oryzae FRR 1675) which confirmed them as A. oryzae species. They were further evaluated for their ability to produce γ-aminobutyric acid (GABA) by cultivating the spore suspension in a broth medium containing 0.4 % (w/v) of glutamic acid as a substrate for GABA production. The results showed that these strains were capable of producing GABA; however, the concentrations differed significantly (P sauce production.

  18. Dynamics of plc gene transcription and α-toxin production during growth of Clostridium perfringens strains with contrasting α-toxin production

    DEFF Research Database (Denmark)

    Abildgaard, Lone; Schramm, Andreas; Rudi, Knut

    2009-01-01

    The aim of the present study was to investigate transcription dynamics of the α-toxin-encoding plc gene relative to two housekeeping genes (gyrA and rplL) in batch cultures of three Clostridium perfringens strains with low, intermediate, and high levels of α-toxin production, respectively. The plc...... transcript level was always low in the low α-toxin producing strain. For the two other strains, plc transcription showed an inducible pattern and reached a maximum level in the late exponential growth phase. The transcription levels were however inversely correlated to α-toxin production for the two strains....... We propose that this discrepancy is due to differences in plc translation rates between the strains and that strain-specific translational rates therefore must be determined before α-toxin production can be extrapolated from transcript levels in C. perfringens....

  19. Biotechnological Screening of Microalgal and Cyanobacterial Strains for Biogas Production and Antibacterial and Antifungal Effects

    Directory of Open Access Journals (Sweden)

    Opayi Mudimu

    2014-05-01

    Full Text Available Microalgae and cyanobacteria represent a valuable natural resource for the generation of a large variety of chemical substances that are of interest for medical research, can be used as additives in cosmetics and food production, or as an energy source in biogas plants. The variety of potential agents and the use of microalgae and cyanobacteria biomass for the production of these substances are little investigated and not exploited for the market. Due to the enormous biodiversity of microalgae and cyanobacteria, they hold great promise for novel products. In this study, we investigated a large number of microalgal and cyanobacterial strains from the Culture Collection of Algae at Göttingen University (SAG with regard to their biomass and biogas production, as well antibacterial and antifungal effects. Our results demonstrated that microalgae and cyanobacteria are able to generate a large number of economically-interesting substances in different quantities dependent on strain type. The distribution and quantity of some of these components were found to reflect phylogenetic relationships at the level of classes. In addition, between closely related species and even among multiple isolates of the same species, the productivity may be rather variable.

  20. Factors influencing production of lipase under metal supplementation by bacterial strain, Bacillus subtilis BDG-8.

    Science.gov (United States)

    Dhevahi, B; Gurusamy, R

    2014-11-01

    Lipases are biocatalyst having wide applications in industries due to their versatile properties. In the present study, a lipolytic bacterial strain, Bacillus subtilis BDG-8 was isolated from an oil based industrial soil. The effect of selenium and nickel as a media supplement on enhancement of lipase production, was studied individually with the isolated strain by varying the concentration of selected metal. 60 μg l(-1) selenium enhanced lipase production to an enzyme activity measuring 7.8 U ml(-1) while 40 μgI(-1) nickel gave the maximum enzyme activity equivalent to 7.5 U ml(-1). However, nickel and selenium together at a range of concentration with an equal w/v ratio, at 60 μg l(-1) each, showed the maximum lipase activity of 8.5 U ml(-1). The effect of pH and temperature on lipase production showed maximum enzyme activity in the presence of each of the metals at pH 7 and 35°C among the other tested ranges. After optimisation of the parameters such as metal concentration, pH and temperature lipase production by Bacillus subtilis BDG-8 had increased several folds. This preliminary investigation may consequently lead as to various industrial applications such as treatment of wastewater contaminated with metal or oil with simultaneous lipase production.

  1. Mini review: Recombinant production of tailored bio-pharmaceuticals in different Bacillus strains and future perspectives.

    Science.gov (United States)

    Lakowitz, Antonia; Godard, Thibault; Biedendieck, Rebekka; Krull, Rainer

    2018-05-01

    Bio-pharmaceuticals like antibodies, hormones and growth factors represent about one-fifth of commercial pharmaceuticals. Host candidates of growing interest for recombinant production of these proteins are strains of the genus Bacillus, long being established for biotechnological production of homologous and heterologous proteins. Bacillus strains benefit from development of efficient expression systems in the last decades and emerge as major industrial workhorses for recombinant proteins due to easy cultivation, non-pathogenicity and their ability to secrete recombinant proteins directly into extracellular medium allowing cost-effective downstream processing. Their broad product portfolio of pharmaceutically relevant recombinant proteins described in research include antibody fragments, growth factors, interferons and interleukins, insulin, penicillin G acylase, streptavidin and different kinases produced in various cultivation systems like microtiter plates, shake flasks and bioreactor systems in batch, fed-batch and continuous mode. To further improve production and secretion performance of Bacillus, bottlenecks and limiting factors concerning proteases, chaperones, secretion machinery or feedback mechanisms can be identified on different cell levels from genomics and transcriptomics via proteomics to metabolomics and fluxomics. For systematical identification of recurring patterns characteristic of given regulatory systems and key genetic targets, systems biology and omics-technology provide suitable and promising approaches, pushing Bacillus further towards industrial application for recombinant pharmaceutical protein production. Copyright © 2017. Published by Elsevier B.V.

  2. Extracellular Phytase Production by the Wine Yeast S. cerevisiae (Finarome Strain) during Submerged Fermentation.

    Science.gov (United States)

    Kłosowski, Grzegorz; Mikulski, Dawid; Jankowiak, Oliwia

    2018-04-08

    One of the key steps in the production of phytases of microbial origin is selection of culture parameters, followed by isolation of the enzyme and evaluation of its catalytic activity. It was found that conditions for S. cerevisiae yeast culture, strain Finarome, giving the reduction in phytic acid concentration of more than 98% within 24 h of incubation were as follows: pH 5.5, 32 °C, continuous stirring at 80 rpm, the use of mannose as a carbon source and aspartic acid as a source of nitrogen. The highest catalytic activity of the isolated phytase was observed at 37 °C, pH 4.0 and using phytate as substrate at concentration of 5.0 mM. The presence of ethanol in the medium at a concentration of 12% v / v reduces the catalytic activity to above 60%. Properties of phytase derived from S. cerevisiae yeast culture, strain Finarome, indicate the possibility of its application in the form of a cell's free crude protein isolate for the hydrolysis of phytic acid to improve the efficiency of alcoholic fermentation processes. Our results also suggest a possibility to use the strain under study to obtain a fusant derived with specialized distillery strains, capable of carrying out a highly efficient fermentation process combined with the utilization of phytates.

  3. Production of anatoxin-a by cyanobacterial strains isolated from Portuguese fresh water systems.

    Science.gov (United States)

    Osswald, Joana; Rellán, Sandra; Gago-Martinez, Ana; Vasconcelos, Vítor

    2009-11-01

    The occurrence of anatoxin-a in several freshwater systems in Portugal and its production by Portuguese cyanobacterial strains, after cultivation in laboratory, were studied. Surface water samples from 9 water bodies, for recreational and human consumption usage, were surveyed for anatoxin-a presence and for obtaining cultures of pure cyanobacterial strains. Anatoxin-a analysis was performed by high performance liquid chromatography (HPLC) with fluorescence detection (FLD) followed by Mass Spectrometry (MS) confirmation. No anatoxin-a was detected in all the natural water samples (limit of detection (LOD) = 25 ng l(-1)) but among the 22 isolated cyanobacterial strains, 13 could produce anatoxin-a in laboratory conditions (LOD = 3 ng g(-1) dw). This proportion of anatoxin-a producing strains (59.1%) in laboratory is discussed considering the hypothesis that anatoxin-a is a more frequent metabolite in cyanobacteria than it was thought before and making its occurrence in Portuguese freshwaters almost certain. Therefore, health and ecological risks caused by anatoxin-a in Portugal, should be seriously considered.

  4. Mutagenesis of Xanthomonas campestris and selection of strains with enhanced Xanthan production

    International Nuclear Information System (INIS)

    Kamal, F.; Mehrgan, H.; Mazaheri, M.; Mortazavi, A. R.

    2003-01-01

    Xanthan gum is microbial polysaccharide of great commercial importance as it has been unusual rheological properties in solution and consequent range of applications. In this study, a series of mutants were isolated from Xanthomonas PTSS 1473 by ethyl methanesulfonate mutagenesis. The polysaccharide yield of one mutant, XC1473E 2 , was 30% better than that of the parent strain. It also showed higher xanthan formation of glucose consumption rates compared to the parent strain. xanthan produced by the mutant and enhanced viscosity, higher pseudo plasticity and larger molecular weight. Since mutant XC1473E 2 appeared white on agar plates, it underwent pigment extraction with methanol. Contrary to the parent strain, the mutant showed no absorption at 443 nm, i.e. the wavelength related to yellow pigment. This finding suggested that yellow pigmentation and normal xanthan biosynthesis are not necessarily concurrent. In general, mutant ZC1473e 2 seems to be a strain with interesting characteristics for use in commercial production of Xanthan

  5. Improvement of xylanase production by a parasexual cross between Aspergillus niger strains

    Directory of Open Access Journals (Sweden)

    Octavio Loera

    2003-03-01

    Full Text Available A diploid strain (D4 isolated via parasexual recombination between two Aspergillus niger xylanase overproducing mutants was characterised in terms of enzyme production and catabolite repression by glucose. This strain increased xylanase production (607 nkat/ml, which was nearly 100% higher than titers achieved by the wild type strain (305 nkat/ml and 28% higher than the best mutant used to induce parasexual cycle. Diploid D4 was also less sensitive to carbon catabolite repression by glucose, since xylanolytic activity was detected under conditions normally repressing production by the wild type strain. No decrease in maximal xylanase levels was observed in the presence of glucose for diploid D4.Um cepa diplóide (D4 isolada por combinação parasexual entre dois Aspergillus niger, mutantes superprodutores de xylanase foi caracterizado através da produção de (607 nkat/ml e repressão catabólica por glicose. Essa cepa aumenta a produção de xylanase em mais de 100% em comparação com uma cepa selvagem (305 nkat/ml e 28% superior do que o melhor mutante usado para induzir o ciclo parasexual. A cepa diplóide D4 foi também menos sensível a repressão catabólica pela glicose, sendo que a atividade xylanolitica foi detectada sob condições normalmente de produção repressiva pela cepa selvagem. Não foi observado um decréscimo na produção máxima de xylanase em presença de glicose para o diplóide D4.

  6. [Construction of high-yield strain by optimizing lycopene cyclase for β-carotene production].

    Science.gov (United States)

    Jin, Yingfu; Han, Li; Zhang, Shasha; Li, Shizhong; Liu, Weifeng; Tao, Yong

    2017-11-25

    To optimize key enzymes, such as to explore the gene resources and to modify the expression level, can maximize metabolic pathways of target products. β-carotene is a terpenoid compound with important application value. Lycopene cyclase (CrtY) is the key enzyme in β-carotene biosynthesis pathway, catalyzing flavin adenine dinucleotide (FAD)-dependent cyclization reaction and β-carotene synthesis from lycopene precursor. We optimized lycopene cyclase (CrtY) to improve the synthesis of β-carotene and determined the effect of CrtY expression on metabolic pathways. Frist, we developed a β-carotene synthesis module by coexpressing the lycopene β-cyclase gene crtY with crtEBI module in Escherichia coli. Then we simultaneously optimized the ribosome-binding site (RBS) intensity and the species of crtY using oligo-linker mediated DNA assembly method (OLMA). Five strains with high β-carotene production capacity were screened out from the OLMA library. The β-carotene yields of these strains were up to 15.79-18.90 mg/g DCW (Dry cell weight), 65% higher than that of the original strain at shake flask level. The optimal strain CP12 was further identified and evaluated for β-carotene production at 5 L fermentation level. After process optimization, the final β-carotene yield could reach to 1.9 g/L. The results of RBS strength and metabolic intermediate analysis indicated that an appropriate expression level of CrtY could be beneficial for the function of the β-carotene synthesis module. The results of this study provide important insight into the optimization of β-carotene synthesis pathway in metabolic engineering.

  7. Cellulase production by two mutant strain of Trichoderma longibranchiatum QM 9414 and Rut C30

    International Nuclear Information System (INIS)

    Blanco, M. J.

    1991-01-01

    Native or pretreated biomass from Onopordum nervosum Boiss, has been examined as candidate feedstock for cellulase production by two mutant strain of Trichoderma Ionqibrachiatum QM9414 and Rut C30. Batch cultivation methods were evaluated and compared with previous experiments using ball-milled, crystalline cellulose (Solka floc). Batch cultivation of T. Ionqibrachiatum Rut C30 on 5% (w/v) acid pretreated O. nervosum biomass yielded enzyme productivities and activities comparable to those obtained on Solka floc. However, the overall enzyme production performance was lower than on Solka floc at comparable cellulose concentrations. This fact may be due to the accumulation of pretreated by products and lignin in the ferment. (Author) 40 refs

  8. Effects of UV-B radiation on microcystin production of a toxic strain of Microcystis aeruginosa and its competitiveness against a non-toxic strain

    International Nuclear Information System (INIS)

    Yang, Zhen; Kong, Fanxiang; Shi, Xiaoli; Yu, Yang; Zhang, Min

    2015-01-01

    Highlights: • UV-B radiation showed higher inhibition to non-toxin producing than toxin-producing strains on growth and photosynthetic activity. • Both intracellular and extracellular MC contents decreased markedly under UV-B radiation. • Higher resistance to UV-B radiation helped toxin-producing M. aeruginosa to predominate in the competition. - Abstract: Microcystins (MCs) produced by toxic cyanobacteria pose a health hazard to humans and animals. Some environmental factors can alter the MC concentrations by affecting the abundance of toxin-producing strains in a cyanobacteria population and/or their toxin production. In this study, we designed a monoculture and competition experiment to investigate the impacts of UV-B radiation on MC production and the competition between toxin and non-toxin producing strains of Microcystis aeruginosa. UV-B radiation resulted in higher inhibition of the growth and photosynthetic activity of the non-toxin producing strain relative to that observed for the toxin-producing strain. Both intracellular and extracellular MC contents decreased markedly when the toxin-producing strain was exposed to UV-B radiation. In addition, a quantitative real-time PCR assay revealed that the ratio of toxin-producing M. aeruginosa under UV-B exposure was higher than that under PAR alone at an early stage of the experiment. However, its abundance under UV-B exposure was lower compared with the PAR alone treatment after day 12. Our study demonstrated that UV-B radiation has a great impact on the abundance of the toxin-producing strain in the Microcystis population and their toxin production, which suggests that the fluctuation of UV-B radiation affects the MC level of cyanobacteria blooms

  9. Effects of UV-B radiation on microcystin production of a toxic strain of Microcystis aeruginosa and its competitiveness against a non-toxic strain

    Energy Technology Data Exchange (ETDEWEB)

    Yang, Zhen, E-mail: zhyang@niglas.ac.cn; Kong, Fanxiang, E-mail: fxkong@niglas.ac.cn; Shi, Xiaoli; Yu, Yang; Zhang, Min

    2015-02-11

    Highlights: • UV-B radiation showed higher inhibition to non-toxin producing than toxin-producing strains on growth and photosynthetic activity. • Both intracellular and extracellular MC contents decreased markedly under UV-B radiation. • Higher resistance to UV-B radiation helped toxin-producing M. aeruginosa to predominate in the competition. - Abstract: Microcystins (MCs) produced by toxic cyanobacteria pose a health hazard to humans and animals. Some environmental factors can alter the MC concentrations by affecting the abundance of toxin-producing strains in a cyanobacteria population and/or their toxin production. In this study, we designed a monoculture and competition experiment to investigate the impacts of UV-B radiation on MC production and the competition between toxin and non-toxin producing strains of Microcystis aeruginosa. UV-B radiation resulted in higher inhibition of the growth and photosynthetic activity of the non-toxin producing strain relative to that observed for the toxin-producing strain. Both intracellular and extracellular MC contents decreased markedly when the toxin-producing strain was exposed to UV-B radiation. In addition, a quantitative real-time PCR assay revealed that the ratio of toxin-producing M. aeruginosa under UV-B exposure was higher than that under PAR alone at an early stage of the experiment. However, its abundance under UV-B exposure was lower compared with the PAR alone treatment after day 12. Our study demonstrated that UV-B radiation has a great impact on the abundance of the toxin-producing strain in the Microcystis population and their toxin production, which suggests that the fluctuation of UV-B radiation affects the MC level of cyanobacteria blooms.

  10. Screening local Lactobacilli from Iran in terms of production of lactic acid and identification of superior strains

    Directory of Open Access Journals (Sweden)

    Fatemeh Soleimanifard

    2015-12-01

    Full Text Available Introduction: Lactobacilli are a group of lactic acid bacteria that their final product of fermentation is lactic acid. The objective of this research is selection of local Lactobacilli producing L (+ lactic acid. Materials and methods: In this research the local strains were screened based on the ability to produce lactic acid. The screening was performed in two stages. The first stage was the titration method and the second stage was the enzymatic method. The superior strains obtained from titration method were selected to do enzymatic test. Finally, the superior strains in the second stage (enzymatic which had the ability to produce L(+ lactic acid were identified by biochemical tests. Then, molecular identification of strains was performed by using 16S rRNA sequencing. Results: In this study, the ability of 79 strains of local Lactobacilli in terms of production of lactic acid was studied. The highest and lowest rates of lactic acid production was 34.8 and 12.4 mg/g. Superior Lactobacilli in terms of production of lactic acid ability of producing had an optical isomer L(+, the highest levels of L(+ lactic acid were with 3.99 and the lowest amount equal to 1.03 mg/g. The biochemical and molecular identification of superior strains showed that strains are Lactobacillus paracasei. Then the sequences of 16S rRNA of superior strains were reported in NCBI with accession numbers KF735654، KF735655، KJ508201and KJ508202. Discussion and conclusion: The amounts of lactic acid production by local Lactobacilli were very different and producing some of these strains on available reports showed more products. The results of this research suggest the use of superior strains of Lactobacilli for production of pure L(+ lactic acid.

  11. Glycerol production by Oenococcus oeni during sequential and simultaneous cultures with wine yeast strains.

    Science.gov (United States)

    Ale, Cesar E; Farías, Marta E; Strasser de Saad, Ana M; Pasteris, Sergio E

    2014-07-01

    Growth and fermentation patterns of Saccharomyces cerevisiae, Kloeckera apiculata, and Oenococcus oeni strains cultured in grape juice medium were studied. In pure, sequential and simultaneous cultures, the strains reached the stationary growth phase between 2 and 3 days. Pure and mixed K. apiculata and S. cerevisiae cultures used mainly glucose, producing ethanol, organic acids, and 4.0 and 0.1 mM glycerol, respectively. In sequential cultures, O. oeni achieved about 1 log unit at 3 days using mainly fructose and L-malic acid. Highest sugars consumption was detected in K. apiculata supernatants, lactic acid being the major end-product. 8.0 mM glycerol was found in 6-day culture supernatants. In simultaneous cultures, total sugars and L-malic acid were used at 3 days and 98% of ethanol and glycerol were detected. This study represents the first report of the population dynamics and metabolic behavior of yeasts and O. oeni in sequential and simultaneous cultures and contributes to the selection of indigenous strains to design starter cultures for winemaking, also considering the inclusion of K. apiculata. The sequential inoculation of yeasts and O. oeni would enhance glycerol production, which confers desirable organoleptic characteristics to wines, while organic acids levels would not affect their sensory profile. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Biosurfactants production potential of native strains of Bacillus cereus and their antimicrobial, cytotoxic and antioxidant activities.

    Science.gov (United States)

    Basit, Madiha; Rasool, Muhammad Hidayat; Naqvi, Syed Ali Raza; Waseem, Muhammad; Aslam, Bilal

    2018-01-01

    Present study was designed to evaluate the biosurfactant production potential by native strains of Bacillus cereus as well as determine their antimicrobial and antioxidant activities. The strains isolated from garden soil were characterized as B. cereus MMIC 1, MMIC 2 and MMIC 3. Biosurfactants were extracted as grey white precipitates. Optimum conditions for biosurfactant production were 37°C, the 7th day of incubation, 0.5% NaCl, pH 7.0. Moreover, corn steep liquor was the best carbon source. Biuret test, Thin Layer Chromatography (TLC), agar double diffusion and Fourier Transform Infrared Spectroscopy (FTIR) characterized the biosurfactants as cationic lipopeptides. Biosurfactants exhibited significant antibacterial and antifungal activity against S. aureus, E. coli, P. aeruginosa, K. pneumoniae, A. niger and C. albicans at 30 mg/ml. Moreover, they also possessed antiviral activity against NDV at 10 mg/ml. Cytotoxicity assay in BHK-21 cell lines revealed 63% cell survival at 10 mg/ml of biosurfactants and thus considered as safe. They also showed very good antioxidant activity by ferric-reducing activity and DPPH scavenging activity at 2 mg/ml. Consequently, the study offers an insight for the exploration of new bioactive molecules from the soil. It was concluded that lipopeptide biosurfactants produced from native strains of B. cereus may be recommended as safe antimicrobial, emulsifier and antioxidant agent.

  13. Effect of Different Carbon Sources on Biosurfactants' Production by Three Strains of Lactobacillus spp.

    Science.gov (United States)

    Mouafo, Tene Hippolyte; Mbawala, Augustin; Ndjouenkeu, Robert

    2018-01-01

    The potential of three indigenous bacterial strains ( Lactobacillus delbrueckii N2, Lactobacillus cellobiosus TM1, and Lactobacillus plantarum G88) for the production of biosurfactants using sugar cane molasses or glycerol as substrates was investigated through emulsifying, surface tension, and antimicrobial activities. The different biosurfactants produced with molasses as substrate exhibited high surface tension reduction from 72 mN/m to values ranged from 47.50 ± 1.78 to 41.90 ± 0.79 mN/m and high emulsification index ranging from 49.89 ± 5.28 to 81.00 ± 1.14%. Whatever the Lactobacillus strain or the substrate used, the biosurfactants produced showed antimicrobial activities against Candida albicans LV1, some pathogenic and/or spoilage Gram-positive and Gram-negative bacteria. The yields of biosurfactants with molasses (2.43 ± 0.09 to 3.03 ± 0.09 g/L) or glycerol (2.32 ± 0.19 to 2.82 ± 0.05 g/L) were significantly ( p biosurfactants reveals that they are mainly glycoproteins and glycolipids with molasses and glycerol as substrate, respectively. Therefore, sugar cane molasses or glycerol can effectively be used by Lactobacillus strains as low-cost substrates to increase their biosurfactants production.

  14. Growth and microcystin production of a Brazilian Microcystis aeruginosa strain (LTPNA 02 under different nutrient conditions

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    Stella Bortoli

    Full Text Available Cyanobacteria are prokaryotic and photosynthetic organisms, which can produce a wide range of bioactive compounds with different properties; including a variety of toxic compounds, also known as cyanotoxins. In this work, we describe the isolation of seven cyanobacterial strains from two reservoirs in São Paulo State, Brazil. Seven different chemical variants of microcystins (MC-RR, MC-LR, MC-YR, MC-LF, MC-LW, and two demethylated variants, dm-MC-RR and dm-MC-LR were detected in three of the ten isolated strains. One particular Microcystis aeruginosa strain (LTPNA 02 was chosen to evaluate its growth by cell count, and its toxin production under seven different nutritional regimes. We observed different growth behaviors in the logarithmic growth period for only three experiments (p < 0.05. The total growth analysis identified four experiments as different from the control (p < 0.01. Three microcystin variants (MC-RR, MC-LR and MC-YR were quantified by liquid chromatography-tandem mass spectrometry. At the experimental end, the toxin content was unchanged when comparing cell growth in ASM-1 (N:P = 1, MLA and BG-11 (N:P = 10 medium. In all other experiments, the lowest microcystin production was observed from cells grown in Bold 3N medium during the exponential growth phase. The highest microcystin content was observed in cultures using BG-11(N:P = 100 medium.

  15. Matrix Production, Pigment Synthesis, and Sporulation in a Marine Isolated Strain of Bacillus pumilus.

    Science.gov (United States)

    Di Luccia, Blanda; Riccio, Antonio; Vanacore, Adele; Baccigalupi, Loredana; Molinaro, Antonio; Ricca, Ezio

    2015-10-21

    The ability to produce an extracellular matrix and form multicellular communities is an adaptive behavior shared by many bacteria. In Bacillus subtilis, the model system for spore-forming bacteria, matrix production is one of the possible differentiation pathways that a cell can follow when vegetative growth is no longer feasible. While in B. subtilis the genetic system controlling matrix production has been studied in detail, it is still unclear whether other spore formers utilize similar mechanisms. We report that SF214, a pigmented strain of Bacillus pumilus isolated from the marine environment, can produce an extracellular matrix relying on orthologs of many of the genes known to be important for matrix synthesis in B. subtilis. We also report a characterization of the carbohydrates forming the extracellular matrix of strain SF214. The isolation and characterization of mutants altered in matrix synthesis, pigmentation, and spore formation suggest that in strain SF214 the three processes are strictly interconnected and regulated by a common molecular mechanism.

  16. Phenotypic and Genotypic Characterization of Staphylococcus aureus Strains from Italian Dairy Products

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    Stefano Morandi

    2009-01-01

    Full Text Available Staphylococcus aureus is a known major cause of foodborne illnesses, and milk and dairy products are often contaminated by enterotoxigenic strains of this bacterium. In the present study, 122 S. aureus isolates collected from different dairy products were characterised by phenotypic properties, by the distribution of genes encoding staphylococcal enterotoxins (sea, sec, sed, seg, seh, sei, sej, and sel and by randomly amplified polymorphic DNA PCR (RAPD-PCR. Moreover, strain resistance to vancomycin and methicillin (oxacillin was studied. The differences in the RAPD-PCR profiles obtained with the primers M13 and AP4 revealed the presence of a great genetic heterogeneity among the different S. aureus strains. Using the primer AP4 and M13, eight groups were distinguished by RAPD-PCR cluster analysis, although, except in few cases, it was not possible to correlate the isolates of different animal species (cow or ovine with the presence of se genes. None of the isolates showed resistance to vancomycin or methicillin.

  17. Molecular characterization, fitness and mycotoxin production of Fusarium graminearum laboratory strains resistant to benzimidazoles.

    Science.gov (United States)

    Sevastos, A; Markoglou, A; Labrou, N E; Flouri, F; Malandrakis, A

    2016-03-01

    Six benzimidazole (BMZ)-resistant Fusarium graminearum strains were obtained after UV mutagenesis and selection on carbendazim (MBC)-amended medium. In vitro bioassays resulted in the identification of two resistant phenotypes that were highly HR (Rf: 40-170, based on EC50) and moderately MR (Rf: 10-20) resistant to carbendazim. Cross resistance studies with other fungicides showed that all mutant strains tested were also resistant to other BMZs, such as benomyl and thiabendazole, but retained their parental sensitivity to fungicides belonging to other chemical groups. A point mutation at codon 6 (His6Asn) was found in the β2-tubulin gene of MR isolates while another mutation at codon 200 (Phe200Tyr) was present in one MR and one HR isolates. Interestingly, low temperatures suppressed MBC-resistance in all isolates bearing the H6N mutation. The three-dimensional homology model of the wild-type and mutants of β-tubulins were constructed, and the possible carbendazim binding site was analyzed. Studies on fitness parameters showed that the mutation(s) for resistance to BMZs did not affect the mycelial growth rate whereas adverse effects were found in sporulation and conidial germination in most of the resistant mutants. Pathogenicity tests on corn cobs revealed that mutants were less or equally aggressive to the wild-type strain but expressed their BMZ-resistance after inoculation on maize cobs treated with MBC. Analysis of mycotoxin production by high performance liquid chromatography revealed that only two HR strains produced zearalenone (ZEA) at concentrations similar to that of the wild-type strain, while no ZEA levels were detected in the rest of the mutants. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Production of lactic acid from sucrose: strain selection, fermentation, and kinetic modeling.

    Science.gov (United States)

    Lunelli, Betânia H; Andrade, Rafael R; Atala, Daniel I P; Wolf Maciel, Maria Regina; Maugeri Filho, Francisco; Maciel Filho, Rubens

    2010-05-01

    Lactic acid is an important product arising from the anaerobic fermentation of sugars. It is used in the pharmaceutical, cosmetic, chemical, and food industries as well as for biodegradable polymer and green solvent production. In this work, several bacterial strains were isolated from industrial ethanol fermentation, and the most efficient strain for lactic acid production was selected. The fermentation was conducted in a batch system under anaerobic conditions for 50 h at a temperature of 34 degrees C, a pH value of 5.0, and an initial sucrose concentration of 12 g/L using diluted sugarcane molasses. Throughout the process, pulses of molasses were added in order to avoid the cell growth inhibition due to high sugar concentration as well as increased lactic acid concentrations. At the end of the fermentation, about 90% of sucrose was consumed to produce lactic acid and cells. A kinetic model has been developed to simulate the batch lactic acid fermentation results. The data obtained from the fermentation were used for determining the kinetic parameters of the model. The developed model for lactic acid production, growth cell, and sugar consumption simulates the experimental data well.

  19. Hydrogen production and metabolic flux analysis of metabolically engineered Escherichia coli strains

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Seohyoung; Seol, Eunhee; Park, Sunghoon [Department of Chemical and Biochemical Engineering, Pusan National University, Busan 609-735 (Korea); Oh, You-Kwan [Bioenergy Research Center, Korea Institute of Energy Research, Daejeon 305-543 (Korea); Wang, G.Y. [Department of Oceanography, University of Hawaii at Manoa Honolulu, HI 96822 (United States)

    2009-09-15

    Escherichia coli can produce H{sub 2} from glucose via formate hydrogen lyase (FHL). In order to improve the H{sub 2} production rate and yield, metabolically engineered E. coli strains, which included pathway alterations in their H{sub 2} production and central carbon metabolism, were developed and characterized by batch experiments and metabolic flux analysis. Deletion of hycA, a negative regulator for FHL, resulted in twofold increase of FHL activity. Deletion of two uptake hydrogenases (1 (hya) and hydrogenase 2 (hyb)) increased H{sub 2} production yield from 1.20 mol/mol glucose to 1.48 mol/mol glucose. Deletion of lactate dehydrogenase (ldhA) and fumarate reductase (frdAB) further improved the H{sub 2} yield; 1.80 mol/mol glucose under high H{sub 2} pressure or 2.11 mol/mol glucose under reduced H{sub 2} pressure. Several batch experiments at varying concentrations of glucose (2.5-10 g/L) and yeast extract (0.3 or 3.0 g/L) were conducted for the strain containing all these genetic alternations, and their carbon and energy balances were analyzed. The metabolic flux analysis revealed that deletion of ldhA and frdAB directed most of the carbons from glucose to the glycolytic pathway leading to H{sub 2} production by FHL, not to the pentose phosphate pathway. (author)

  20. Increasing antibiotic resistance in preservative-tolerant bacterial strains isolated from cosmetic products.

    Science.gov (United States)

    Orús, Pilar; Gomez-Perez, Laura; Leranoz, Sonia; Berlanga, Mercedes

    2015-03-01

    To ensure the microbiological quality, consumer safety and organoleptic properties of cosmetic products, manufacturers need to comply with defined standards using several preservatives and disinfectants. A drawback regarding the use of these preservatives is the possibility of generating cross-insusceptibility to other disinfectants or preservatives, as well as cross resistance to antibiotics. Therefore, the objective of this study was to understand the adaptive mechanisms of Enterobacter gergoviae, Pseudomonas putida and Burkholderia cepacia that are involved in recurrent contamination in cosmetic products containing preservatives. Diminished susceptibility to formaldehyde-donors was detected in isolates but not to other preservatives commonly used in the cosmetics industry, although increasing resistance to different antibiotics (β-lactams, quinolones, rifampicin, and tetracycline) was demonstrated in these strains when compared with the wild-type strain. The outer membrane protein modifications and efflux mechanism activities responsible for the resistance trait were evaluated. The development of antibiotic-resistant microorganisms due to the selective pressure from preservatives included in cosmetic products could be a risk for the emergence and spread of bacterial resistance in the environment. Nevertheless, the large contribution of disinfection and preservation cannot be denied in cosmetic products. Copyright© by the Spanish Society for Microbiology and Institute for Catalan Studies.

  1. Analysis of β-galactosidase production and their genes of two strains of Lactobacillus bulgaricus.

    Science.gov (United States)

    Zhang, Wen; Wang, Chuan; Huang, Cheng-Yu; Yu, Qian; Liu, Heng-Chuan; Zhang, Chao-Wu; Pei, Xiao-Fang; Xu, Xin; Wang, Guo-Qing

    2012-06-01

    A bacterial β-galactosidase delivery system is a potential therapy for lactose intolerance. Currently, two Lactobacillus bulgaricus strains with different biological characteristics are under consideration as potential sources. However, differences in these β-galactosidase genes and their resulting production levels are poorly characterized. The β-galactosidase ORF of L. bulgaricus yogurt isolate had high variability and was terminated at site 1924 due to a stop codon. However, the full 114 kDa β-galactosidase band was still resolved by SDS-PAGE, which may indicate that the interrupted ORF was translated into more than one peptide, and they together were folded into the complete enzyme protein that showed much higher β-galactosidase activity (6.2 U/mg protein) than the enzyme generated from L. bulgaricus reference strain (2.5 U/mg protein).

  2. Production of volatile and sulfur compounds by ten Saccharomyces cerevisiae strains inoculated in Trebbiano must

    Directory of Open Access Journals (Sweden)

    Francesca ePatrignani

    2016-03-01

    Full Text Available In wines, the presence of sulphur compounds is the resulting of several contributions among which yeast metabolism. The characterization of the starter Saccharomyces cerevisiae needs to be performed also taking into account this ability even if evaluated together with the overall metabolic profile. In this perspective, principal aim of this experimental research was the evaluation of the volatile profiles, throughout GC/MS technique coupled with solid phase micro extraction, of wines obtained throughout the fermentation of 10 strains of Saccharomyces cerevisiae. In addition, the production of sulphur compounds was further evaluated by using a gas-chromatograph coupled with a Flame Photometric Detector. Specifically, the ten strains were inoculated in Trebbiano musts and the fermentations were monitored for 19 days. In the produced wines, volatile and sulphur compounds as well as amino acid concentrations were investigated. Also the physico-chemical characteristics of the wines and their electronic nose profiles were evaluated.

  3. Improved hydrogen production by uptake hydrogenase deficient mutant strain of Rhodobacter sphaeroides O.U.001

    Energy Technology Data Exchange (ETDEWEB)

    Kars, Goekhan; Guenduez, Ufuk; Yuecel, Meral [Department of Biological Sciences, Middle East Technical University, 06531 Ankara (Turkey); Rakhely, Gabor; Kovacs, Kornel L. [Institute of Biophysics, Biological Research Centre, Hungarian Academy of Sciences, Szeged (Hungary); Eroglu, Inci [Department of Chemical Engineering, Middle East Technical University, 06531 Ankara (Turkey)

    2008-06-15

    Rhodobacter sphaeroides O.U.001 is a purple non-sulfur bacterium producing hydrogen under photoheterotrophic conditions. Hydrogen is produced by Mo-nitrogenase enzyme and substantial amount of H{sub 2} is reoxidized by a membrane-bound uptake hydrogenase in the wild type strain. To improve the hydrogen producing capacity of the cells, a suicide vector containing a gentamicin cassette in the hupSL genes was introduced into R. sphaeroiodes O.U.001 and the uptake hydrogenase genes were destroyed by site directed mutagenesis. The correct integration of the construct was confirmed by uptake hydrogenase activity measurement, PCR and subsequent sequence analysis. The wild type and the mutant cells showed similar growth patterns but the total volume of hydrogen gas evolved by the mutant was 20% higher than that of the wild type strain. This result demonstrated that the hydrogen produced by the nitrogenase was not consumed by uptake hydrogenase leading to higher hydrogen production. (author)

  4. Fusarium verticillioides strains isolated from corn feed: characterization by fumonisin production and RAPD fingerprinting

    Directory of Open Access Journals (Sweden)

    Elisabete Yurie Sataque Ono

    2010-08-01

    Full Text Available In this study a total of 16 Fusarium verticillioides strains isolated from corn feed samples were characterized by fumonisin (FB production and random amplified polymorphic DNA (RAPD. All the strains produced FB1 and FB2 with levels ranging from 2.41 to 3996.36 µg/g, and from 1.18 to 1209.91 µg/g, respectively. From the 16 F. verticillioides strains, four were identified as low (3.59 to 1289.84 µg/g, eight as intermediate (>1289.84 to 3772.44 µg/g and four strains as high (>3772.44 µg/g fumonisin producers. From the total of 105 loci amplified, 60 (57.14% were polymorphic. RAPD analysis showed very similar patterns among low, moderate and high fumonisin-producing strains. Although RAPD markers were capable of discriminating the different F. verticillioides strains, there was no clear association between these makers and fumonisin production.Neste estudo, 16 cepas de F. verticillioides isoladas de amostras de ração de milho foram caracterizadas com base na produção de fumonisinas (FB e em marcadores de polimorfismos de DNA amplificado ao acaso (RAPD. Todas as cepas produziram FB1 e FB2, com níveis variando, respectivamente, de 2,41 a 3996,36 µg/g e 1,18 a 1209,91 µg/g. De acordo com a produção de fumonisinas totais (FB1 + FB2 e a distribuição por análise de quartis, do total de 16 cepas de F. verticillioides, quatro foram identificadas como baixas produtoras de fumonisinas (3,59 a 1289,84 µg/g, oito como intermediárias (>1289,84 a 3772,44 µg/g e quatro como altas produtoras de fumonisinas (>3772,44 µg/g. Os 10 primers utilizados amplificaram 105 locos, 60 (57,14% dos quais foram polimórficos. As análises de RAPD mostraram padrões muito similares entre as cepas baixas, médias e altas produtoras de fumonisinas. Embora os marcadores RAPD tenham se mostrado capazes de discriminar as diferentes cepas de F. verticillioides, não foi detectada nenhuma associação entre estes marcadores e a produção de fumonisinas.

  5. Production of cellobionate from cellulose using an engineered Neurospora crassa strain with laccase and redox mediator addition

    Science.gov (United States)

    We report a novel production process for cellobionic acid from cellulose using an engineered fungal strain with the exogenous addition of laccase and a redox mediator. A previously engineered strain of Neurospora crassa (F5'ace-1'cre-1'ndvB) was shown to produce cellobionate directly from cellulose ...

  6. Strain improvement and statistical optimization as a combined strategy for improving fructosyltransferase production by Aureobasidium pullulans NAC8

    Directory of Open Access Journals (Sweden)

    Adedeji Nelson Ademakinwa

    2017-12-01

    A relatively low FTase-producing strain of Aureobasidium pullulans NAC8 was enhanced for optimum production using a two-pronged approach involving mutagenesis and statistical optimization. The improved mutant strain also had remarkable biotechnological properties that make it a suitable alternative than the wild-type.

  7. Characterization of genome-reduced Bacillus subtilis strains and their application for the production of guanosine and thymidine.

    Science.gov (United States)

    Li, Yang; Zhu, Xujun; Zhang, Xueyu; Fu, Jing; Wang, Zhiwen; Chen, Tao; Zhao, Xueming

    2016-06-03

    Genome streamlining has emerged as an effective strategy to boost the production efficiency of bio-based products. Many efforts have been made to construct desirable chassis cells by reducing the genome size of microbes. It has been reported that the genome-reduced Bacillus subtilis strain MBG874 showed clear advantages for the production of several heterologous enzymes including alkaline cellulase and protease. In addition to enzymes, B. subtilis is also used for the production of chemicals. To our best knowledge, it is still unknown whether genome reduction could be used to optimize the production of chemicals such as nucleoside products. In this study, we constructed a series of genome-reduced strains by deleting non-essential regions in the chromosome of B. subtilis 168. These strains with genome reductions ranging in size from 581.9 to 814.4 kb displayed markedly decreased growth rates, sporulation ratios, transformation efficiencies and maintenance coefficients, as well as increased cell yields. We re-engineered the genome-reduced strains to produce guanosine and thymidine, respectively. The strain BSK814G2, in which purA was knocked out, and prs, purF and guaB were co-overexpressed, produced 115.2 mg/L of guanosine, which was 4.4-fold higher compared to the control strain constructed by introducing the same gene modifications into the parental strain. We also constructed a thymidine producer by deleting the tdk gene and overexpressing the prs, ushA, thyA, dut, and ndk genes from Escherichia coli in strain BSK756, and the resulting strain BSK756T3 accumulated 151.2 mg/L thymidine, showing a 5.2-fold increase compared to the corresponding control strain. Genome-scale genetic manipulation has a variety of effects on the physiological characteristics and cell metabolism of B. subtilis. By introducing specific gene modifications related to guanosine and thymidine accumulation, respectively, we demonstrated that genome-reduced strains had greatly improved

  8. Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan.

    Science.gov (United States)

    Fujita, M; Nomura, K; Hong, K; Ito, Y; Asada, A; Nishimuro, S

    1993-12-30

    A strong fibrinolytic enzyme (nattokinase) was purified from the vegetable cheese natto. Nattokinase was extracted from natto with saline and isolated by sequential use of hydrophobic chromatography on Butyl-Toyopearl, ion-exchange chromatography on CM-Toyopearl, and gel-filtration on Sephadex G-50. The isolated protein gave a single sharp band on SDS-PAGE either before or after reduction. The sequence, as determined by automated Edman degradation of the uncleaved molecule and its enzymatically derived peptide, consisted of a total 275 amino acid residues (M.W = 27,728) and exhibited a high homology with the subtilisins. The purified nattokinase digested not only fibrin but also several synthetic substrates. Among the synthetic substrates, the most sensitive substrate was Suc-Ala-Ala-Pro-Phe-pNA for subtilisin. PMSF inhibited both the fibrinolytic activity and the amidolytic activity. The results indicate that nattokinase is a subtilisin-like serine protease.

  9. Natto and viscous vegetables in a Japanese-style breakfast improved insulin sensitivity, lipid metabolism and oxidative stress in overweight subjects with impaired glucose tolerance.

    Science.gov (United States)

    Taniguchi-Fukatsu, Akiko; Yamanaka-Okumura, Hisami; Naniwa-Kuroki, Yuko; Nishida, Yuka; Yamamoto, Hironori; Taketani, Yutaka; Takeda, Eiji

    2012-04-01

    We previously suggested that the consumption of natto and viscous vegetables as part of a Japanese-style meal based on white rice (WR) reduced postprandial glucose and insulin levels in healthy subjects. The aim of the present study was to assess whether a single breakfast of natto and viscous vegetables or the same breakfast consumed for 2 weeks could improve glucose control, insulin sensitivity, lipid metabolism and oxidative stress in overweight subjects with impaired glucose tolerance (IGT). A total of eleven free-living subjects with IGT followed a randomised, crossover breakfast intervention for 2 weeks. The test meal included boiled WR with natto (viscous fermented soyabeans), Japanese yam and okra. The control meal included WR with non-viscous boiled soyabeans, potatoes and broccoli. Both meals contained comparable amounts of carbohydrate, fat, protein and fibre. The test meal reduced acute glucose and insulin responses compared to the control meal in the study participants. Insulin sensitivity was assessed using the composite insulin sensitivity index (CISI) after both the test and control meal periods. The test meal resulted in improvements in CISI compared to the baseline, whereas no significant changes were observed after the control meal period. Serum levels of both total and LDL-cholesterol were assessed before and after the test meal period and found to decrease significantly. There was also a tendency towards reduced serum malondialdehyde-modified LDL and N(ɛ)-carboxymethyllysine. No differences were observed in the measures of chronic glycaemic control. Thus, we conclude that a breakfast of natto and viscous vegetables consumed for 2 weeks improves insulin sensitivity, serum lipid and oxidative stress.

  10. [Hydrogen production and enzyme activity of acidophilic strain X-29 at different C/N ratio].

    Science.gov (United States)

    Li, Qiu-bo; Xing, De-feng; Ren, Nan-qi; Zhao, Li-hua; Song, Ye-ying

    2006-04-01

    Some fermentative bacteria can produce hydrogen by utilizing carbohydrate and other kinds of organic compounds as substrates. Hydrogen production was also determined by both the limiting of growth and related enzyme activity in energy metabolism. Carbon and nitrogen are needed for the growth and metabolism of microorganisms. In addition, the carbon/nitrogen (C/N) ratio can influence the material metabolized and the energy produced. In order to improve the hydrogen production efficiency of the bacteria, we analyzed the effect of different C/N ratios on hydrogen production and the related enzyme activities in the acidophilic strain X-29 using batch test. The results indicate that the differences in the metabolism level and enzyme activity are obvious at different C/N ratios. Although the difference in liquid fermentative products produced per unit of biomass is not obvious, hydrogen production is enhanced at a specifically determined ratio. At a C/N ratio of 14 the accumulative hydrogen yield of strain X-29 reaches the maximum, 2210.9 mL/g. At different C/N ratios, the expression of hydrogenase activity vary; the activity of hydrogenase decrease quickly after reaching a maximum along with the fermentation process, but the time of expression is short. The activity of alcohol dehydrogenase (ADH) tend to stabilize after reaching a peak along with the fermentation process, the difference in expression activity is little, and the expression period is long at different C/N ratios. At a C/N ratio of 14 hydrogenase and ADH reach the maximum 2.88 micromol x (min x mg)(-1) and 33.2 micromol x (min x mg)(-1), respectively. It is shown that the C/N ratio has an important effect on enhancing hydrogen production and enzyme activity.

  11. Metabolic Engineering of the Actinomycete Amycolatopsis sp. Strain ATCC 39116 towards Enhanced Production of Natural Vanillin.

    Science.gov (United States)

    Fleige, Christian; Meyer, Florian; Steinbüchel, Alexander

    2016-06-01

    The Gram-positive bacterium Amycolatopsis sp. ATCC 39116 is used for the fermentative production of natural vanillin from ferulic acid on an industrial scale. The strain is known for its outstanding tolerance to this toxic product. In order to improve the productivity of the fermentation process, the strain's metabolism was engineered for higher final concentrations and molar yields. Degradation of vanillin could be decreased by more than 90% through deletion of the vdh gene, which codes for the central vanillin catabolism enzyme, vanillin dehydrogenase. This mutation resulted in improvement of the final concentration of vanillin by more than 2.2 g/liter, with a molar yield of 80.9%. Further improvement was achieved with constitutive expression of the vanillin anabolism genes ech and fcs, coding for the enzymes feruloyl-coenzyme A (CoA) synthetase (fcs) and enoyl-CoA hydratase/aldolase (ech). The transcription of both genes was shown to be induced by ferulic acid, which explains the unwanted adaptation phase in the fermentation process before vanillin was efficiently produced by the wild-type cells. Through the constitutive and enhanced expression of the two genes, the adaptation phase was eliminated and a final vanillin concentration of 19.3 g/liter, with a molar yield of 94.9%, was obtained. Moreover, an even higher final vanillin concentration of 22.3 g/liter was achieved, at the expense of a lower molar yield, by using an improved feeding strategy. This is the highest reported vanillin concentration reached in microbial fermentation processes without extraction of the product. Furthermore, the vanillin was produced almost without by-products, with a molar yield that nearly approached the theoretical maximum. Much effort has been put into optimization of the biotechnological production of natural vanillin. The demand for this compound is growing due to increased consumer concerns regarding chemically produced food additives. Since this compound is toxic to most

  12. Construction of novel Saccharomyces cerevisiae strains for bioethanol active dry yeast (ADY) production.

    Science.gov (United States)

    Zheng, Daoqiong; Zhang, Ke; Gao, Kehui; Liu, Zewei; Zhang, Xing; Li, Ou; Sun, Jianguo; Zhang, Xiaoyang; Du, Fengguang; Sun, Peiyong; Qu, Aimin; Wu, Xuechang

    2013-01-01

    The application of active dry yeast (ADY) in bioethanol production simplifies operation processes and reduces the risk of bacterial contamination. In the present study, we constructed a novel ADY strain with improved stress tolerance and ethanol fermentation performances under stressful conditions. The industrial Saccharomyces cerevisiae strain ZTW1 showed excellent properties and thus subjected to a modified whole-genome shuffling (WGS) process to improve its ethanol titer, proliferation capability, and multiple stress tolerance for ADY production. The best-performing mutant, Z3-86, was obtained after three rounds of WGS, producing 4.4% more ethanol and retaining 2.15-fold higher viability than ZTW1 after drying. Proteomics and physiological analyses indicated that the altered expression patterns of genes involved in protein metabolism, plasma membrane composition, trehalose metabolism, and oxidative responses contribute to the trait improvement of Z3-86. This work not only successfully developed a novel S. cerevisiae mutant for application in commercial bioethanol production, but also enriched the current understanding of how WGS improves the complex traits of microbes.

  13. Construction of novel Saccharomyces cerevisiae strains for bioethanol active dry yeast (ADY production.

    Directory of Open Access Journals (Sweden)

    Daoqiong Zheng

    Full Text Available The application of active dry yeast (ADY in bioethanol production simplifies operation processes and reduces the risk of bacterial contamination. In the present study, we constructed a novel ADY strain with improved stress tolerance and ethanol fermentation performances under stressful conditions. The industrial Saccharomyces cerevisiae strain ZTW1 showed excellent properties and thus subjected to a modified whole-genome shuffling (WGS process to improve its ethanol titer, proliferation capability, and multiple stress tolerance for ADY production. The best-performing mutant, Z3-86, was obtained after three rounds of WGS, producing 4.4% more ethanol and retaining 2.15-fold higher viability than ZTW1 after drying. Proteomics and physiological analyses indicated that the altered expression patterns of genes involved in protein metabolism, plasma membrane composition, trehalose metabolism, and oxidative responses contribute to the trait improvement of Z3-86. This work not only successfully developed a novel S. cerevisiae mutant for application in commercial bioethanol production, but also enriched the current understanding of how WGS improves the complex traits of microbes.

  14. Ethanol production from xylose in engineered Saccharomyces cerevisiae strains. Current state and perspectives

    Energy Technology Data Exchange (ETDEWEB)

    Matsushika, Akinori; Inoue, Hiroyuki; Sawayama, Shigeki [National Inst. of Advanced Industrial Science and Technology (AIST), Hiroshima (JP). Biomass Technology Research Center (BTRC); Kodaki, Tsutomu [Kyoto Univ. (Japan). Inst. of Advanced Energy

    2009-08-15

    Bioethanol production from xylose is important for utilization of lignocellulosic biomass as raw materials. The research on yeast conversion of xylose to ethanol has been intensively studied especially for genetically engineered Saccharomyces cerevisiae during the last 20 years. S. cerevisiae, which is a very safe microorganism that plays a traditional and major role in industrial bioethanol production, has several advantages due to its high ethanol productivity, as well as its high ethanol and inhibitor tolerance. However, this yeast cannot ferment xylose, which is the dominant pentose sugar in hydrolysates of lignocellulosic biomass. A number of different strategies have been applied to engineer yeasts capable of efficiently producing ethanol from xylose, including the introduction of initial xylose metabolism and xylose transport, changing the intracellular redox balance, and overexpression of xylulokinase and pentose phosphate pathways. In this review, recent progress with regard to these studies is discussed, focusing particularly on xylose-fermenting strains of S. cerevisiae. Recent studies using several promising approaches such as host strain selection and adaptation to obtain further improved xylose-utilizing S. cerevisiae are also addressed. (orig.)

  15. Pectinase production by fungal strains in solid-state fermentation using agro-industrial bioproduct

    OpenAIRE

    Martin,Natalia; Souza,Simone Regina de; Silva,Roberto da; Gomes,Eleni

    2004-01-01

    Pectin lyase and polygalacturonase production by newly isolated fungal strains was carried out in solid-state fermentation. Moniliella SB9 and Penicillium sp EGC5 produced polygalcturonase (PG) and pectin lyase (PL) on mixture of orange bagasse, sugar cane bagasse and wheat bran as substrate. PG and PL produced by Moniliella presented optimum activity at pH 4.5 and 10.0 and at 55 and 45°C, respectively, while these enzymes from Penicillium sp presented optimum activity at pH 4.5-5.0 and 9.0 a...

  16. Development of over-production strain of saccharification enzyme and biomass pretreatment by proton beam irradiation

    International Nuclear Information System (INIS)

    Kim, S. W.; Lee, J. Y.; Song, Y. S.; Lee, S. J.; Shin, H. Y.; Kim, S. B.

    2010-04-01

    When lignocellulosic biomass converts to ethanol, enzyme takes lots of part of whole cost. Therefore, cellulase production is one of the important processes for the successful enzymatic conversion of cellulosic biomass to ethanol. Among cellulolytic enzymes, cellulase is multi-complex enzyme containing endo-glucanase, exo-glucanase and β-glucosidase. Cellulolyticfungi, Trichodema reesei is well known to produce the highest yields of cellulase. Especially, suitable cellulase composition was important for the effective saccharification of lignocellulosic biomass and strain having high level production of cellulase should be developed for hydrolysis. For efficient ethanol production, hemicellullase of Aspergillus also develop to use xylose generated from saccharification of biomass. In this study, pretreatment process of rice straw using proton beam irradiation (PBI) was carried out for enhancement of enzyme digestibility at different proton beam doses. Also, PBI pretreatment on ammonia soaking treated (SAA, Soaking aqueous ammonia) rice straw was conducted to solve the problem that is micro-structural inhibition of rice straw. Optimal dosages of proton beam on rice straw and SAA treated rice straw for efficient recovery of sugar were 15 KGy and 3 KGy, respectively. Enzymatic saccharification of PBI treated rice straw and SAA rice straw was conducted for the guidance of NREL standard procedure. Analysis using X-ray diffractometry (XRD) for crystallinity index was carried out and CrI found to be 33.38% of control and 35.72% of 15 KGy. Also, CrI was determined to be 67.11% of control and approximately 65.58% of 3 kGy dose in PBI pretreatment on SAA treated rice straw. The result of sugar recovery of both was approximately 70 % and 91 % of theoretical glucose contents, respectively. The initial reaction rate was increased from 7.610 -4 g·l -1 ·s -1 of 15 KGy (PBI pretreated rice straw) to 9.710 -4 g·l -1 ·s -1 (3 KGy PBI pretreated SAA rice straw). The selection of

  17. Seaweed as source of energy. 1: effect of a specific bacterial strain on biogas production

    Energy Technology Data Exchange (ETDEWEB)

    Sreenivasa R.P.; Tarwade, S.J.; Sarma, K.S.R.

    1980-09-01

    Only certain marine bacteria capable of digesting the special type of polysaccharide - agar and alginic acid can bring about the biodegradation of these substances and utilise them as carbon source to produce the organics which will be utilised by the methane bacteria to produce methane. When bacterial strain was used in conjunction with cowdung as a source of methane bacteria in seaweed digester, production of biogas from seaweed was accelerated. Adding of small amount of Ulva to seaweed digester increased the output of gas. (Refs. 4).

  18. Inhibition of ochratoxigenic moulds by Debaryomyces hansenii strains for biopreservation of dry-cured meat products.

    Science.gov (United States)

    Andrade, Maria J; Thorsen, Line; Rodríguez, Alicia; Córdoba, Juan J; Jespersen, Lene

    2014-01-17

    The ability of the osmotolerant yeast Debaryomyces hansenii to inhibit Penicillium nordicum, the most common ochratoxigenic mould encountered in dry-cured meat products, was evaluated. The antagonistic effect of ten D. hansenii strains isolated from dry-cured ham was screened in vitro using malt extract media and meat extract peptone media with the water activity (a(w)) adjusted to 0.97 and 0.90. A significant inhibition of the two tested P. nordicum strains by D. hansenii cells and cell-free supernatants was observed. At 0.97 a(w), increasing D. hansenii inoculum concentrations significantly improved the inhibition of mould growth on solid medium, whereas at 0.90 a(w) this was not always the case. As observed by bright field microscopy, most D. hansenii strains were able to delay P. nordicum spore germination when co-cultured in malt extract broth. D. hansenii FHSCC 253H showed the highest overall in vitro inhibition of ochratoxigenic mould growth, and was therefore chosen for co-cultivation assays in dry-cured ham slices incubated at 0.94 and 0.84 a(w) simulating ham ripening. Regardless of the experimental conditions tested, lower levels of the inoculated P. nordicum strain were detected in co-cultivation batches compared with batches without D. hansenii. The highest level of mould growth inhibition was observed in batches at 0.94 a(w). Ochratoxin A (OTA) production in ham samples was detected by HPLC-MS. Co-culturing of P. nordicum with D. hansenii FHSCC 253H resulted in lower OTA levels compared with control samples without D. hansenii. The decrease of the mycotoxin presence due to D. hansenii FHSCC 253H was more efficient at 0.94 a(w) (OTA was below the detection limit). In conclusion, D. hansenii is potentially suitable as a biopreservative agent for preventing ochratoxigenic mould growth and OTA accumulation in dry-cured meat products. The inoculation of D. hansenii should be made at the beginning of processing (at the end of post salting) when the a(w) of

  19. Production of multiple bacteriocins from a single locus by gastrointestinal strains of Lactobacillus salivarius.

    Science.gov (United States)

    O'Shea, Eileen F; O'Connor, Paula M; Raftis, Emma J; O'Toole, Paul W; Stanton, Catherine; Cotter, Paul D; Ross, R Paul; Hill, Colin

    2011-12-01

    Bacteriocins produced by Lactobacillus salivarius isolates derived from a gastrointestinal origin have previously demonstrated efficacy for in vivo protection against Listeria monocytogenes infection. In this study, comparative genomic analysis was employed to investigate the intraspecies diversity of seven L. salivarius isolates of human and porcine intestinal origin, based on the genome of the well-characterized bacteriocin-producing strain L. salivarius UCC118. This revealed a highly conserved megaplasmid-borne gene cluster in these strains involved in the regulation and secretion of two-component class IIb bacteriocins. However, considerable intraspecific variation was observed in the structural genes encoding the bacteriocin peptides. They ranged from close relatives of abp118, such as salivaricin P, which differs by 2 amino acids, to completely novel bacteriocins, such as salivaricin T, which is characterized in this study. Salivaricin T inhibits closely related lactobacilli and bears little homology to previously characterized salivaricins. Interestingly, the two peptides responsible for salivaricin T activity, SalTα and SalTβ, share considerable identity with the component peptides of thermophilin 13, a bacteriocin produced by Streptococcus thermophilus. Furthermore, the salivaricin locus of strain DPC6488 also encodes an additional novel one-component class IId anti-listerial bacteriocin, salivaricin L. These findings suggest a high level of redundancy in the bacteriocins that can be produced by intestinal L. salivarius isolates using the same enzymatic production and export machinery. Such diversity may contribute to their ability to dominate and compete within the complex microbiota of the mammalian gut.

  20. Lactic Acid Production from Pretreated Hydrolysates of Corn Stover by a Newly Developed Bacillus coagulans Strain

    Science.gov (United States)

    Jiang, Ting; Qiao, Hui; Zheng, Zhaojuan; Chu, Qiulu; Li, Xin; Yong, Qiang; Ouyang, Jia

    2016-01-01

    An inhibitor-tolerance strain, Bacillus coagulans GKN316, was developed through atmospheric and room temperature plasma (ARTP) mutation and evolution experiment in condensed dilute-acid hydrolysate (CDH) of corn stover. The fermentabilities of other hydrolysates with B. coagulans GKN316 and the parental strain B. coagulans NL01 were assessed. When using condensed acid-catalyzed steam-exploded hydrolysate (CASEH), condensed acid-catalyzed liquid hot water hydrolysate (CALH) and condensed acid-catalyzed sulfite hydrolysate (CASH) as substrates, the concentration of lactic acid reached 45.39, 16.83, and 18.71 g/L by B. coagulans GKN316, respectively. But for B. coagulans NL01, only CASEH could be directly fermented to produce 15.47 g/L lactic acid. The individual inhibitory effect of furfural, 5-hydroxymethylfurfural (HMF), vanillin, syringaldehyde and p-hydroxybenzaldehyde (pHBal) on xylose utilization by B. coagulans GKN316 was also studied. The strain B. coagulans GKN316 could effectively convert these toxic inhibitors to the less toxic corresponding alcohols in situ. These results suggested that B. coagulans GKN316 was well suited to production of lactic acid from undetoxified lignocellulosic hydrolysates. PMID:26863012

  1. Comparisons of five Saccharomyces cerevisiae strains for ethanol production from SPORL-pretreated lodgepole pine.

    Science.gov (United States)

    Zhou, Haifeng; Lan, Tianqing; Dien, Bruce S; Hector, Ronald E; Zhu, J Y

    2014-01-01

    The performances of five yeast strains under three levels of toxicity were evaluated using hydrolysates from lodgepole pine pretreated by Sulfite Pretreatment to Overcome the Recalcitrance of Lignocelluloses (SPORL). The highest level of toxicity was represented by the whole pretreated biomass slurry, while intermediate toxicity was represented by the hydrolysate with partial loading of pretreatment spent liquor. The zero toxicity was represented using the enzymatic hydrolysate produced from thoroughly washed SPORL lodgepole pine solids. The results indicate that strains D5A and YRH400 can tolerate the whole pretreated biomass slurry to produce 90.1 and 73.5% theoretical ethanol yield. Strains Y1528, YRH403, and FPL450 did not grow in whole hydrolysate cultures and were observed to have lower ethanol productivities than D5A and YRH400 on the hydrolysate with intermediate toxicity. Both YRH400 and YRH403 were genetically engineered for xylose fermentation but were not able to consume xylose efficiently in hydrolysate. © 2014 American Institute of Chemical Engineers.

  2. Lactic Acid Production from Pretreated Hydrolysates of Corn Stover by a Newly Developed Bacillus coagulans Strain.

    Science.gov (United States)

    Jiang, Ting; Qiao, Hui; Zheng, Zhaojuan; Chu, Qiulu; Li, Xin; Yong, Qiang; Ouyang, Jia

    2016-01-01

    An inhibitor-tolerance strain, Bacillus coagulans GKN316, was developed through atmospheric and room temperature plasma (ARTP) mutation and evolution experiment in condensed dilute-acid hydrolysate (CDH) of corn stover. The fermentabilities of other hydrolysates with B. coagulans GKN316 and the parental strain B. coagulans NL01 were assessed. When using condensed acid-catalyzed steam-exploded hydrolysate (CASEH), condensed acid-catalyzed liquid hot water hydrolysate (CALH) and condensed acid-catalyzed sulfite hydrolysate (CASH) as substrates, the concentration of lactic acid reached 45.39, 16.83, and 18.71 g/L by B. coagulans GKN316, respectively. But for B. coagulans NL01, only CASEH could be directly fermented to produce 15.47 g/L lactic acid. The individual inhibitory effect of furfural, 5-hydroxymethylfurfural (HMF), vanillin, syringaldehyde and p-hydroxybenzaldehyde (pHBal) on xylose utilization by B. coagulans GKN316 was also studied. The strain B. coagulans GKN316 could effectively convert these toxic inhibitors to the less toxic corresponding alcohols in situ. These results suggested that B. coagulans GKN316 was well suited to production of lactic acid from undetoxified lignocellulosic hydrolysates.

  3. Lactic Acid Production from Pretreated Hydrolysates of Corn Stover by a Newly Developed Bacillus coagulans Strain.

    Directory of Open Access Journals (Sweden)

    Ting Jiang

    Full Text Available An inhibitor-tolerance strain, Bacillus coagulans GKN316, was developed through atmospheric and room temperature plasma (ARTP mutation and evolution experiment in condensed dilute-acid hydrolysate (CDH of corn stover. The fermentabilities of other hydrolysates with B. coagulans GKN316 and the parental strain B. coagulans NL01 were assessed. When using condensed acid-catalyzed steam-exploded hydrolysate (CASEH, condensed acid-catalyzed liquid hot water hydrolysate (CALH and condensed acid-catalyzed sulfite hydrolysate (CASH as substrates, the concentration of lactic acid reached 45.39, 16.83, and 18.71 g/L by B. coagulans GKN316, respectively. But for B. coagulans NL01, only CASEH could be directly fermented to produce 15.47 g/L lactic acid. The individual inhibitory effect of furfural, 5-hydroxymethylfurfural (HMF, vanillin, syringaldehyde and p-hydroxybenzaldehyde (pHBal on xylose utilization by B. coagulans GKN316 was also studied. The strain B. coagulans GKN316 could effectively convert these toxic inhibitors to the less toxic corresponding alcohols in situ. These results suggested that B. coagulans GKN316 was well suited to production of lactic acid from undetoxified lignocellulosic hydrolysates.

  4. Gallic acid production under anaerobic submerged fermentation by two bacilli strains.

    Science.gov (United States)

    Aguilar-Zárate, Pedro; Cruz, Mario A; Montañez, Julio; Rodríguez-Herrera, Raúl; Wong-Paz, Jorge E; Belmares, Ruth E; Aguilar, Cristóbal N

    2015-12-30

    Tannase is an enzyme that catalyses the breakdown of ester bonds in gallotannins such as tannic acid. In recent years, the interest on bacterial tannases has increased because of its wide applications. The lactic acid bacteria (LAB) plays an important role in food tannin biotransformation, it has the ability of hydrolyse tannins in ruminants intestine. The finding of tannin hydrolysis by LAB has sparked their use as tannase producer. The bacterial strains used in the present work were identified as Bacillus subtilis AM1 and Lactobacillus plantarum CIR1. The maximal tannase production levels were 1400 and 1239 U/L after 32 and 36 h of fermentation respectively, for B. subtilis AM1 and L. plantarum CIR1. Maximum gallic acid release was 24.16 g/L for B. subtilis AM1 and 23.73 g/L for L. plantarum CIR1. HPLC analysis showed the formation of another peaks in the retention time range of 9-14 min, which could be attributed to the formation of di or tri-galloyl glucose. According to database, the strains were identified as Bacillus subtilis AM1 and Lactobacillus plantarum CIR1. In conclusion, both strains had the capability to produce good titres of extracellular tannase and release gallic acid.

  5. Lipase production by recombinant strains of Aspergillus niger expressing a lipase-encoding gene from Thermomyces lanuginosus

    DEFF Research Database (Denmark)

    Prathumpai, Wai; Flitter, S.J.; Mcintyre, Mhairi

    2004-01-01

    Two recombinant strains of Aspergillus niger (NW 297-14 and NW297-24) producing a heterologous lipase from Thermomyces lanuginosus were constructed. The heterologous lipase was expressed using the TAKA amylase promoter from Aspergillus oryzae. The production kinetics of the two strains on different...... shows that it is possible to obtain high productivities of heterologous fungal enzymes in A. niger. However, SDS-PAGE analysis showed that most of the produced lipase was bound to the cell wall....

  6. Enhancement of Exochitinase Production by Bacillus licheniformis AT6 Strain and Improvement of N-Acetylglucosamine Production.

    Science.gov (United States)

    Aounallah, Mohamed Amine; Slimene-Debez, Imen Ben; Djebali, Kais; Gharbi, Dorra; Hammami, Majdi; Azaiez, Sana; Limam, Ferid; Tabbene, Olfa

    2017-02-01

    A strain producing chitinase, isolated from potato stem tissue, was identified as Bacillus licheniformis by biochemical properties and 16S RNA sequence analysis. Statistical experimental designs were used to optimize nine independent variables for chitinase production by B. licheniformis AT6 strain in submerged fermentation. Using Plackett-Burman design, (NH 4 ) 2 SO 4 , MgSO 4 .7H 2 O, colloidal chitin, MnCl 2 2H 2 O, and temperature were found to influence chitinase production significantly. According to Box-Behnken response surface methodology, the optimal fermentation conditions allowing maximum chitinase production were (in gram per liter): (NH 4 ) 2 SO 4 , 7; K 2 HPO 4 , 1; NaCl, 1; MgSO 4 .7H 2 O, 0.1; yeast extract, 0.5; colloidal chitin, 7.5; MnCl 2 .2H 2 O, 0.2; temperature 35 °C; pH medium 7. The optimization strategy led to a 10-fold increase in chitinase activity (505.26 ± 22.223 mU/mL versus 50.35 ± 19.62 mU/mL for control basal medium). A major protein band with a molecular weight of 61.9 kDa corresponding to chitinase activity was clearly detected under optimized conditions. Chitinase activity produced in optimized medium mainly releases N-acetyl glucosamine (GlcNAc) monomer from colloidal chitin. This enzyme also acts as an exochitinase with β-N-acetylglucosaminidase. These results suggest that B. licheniformis AT6 secreting exochitinase is highly efficient in GlcNAc production which could in turn be envisaged as a therapeutic agent or as a conservator against the alteration of several ailments.

  7. Matrix production and organization by endothelial colony forming cells in mechanically strained engineered tissue constructs.

    Directory of Open Access Journals (Sweden)

    Nicky de Jonge

    Full Text Available AIMS: Tissue engineering is an innovative method to restore cardiovascular tissue function by implanting either an in vitro cultured tissue or a degradable, mechanically functional scaffold that gradually transforms into a living neo-tissue by recruiting tissue forming cells at the site of implantation. Circulating endothelial colony forming cells (ECFCs are capable of differentiating into endothelial cells as well as a mesenchymal ECM-producing phenotype, undergoing Endothelial-to-Mesenchymal-transition (EndoMT. We investigated the potential of ECFCs to produce and organize ECM under the influence of static and cyclic mechanical strain, as well as stimulation with transforming growth factor β1 (TGFβ1. METHODS AND RESULTS: A fibrin-based 3D tissue model was used to simulate neo-tissue formation. Extracellular matrix organization was monitored using confocal laser-scanning microscopy. ECFCs produced collagen and also elastin, but did not form an organized matrix, except when cultured with TGFβ1 under static strain. Here, collagen was aligned more parallel to the strain direction, similar to Human Vena Saphena Cell-seeded controls. Priming ECFC with TGFβ1 before exposing them to strain led to more homogenous matrix production. CONCLUSIONS: Biochemical and mechanical cues can induce extracellular matrix formation by ECFCs in tissue models that mimic early tissue formation. Our findings suggest that priming with bioactives may be required to optimize neo-tissue development with ECFCs and has important consequences for the timing of stimuli applied to scaffold designs for both in vitro and in situ cardiovascular tissue engineering. The results obtained with ECFCs differ from those obtained with other cell sources, such as vena saphena-derived myofibroblasts, underlining the need for experimental models like ours to test novel cell sources for cardiovascular tissue engineering.

  8. Effect of irradiation on protease production by a Philippine strain of Aspergillus oryzae (ahlburg) cohn

    International Nuclear Information System (INIS)

    Anglo, P.G.

    1974-03-01

    The Philippine strain of Aspergillus oryzae (ahlburg) cohn. was exposed to ultraviolet rays and ionizing radiation from cobalt-60 for the purpose of obtaining possible mutants or resistant strains which produce powerful proteolytic enzymes. Out of 58 isolates, only 3 gave significant proteolytic values (PV) high enough to merit further investigation. The isolates, G-10, G-110, and 23-110, were picked from plates exposed to gamma rays from cobalt-60. Optimum incubation temperature for these isolates for highest percentage of active protease was 24 0 -27 0 C. The isolates were found capable of producing active protease from the second day of incubation up to the fifth day, whereas the activity of the parent strain was retained the fourth day only. The isolates showed maximum digestive ability at 25 0 -55 0 C, giving proteolytic values of 833. The pH activity curves showed that the enzyme produced by the irradiated isolates G-10 and G-110 were very active at pH 9.0-10.0, and isolate 23-110 at pH 6.0-10.0. The parent strain revealed two pH optima, one at pH 7.5-8.5 and the other at pH 9.0-9.5. Crude enzyme powder gave activities comparable to alkalase and maxatase, commercial proteolytic enzymes imported from Belgium and Netherlands being used as component of laundry detergents by some manufacturing companies in the Philippines. The results obtained give valuable information for the commercial application of the enzyme. Since the organism can produce high yields of protease from copra meal, a by-product of the coconut industry, commerical feasibility may be envisioned in the near future

  9. Production of α-amylase from Streptomyces sp. SLBA-08 strain using agro-industrial by-products

    Directory of Open Access Journals (Sweden)

    Édilla Ribeiro dos Santos

    2012-10-01

    Full Text Available Approximately 1.5 trillion tons are the estimated yearly biomass production, making it an essentially unlimited source of raw material for environmentally friendly and biocompatible products transformed by microorganism, specially fungi and actinomycetes. Several lignocellulosic residues, such as sisal waste and sugarcane bagasse contain starch in their structures which could become important sources for the production of amylases. This study evaluated the production of amylolytic enzymes using Streptomyces sp. SLBA-08 strain, isolated from a semi-arid soil, according to their ability to grow on soluble starch as the sole carbon source. The effect of the carbon source (sisal waste and sugarcane bagasse on α-amylase production was studied using submerged cultivations at 30 ºC. The highest level of α-amylase activity corresponded to 10.1 U. mL-1 and was obtained using sisal waste (2.7% and urea (0.8% in submerged fermentation after 3 days of cultivation. The partial characterization showed the best α-amylase activity at 50ºC and pH 7.0. These results are of great importance for the use of sisal waste as a substrate for biotechnological proposes.

  10. Enhanced production of protease by mutagenized strain of aspergillus oryzae in solid substrate fermentation of rice bran

    International Nuclear Information System (INIS)

    Yousif, M.; Irfan, M.; Baig, S.; Iqbal, A.

    2010-01-01

    Neutral protease activity of parent strain of Aspergellus oryzae was enhanced by UV and chemical mutagenization with ethyl methane sulphonate (EMS). After screening, a hyper producing strain was isolated and found effective for tile production of neutral protease as compared to the parent strain of Aspergellus oryzae. Solid substrate fermentation was carried out in 250ml conical flask with 45 % initial moisture contents at a temperature of 30 deg. C for 72 flours. Under the optimum conditions maximum yield of neutral protease obtained was 662.61+-0.36 U/gds, Almost all the organic nitrogen supplements favored the enzyme production while sucrose proved as a best carbon source. (author)

  11. Induction of bacteriocin production by coculture is widespread among plantaricin-producing Lactobacillus plantarum strains with different regulatory operons.

    Science.gov (United States)

    Maldonado-Barragán, Antonio; Caballero-Guerrero, Belén; Lucena-Padrós, Helena; Ruiz-Barba, José Luis

    2013-02-01

    We describe the bacteriocin-production phenotype in a group of eight singular bacteriocinogenic Lactobacillus plantarum strains with three distinct genotypes regarding the plantaricin locus. Genotyping of these strains revealed the existence of two different plantaricin-production regulatory operons, plNC8-plNC8HK-plnD or plnABCD, involving three-component systems controlled each of them by a specific autoinducer peptide (AIP), i.e. PLNC8IF or PlnA. While all of the strains produced antimicrobial activity when growing on solid medium, most of them halted this production when cultured in broth, thus reflecting the functionality of regulatory mechanisms. Antimicrobial activity in broth cultures was re-established or enhanced when the specific AIP was added to the culture or by coculturing with specific bacterial strains. The latter trait appeared to be widespread in bacteriocinogenic L. plantarum strains independently of the regulatory system used to regulate bacteriocin production or the specific bacteriocins produced. The induction spectrum through coculture, i.e. the pattern of bacterial strains able to induce bacteriocin production, was characteristic of each individual L. plantarum strain. Also, the ability of some bacteria to induce bacteriocin production in L. plantarum by coculture appeared to be strain specific. The fact that induction of bacteriocin production by coculturing appeared to be a common feature in L. plantarum can be exploited accordingly to enhance the viability of this species in food and feed fermentations, as well as to contribute to probiotic functionality when colonising the gastrointestinal tract. Copyright © 2012 Elsevier Ltd. All rights reserved.

  12. Production and characterization of a group of bioemulsifiers from the marine Bacillus velezensis strain H3.

    Science.gov (United States)

    Liu, Xiangyang; Ren, Biao; Chen, Ming; Wang, Haibin; Kokare, Chandrakant R; Zhou, Xianlong; Wang, Jidong; Dai, Huanqin; Song, Fuhang; Liu, Mei; Wang, Jian; Wang, Shujin; Zhang, Lixin

    2010-08-01

    Marine microbes are a rich source of bioactive compounds, such as drugs, enzymes, and biosurfactants. To explore the bioactive compounds from our marine natural product library, an oil emulsification assay was applied to discover biosurfactants and bioemulsifiers. A spore-forming bacterial strain from sea mud was found to produce bioemulsifiers with good biosurfactant activity and a broad spectrum of antimicrobial properties. It was identified as Bacillus velezensis H3 using genomic and phenotypic data analysis. This strain was able to produce biosurfactants with an optimum emulsification activity at pH 6.0 and 2% NaCl by using starch as the carbon source and ammonium sulfate as the nitrogen source. The emulsification-guided isolation and purification procedure led to the discovery of the biosurfactant components, which were mainly composed of nC(14)-surfactin and anteisoC(15)-surfactin as determined by NMR and MS spectra. These compounds can reduce the surface tension of phosphate-buffered saline (PBS) from 71.8 to 24.8 mN/m. The critical micelle concentrations (CMCs) of C(14)-surfactin and C(15)-surfactin in 0.1 M PBS (pH 8.0) were determined to be 3.06 x 10(-5) and 2.03 x 10(-5) mol/L, respectively. The surface tension values at CMCs for C(14)-surfactin and C(15)-surfactin were 25.7 and 27.0 mM/m, respectively. In addition, the H3 biosurfactant exhibited antimicrobial activities against Staphyloccocus aureus, Mycobacterium, Klebsiella peneumoniae, Pseudomonas aeruginosa, and Candida albicans. Thus B. velezensis H3 is an alternative surfactin producer with potential application as an industrial strain for the lipopeptide production.

  13. Effect of Different Carbon Sources on Biosurfactants’ Production by Three Strains of Lactobacillus spp.

    Directory of Open Access Journals (Sweden)

    Tene Hippolyte Mouafo

    2018-01-01

    Full Text Available The potential of three indigenous bacterial strains (Lactobacillus delbrueckii N2, Lactobacillus cellobiosus TM1, and Lactobacillus plantarum G88 for the production of biosurfactants using sugar cane molasses or glycerol as substrates was investigated through emulsifying, surface tension, and antimicrobial activities. The different biosurfactants produced with molasses as substrate exhibited high surface tension reduction from 72 mN/m to values ranged from 47.50 ± 1.78 to 41.90 ± 0.79 mN/m and high emulsification index ranging from 49.89 ± 5.28 to 81.00 ± 1.14%. Whatever the Lactobacillus strain or the substrate used, the biosurfactants produced showed antimicrobial activities against Candida albicans LV1, some pathogenic and/or spoilage Gram-positive and Gram-negative bacteria. The yields of biosurfactants with molasses (2.43 ± 0.09 to 3.03 ± 0.09 g/L or glycerol (2.32 ± 0.19 to 2.82 ± 0.05 g/L were significantly (p<0.05 high compared to those obtained with MRS broth as substrate (0.30 ± 0.02 to 0.51 ± 0.09 g/L. Preliminary characterization of crude biosurfactants reveals that they are mainly glycoproteins and glycolipids with molasses and glycerol as substrate, respectively. Therefore, sugar cane molasses or glycerol can effectively be used by Lactobacillus strains as low-cost substrates to increase their biosurfactants production.

  14. Galactooligosaccharide Production from Pantoea anthophila Strains Isolated from “Tejuino”, a Mexican Traditional Fermented Beverage

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    Claudia V. Yañez-Ñeco

    2017-08-01

    Full Text Available Two Pantoea anthophila bacterial strains were isolated from “tejuino”, a traditional Mexican beverage, and studied as β-galactosidase producers for galactooligosaccharides synthesis. Using 400 g/L of lactose, 50 °C, and 15 U/mL of β-galactosidase activity with ethanol-permeabilized cells, the maximum galactooligosaccharides (GOS yield determined by High performance anion exchange chromatography with pulse amperometric detection (HPAEC-PAD was 136 g/L (34% w/w of total sugars at 96% of lactose conversion for Bac 55.2 and 145 g/L (36% w/w of total sugars at 94% of lactose conversion for Bac 69.1. The main synthesized products were the disaccharides allolactose [Gal-β(1 → 6-Glc] and 6-galactobiose [Gal-β(1 → 6-Gal], as well as the trisaccharides 3′-galactosyl-lactose [Gal-β(1 → 3-Gal-β(1 → 4-Glc], 6-galactotriose [Gal-β(1 → 6-Gal-β(1 → 6-Gal], 3′-galactosyl-allolactose [Gal-β(1 → 3-Gal-β(1 → 6-Glc], and 6′-galactosyl-lactose [Gal-β(1 → 6-Gal-β(1 → 4-Glc]. The β-galactosidases present in both strains showed a high transgalactosylation activity and formed principally β(1 → 3 and β(1 → 6 linkages. Considering the stability and bifidogenic properties of GOS containing such types of bonds, P. anthophila strains Bac 55.2 and Bac 69.1 possess a high potential as novel biocatalysts for prebiotic industrial production.

  15. Investigation of Antibacterial Properties of Yeast Strains Isolated from Iranian Richal and Traditional Dairy Products in Armenia

    Directory of Open Access Journals (Sweden)

    F Karimpour

    2016-09-01

    Full Text Available Background & aim:The use of bio preservative or strains as sources are interesting for food bioprocessing technologist,   and is one of the latest methods to increase the shelf life of food by the health authorities . The present study aimed to investigate the antibacterial activity of supernatants of yeasts isolated from Richal as a traditional dairy product and fermented dairy products in Armenia. Methods: In the present experimental study, the purified supernatant of 77 strains of Armenian yeast products and 12 strains from Iranian Richal were isolated. The purified supernatant were tested against three strains as food spoilages bacteria includes: B. subtilis 17-89, B. Thuringensis17-89, S.typhimuium G-38 , on 3media in 2 condition as aerobic and anaerobic. The inhibition zone of the supernatant were measured   and reported as antibacterial activity. Data were analyzed using statistical tests. Result: A total of 89 strains of yeasts, three species of Rachel and 9 strains of Armenian products (13.5% percent had demonstrated antibacterial activity. T86 strains of Armenian yeasts and FA1 (25 of Rachel had shown more ZOI and antibacterial activity on three media at both aerobic and anaerobic conditions. Comparing the mean of ZOI upon three corruption factors, Rachel strains were significantly different (p <0.05. The highest and lowest effect was observed on Bacillus subtilis effect and Salmonella typhimurium respectively. Conclusion: The results indicated that the yeast strains isolated in anaerobic and aerobic conditions on spoilage bacteria had antibacterial activity effect. Thus, it could be concluded that adding the yeast or its supernatant to food as a bio preservative, may introduce a operative product to the food industry.

  16. Under pressure: evolutionary engineering of yeast strains for improved performance in fuels and chemicals production.

    Science.gov (United States)

    Mans, Robert; Daran, Jean-Marc G; Pronk, Jack T

    2018-04-01

    Evolutionary engineering, which uses laboratory evolution to select for industrially relevant traits, is a popular strategy in the development of high-performing yeast strains for industrial production of fuels and chemicals. By integrating whole-genome sequencing, bioinformatics, classical genetics and genome-editing techniques, evolutionary engineering has also become a powerful approach for identification and reverse engineering of molecular mechanisms that underlie industrially relevant traits. New techniques enable acceleration of in vivo mutation rates, both across yeast genomes and at specific loci. Recent studies indicate that phenotypic trade-offs, which are often observed after evolution under constant conditions, can be mitigated by using dynamic cultivation regimes. Advances in research on synthetic regulatory circuits offer exciting possibilities to extend the applicability of evolutionary engineering to products of yeasts whose synthesis requires a net input of cellular energy. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

  17. Optimizing Carbon/Nitrogen Ratio for Biosurfactant Production by a Bacillus subtilis Strain

    Science.gov (United States)

    Fonseca, R. R.; Silva, A. J. R.; de Franca, F. P.; Cardoso, V. L.; Sérvulo, E. F. C.

    A Bacillus subtilis strain isolated from contaminated soil from a refinery has been screened for biosurfactant production in crystal sugar (sucrose) with different nitrogen sources (NaNO3' (NH4)2SO4' urea, and residual brewery yeast). The highest reduction in surface tension was achieved with a 48-h fermentation of crystal sugar and ammonium nitrate. Optimization of carbon/nitrogen ratio (3,9, and 15) and agitation rate (50, 150, and 250 rpm) for biosurfactant production was carried out using complete factorial design and response surface analysis. The condition of C/N 3 and 250 rpm allowed the maximum increase in surface activity of biosurfactant. A suitable model has been developed, having presented great accordance experimental data. Preliminary characterization of the bioproduct suggested it to be a lipopeptide with some isomers differing from those of a commercial surfactin.

  18. Physiological and Biochemical characterization of Chlamydomonas sp. the Hydrogen Production's Strain

    International Nuclear Information System (INIS)

    Chader, S.; Belhamel, M.; H Hacene

    2006-01-01

    The hydrogen produced by biological way became, one of the most interesting subjects of research relating to development the energy system starting from renewable sources. This study describes the closed relation between the physiological behaviour, biochemical and rate of gases produced by Chlamydomonas sp. strain AT14, isolated in the area of Touat (the Sahara Algerian) and cultivated in a toric photo-bioreactor. A considerable growth was noted, where the concentration of the biomass double in only two days after incubation. The micro-algal cells present a 100% of viability, which relocate has satisfactory behaviour in the toric engine. In addition, the displacement water level in the system of measurement implies has gas production (0.1 ml) in coordination with the anaerobic period of the reactional enclosure. The yield of this way of hydrogen production is depending on the species used, the light intensity, and the conditions of culture. (authors)

  19. Optimization of lactic acid production from glucose using geobacillus stearothermophilus strain 15

    Science.gov (United States)

    Kunasundari, Balakrishnan; Naresh, Sandrasekaran; Safie, Mohammad Farhan Mohd

    2017-09-01

    This study investigated the conversion efficiency of glucose to lactic acid by Geobacillus stearothermophilus strain 15. Six parameters (temperature, pH, incubation time, agitation speed, carbon and nitrogen concentrations) were screened to identify the most significant factors in affecting lactic acid production using glucose. Three most significant factors (temperature, pH and incubation time) were further optimized in this experiment to determine the optimal production of lactic acid. Numerical optimization gave the point prediction of lactic acid concentration produced at 9.95 g/L with the desirability of 0.979 at 40°C, pH 8.5, 24 h, 100 rpm with 5% glucose and 3% yeast extract.

  20. The use of a thermotolerant fermentative Kluyveromyces marxianus IMB3 yeast strain for ethanol production

    Energy Technology Data Exchange (ETDEWEB)

    Banat, I.M. [Univ. of the United Arab Emirates, Al-Ain (United Arab Emirates). Dept. of Biolology; Singh, D. [Haryana Agriculture Univ., Hisar (India). Dept. of Microbiology; Marchant, R. [Ulster Univ. (United Kingdom). School of Applied Biological and Chemical Sciences

    1996-12-31

    An investigation was carried out on the growth and ethanol production of a novel thermotolerant ethanol-producing Kluyveromyces marxianus IMB3 yeast strain. It grew aerobically on glucose, lactose, cellobiose, xylose and whey permeate and fermented all the above carbon sources to ethanol at 45 C. This strain was capable of growing under anaerobic chemostat fermentation conditions at 45 C and a dilution rate of 0.15 h{sup -1} and produced {<=}0.9 g/l biomass and 1.8% (v/v) ethanol. An increase in biomass (up to 10.0 g/l) and ethanol (up to 4.3% v/v at 45 C and 7.7% v/v at 40 C) were achieved by applying a continuous two-stage fermentation in sequence (one aerobic and one anerobic stage) or a two-stage anaerobic fermentation with cell recycling. Potential applications, involving alcohol production systems, for use in dairy and wood related industries, were discussed. (orig.)

  1. Fungal Competitors Affect Production of Antimicrobial Lipopeptides in Bacillus subtilis Strain B9-5.

    Science.gov (United States)

    DeFilippi, Stefanie; Groulx, Emma; Megalla, Merna; Mohamed, Rowida; Avis, Tyler J

    2018-04-01

    Bacillus subtilis has shown success in antagonizing plant pathogens where strains of the bacterium produce antimicrobial cyclic lipopeptides (CLPs) in response to microbial competitors in their ecological niche. To gain insight into the inhibitory role of these CLPs, B. subtilis strain B9-5 was co-cultured with three pathogenic fungi. Inhibition of mycelial growth and spore germination was assessed and CLPs produced by B. subtilis B9-5 were quantified over the entire period of microbial interaction. B. subtilis B9-5 significantly inhibited mycelial growth and spore germination of Fusarium sambucinum and Verticillium dahliae, but not Rhizopus stolonifer. LC-MS analysis revealed that B. subtilis differentially produced fengycin and surfactin homologs depending on the competitor. CLP quantification suggested that the presence of Verticillium dahliae, a fungus highly sensitive to the compounds, caused an increase followed by a decrease in CLP production by the bacterium. In co-cultures with Fusarium sambucinum, a moderately sensitive fungus, CLP production increased more gradually, possibly because of its slower rate of spore germination. With co-cultures of the tolerant fungus Rhizopus stolonifer, B. subtilis produced high amounts of CLPs (per bacterial cell) for the duration of the interaction. Variations in CLP production could be explained, in part, by the pathogens' overall sensitivities to the bacterial lipopeptides and/or the relative growth rates between the plant pathogen and B. subtilis. CLP production varied substantially temporally depending on the targeted fungus, which provides valuable insight concerning the effectiveness of B. subtilis B9-5 protecting its ecological niche against the ingress of these pathogens.

  2. Genome-scale model-driven strain design for dicarboxylic acid production in Yarrowia lipolytica.

    Science.gov (United States)

    Mishra, Pranjul; Lee, Na-Rae; Lakshmanan, Meiyappan; Kim, Minsuk; Kim, Byung-Gee; Lee, Dong-Yup

    2018-03-19

    Recently, there have been several attempts to produce long-chain dicarboxylic acids (DCAs) in various microbial hosts. Of these, Yarrowia lipolytica has great potential due to its oleaginous characteristics and unique ability to utilize hydrophobic substrates. However, Y. lipolytica should be further engineered to make it more competitive: the current approaches are mostly intuitive and cumbersome, thus limiting its industrial application. In this study, we proposed model-guided metabolic engineering strategies for enhanced production of DCAs in Y. lipolytica. At the outset, we reconstructed genome-scale metabolic model (GSMM) of Y. lipolytica (iYLI647) by substantially expanding the previous models. Subsequently, the model was validated using three sets of published culture experiment data. It was finally exploited to identify genetic engineering targets for overexpression, knockout, and cofactor modification by applying several in silico strain design methods, which potentially give rise to high yield production of the industrially relevant long-chain DCAs, e.g., dodecanedioic acid (DDDA). The resultant targets include (1) malate dehydrogenase and malic enzyme genes and (2) glutamate dehydrogenase gene, in silico overexpression of which generated additional NADPH required for fatty acid synthesis, leading to the increased DDDA fluxes by 48% and 22% higher, respectively, compared to wild-type. We further investigated the effect of supplying branched-chain amino acids on the acetyl-CoA turn-over rate which is key metabolite for fatty acid synthesis, suggesting their significance for production of DDDA in Y. lipolytica. In silico model-based strain design strategies allowed us to identify several metabolic engineering targets for overproducing DCAs in lipid accumulating yeast, Y. lipolytica. Thus, the current study can provide a methodological framework that is applicable to other oleaginous yeasts for value-added biochemical production.

  3. Virulence factors, serogroups and antimicrobial resistance properties of Escherichia coli strains in fermented dairy products.

    Science.gov (United States)

    Dehkordi, Farhad Safarpoor; Yazdani, Farshad; Mozafari, Jalal; Valizadeh, Yousef

    2014-04-07

    From a clinical perspective, it is essential to know the microbial safety of fermented dairy products. Doogh and kashk are fermented dairies. These products are used by millions of people but their microbial qualities are unknown. Shiga toxin producing Escherichia coli (STEC) is one of the most commonly detected pathogens in the cases of food poisoning and food-borne illnesses. The present investigation was carried out in order to study the molecular characterization and antimicrobial resistance properties of STEC strains isolated from fermented dairy products. Six hundred fermented dairy samples were collected and immediately transferred to the laboratory. All samples were cultured immediately and those that were E. coli-positive were analyzed for the presence of O157 , O26, O103, O111, O145, O45, O91, O113, O121 and O128 STEC serogroups, tetA, tetB, blaSHV, CITM, cmlA, cat1, aadA1, dfrA1, qnr, aac (3)-IV, sul1 and ereA antibiotic resistance genes and stx1, stx2, eaeA, ehly, cnf1, cnf2, iutA, cdtB, papA, traT, sfaS and fyuA virulence factors using PCR. Antimicrobial susceptibility testing was performed also using disk diffusion methodology with Mueller-Hinton agar. Fifty out of 600 (8.33%) dairy samples harbored E. coli. In addition, yoghurt was the most commonly contaminated dairy. O157 (26%) and O26 (12%) were the most commonly detected serogroups. A significant difference was found between the frequency of Attaching and Effacing E. coli and Enterohaemorrhagic E. coli (P Fermented dairy products can easily become contaminated by antibiotic resistant STEC strains. Our findings should raise awareness about antibiotic resistance in Iran. Clinicians should exercise caution when prescribing antibiotics, especially in veterinary treatments.

  4. Efficient production of 2,3-butanediol from corn stover hydrolysate by using a thermophilic Bacillus licheniformis strain.

    Science.gov (United States)

    Li, Lixiang; Li, Kun; Wang, Kai; Chen, Chao; Gao, Chao; Ma, Cuiqing; Xu, Ping

    2014-10-01

    In this study, a thermophilic Bacillus licheniformis strain X10 was newly isolated for 2,3-butanediol (2,3-BD) production from lignocellulosic hydrolysate. Strain X10 could utilize glucose and xylose simultaneously without carbon catabolite repression. In addition, strain X10 possesses high tolerance to fermentation inhibitors including furfural, vanillin, formic acid, and acetic acid. In a fed-batch fermentation, 74.0g/L of 2,3-BD was obtained from corn stover hydrolysate, with a productivity of 2.1g/Lh and a yield of 94.6%. Thus, this thermophilic B. licheniformis strain is a candidate for the development of efficient industrial production of 2,3-BD from corn stover hydrolysate. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Enhanced biohydrogen production by the N{sub 2}-fixing cyanobacterium Anabaena siamensis strain TISTR 8012

    Energy Technology Data Exchange (ETDEWEB)

    Khetkorn, Wanthanee [Program of Biotechnology, Faculty of Science, Chulalongkorn University, Bangkok, 10330 (Thailand); Laboratory of Cyanobacterial Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Bangkok, 10330 (Thailand); Department of Photochemistry and Molecular Science, Uppsala University, Box 523, SE-75120, Uppsala (Sweden); Lindblad, Peter [Department of Photochemistry and Molecular Science, Uppsala University, Box 523, SE-75120, Uppsala (Sweden); Incharoensakdi, Aran [Laboratory of Cyanobacterial Biotechnology, Department of Biochemistry, Faculty of Science, Chulalongkorn University, Phayathai Road, Bangkok, 10330 (Thailand)

    2010-12-15

    The efficiency of hydrogen production depends on several factors. We focused on external conditions leading to enhanced hydrogen production when using the N{sub 2}-fixing cyanobacterium Anabaena siamensis TISTR 8012, a novel strain isolated from a rice paddy field in Thailand. In this study, we controlled key factors affecting hydrogen production such as cell age, light intensity, time of light incubation and source of carbon. Our results showed an enhanced hydrogen production when cells, at log phase, were adapted under N{sub 2}-fixing condition using 0.5% fructose as carbon source and a continuous illumination of 200 {mu}E m{sup -2} s{sup -1} for 12 h under anaerobic incubation. The maximum hydrogen production rate was 32 {mu}mol H{sub 2} mg chl a{sup -1} h{sup -1}. This rate was higher than that observed in the model organisms Anabaena PCC 7120, Nostoc punctiforme ATCC 29133 and Synechocystis PCC 6803. This higher production was likely caused by a higher nitrogenase activity since we observed an upregulation of nifD. The production did not increase after 12 h which was probably due to an increased activity of the uptake hydrogenase as evidenced by an increased hupL transcript level. Interestingly, a proper adjustment of light conditions such as intensity and duration is important to minimize both the photodamage of the cells and the uptake hydrogenase activity. Our results indicate that A. siamensis TISTR 8012 has a high potential for hydrogen production with the ability to utilize sugars as substrate to produce hydrogen. (author)

  6. Performance in nondairy drinks of probiotic L. casei strains usually employed in dairy products.

    Science.gov (United States)

    Céspedes, Mario; Cárdenas, Pamela; Staffolani, Martín; Ciappini, María C; Vinderola, Gabriel

    2013-05-01

    The increase in vegetarianism as dietary habit and the increased allergy episodes against dairy proteins fuel the demand for probiotics in nondairy products. Lactose intolerance and the cholesterol content of dairy products can also be considered two additional reasons why some consumers are looking for probiotics in other foods. We aimed at determining cell viability in nondairy drinks and resistance to simulated gastric digestion of commercial probiotic lactobacilli commonly used in dairy products. Lactobacillus casei LC-01 and L. casei BGP 93 were added to different commercial nondairy drinks and viability and resistance to simulated gastric digestion (pH 2.5, 90 min, 37 °C) were monitored along storage (5 and 20 °C). For both strains, at least one nondairy drink was found to offer cell counts around 7 log orders until the end of the storage period. Changes in resistance to simulated gastric digestion were observed as well. Commercial probiotic cultures of L. casei can be added to commercial fruit juices after a carefull selection of the product that warrants cell viability. The resistance to simulated gastric digestion is an easy-to-apply in vitro tool that may contribute to product characterization and may help in the choice of the food matrix when no changes in cell viability are observed along storage. Sensorial evaluation is mandatory before marketing since the product type and storage conditions might influence the sensorial properties of the product due to the possibility of growth and lactic acid production by probiotic bacteria. © 2013 Institute of Food Technologists®

  7. Impact of growth temperature on exopolysaccharide production and probiotic properties of Lactobacillus paracasei strains isolated from kefir grains.

    Science.gov (United States)

    Bengoa, Ana A; Llamas, M Goretti; Iraporda, Carolina; Dueñas, M Teresa; Abraham, Analía G; Garrote, Graciela L

    2018-02-01

    EPS-producing LAB are widely used in the dairy industry since these polymers improve the viscosity and texture of the products. Besides, EPS might be responsible for several health benefits attributed to probiotic strains. However, growth conditions (culture media, temperature, pH) could modify EPS production affecting both technological and probiotic properties. In this work, the influence of growth temperature on EPS production was evaluated, as well as the consequences of these changes in the probiotic properties of the strains. All Lactobacillus paracasei strains used in the study showed changes in EPS production caused by growth temperature, evidenced by the appearance of a high molecular weight fraction and an increment in the total amount of produced EPS at lower temperature. Nevertheless, these changes do not affect the probiotic properties of the strains; L. paracasei strains grown at 20 °C, 30 °C and 37 °C were able to survive in simulated gastrointestinal conditions, to adhere to Caco-2 cells after that treatment and to modulate the epithelial innate immune response. The results suggest that selected L. paracasei strains are new probiotic candidates that can be used in a wide range of functional foods in which temperature could be used as a tool to improve the technological properties of the product. Copyright © 2017 Elsevier Ltd. All rights reserved.

  8. Benz[a]anthracene Biotransformation and Production of Ring Fission Products by Sphingobium sp. Strain KK22

    Science.gov (United States)

    Kunihiro, Marie; Ozeki, Yasuhiro; Nogi, Yuichi; Hamamura, Natsuko

    2013-01-01

    A soil bacterium, designated strain KK22, was isolated from a phenanthrene enrichment culture of a bacterial consortium that grew on diesel fuel, and it was found to biotransform the persistent environmental pollutant and high-molecular-weight polycyclic aromatic hydrocarbon (PAH) benz[a]anthracene. Nearly complete sequencing of the 16S rRNA gene of strain KK22 and phylogenetic analysis revealed that this organism is a new member of the genus Sphingobium. An 8-day time course study that consisted of whole-culture extractions followed by high-performance liquid chromatography (HPLC) analyses with fluorescence detection showed that 80 to 90% biodegradation of 2.5 mg liter−1 benz[a]anthracene had occurred. Biodegradation assays where benz[a]anthracene was supplied in crystalline form (100 mg liter−1) confirmed biodegradation and showed that strain KK22 cells precultured on glucose were equally capable of benz[a]anthracene biotransformation when precultured on glucose plus phenanthrene. Analyses of organic extracts from benz[a]anthracene biodegradation by liquid chromatography negative electrospray ionization tandem mass spectrometry [LC/ESI(−)-MS/MS] revealed 10 products, including two o-hydroxypolyaromatic acids and two hydroxy-naphthoic acids. 1-Hydroxy-2- and 2-hydroxy-3-naphthoic acids were unambiguously identified, and this indicated that oxidation of the benz[a]anthracene molecule occurred via both the linear kata and angular kata ends of the molecule. Other two- and single-aromatic-ring metabolites were also documented, including 3-(2-carboxyvinyl)naphthalene-2-carboxylic acid and salicylic acid, and the proposed pathways for benz[a]anthracene biotransformation by a bacterium were extended. PMID:23686261

  9. Identification and characterization of Lactococcus starter strains in milk-based traditional fermented products in the region of Iran

    Directory of Open Access Journals (Sweden)

    Farzad Rahmati

    2018-02-01

    Full Text Available The aim of the present research was identification and investigation of technological attributes of Lactococcus starter strains from traditional dairy products collected from the countryside of Boroujerd in Iran. 33 samples were cultured on selective media M17 and typical colonies surveyed for morphological properties. Totally, 37 strains were isolated based on the diversity in cell morphology and identified using API galleries and carbohydrate fermentation includes 17 strains of Lactococcus lactis (45.96%, 12 strains of Lactococcus garvieae (32.43% and 8 strains of Lactococcusplantarum (21.62%. Strains were appraised for hydrolysis of L-arginine, casein and starch. Furthermore, strains were evaluated for the ability to grow at temperature 10 °C, 45 °C and presence of 4% and 6.5% NaCl, antibiotic sensitivity, acidification ability, proteolytic and lipolytic activities. Generally, 3 strains of Lc.garvieae (GYLC1, BWLC1, DCLC1 and 7 strains of Lc. lactis (GCLC4, GWLC2, GWLC3, SWLC1, SWLC3, BCLC5, DYLC1 exposed the highest levels of technological properties in order to use as starter cultures.

  10. Production of fermentation aroma compounds by Saccharomyces cerevisiae wine yeasts: effects of yeast assimilable nitrogen on two model strains.

    Science.gov (United States)

    Carrau, Francisco M; Medina, Karina; Farina, Laura; Boido, Eduardo; Henschke, Paul A; Dellacassa, Eduardo

    2008-11-01

    The contribution of yeast fermentation metabolites to the aromatic profile of wine is well documented; however, the biotechnological application of this knowledge, apart from strain selection, is still rather limited and often contradictory. Understanding and modeling the relationship between nutrient availability and the production of desirable aroma compounds by different strains must be one of the main objectives in the selection of industrial yeasts for the beverage and food industry. In order to overcome the variability in the composition of grape juices, we have used a chemically defined model medium for studying yeast physiological behavior and metabolite production in response to nitrogen supplementation so as to identify an appropriate yeast assimilable nitrogen level for strain differentiation. At low initial nitrogen concentrations, strain KU1 produced higher quantities of esters and fatty acids whereas M522 produced higher concentrations of isoacids, gamma-butyrolactone, higher alcohols and 3-methylthio-1-propanol. We propose that although strains KU1 and M522 have a similar nitrogen consumption profile, they represent useful models for the chemical characterization of wine strains in relation to wine quality. The differential production of aroma compounds by the two strains is discussed in relation to their capacity for nitrogen usage and their impact on winemaking. The results obtained here will help to develop targeted metabolic footprinting methods for the discrimination of industrial yeasts.

  11. Isolation of Penicillium nalgiovense strains impaired in penicillin production by disruption of the pcbAB gene and application as starters on cured meat products.

    Science.gov (United States)

    Laich, Federico; Fierro, Francisco; Martin, Juan F

    2003-06-01

    The presence of some fungi on a variety of food products, like cheeses or cured meat products, is beneficial for the ripening of the product and for the development of specific flavour features. The utilization of these fungi as starters, which are inoculated normally as asexual spores on the food products at the beginning of the ripening process, is becoming a usual procedure in the food industry. The starter culture also prevents undesirable fungi or bacteria from growing on the product. Penicillium nalgiovense is the most frequently used starter for cured and fermented meat products, but the fact that this fungus can secrete penicillin to the meat product makes it important to get strains unable to synthesize this antibiotic. In this work we report that P. nalgiovense strains impaired in penicillin production can be obtained by disruption of the pcbAB gene (the first gene of the penicillin biosynthetic pathway). When applied as starter on cecina (a salted, smoke-cured beef meat product from the region of León, Spain), the pcbAB-disrupted strain showed no differences with respect to the parental penicillin-producing strain in its ability to colonize the meat pieces and to control their normal mycoflora. Both strains exerted a similar control on the presence of bacteria in cecina. A similar proportion of penicillin-sensitive and penicillin-resistant bacteria were isolated from pieces inoculated with the penicillin-producing or the non-producing P. nalgiovense strains. The decrease of the bacterial population on the surface of cecina seems to be due to the higher competition for nutrients as a consequence of the inoculation and development of the P. nalgiovense mycelium and not due to the production of penicillin by this fungus. Penicillin production was less affected than growth in a solid medium with high NaCl concentrations; this suggests that the high salt concentration present in cecina is not a limiting factor for penicillin production by P. nalgiovense.

  12. Glycogen production for biofuels by the euryhaline cyanobacteria Synechococcus sp. strain PCC 7002 from an oceanic environment.

    Science.gov (United States)

    Aikawa, Shimpei; Nishida, Atsumi; Ho, Shih-Hsin; Chang, Jo-Shu; Hasunuma, Tomohisa; Kondo, Akihiko

    2014-01-01

    Oxygenic photosynthetic microorganisms such as cyanobacteria and microalgae have attracted attention as an alternative carbon source for the next generation of biofuels. Glycogen abundantly accumulated in cyanobacteria is a promising feedstock which can be converted to ethanol through saccharification and fermentation processes. In addition, the utilization of marine cyanobacteria as a glycogen producer can eliminate the need for a freshwater supply. Synechococcus sp. strain PCC 7002 is a fast-growing marine coastal euryhaline cyanobacteria, however, the glycogen yield has not yet been determined. In the present study, the effects of light intensity, CO2 concentration, and salinity on the cell growth and glycogen content were investigated in order to maximize glycogen production in Synechococcus sp. strain PCC 7002. The optimal culture conditions for glycogen production in Synechococcus sp. strain PCC 7002 were investigated. The maximum glycogen production of 3.5 g L(-1) for 7 days (a glycogen productivity of 0.5 g L(-1) d(-1)) was obtained under a high light intensity, a high CO2 level, and a nitrogen-depleted condition in brackish water. The glycogen production performance in Synechococcus sp. strain PCC 7002 was the best ever reported in the α-polyglucan (glycogen or starch) production of cyanobacteria and microalgae. In addition, the robustness of glycogen production in Synechococcus sp. strain PCC 7002 to salinity was evaluated in seawater and freshwater. The peak of glycogen production of Synechococcus sp. strain PCC 7002 in seawater and freshwater were 3.0 and 1.8 g L(-1) in 7 days, respectively. Glycogen production in Synechococcus sp. strain PCC 7002 maintained the same level in seawater and half of the level in freshwater compared with the optimal result obtained in brackish water. We conclude that Synechococcus sp. strain PCC 7002 has high glycogen production activity and glycogen can be provided from coastal water accompanied by a fluctuation

  13. Nitrate salts suppress sporulation and production of enterotoxin in Clostridium perfringens strain NCTC8239.

    Science.gov (United States)

    Yasugi, Mayo; Otsuka, Keisuke; Miyake, Masami

    2016-10-01

    Clostridium perfringens type A is a common source of food-borne illness in humans. Ingested vegetative cells sporulate in the small intestinal tract and in the process produce C. perfringens enterotoxin (CPE). Although sporulation plays a critical role in the pathogenesis of food-borne illness, the molecules triggering/inhibiting sporulation are still largely unknown. It has previously been reported by our group that sporulation is induced in C. perfringens strain NCTC8239 co-cultured with Caco-2 cells in Dulbecco's Modified Eagle Medium (DMEM). In contrast, an equivalent amount of spores was not observed when bacteria were co-cultured in Roswell Park Memorial Institute-1640 medium (RPMI). In the present study it was found that, when these two media are mixed, RPMI inhibits sporulation and CPE production induced in DMEM. When a component of RPMI was added to DMEM, it was found that calcium nitrate (Ca[NO 3 ] 2 ) significantly inhibits sporulation and CPE production. The number of spores increased when Ca(NO 3 ) 2 -deficient RPMI was used. The other nitrate salts significantly suppressed sporulation, whereas the calcium salts used did not. qPCR revealed that nitrate salts increased expression of bacterial nitrate/nitrite reductase. Furthermore, it was found that nitrite and nitric oxide suppress sporulation. In the sporulation stages, Ca(NO 3 ) 2 down-regulated the genes controlled by Spo0A, a master regulator of sporulation, but not spo0A itself. Collectively, these results indicate that nitrate salts suppress sporulation and CPE production by down-regulating Spo0A-regulated genes in C. perfringens strain NCTC8239. Nitrate reduction may be associated with inhibition of sporulation. © 2016 The Societies and John Wiley & Sons Australia, Ltd.

  14. Cholesterol Assimilation with Isolated lactobacilli Strains of Fars’ Local Dairy Products

    Directory of Open Access Journals (Sweden)

    A Emami

    2008-12-01

    Full Text Available ABSTRACT: Introduction & Objective: Cholesterol is an important compound in most of the biological reactions which the excess of it can be seen as a harmful compound of causing heart diseases. The aim of the present study was to evaluate the cholesterol removal property and also its pathway by dairy lactobacillus in in vitro condition under different bile salts concentration. Materials & Methods: After isolation of lactobacillus strains from dairy products, they were identified with chemical tests and their growths were evaluated under presence of cholesterol and bile salts. The method of action of the bacillus in cholesterol removal was assayed by spectrophotometer method. Collected data was analyzed by SPSS software. Results: result of this study showed that any strains of the bacteria had the ability of cholesterol removal (7.82-34.69 µg/ml. L.casei had more competence for removal of cholesterol in compare to the rest of bacilli. The evaluation of cholesterol cell wall attachment revealed that most of removed cholesterols have been changed to the other products. Conclusion: Considering the result of this study, it can be concluded that cholesterol removal has a direct association with growth of bacteria where the L. casei with high growth rate had more capability of cholesterol removal. Whereas the Lactobacillus can remove the cholesterol with different methods, results of this study showed that dairy products, especially yogurt, can remove the harmful substances such as cholesterol using non chemical methods. The results of this study could be expanded on human use if more study and research could be carried out.

  15. Optimization of trehalose production by a novel strain Brevibacterium sp. SY361.

    Science.gov (United States)

    Wang, Lei; Huang, Rui; Gu, Guanbin; Fang, Hongying

    2008-10-01

    Trehalose production by a novel strain of Brevibacterium sp. SY361 was optimized in submerged fermentation. Different chemical and physical parameters such as carbon and nitrogen sources, inoculum level, initial pH, incubation temperature, aeration and time-course of fermentation, were studied in order to increase trehalose productivity. An optimal production medium containing 3% (w/v) glucose, 0.9% (v/v) corn steep liquor, 0.5% (w/v) KH(2)PO(4) and 0.4% (w/v) MgSO(4).7 H(2)O was found suitable for trehalose production. An optimal volume of medium in a 500 ml flask was 80 ml. The optimal levels of other parameters were 4.0% (v/v) of inoculum, initial pH of 6.0, incubation temperature of 28-32 degrees C and time-course of 60 h. Optimized parameters gave a maximum trehalose of 12.2 mg/ml with a conversion rate of 58.4%. (c) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. Production of pullulan from raw potato starch hydrolysates by a new strain of Auerobasidium pullulans.

    Science.gov (United States)

    Wu, Shengjun; Lu, Mingsheng; Chen, Jing; Fang, Yaowei; Wu, Leilei; Xu, Yan; Wang, Shujun

    2016-01-01

    In the present study, hydrolysis of potato starch with marine cold-adapted α-amylase and pullulan production from the hydrolysates by a new strain of Auerobasidium pullulans isolated from sea mud were conducted. The hydrolysis conditions were optimized as follows: reaction time 2h, pH 6.5, temperature 20°C, and α-amylase amount 12 U/g. Under these optimum hydrolysis conditions, the DE value of the potato starch hydrolysates reached to 49.56. The potato starch hydrolysates consist of glucose, maltose, isomaltose, maltotriose, and trace of other maltooligosaccharides with degree of polymerization ranged 4-7. The maximum production of pullulan at 96 h from the hydrolysate of potato starch was 36.17 g/L, which was higher than those obtained from glucose (22.07 g/L, p<0.05) and sucrose (31.42 g/L, p<0.05). Analysis of the high performance liquid chromatography of the hydrolysates of the pullulan product with pullulanase indicated that the main composition is maltotriose, thus confirming the pullulan structure of this pullulan product. Copyright © 2015 Elsevier B.V. All rights reserved.

  17. The Impact of Single Amino Acids on Growth and Volatile Aroma Production by Saccharomyces cerevisiae Strains

    Directory of Open Access Journals (Sweden)

    Samantha Fairbairn

    2017-12-01

    Full Text Available Nitrogen availability and utilization by Saccharomyces cerevisiae significantly influence fermentation kinetics and the production of volatile compounds important for wine aroma. Amino acids are the most important nitrogen source and have been classified based on how well they support growth. This study evaluated the effect of single amino acids on growth kinetics and major volatile production of two phenotypically different commercial wine yeast strains in synthetic grape must. Four growth parameters, lag phase, maximum growth rate, total biomass formation and time to complete fermentation were evaluated. In contrast with previous findings, in fermentative conditions, phenylalanine and valine supported growth well and asparagine supported it poorly. The four parameters showed good correlations for most amino acid treatments, with some notable exceptions. Single amino acid treatments resulted in the predictable production of aromatic compounds, with a linear correlation between amino acid concentration and the concentration of aromatic compounds that are directly derived from these amino acids. With the increased complexity of nitrogen sources, linear correlations were lost and aroma production became unpredictable. However, even in complex medium minor changes in amino acid concentration continued to directly impact the formation of aromatic compounds, suggesting that the relative concentration of individual amino acids remains a predictor of aromatic outputs, independently of the complexity of metabolic interactions between carbon and nitrogen metabolism and between amino acid degradation and utilization pathways.

  18. Constitutive cellulase production from glucose using the recombinant Trichoderma reesei strain overexpressing an artificial transcription activator.

    Science.gov (United States)

    Zhang, Xiaoyue; Li, Yonghao; Zhao, Xinqing; Bai, Fengwu

    2017-01-01

    The high cost of cellulase production presents biggest challenge in biomass deconstruction. Cellulase production by Trichoderma reesei using low cost carbon source is of great interest. In this study, an artificial transcription activator containing the Cre1 binding domain linked to the Xyr1 effector and binding domains was designed and constitutively overexpressed in T. reesei RUT C30. The recombinant strain T. reesei zxy-2 displayed constitutive cellulase production using glucose as a sole carbon source, and the production titer was 12.75-fold of that observed with T. reesei RUT C30 in shake flask culture. Moreover, FPase and xylanase titers of 2.63 and 108.72IU/mL, respectively, were achieved using glucose as sole carbon source within 48h in a 7-L fermenter by batch fermentation using T. reesei zxy-2. The crude enzyme obtained was used to hydrolyze alkali pretreated corn stover, and a high glucose yield of 99.18% was achieved. Copyright © 2016. Published by Elsevier Ltd.

  19. Production of xylanases by an Aspergillus niger strain in wastes grain

    Directory of Open Access Journals (Sweden)

    Simone Cristine Izidoro

    2014-08-01

    Full Text Available Many fungi are used in order to extract products from their metabolism through bioprocesses capable of minimizing adverse effects caused by agro-industrial wastes in the environment. The aim of this study was to evaluate the xylanase production by an Aspergillus niger strain, using agro-industrial wastes as substrate. Brewer’s spent grain was the best inducer of xylanase activity. Higher levels of xylanase were obtained when the fungus was grown in liquid Vogel medium, pH 5.0, at 30ºC, during 5 days. The temperature for optimum activity was 50ºC and optimum pH 5.0. The enzyme was stable at 50ºC, with a half-life of 240 min. High pH stability was verified from pH 4.5 to 7.0. These characteristics exhibited by A. niger xylanase turn this enzyme attractive for some industrial applications, such as in feed and food industries. Additionally, the use of brewer’s spent grain, an abundantly available and low-cost residue, as substrate for xylanase production can not only add value and decrease the amount of this waste, but also reduce xylanase production cost.

  20. Biomass production by Antarctic yeast strains: an investigation on the lipid composition

    International Nuclear Information System (INIS)

    Zlatanov, M.; Antova, G.; Angelova-Romova, M.; Pavlova, K.; Georgieva, K.; Rousenova-Videva, S.

    2010-01-01

    Psychrophilic yeast strains Rhodotorula glutinis AL_1_0_7, Sporobolomyces roseus AL_1_0_8, Cryptococcus albidus AL_5_5, Cryptococcus laurentii AL_5_6 and Cryptococcus laurentii AL_5_8 isolated from soil sample taken from the region of the Bulgarien base on Livingston Island, Antarctica, were studied. The biomass production was followed after cultivation of the yeasts in a medium with pH 5.3 at 15°C for 120 h. The biomass concentration by psychrophilic yeast strains was: R. glutinis AL_1_0_7-6.05 g/l, S. roseus AL108-5.78 g/l, Cr. albidus AL_5_5, Cr. laurentii AL_5_6 and Cr. laurentii AL_5_8-6.52 g/l, 6.84 g/l and 6.24 g/l, respectively. The extracted and separated lipids from the samples were supplied to analysis and the compositions of fatty acids, phospholipids, sterols as well as tocopherols were determined. Unsaturated fatty acids, mainly oleic (58.6-63.5%) and of saturated palmitic (18.2-24.5%), predominated in triacylglycerols. Sterols (0.1-0.3%) were valued in the dry yeast biomass. The content of phospholipids, mainly phosphatidylcholine, phosphatidylinositole and phosphatidylethanolamine was found to be in the range of 0.2-1.6%. The quantity of tocopherols was 0-26.3 mg/kg. All of tocopherol classes were established.

  1. Construction and application of recombinant strain for the production of an alkaline protease from Bacillus licheniformis.

    Science.gov (United States)

    Lin, Songyi; Zhang, Meishuo; Liu, Jingbo; Jones, Gregory S

    2015-03-01

    The alkaline protease gene, Apr, from Bacillus licheniformis 2709 was cloned into an expression vector pET - 28b (+), to yield the recombinant plasmid pET-28b (+) - Apr. The pET-28b (+) - Apr was expressed in a high expression strain E. coli BL21. The amino acid sequence deduced from the DNA sequence analysis revealed a 98% identity to that of Bacillus licheniformis 2709. Sodium salt-Polyacrylamide gel electrophoresis (SDS-PAGE) was used to access the protein expression. SDS-PAGE analysis indicated a protein of Mr of 38.8 kDa. The medium components and condition of incubation were optimized for the growth state of a recombinant strain. The optimal composition of production medium was composed of glucose 8 g/L, peptone 8 g/L and salt solution 10 mL. The samples were incubated on a rotary shaker of 180 r/min at 37°C for 24 h. Copyright © 2014. Published by Elsevier B.V.

  2. INVERTASE FROM A CANDIDA STELLATA STRAIN ISOLATED FROM GRAPE: PRODUCTION AND PHYSICO-CHEMICAL CHARACTERIZATION

    Directory of Open Access Journals (Sweden)

    Cristiane Abe Gargel

    2014-08-01

    Full Text Available Invertases are enzymes which hydrolyze the sucrose and are widely employed in food and pharmaceutical industries. In this work, the screening of autochthonous grape yeasts from Brazil was carried out in order to investigate their invertase production potential. Yeasts belonging to Saccharomyces, Hanseniaspora, Sporidiobolus, Issatchenkia, Candida, Cryptococcus and Pichia genera were analyzed by submerged fermentation (SbmF using sucrose as substrate. Among them, Candida stellata strain (N5 strain was selected as the best producer (10.6 U/ml after 48 hours of SbmF. This invertase showed optimal activity at pH 3.0 and 55°C, demonstrating appropriate characters for application in several industrial processes, which includes high temperatures and acid pHs. In addition, this invertase extract presented tolerance to low concentrations of ethanol, suggesting that it could also be suitable for application at the beginning of alcoholic fermentation. These data provide promising prospects of the use of this new invertase in food and ethanol industry.

  3. A Mutated Yeast Strain with Enhanced Ethanol Production Efficiency and Stress Tolerance

    Directory of Open Access Journals (Sweden)

    Naghmeh Hemmati1*, David A. Lightfoot1,2, and Ahmed Fakhoury3

    2012-05-01

    Full Text Available One of the strategies to improve and optimize bio-ethanolproduction from new feed stocks is to develop new strainsof Saccharomyces cerevisiae with tolerance to stresses. Themain objectives here were to; generate S. cerevisiae mutantstolerant to high ethanol concentrations; test for their abilityto ferment maize starch; and partially characterize the mutationsresponsible for the new phenotypes. A combinationof mutagenesis, selection and cross-stress protection methodswere used. EMS (ethyl methanesulfonate was used tomutagenize one S. cerevisiae strain. The mutagenized yeaststrain was exposed to high concentrations of ethanol andtolerant mutants were isolated. Mutants showed improvedethanol yield (0.02-0.03 g/g of maize and fermentation efficiency(3-5%. Finally, AFLP (Amplified Fragment LengthPolymorphism was performed to identify polymorphisms inthe mutants that might underlie the strains ethanol tolerance.The best performing mutant isolate had four altered genetranscripts encoding; an arginine uptake and canavanine resistanceprotein (CAN1; mitochondrial membrane proteins(SLS1; a putative membrane glycoprotein (VTH1; and cytochromeC oxidase (COX6; EC 1.9.3.1 among about 1,000tested. It was concluded these mutations might underlie theimproved ethanol production efficiency and stress tolerance.

  4. Glucoamylase production in batch, chemostat and fed-batch cultivations by an industrial strain of Aspergillus niger

    DEFF Research Database (Denmark)

    Pedersen, Henrik; Beyer, Michael; Nielsen, Jens

    2000-01-01

    The Aspergillus niger strain BO-1 was grown in batch, continuous (chemostat) and fed-batch cultivations in order to study the production of the extracellular enzyme glucoamylase under different growth conditions. In the pH range 2.5-6.0, the specific glucoamylase productivity and the specific...

  5. Xylanase production from marine derived Trichoderma pleuroticola 08ÇK001 strain isolated from Mediterranean coastal sediments.

    Science.gov (United States)

    Korkmaz, Melih N; Ozdemir, Sennur C; Uzel, Ataç

    2017-10-01

    Xylanases constitutes one the most important enzymes with diverse applications in different industries such as bioethanol production, animal feedstock production, production of xylo-oligosaccharides, baking industry, paper and pulp industry, xylitol production, fruit juice, and beer finishing, degumming, and agriculture. Currently, industrial xylanases are mainly produced by Aspergillus and Trichoderma members. Since the marine environments are less studied compared to terrestrial environments and harbors great microbial diversity we aimed to investigate the xylanase production of 88 marine-derived filamentous fungal strains. These strains are semi-quantitatively screened for their extracellular xylanase production and Trichoderma pleuroticola 08ÇK001 xylanase activity was further characterized. Optimum pH and temperature was determined as 5.0 and 50 °C, respectively. The enzyme preparation retained 53% of its activity at pH 5.0 after 1 h and have found resistant against several ions and compounds such as K + , Ba 2+ , Na + , β-mercaptoethanol, Triton X-100 and toluene. This study demonstrates that marine-derived fungal strains are prolific sources for xylanase production and presents the first report about the production and characterization of xylanase from a marine derived T. pleuroticola strain. The characteristics of T. pleuroticola 08ÇK001 xylanase activity indicate possible employment in some industrial processes such as animal feed, juice and wine industries or paper pulping applications. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Simultaneous production of acetic and gluconic acids by a thermotolerant Acetobacter strain during acetous fermentation in a bioreactor.

    Science.gov (United States)

    Mounir, Majid; Shafiei, Rasoul; Zarmehrkhorshid, Raziyeh; Hamouda, Allal; Ismaili Alaoui, Mustapha; Thonart, Philippe

    2016-02-01

    The activity of bacterial strains significantly influences the quality and the taste of vinegar. Previous studies of acetic acid bacteria have primarily focused on the ability of bacterial strains to produce high amounts of acetic acid. However, few studies have examined the production of gluconic acid during acetous fermentation at high temperatures. The production of vinegar at high temperatures by two strains of acetic acid bacteria isolated from apple and cactus fruits, namely AF01 and CV01, respectively, was evaluated in this study. The simultaneous production of gluconic and acetic acids was also examined in this study. Biochemical and molecular identification based on a 16s rDNA sequence analysis confirmed that these strains can be classified as Acetobacter pasteurianus. To assess the ability of the isolated strains to grow and produce acetic acid and gluconic acid at high temperatures, a semi-continuous fermentation was performed in a 20-L bioreactor. The two strains abundantly grew at a high temperature (41°C). At the end of the fermentation, the AF01 and CV01 strains yielded acetic acid concentrations of 7.64% (w/v) and 10.08% (w/v), respectively. Interestingly, CV01 was able to simultaneously produce acetic and gluconic acids during acetic fermentation, whereas AF01 mainly produced acetic acid. In addition, CV01 was less sensitive to ethanol depletion during semi-continuous fermentation. Finally, the enzymatic study showed that the two strains exhibited high ADH and ALDH enzyme activity at 38°C compared with the mesophilic reference strain LMG 1632, which was significantly susceptible to thermal inactivation. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  7. Proteomic Analysis of Anti-Cancerous Scopularide Production by a Marine Microascus brevicaulis Strain and Its UV Mutant.

    Directory of Open Access Journals (Sweden)

    Annemarie Kramer

    Full Text Available The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularides A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing faster growth and differences in pellet formation besides higher production levels. Here, we show the first proteome study of a marine fungus. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of the wild type strain and its mutant. For this purpose, an optimised protein extraction protocol was established. In total, 4759 proteins were identified. The central metabolic pathway of strain LF580 was mapped using the KEGG pathway analysis and GO annotation. Employing iTRAQ labelling, 318 proteins were shown to be significantly regulated in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to limited nutrient availability in the wild type strain due to a strong pellet formation. This information can be applied for optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum.

  8. Proteomic Analysis of Anti-Cancerous Scopularide Production by a Marine Microascus brevicaulis Strain and Its UV Mutant.

    Science.gov (United States)

    Kramer, Annemarie; Beck, Hans Christian; Kumar, Abhishek; Kristensen, Lars Peter; Imhoff, Johannes F; Labes, Antje

    2015-01-01

    The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularides A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing faster growth and differences in pellet formation besides higher production levels. Here, we show the first proteome study of a marine fungus. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of the wild type strain and its mutant. For this purpose, an optimised protein extraction protocol was established. In total, 4759 proteins were identified. The central metabolic pathway of strain LF580 was mapped using the KEGG pathway analysis and GO annotation. Employing iTRAQ labelling, 318 proteins were shown to be significantly regulated in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to limited nutrient availability in the wild type strain due to a strong pellet formation. This information can be applied for optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum.

  9. Proteomic Analysis of Anti-Cancerous Scopularide Production by a Marine Microascus brevicaulis Strain and Its UV Mutant

    Science.gov (United States)

    Kramer, Annemarie; Beck, Hans Christian; Kumar, Abhishek; Kristensen, Lars Peter; Imhoff, Johannes F.; Labes, Antje

    2015-01-01

    The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularides A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing faster growth and differences in pellet formation besides higher production levels. Here, we show the first proteome study of a marine fungus. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of the wild type strain and its mutant. For this purpose, an optimised protein extraction protocol was established. In total, 4759 proteins were identified. The central metabolic pathway of strain LF580 was mapped using the KEGG pathway analysis and GO annotation. Employing iTRAQ labelling, 318 proteins were shown to be significantly regulated in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to limited nutrient availability in the wild type strain due to a strong pellet formation. This information can be applied for optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum. PMID:26460745

  10. Metabolic engineering of strains: from industrial-scale to lab-scale chemical production.

    Science.gov (United States)

    Sun, Jie; Alper, Hal S

    2015-03-01

    A plethora of successful metabolic engineering case studies have been published over the past several decades. Here, we highlight a collection of microbially produced chemicals using a historical framework, starting with titers ranging from industrial scale (more than 50 g/L), to medium-scale (5-50 g/L), and lab-scale (0-5 g/L). Although engineered Escherichia coli and Saccharomyces cerevisiae emerge as prominent hosts in the literature as a result of well-developed genetic engineering tools, several novel native-producing strains are gaining attention. This review catalogs the current progress of metabolic engineering towards production of compounds such as acids, alcohols, amino acids, natural organic compounds, and others.

  11. Seaweed as source of energy. I: effect of a specific bacterial strain on biogas production

    Energy Technology Data Exchange (ETDEWEB)

    Rao, P.S.; Tarwade, S.J.; Sarma, K.S.R.

    1980-01-01

    Biogas was produced from seaweed by making use of alginate-digesting marine bacteria that were isolated from decomposing seaweed and can digest seaweed carbohydrates (agar and alginic acid). Laboratory digesters containing 100 g seaweed were inoculated with 50 mL broth cultures of different seaweed-derived bacterial strains, and the maximum amount of degradation obtained was 28% (compared with 13% for a bacteria-free digestion). Cow dung was added as a source of methanogenic bacteria, and the amount of biogas produced was more than double the amount obtained when seaweed and cow dung were digested in the absence of the seaweed-derived bacteria. Adding a small amount of Ulva to the seaweed digester increased the production of biogas.

  12. Enhancing cellulase production by overexpression of xylanase regulator protein gene, xlnR, in Talaromyces cellulolyticus cellulase hyperproducing mutant strain.

    Science.gov (United States)

    Okuda, Naoyuki; Fujii, Tatsuya; Inoue, Hiroyuki; Ishikawa, Kazuhiko; Hoshino, Tamotsu

    2016-10-01

    We obtained strains with the xylanase regulator gene, xlnR, overexpressed (HXlnR) and disrupted (DXlnR) derived from Talaromyces cellulolyticus strain C-1, which is a cellulase hyperproducing mutant. Filter paper degrading enzyme activity and cellobiohydrolase I gene expression was the highest in HXlnR, followed by C-1 and DXlnR. These results indicate that the enhancement of cellulase productivity was succeeded by xlnR overexpression.

  13. Correlation between oxalic acid production and tolerance of Tyromyces palustris strain TYP-6137 to N',N-naphthaloylhydroxamine

    Science.gov (United States)

    Rachel A. Arango; Patricia K. Lebow; Frederick III Green

    2009-01-01

    Eleven strains of T. palustris were evaluated for mass loss and production of phosphate buffer soluble oxalic acid on pine wood blocks treated with 0.5% N’,N-naphthaloylhydroxamine (NHA) in a soil-block test. After 12 weeks higher percentage mass loss was observed in control groups for 10 strains, while TYP-6137 was shown to be tolerant with no difference between the...

  14. MD simulations to evaluate effects of applied tensile strain on irradiation-induced defect production at various PKA energies

    International Nuclear Information System (INIS)

    Miyashiro, S.; Fujita, S.; Okita, T.; Okuda, H.

    2012-01-01

    Highlights: ► Strain effects on defect formation were evaluated at various PKA energies by MD. ► Radiation-induced defects were increased numerically by external strain. ► Enhanced formation of larger clusters causes the numerical increase of defects. ► Strain influence on the number of defects was greatest at about 20 keV PKA. ► Cluster size, which is mostly affected by strain, was greater with higher PKA energy. - Abstract: Molecular Dynamics (MD) simulations were conducted to investigate the influence of applied tensile strain on defect production during cascade damages at various Primary Knock-on Atom (PKA) energies of 1–30 keV. When 1% strain was applied, the number of surviving defects increased at PKA energies higher than 5 keV, although they did not increase at 1 keV. The rate of increase by strain application was higher with higher PKA energy, and attained the maximum at 20 keV PKA energy with a subsequent gradual decrease at 30 keV PKA energy The cluster size, mostly affected by strain, was larger with higher PKA energy, although clusters with fewer than seven interstitials did not increase in number at any PKA energy.

  15. Isolation and identification of new lactobacilli from goatling stomach and investigation of reuterin production in Lactobacillus reuteri strains.

    Science.gov (United States)

    Kiňová Sepová, Hana; Bilková, Andrea

    2013-01-01

    Five new strains of lactobacilli isolated from goatling's stomach were identified by molecular-biological approaches. Profiles of fermentable saccharides, Gram staining, and cell morphology were also determined. They were identified as Lactobacillus reuteri (strains KO4b, KO4m, KO5) and as Lactobacillus plantarum (strains KG1z, KG4). In DNA samples of all newly isolated L. reuteri strains as well as in L. reuteri E (Lreu E; originated from lamb), the part of gldC gene, coding large subunit of glycerol dehydratase, that is necessary for 3-hydroxypropionaldehyde (3-HPA; reuterin) production, was amplified using two designed primer sets. However, the 3-HPA production was revealed only in the strain Lreu E. It produced five- or ten-fold lower amount of 3-HPA in comparison with probiotic L. reuteri ATCC 55730 in aerobic or anaerobic conditions, respectively. Moreover, Lreu E completely lost its production ability after ca. five passages in MRS medium. The co-incubation of Lreu E, but not other L. reuteri isolates, with Escherichia coli re-induced 3-HPA production. In the case of L. reuteri ATCC 55730, the 3-HPA production increased more than four times after co-incubation with E. coli.

  16. Aerobic biotransformation of 3-methylindole to ring cleavage products by Cupriavidus sp. strain KK10.

    Science.gov (United States)

    Fukuoka, Kimiko; Ozeki, Yasuhiro; Kanaly, Robert A

    2015-09-01

    3-Methylindole, also referred to as skatole, is a pollutant of environmental concern due to its persistence, mobility and potential health impacts. Petroleum refining, intensive livestock production and application of biosolids to agricultural lands result in releases of 3-methylindole to the environment. Even so, little is known about the aerobic biodegradation of 3-methylindole and comprehensive biotransformation pathways have not been established. Using glycerol as feedstock, the soil bacterium Cupriavidus sp. strain KK10 biodegraded 100 mg/L of 3-methylindole in 24 h. Cometabolic 3-methylindole biodegradation was confirmed by the identification of biotransformation products through liquid chromatography electrospray ionization tandem mass spectrometry analyses. In all, 14 3-methylindole biotransformation products were identified which revealed that biotransformation occurred through different pathways that included carbocyclic aromatic ring-fission of 3-methylindole to single-ring pyrrole carboxylic acids. This work provides first comprehensive evidence for the aerobic biotransformation mechanisms of 3-methylindole by a soil bacterium and expands our understanding of the biodegradative capabilities of members of the genus Cupriavidus towards heteroaromatic pollutants.

  17. Continuous ethanol production from sugar beet molasses using an osmotolerant mutant strain of zymomonas mobilis

    Energy Technology Data Exchange (ETDEWEB)

    Park, S.C.; Baratti, J.C. (Univ. de Provence, Marseille (France). Centre National de la Recherche Scientifique)

    1992-01-25

    In conventional alcohol fermentation processes using yeast species, the substrate cost represents a major fraction of the total production cost. Therefore, it may be very attractive to use the bacterium Zymomonas mobilis, since it has shown higher ethanol yields than yeasts when grown on a glucose-based medium. A report is made on the use of mutant strain of Zymomonas mobilis for ethanol production from hydrolyzed sugar beet molasses in a two-stage continuous culture which showed high ethanol yield and an ethanol concentration sufficiently high for economical recovery. A single stage continuous culture was first operated in an attempt to reduce the formation of sorbitol. Further on, a second fermentor was added with additional substrate feeding to increase the effluent ethanol concentration. An ethanol concentration of 59.9g/l was obtained at 97% sugar conversion and at high ethanol yield. The volumetric ethanol productivity was superior to that of batch fermentation but inferior to that of a single-stage continuous system with the same medium. However, the ethanol concentration was increased to a level acceptable for economical recovery. 18 refs., 3 figs., 5 tabs.

  18. Molecular Detection of Two Potential Probiotic Lactobacilli Strains and Evaluation of Their Performance as Starter Adjuncts in Yogurt Production.

    Science.gov (United States)

    Saxami, Georgia; Papadopoulou, Olga S; Chorianopoulos, Nikos; Kourkoutas, Yiannis; Tassou, Chrysoula C; Galanis, Alex

    2016-05-04

    A molecular method for efficient and accurate detection and identification of two potential probiotic lactobacilli strains isolated from fermented olives, namely Lactobacillus pentosus B281 and Lb. plantarum B282, was developed in the present study. Random Amplified Polymorphic DNA (RAPD) analysis was performed, and strain specific primers were designed and applied in a multiplex polymerase chain reaction (PCR) assay. The specificity of the assay was tested and successfully confirmed in 27 and 22 lactobacilli strains for Lb. pentosus B281 and Lb. plantarum B282, respectively. Moreover, the two strains were used as starter cultures in yogurt production. Cell enumeration followed by multiplex PCR analysis demonstrated that the two strains were present in yogurt samples at levels ≥6 log CFU/g even after 35 days of storage at 4 °C. Microbiological analysis showed that lactobacilli and streptococci were present within usual levels, whereas enterobacteriaceae and yeast/mold counts were not detected as expected. Although the pH values of the novel products were slightly lower than the control ones, the yogurt containing the probiotic cultures scored similar values compared to the control in a series of sensory tests. Overall, these results demonstrated the possible use of the two strains as starter adjuncts in the production of yogurt with potential probiotic properties.

  19. Thermotolerant Kluyveromyces marxianus and Saccharomyces cerevisiae strains representing potentials for bioethanol production from Jerusalem artichoke by consolidated bioprocessing.

    Science.gov (United States)

    Hu, Nan; Yuan, Bo; Sun, Juan; Wang, Shi-An; Li, Fu-Li

    2012-09-01

    Thermotolerant inulin-utilizing yeast strains are desirable for ethanol production from Jerusalem artichoke tubers by consolidated bioprocessing (CBP). To obtain such strains, 21 naturally occurring yeast strains isolated by using an enrichment method and 65 previously isolated Saccharomyces cerevisiae strains were investigated in inulin utilization, extracellular inulinase activity, and ethanol fermentation from inulin and Jerusalem artichoke tuber flour at 40 °C. The strains Kluyveromyces marxianus PT-1 (CGMCC AS2.4515) and S. cerevisiae JZ1C (CGMCC AS2.3878) presented the highest extracellular inulinase activity and ethanol yield in this study. The highest ethanol concentration in Jerusalem artichoke tuber flour fermentation (200 g L(-1)) at 40 °C achieved by K. marxianus PT-1 and S. cerevisiae JZ1C was 73.6 and 65.2 g L(-1), which corresponded to the theoretical ethanol yield of 90.0 and 79.7 %, respectively. In the range of 30 to 40 °C, temperature did not have a significant effect on ethanol production for both strains. This study displayed the distinctive superiority of K. marxianus PT-1 and S. cerevisiae JZ1C in the thermotolerance and utilization of inulin-type oligosaccharides reserved in Jerusalem artichoke tubers. It is proposed that both K. marxianus and S. cerevisiae have considerable potential in ethanol production from Jerusalem artichoke tubers by a high temperature CBP.

  20. Thermotolerant Kluyveromyces marxianus and Saccharomyces cerevisiae strains representing potentials for bioethanol production from Jerusalem artichoke by consolidated bioprocessing

    Energy Technology Data Exchange (ETDEWEB)

    Hu, Nan [Agricultural Univ., Qingdao, SD (China). College of Animal Science and Technology; Chinese Academy of Sciences, Qingdao, SD (China). Key Lab. of Biofuels; Yuan, Bo; Wang, Shi-An; Li, Fu-Li [Chinese Academy of Sciences, Qingdao, SD (China). Key Lab. of Biofuels; Sun, Juan [Agricultural Univ., Qingdao, SD (China). College of Animal Science and Technology

    2012-09-15

    Thermotolerant inulin-utilizing yeast strains are desirable for ethanol production from Jerusalem artichoke tubers by consolidated bioprocessing (CBP). To obtain such strains, 21 naturally occurring yeast strains isolated by using an enrichment method and 65 previously isolated Saccharomyces cerevisiae strains were investigated in inulin utilization, extracellular inulinase activity, and ethanol fermentation from inulin and Jerusalem artichoke tuber flour at 40 C. The strains Kluyveromyces marxianus PT-1 (CGMCC AS2.4515) and S. cerevisiae JZ1C (CGMCC AS2.3878) presented the highest extracellular inulinase activity and ethanol yield in this study. The highest ethanol concentration in Jerusalem artichoke tuber flour fermentation (200 g L{sup -1}) at 40 C achieved by K. marxianus PT-1 and S. cerevisiae JZ1C was 73.6 and 65.2 g L{sup -1}, which corresponded to the theoretical ethanol yield of 90.0 and 79.7 %, respectively. In the range of 30 to 40 C, temperature did not have a significant effect on ethanol production for both strains. This study displayed the distinctive superiority of K. marxianus PT-1 and S. cerevisiae JZ1C in the thermotolerance and utilization of inulin-type oligosaccharides reserved in Jerusalem artichoke tubers. It is proposed that both K. marxianus and S. cerevisiae have considerable potential in ethanol production from Jerusalem artichoke tubers by a high temperature CBP. (orig.)

  1. Molecular Detection of Two Potential Probiotic Lactobacilli Strains and Evaluation of Their Performance as Starter Adjuncts in Yogurt Production

    Directory of Open Access Journals (Sweden)

    Georgia Saxami

    2016-05-01

    Full Text Available A molecular method for efficient and accurate detection and identification of two potential probiotic lactobacilli strains isolated from fermented olives, namely Lactobacillus pentosus B281 and Lb. plantarum B282, was developed in the present study. Random Amplified Polymorphic DNA (RAPD analysis was performed, and strain specific primers were designed and applied in a multiplex polymerase chain reaction (PCR assay. The specificity of the assay was tested and successfully confirmed in 27 and 22 lactobacilli strains for Lb. pentosus B281 and Lb. plantarum B282, respectively. Moreover, the two strains were used as starter cultures in yogurt production. Cell enumeration followed by multiplex PCR analysis demonstrated that the two strains were present in yogurt samples at levels ≥6 log CFU/g even after 35 days of storage at 4 °C. Microbiological analysis showed that lactobacilli and streptococci were present within usual levels, whereas enterobacteriaceae and yeast/mold counts were not detected as expected. Although the pH values of the novel products were slightly lower than the control ones, the yogurt containing the probiotic cultures scored similar values compared to the control in a series of sensory tests. Overall, these results demonstrated the possible use of the two strains as starter adjuncts in the production of yogurt with potential probiotic properties.

  2. Functional properties of Lactobacillus plantarum strains isolated from Maasai traditional fermented milk products in Kenya.

    Science.gov (United States)

    Mathara, Julius Maina; Schillinger, Ulrich; Kutima, Phillip M; Mbugua, Samuel K; Guigas, Claudia; Franz, Charles; Holzapfel, Wilhelm H

    2008-04-01

    Lactobacillus plantarum was the major species among the lactic acid bacterial strains isolated from traditional fermented milk of the Maasai in Kenya. Selected strains were characterized for their functional properties using in vitro standard procedures. All strains expressed acid tolerance at pH 2.0 after 2-h exposure of values that ranged from 1% to 100%, while bile tolerance of acid-stressed cells at 0.3% oxgal varied from 30% to 80%. In vitro adhesion to the mucus-secreting cell line HT 29 MTX and binding capacity to extracellular protein matrices was demonstrated for several strains. The four strains tested in a simulated stomach duodenum passage survived with recovery rates ranging from 17% to 100%. Strains were intrinsically resistant to several antibiotics tested. From these in vitro studies, a number of Lb. plantarum strains isolated from the Maasai traditional fermented milk showed probiotic potential. The strains are good candidates for multifunctional starter culture development.

  3. High-yield production of herbicidal thaxtomins and analogs in a nonpathogenic Streptomyces strain.

    Science.gov (United States)

    Jiang, Guangde; Zhang, Yucheng; Powell, Magan M; Zhang, Peilan; Zuo, Ran; Zhang, Yi; Kallifidas, Dimitrios; Tieu, Albert M; Luesch, Hendrik; Loria, Rosemary; Ding, Yousong

    2018-03-30

    Thaxtomins are virulence factors of most plant pathogenic Streptomyces strains. Due to their potent herbicidal activity, attractive environmental compatibility and inherent biodegradability, thaxtomins are key active ingredients of bioherbicides approved by the United States Environmental Protection Agency. However, the low yield of thaxtomins in native Streptomyces producers limits their wide agricultural applications. Here, we describe the high-yield production of thaxtomins in a heterologous host. The thaxtomin gene cluster from S. scabiei 87.22 was cloned and expressed in S. albus J1074 after chromosomal integration. The production of thaxtomins and nitro-tryptophan analogs were observed using LC-MS analysis. When culturing the engineered S. albus J1074 in the minimal medium TMDc, the yield of the most abundant and herbicidal analog, thaxtomin A, was 10 times higher than S. scabiei 87.22, and optimization of the medium resulted in the highest yield of thaxtomin analogs at about 222 mg/L. Further engineering of the thaxtomin biosynthetic gene cluster through gene deletion led to the production of multiple biosynthetic intermediates important to the chemical synthesis of new analogs. Additionally, the versatility of the thaxtomin biosynthetic system in S. albus J1074 was capitalized to produce one unnatural fluorinated analog 5-F-thaxtomin A, whose structure was elucidated by a combination of MS and 1D and 2D NMR analyses. Natural and unnatural thaxtomins demonstrated potent herbicidal activity in radish seedling assays. These results indicated that S. albus J1074 has the potential to produce thaxtomins and thereof with high yield, fostering their agricultural applications. IMPORTANCE Thaxtomins are agriculturally valuable herbicidal natural products but the productivity of native producers is limiting. Heterologous expression of thaxtomin gene cluster in S. albus J1074 resulted in the highest yield of thaxtomins ever reported, representing a significant leap

  4. Demonstration of persistent contamination of a cooked egg product production facility with Salmonella enterica serovar Tennessee and characterization of the persistent strain.

    Science.gov (United States)

    Jakočiūnė, D; Bisgaard, M; Pedersen, K; Olsen, J E

    2014-08-01

    The aim of this study was to investigate whether continuous contamination of light pasteurized egg products with Salmonella enterica serovar Tennessee (S. Tennessee) at a large European producer of industrial egg products was caused by persistent contamination of the production facility and to characterize the persistent strains. Seventy-three S. Tennessee isolates collected from products over a 3-year period with intermittent contamination, and 15 control strains were compared by pulsed field gel electrophoresis (PFGE) using two enzymes. Forty-five case isolates distributed throughout the full period were shown to belong to one profile type. Isolates representing different PFGE profiles were all assigned to ST 319 by multilocus sequence typing (MLST). The case isolates did not show a higher ability to form biofilm on a plastic surface than noncase isolates. Characteristically, members of the persistent clone were weak producers of H2 S in laboratory medium. S. Tennessee isolated from the case was able to grow better in pasteurized egg product compared with other serovars investigated. It was concluded that the contamination was caused by a persistent strain in the production facility and that this strain apparently had adapted to grow in the relevant egg product. S. Tennessee has previously been associated with persistence in hatching facilities. This is the first report of persistent contamination of an egg production facility with this serovar. © 2014 The Society for Applied Microbiology.

  5. Demonstration of persistent contamination of a cooked egg product production facility with Salmonella enterica serovar Tennessee and characterization of the persistent strain

    DEFF Research Database (Denmark)

    Jakociune, D.; Bisgaard, M.; Pedersen, Karl

    2014-01-01

    Aims: The aim of this study was to investigate whether continuous contamination of light pasteurized egg products with Salmonella enterica serovar Tennessee (S. Tennessee) at a large European producer of industrial egg products was caused by persistent contamination of the production facility......, members of the persistent clone were weak producers of H2S in laboratory medium. S. Tennessee isolated from the case was able to grow better in pasteurized egg product compared with other serovars investigated. Conclusions: It was concluded that the contamination was caused by a persistent strain...... in the production facility and that this strain apparently had adapted to grow in the relevant egg product. Significance and Impact of the Study: S. Tennessee has previously been associated with persistence in hatching facilities. This is the first report of persistent contamination of an egg production facility...

  6. Production traits of broiler chicken strains fed Ad Libitum and raised ...

    African Journals Online (AJOL)

    ... of commercial broiler chickens reared on full-feeding, and under the same housing, feeding regime, agro-climatic zone and management practices. A total number of 150 broiler dayold chicks, that, is 50 chicks each of Arbor Acres (strain A), Hubbard (strain B) and Marshall (strain C) were purchased from local hatcheries, ...

  7. Light affects fumonisin production in strains of Fusarium fujikuroi, Fusarium proliferatum, and Fusarium verticillioides isolated from rice.

    Science.gov (United States)

    Matić, Slavica; Spadaro, Davide; Prelle, Ambra; Gullino, Maria Lodovica; Garibaldi, Angelo

    2013-09-16

    Three Fusarium species associated with bakanae disease of rice (Fusarium fujikuroi, Fusarium proliferatum, and Fusarium verticillioides) were investigated for their ability to produce fumonisins (FB1 and FB2) under different light conditions, and for pathogenicity. Compared to darkness, the conditions that highly stimulated fumonisin production were yellow and green light in F. verticillioides strains; white and blue light, and light/dark alternation in F. fujikuroi and F. proliferatum strains. In general, all light conditions positively influenced fumonisin production with respect to the dark. Expression of the FUM1 gene, which is necessary for the initiation of fumonisin production, was in accordance with the fumonisin biosynthetic profile. High and low fumonisin-producing F. fujikuroi strains showed typical symptoms of bakanae disease, abundant fumonisin-producing F. verticillioides strains exhibited chlorosis and stunting of rice plants, while fumonisin-producing F. proliferatum strains were asymptomatic on rice. We report that F. fujikuroi might be an abundant fumonisin producer with levels comparable to that of F. verticillioides and F. proliferatum, highlighting the need of deeper mycotoxicological analyses on rice isolates of F. fujikuroi. Our results showed for the first time the influence of light on fumonisin production in isolates of F. fujikuroi, F. proliferatum, and F. verticillioides from rice. © 2013 Elsevier B.V. All rights reserved.

  8. Gene expression cross-profiling in genetically modified industrial Saccharomyces cerevisiae strains during high-temperature ethanol production from xylose.

    Science.gov (United States)

    Ismail, Ku Syahidah Ku; Sakamoto, Takatoshi; Hatanaka, Haruyo; Hasunuma, Tomohisa; Kondo, Akihiko

    2013-01-10

    Production of ethanol from xylose at high temperature would be an economical approach since it reduces risk of contamination and allows both the saccharification and fermentation steps in SSF to be running at elevated temperature. Eight recombinant xylose-utilizing Saccharomyces cerevisiae strains developed from industrial strains were constructed and subjected to high-temperature fermentation at 38 °C. The best performing strain was sun049T, which produced up to 15.2 g/L ethanol (63% of the theoretical production), followed by sun048T and sun588T, both with 14.1 g/L ethanol produced. Via transcriptomic analysis, expression profiling of the top three best ethanol producing strains compared to a negative control strain, sun473T, led to the discovery of genes in common that were regulated in the same direction. Identification of the 20 most highly up-regulated and the 20 most highly down-regulated genes indicated that the cells regulate their central metabolism and maintain the integrity of the cell walls in response to high temperature. We also speculate that cross-protection in the cells occurs, allowing them to maintain ethanol production at higher concentration under heat stress than the negative controls. This report provides further transcriptomics information in the interest of producing a robust microorganism for high-temperature ethanol production utilizing xylose. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Mechanism of protection of transepithelial barrier function by Lactobacillus salivarius: strain dependence and attenuation by bacteriocin production.

    Science.gov (United States)

    Miyauchi, Eiji; O'Callaghan, John; Buttó, Ludovica F; Hurley, Gráinne; Melgar, Silvia; Tanabe, Soichi; Shanahan, Fergus; Nally, Kenneth; O'Toole, Paul W

    2012-11-01

    Enhanced barrier function is one mechanism whereby commensals and probiotic bacteria limit translocation of foreign antigens or pathogens in the gut. However, barrier protection is not exhibited by all probiotic or commensals and the strain-specific molecules involved remain to be clarified. We evaluated the effects of 33 individual Lactobacillus salivarius strains on the hydrogen peroxide (H(2)O(2))-induced barrier impairment in human epithelial Caco-2 cells. These strains showed markedly different effects on H(2)O(2)-induced reduction in transepithelial resistance (TER). The effective strains such as UCC118 and CCUG38008 attenuated H(2)O(2)-induced disassembly and relocalization of tight junction proteins, but the ineffective strain AH43324 did not. Strains UCC118 and CCUG38008 induced phosphorylation of extracellular signal-regulated kinase (ERK) in Caco-2 cells, and the ERK inhibitor U0126 attenuated the barrier-protecting effect of these strains. In contrast, the AH43324 strain induced phosphorylation of Akt and p38, which was associated with an absence of a protective effect. Global transcriptome analysis of UCC118 and AH43324 revealed that some genes in a bacteriocin gene cluster were upregulated in AH43324 under TER assay conditions. A bacteriocin-negative UCC118 mutant displayed significantly greater suppressive effect on H(2)O(2)-induced reduction in TER compared with wild-type UCC118. The wild-type strain augmented H(2)O(2)-induced phosphorylation of Akt and p38, whereas a bacteriocin-negative UCC118 mutant did not. These observations indicate that L. salivarius strains are widely divergent in their capacity for barrier protection, and this is underpinned by differences in the activation of intracellular signaling pathways. Furthermore, bacteriocin production appears to have an attenuating influence on lactobacillus-mediated barrier protection.

  10. Population Genetic Structure of Listeria monocytogenes Strains Isolated From the Pig and Pork Production Chain in France

    Directory of Open Access Journals (Sweden)

    Benjamin Félix

    2018-04-01

    Full Text Available Listeria monocytogenes is an ubiquitous pathogenic bacterium, transmissible to humans through the consumption of contaminated food. The pork production sector has been hit hard by a series of L. monocytogenes-related food poisoning outbreaks in France. An overview of the diversity of strains circulating at all levels of the pork production chain, from pig farming (PF to finished food products (FFP, is needed to identify the contamination routes and improve food safety. Until now, no typing data has been available on strains isolated across the entire pig and pork production chain. Here, we analyzed the population genetic structure of 687 L. monocytogenes strains isolated over the last 20 years in virtually all the French départements from three compartments of this production sector: PF, the food processing environment (FPE, and FFP. The genetic structure was described based on Multilocus sequence typing (MLST clonal complexes (CCs. The CCs were obtained by mapping the PFGE profiles of the strains. The distribution of CCs was compared firstly between the three compartments and then with CCs obtained from 1106 strains isolated from other food production sectors in France. The predominant CCs of pig and pork strains were not equally distributed among the three compartments: the CC37, CC59, and CC77 strains, rarely found in FPE and FFP, were prevalent in PF. The two most prevalent CCs in the FPE and FFP compartments, CC9 and CC121, were rarely or never detected in PF. No CC was exclusively associated with the pork sector. Three CCs (CC5, CC6, and CC2 were considered ubiquitous, because they were observed in comparable proportions in all food production sectors. The two most prevalent CCs in all sectors were CC9 and CC121, but their distribution was disparate. CC9 was associated with meat products and food products combining several food categories, whereas CC121 was not associated with any given sector. Based on these results, CC121 is likely able

  11. Population Genetic Structure of Listeria monocytogenes Strains Isolated From the Pig and Pork Production Chain in France

    Science.gov (United States)

    Félix, Benjamin; Feurer, Carole; Maillet, Aurelien; Guillier, Laurent; Boscher, Evelyne; Kerouanton, Annaëlle; Denis, Martine; Roussel, Sophie

    2018-01-01

    Listeria monocytogenes is an ubiquitous pathogenic bacterium, transmissible to humans through the consumption of contaminated food. The pork production sector has been hit hard by a series of L. monocytogenes-related food poisoning outbreaks in France. An overview of the diversity of strains circulating at all levels of the pork production chain, from pig farming (PF) to finished food products (FFP), is needed to identify the contamination routes and improve food safety. Until now, no typing data has been available on strains isolated across the entire pig and pork production chain. Here, we analyzed the population genetic structure of 687 L. monocytogenes strains isolated over the last 20 years in virtually all the French départements from three compartments of this production sector: PF, the food processing environment (FPE), and FFP. The genetic structure was described based on Multilocus sequence typing (MLST) clonal complexes (CCs). The CCs were obtained by mapping the PFGE profiles of the strains. The distribution of CCs was compared firstly between the three compartments and then with CCs obtained from 1106 strains isolated from other food production sectors in France. The predominant CCs of pig and pork strains were not equally distributed among the three compartments: the CC37, CC59, and CC77 strains, rarely found in FPE and FFP, were prevalent in PF. The two most prevalent CCs in the FPE and FFP compartments, CC9 and CC121, were rarely or never detected in PF. No CC was exclusively associated with the pork sector. Three CCs (CC5, CC6, and CC2) were considered ubiquitous, because they were observed in comparable proportions in all food production sectors. The two most prevalent CCs in all sectors were CC9 and CC121, but their distribution was disparate. CC9 was associated with meat products and food products combining several food categories, whereas CC121 was not associated with any given sector. Based on these results, CC121 is likely able to colonize a

  12. Penicillium salamii strain ITEM 15302: A new promising fungal starter for salami production.

    Science.gov (United States)

    Magistà, D; Ferrara, M; Del Nobile, M A; Gammariello, D; Conte, A; Perrone, G

    2016-08-16

    Traditional sausages are often considered of superior quality to sausages inoculated with commercial starter cultures and this is partially due to the action of the typical house microflora. Penicillium nalgiovense is the species commonly used as starter culture for dry-cured meat production. Recently a new species, Penicillium salamii, was described as typical colonizer during salami seasoning. In order to understand its contribution to the seasoning process, two different experiments on curing of fresh pork sausages were conducted using P. salamii ITEM 15302 in comparison with P. nalgiovense ITEM 15292 at small and industrial scale, and the dry-cured sausages were subjected to sensory analyses. Additionally, proteolytic and lipolytic in vitro assays were performed on both strains. P. salamii ITEM 15302 proved to be a fast growing mould on dry-cured sausage casings, well adapted to the seasoning process, with high lipolytic and proteolytic enzymatic activity that confers typical sensory characteristics to meat products. Therefore, P. salamii ITEM 15302 was shown to be a good candidate as new starter for meat industry. Copyright © 2015 Elsevier B.V. All rights reserved.

  13. 'Synthetic lipase' production from a newly isolated Sporidiobolus pararoseus strain by submerged fermentation

    Directory of Open Access Journals (Sweden)

    Alessandra Smaniotto

    2012-12-01

    Full Text Available The lipase produced by a newly isolate Sporidiobolus pararoseus strain has potential catalysis ability for esterification reactions. In order to improve its synthetic activity, this work aimed at optimizing 'synthetic lipase' production by submerged fermentation of a conventional media based on peptone, yeast extract, NaCl and olive oil using experimental design technique. According to the results obtained in the first experimental design (2(4-1, yeast extract and NaCl concentrations were tested to further optimization by response surface methodology. The maximum 'synthetic lipase' activity obtained was 26.9 U/mL in the optimized media (5.0, 6.8, 7.0 and 1.0% (wt/v of peptone, yeast extract, NaCl and olive oil, respectively, representing a 6.36-fold increase compared to the initial medium. The time course of 'synthetic lipase' production in the optimized condition was evaluated in terms of synthetic activity, protease activity, biomass and total carbon and the maximum synthetic activity was observed during the stationary phase of growth.

  14. Engineering biotin prototrophic Corynebacterium glutamicum strains for amino acid, diamine and carotenoid production.

    Science.gov (United States)

    Peters-Wendisch, P; Götker, S; Heider, S A E; Komati Reddy, G; Nguyen, A Q; Stansen, K C; Wendisch, V F

    2014-12-20

    The Gram-positive Corynebacterium glutamicum is auxotrophic for biotin. Besides the biotin uptake system BioYMN and the transcriptional regulator BioQ, this bacterium possesses functional enzymes for the last three reactions of biotin synthesis starting from pimeloyl-CoA. Heterologous expression of bioF from the Gram-negative Escherichia coli enabled biotin synthesis from pimelic acid added to the medium, but expression of bioF together with bioC and bioH from E. coli did not entail biotin prototrophy. Heterologous expression of bioWAFDBI from Bacillus subtilis encoding another biotin synthesis pathway in C. glutamicum allowed for growth in biotin-depleted media. Stable growth of the recombinant was observed without biotin addition for eight transfers to biotin-depleted medium while the empty vector control stopped growth after the first transfer. Expression of bioWAFDBI from B. subtilis in C. glutamicum strains overproducing the amino acids l-lysine and l-arginine, the diamine putrescine, and the carotenoid lycopene, respectively, enabled formation of these products under biotin-depleted conditions. Thus, biotin-prototrophic growth and production by recombinant C. glutamicum were achieved. Copyright © 2014 Elsevier B.V. All rights reserved.

  15. Study of itaconic acid production by Aspergillus terrus MJL05 strain with different variable

    Directory of Open Access Journals (Sweden)

    Mariana Juy

    2010-07-01

    Full Text Available Título en español: Estudio de la producción de ácido itacónico con Aspergillus terreus de la cepa MJL05 con diferentes variables Abstract Itaconic acid (IA production by Aspergillus terreus MJL05 strain was investigated in submerged batch fermentation in a stirred bioreactor to determine the effect of varying the nitrogen, phosphorous and carbon sources concentrations in the production medium. Glycerol, a biodiesel by-product was reported as an efficient substrate to achieve high itaconic acid productivities. This was used as the sole carbon source. The resulting C:N 18, N:P 10.8 and C:P 195 ratios were selected as the best and allowed to improve IA concentration from 11.0 to 27.6 g/l with a volumetric productivity of 0.192 g IA/l.h and a specific productivity 0.013 g IA/g biomass.h. Bioprocess yields, Yx:s 0.27 g d.w. biomass/g substrate; Yp:x 1.63 g IA/g d.w. biomass and Yp:s 0.44 g IA/g substrate, allowed to assume the feasibility of using this strain for IA production. Key words: Itaconic acid; Aspergillus terreus; glycerol. Resumen La producción de ácido itacónico (AI con Aspergillus terreus MJL05 se realizó en fermentación sumergida en lote en un biorreactor agitado para estudiar el efecto de la variación de las concentraciones de nitrógeno, fósforo y carbono en el medio de producción. El glicerol, subproducto del biodiesel, fue reportado como un sustrato eficiente para obtener altas productividades de AI. Este fue utilizado como única fuente de carbono. Las relaciones entre nutrientes, C:N 18, N:P 10,8 y C:P 195 fueron seleccionadas como las mejores para aumentar la concentración de AI de 11,0 a 27,6 g/l con un a productividad volumétrica de 0,192 g IA/l.h, y una productividad específica de 0,013 g IA/g biomasa.h. Los rendimientos del bioproceso obtenidos fueron de Yx:s 0,27 g p.s. biomasa/g sustrato; Yp:x 1,63 g IA/g p.s. biomasa y Yp:s 0,44 g IA/g sustrato, lo que permite asumir la factibilidad de usar esta cepa para

  16. Recent advances and strategies in process and strain engineering for the production of butyric acid by microbial fermentation.

    Science.gov (United States)

    Luo, Hongzhen; Yang, Rongling; Zhao, Yuping; Wang, Zhaoyu; Liu, Zheng; Huang, Mengyu; Zeng, Qingwei

    2018-04-01

    Butyric acid is an important platform chemical, which is widely used in the fields of food, pharmaceutical, energy, etc. Microbial fermentation as an alternative approach for butyric acid production is attracting great attention as it is an environmentally friendly bioprocessing. However, traditional fermentative butyric acid production is still not economically competitive compared to chemical synthesis route, due to the low titer, low productivity, and high production cost. Therefore, reduction of butyric acid production cost by utilization of alternative inexpensive feedstock, and improvement of butyric acid production and productivity has become an important target. Recently, several advanced strategies have been developed for enhanced butyric acid production, including bioprocess techniques and metabolic engineering methods. This review provides an overview of advances and strategies in process and strain engineering for butyric acid production by microbial fermentation. Additionally, future perspectives on improvement of butyric acid production are also proposed. Copyright © 2018 Elsevier Ltd. All rights reserved.

  17. Genetic and phenotypic diversity of naturally isolated wild strains of Aspergillus niger with hyper glucose oxidase production

    Directory of Open Access Journals (Sweden)

    MAHMOUD EL-HARIRI

    2015-12-01

    Full Text Available Glucose oxidase (GOx is the basic stone for many of biological industry worldwide. The improvement of GOx production basically depends on selection of hyper producer strain of Aspergillus niger. Selective isolation and screening for natural hyper producer strains of A. niger and sequence analysis of the GOD gene, which is responsible for production of the enzyme, are very effective approaches to investigate the naturally modified strains of A. niger with hyper productive capacity of GOx enzyme. The aims of the current study were selective isolation of naturally hyper GOx producing strains of A. niger and evaluation of their GOx activities under optimized parameters in the laboratory. Five wild Egyptian isolates of A. niger were screened for GOx and catalase activity using two types of modified basal liquid media. The GOx activity was evaluated by high throughout liquid phase system. The isolates showed a variable activity for GOx production ranged from 0 to 28.7 U.ml-1. One isolate coded Strain 7 was negative GOx producer on Vogel's broth medium in comparison to other isolates, while its GOx activity on Cazpek Dox was considered as positive (7.28 U.ml-1. It was concluded that GOx production is affected by three controllable factors – the basal media components, time of incubation, and the strain with its adaption to the media components‎. Also, the catalase activity was tested and it was produced with a different degree of variability, which may be reflected on GOx stability. GOD genes of these wild variant of A. niger were cloned and sequenced to determine intraspecies diversity of GOD between the wild variants. The comparison of isolated wild variants to other reference hyper GOx producer strains of A. niger was performed to determine if the GOD sequence analysis of these strains can be distinguished based on their GOx activity. This is the first report for isolation and detection of naturally A. niger hyper GOx-producer strains with

  18. Proteomic Analysis of Anti-Cancerous Scopularide Production by a Marine Microascus brevicaulis Strain and Its UV Mutant

    DEFF Research Database (Denmark)

    Kramer, Annemarie; Beck, Hans Christian; Kumar, Abhishek

    2015-01-01

    The marine fungus Microascus brevicaulis strain LF580 is a non-model secondary metabolite producer with high yields of the two secondary metabolites scopularides A and B, which exhibit distinct activities against tumour cell lines. A mutant strain was obtained using UV mutagenesis, showing faster...... for optimisation on strain and process level. The linkage between nutrient limitation and pellet formation in the non-model fungus M. brevicaulis is in consensus with the knowledge on model organisms like Aspergillus niger and Penicillium chrysogenum....... growth and differences in pellet formation besides higher production levels. Here, we show the first proteome study of a marine fungus. Comparative proteomics were applied to gain deeper understanding of the regulation of production and of the physiology of the wild type strain and its mutant...... in the mutant strain: 189 were down- and 129 upregulated. Proteomics are a powerful tool for the understanding of regulatory aspects: The differences on proteome level could be attributed to limited nutrient availability in the wild type strain due to a strong pellet formation. This information can be applied...

  19. Degradation of 2,4-dichlorophenoxyacetic acid by a halotolerant strain of Penicillium chrysogenum: antibiotic production.

    Science.gov (United States)

    Ferreira-Guedes, Sumaya; Mendes, Benilde; Leitão, Ana Lúcia

    2012-01-01

    The extensive use of pesticides in agriculture has prompted intensive research on chemical and biological methods in order to protect contamination of water and soil resources. In this paper the degradation of the pesticide 2,4-dichlorophenoxyacetic acid by a Penicillium chrysogenum strain previously isolated from a salt mine was studied in batch cultures. Co-degradation of 2,4-dichlorophenoxyacetic acid with additives such as sugar and intermediates of pesticide metabolism was also investigated. Penicillium chrysogenum in solid medium was able to grow at concentrations up to 1000 mg/L of 2,4-dichlorophenoxyacetic acid (2,4-D) with sucrose. Meanwhile, supplementation of the solid medium with glucose and lactose led to fungal growth at concentrations up to 500 mg/L of herbicide. Batch cultures of 2,4-D at 100 mg/L were developed under aerobic conditions with the addition of glucose, lactose and sucrose, showing sucrose as the best additional carbon source. The 2,4-D removal was quantified by liquid chromatography. The fungus was able to use 2,4-D as the sole carbon and energy source under 0%, 2% and 5.9% NaCl. The greatest 2,4-D degradation efficiency was found using alpha-ketoglutarate and ascorbic acid as co-substrates under 2% NaCl at pH 7. Penicillin production was evaluated in submerged cultures by bioassay, and higher amounts of beta-lactam antibiotic were produced when the herbicide was alone. Taking into account the ability of P. chrysogenum CLONA2 to degrade aromatic compounds, this strain could be an interesting tool for 2,4-D herbicide remediation in saline environments.

  20. Genetic strain and diet effects on grazing behavior, pasture intake, and milk production.

    Science.gov (United States)

    Sheahan, A J; Kolver, E S; Roche, J R

    2011-07-01

    Understanding how dairy cows adjust their grazing behavior in response to feed supplements is important for the development of management strategies that optimize profit from supplementation. New Zealand Holstein-Friesian (HF) cows have been selected for milk production on a predominantly pasture-based diet; in comparison, HF cows of North American (NA) ancestry have been selected almost exclusively for milk yield and fed diets high in nonfiber carbohydrates (NFC). We hypothesized, therefore, that supplementation would have differing effects on grazing behavior, pasture dry matter intake (DMI), and milk production in these genetic strains at peak, mid, and late lactation. A study was conducted over 2 consecutive lactations, with NA and NZ cows randomly allocated at calving to 0, 3, or 6 kg of dry matter/day concentrate plus unrestricted access to pasture. Pasture DMI, milk production, and grazing behavior were recorded at peak, mid, and late lactation. Concentrates were fed in equal amounts at morning and afternoon milking. The NA cows produced more milk and milk components, and had a greater pasture DMI, despite spending less time grazing. Declines in time spent grazing and pasture DMI were associated with increasing concentrate DMI. Grazing behavior following morning supplementation was different from that recorded following afternoon supplementation. Grazing ceased following morning supplementation before rumen fill could be a limiting factor, and the length of the grazing interval was inversely proportional to the amount of concentrate offered; these results suggest that physiological rather than physical stimuli were responsible for grazing cessation. The decrease in time spent grazing with increasing concentrate DMI is consistent with changes in neuroendocrine factors secreted in response to the presence of food in the digestive tract or with circulating products of digestion. After afternoon supplementation, sunset signaled the end of grazing irrespective of

  1. Employing a Recombinant Strain of Advenella mimigardefordensis for Biotechnical Production of Homopolythioesters from 3,3′-Dithiodipropionic Acid

    Science.gov (United States)

    Xia, Yongzhen; Wübbeler, Jan Hendrik; Qi, Qingsheng

    2012-01-01

    Advenella mimigardefordensis strain DPN7T was genetically modified to produce poly(3-mercaptopropionic acid) (PMP) homopolymer by exploiting the recently unraveled process of 3,3′-dithiodipropionic acid (DTDP) catabolism. Production was achieved by systematically engineering the metabolism of this strain as follows: (i) deletion of its inherent 3MP dioxygenase-encoding gene (mdo), (ii) introduction of the buk-ptb operon (genes encoding the butyrate kinase, Buk, and the phosphotransbutyrylase, Ptb, from Clostridium acetobutylicum), and (iii) overexpression of its own polyhydroxyalkanoate synthase (phaCAm). These measures yielded the potent PMP production strain A. mimigardefordensis strain SHX22. The deletion of mdo was required for adequate synthesis of PMP due to the resulting accumulation of 3MP during utilization of DTDP. Overexpression of the plasmid-borne buk-ptb operon caused a severe growth repression. This effect was overcome by inserting this operon into the genome. Polyhydroxyalkanoate (PHA) synthases from different origins were compared. The native PHA synthase of A. mimigardefordensis (phaCAm) was obviously the best choice to establish homopolythioester production in this strain. In addition, the cultivation conditions, including an appropriate provision of the carbon source, were further optimized to enhance PMP production. The engineered strain accumulated PMP up to approximately 25% (wt/wt) of the cell dry weight when cultivated in mineral salts medium containing glycerol as the carbon source in addition to DTDP as the sulfur-providing precursor. According to our knowledge, this is the first report of PMP homopolymer production by a metabolically engineered bacterium using DTDP, which is nontoxic, as the precursor substrate. PMID:22344658

  2. Novel thermostable clostridial strains through protoplast fusion for enhanced biobutanol production at higher temperature—preliminary study

    Directory of Open Access Journals (Sweden)

    Muhammad Ferhan

    2016-01-01

    Full Text Available The objective of this study is to improve the thermal stability of clostridium strains for enhanced biobutanol production. Thermostable clostridia species were developed through protoplast fusion between mesophilic clostridial species (i.e., Clostridium beijerinckii and Clostridium acetobutylicum and thermophilic clostridial species (i.e., Clostridium thermocellum. Production of biobutanol was examined in the present preliminary study using the clostridium strains and their protoplast fusants using sugar mixture with composition identical to that of wheat straw acid hydrolysate. Maximum biobutanol production of 9.4 g/L was achieved by a fused strain at 45 °C with total sugar consumption of 66% compared to that at 35 °C (i.e., 8.4 g/L production and 64% total sugar consumption. Glucose and xylose uptake rates were generally higher compared to all other individual sugars in the feedstock. In general, average cell concentrations were in close proximity for all parenting and fused strains at 35 °C; i.e., in the range of 5.12 × 107 to 5.49 × 107 cells/mL. Average cell concentration of fusants between the mesophilic clostridial species and the thermophilic clostridial species slightly increased to ~ 5.62 × 107 cells/mL at a higher temperature of 45 °C. These results, in addition to the ones obtained for the butanol production, demonstrate enhanced thermal stability of both fusants at a higher temperature (45 °C.

  3. DNAs from Brucella strains activate efficiently murine immune system with production of cytokines, reactive oxygen and nitrogen species.

    Science.gov (United States)

    Tavakoli, Zahra; Ardestani, Sussan K; Lashkarbolouki, Taghi; Kariminia, Amina; Zahraei Salehi, Taghi; Tavassoli, Nasser

    2009-09-01

    Brucellosis is an infectious disease with high impact on innate immune responses which is induced partly by its DNA. In the present study the potential differences of wild type and patients isolates versus attenuated vaccine strains in terms of cytokines, ROS and NO induction on murine splenocytes and peritoneal macrophages were investigated. This panel varied in base composition and included DNA from B. abortus, B. melitensis, B.abortus strain S19 and melitensis strain Rev1, as attenuated live vaccine. Also we included Escherichia coli DNA, calf thymus DNA (a mammalian DNA), as controls. These DNA were evaluated for their ability to stimulate IL-12, TNF-alpha, IL-10, IFN-gamma and ROS production from spleenocytes as well as NO production from peritoneal macrophages. Spleen cells were cultured in 24 well at a concentration of 106 cells/ ml with subsequent addition of 10 microg/ml of Brucella or Ecoli DNAs. These cultures were incubated at 37 degrees C with 5% CO2 for 5 days. Supernatants were harvested and cytokines, ROS and NOx were evaluated. It was observed that TNF-alpha was induced in days 1,3,5 by all Brucella strains DNAs and E. coli DNA, IL-10 only was induced in day 1, IFN- gamma was induced only in day 5 and IL-12 not induced. ROS and NOx were produced by all strains; however, we observed higher production of NOx which were stimulated by DNA of B. melitensis.

  4. Characterization of antimicrobial resistance in Salmonella enterica strains isolated from Brazilian poultry production.

    Science.gov (United States)

    Mattiello, Samara P; Drescher, Guilherme; Barth, Valdir C; Ferreira, Carlos A S; Oliveira, Sílvia D

    2015-11-01

    Antimicrobial resistance profiles and presence of resistance determinants and integrons were evaluated in Salmonella enterica strains from Brazilian poultry. The analysis of 203 isolates showed that those from the poultry environment (88 isolates) were significantly more resistant to antimicrobials than isolates from other sources, particularly those isolated from poultry by-product meal (106 isolates). Thirty-seven isolates were resistant to at least three antimicrobial classes. Class 1 integrons were detected in 26 isolates, and the analysis of the variable region between the 5' conserved segment (CS) and 3' CS of each class 1 integron-positive isolate showed that 13 contained a typical 3' CS and 14 contained an atypical 3' CS. One Salmonella Senftenberg isolate harbored two class 1 integrons, showing both typical and atypical 3' CSs. The highest percentage of resistance was found to sulfonamides, and sul genes were detected in the majority of the resistant isolates. Aminoglycoside resistance was detected in 50 isolates, and aadA and aadB were present in 28 and 32 isolates, respectively. In addition, strA and strB were detected in 78.1 and 65.6% isolates resistant to streptomycin, respectively. Twenty-one isolates presented reduced susceptibility to β-lactams and harbored bla(TEM), bla(CMY), and/or bla(CTX-M). Forty isolates showed reduced susceptibility to tetracycline, and most presented tet genes. These results highlight the importance of the environment as a reservoir of resistant Salmonella, which may enable the persistence of resistance determinants in the poultry production chain, contributing, therefore, to the debate regarding the impacts that antimicrobial use in animal production may exert in human health.

  5. Acetone-butanol-ethanol production from substandard and surplus dates by Egyptian native Clostridium strains.

    Science.gov (United States)

    Abd-Alla, Mohamed Hemida; Zohri, Abdel-Naser Ahmed; El-Enany, Abdel-Wahab Elsadek; Ali, Shimaa Mohamed

    2015-04-01

    One hundred and seven mesophilic isolates of Clostridium were isolated from agricultural soils cultivated with different plants in Assuit Governorate, Egypt. Eighty isolates (out of 107) showed the ability to produce ABE (Acetone, butanol and ethanol) on T6 medium ranging from 0.036 to 31.89 g/L. The highest numbers of ABE producing isolates were obtained from soil samples of potato contributing 27 isolates, followed by 18 isolates from wheat and 10 isolates from onion. On the other hand, there were three native isolates that produced ABE more than those produced by the reference isolate Clostridium acetobutylicum ATCC 824 (11.543 g/L). The three isolates were identified based on phenotypic and gene encoding 16S rRNA as Clostridium beijerinckii ASU10 (KF372577), Clostridium chauvoei ASU55 (KF372580) and Clostridium roseum ASU58 (KF372581). The highest ABE level from substandard and surplus dates was produced by C. beijerinckii ASU10 (24.07 g/L) comprising butanol 67.15% (16.16 g/L), acetone 30.73% (7.4 g/L) and ethanol 2.12% (0.51 g/L), while C. roseum ASU58 and C. chauvoei ASU55 produced ABE contributing 20.20 and 13.79 g/L, respectively. ABE production by C. acetobutylicum ATCC 824 was 15.01 g/L. This study proved that the native strains C. beijerinckii ASU10 and C. roseum ASU58 have high competitive efficacy on ABE production from economical substrate as substandard and surplus date fruits. Additionally, using this substrate without any nutritional components is considered to be a commercial substrate for desired ABE production. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Excessive by-product formation: A key contributor to low isobutanol yields of engineered Saccharomyces cerevisiae strains.

    Science.gov (United States)

    Milne, N; Wahl, S A; van Maris, A J A; Pronk, J T; Daran, J M

    2016-12-01

    It is theoretically possible to engineer Saccharomyces cerevisiae strains in which isobutanol is the predominant catabolic product and high-yielding isobutanol-producing strains are already reported by industry. Conversely, isobutanol yields of engineered S. cerevisiae strains reported in the scientific literature typically remain far below 10% of the theoretical maximum. This study explores possible reasons for these suboptimal yields by a mass-balancing approach. A cytosolically located, cofactor-balanced isobutanol pathway, consisting of a mosaic of bacterial enzymes whose in vivo functionality was confirmed by complementation of null mutations in branched-chain amino acid metabolism, was expressed in S. cerevisiae . Product formation by the engineered strain was analysed in shake flasks and bioreactors. In aerobic cultures, the pathway intermediate isobutyraldehyde was oxidized to isobutyrate rather than reduced to isobutanol. Moreover, significant concentrations of the pathway intermediates 2,3-dihydroxyisovalerate and α-ketoisovalerate, as well as diacetyl and acetoin, accumulated extracellularly. While the engineered strain could not grow anaerobically, micro-aerobic cultivation resulted in isobutanol formation at a yield of 0.018±0.003 mol/mol glucose. Simultaneously, 2,3-butanediol was produced at a yield of 0.649±0.067 mol/mol glucose. These results identify massive accumulation of pathway intermediates, as well as overflow metabolites derived from acetolactate, as an important, previously underestimated contributor to the suboptimal yields of 'academic' isobutanol strains. The observed patterns of by-product formation is consistent with the notion that in vivo activity of the iron-sulphur-cluster-requiring enzyme dihydroxyacid dehydratase is a key bottleneck in the present and previously described 'academic' isobutanol-producing yeast strains.

  7. Dinitrogenase-Driven Photobiological Hydrogen Production Combats Oxidative Stress in Cyanothece sp. Strain ATCC 51142

    Energy Technology Data Exchange (ETDEWEB)

    Sadler, Natalie C.; Bernstein, Hans C.; Melnicki, Matthew R.; Charania, Moiz A.; Hill, Eric A.; Anderson, Lindsey N.; Monroe, Matthew E.; Smith, Richard D.; Beliaev, Alexander S.; Wright, Aaron T.; Nojiri, H.

    2016-10-14

    ABSTRACT

    Photobiologically synthesized hydrogen (H2) gas is carbon neutral to produce and clean to combust, making it an ideal biofuel.Cyanothecesp. strain ATCC 51142 is a cyanobacterium capable of performing simultaneous oxygenic photosynthesis and H2production, a highly perplexing phenomenon because H2evolving enzymes are O2sensitive. We employed a system-levelin vivochemoproteomic profiling approach to explore the cellular dynamics of protein thiol redox and how thiol redox mediates the function of the dinitrogenase NifHDK, an enzyme complex capable of aerobic hydrogenase activity. We found that NifHDK responds to intracellular redox conditions and may act as an emergency electron valve to prevent harmful reactive oxygen species formation in concert with other cell strategies for maintaining redox homeostasis. These results provide new insight into cellular redox dynamics useful for advancing photolytic bioenergy technology and reveal a new understanding for the biological function of NifHDK.

    IMPORTANCEHere, we demonstrate that high levels of hydrogen synthesis can be induced as a protection mechanism against oxidative stress via the dinitrogenase enzyme complex inCyanothecesp. strain ATCC 51142. This is a previously unknown feature of cyanobacterial dinitrogenase, and we anticipate that it may represent a strategy to exploit cyanobacteria for efficient and scalable hydrogen production. We utilized a chemoproteomic approach to capture thein situdynamics of reductant partitioning within the cell, revealing proteins and reactive thiols that may be involved in redox sensing and signaling. Additionally, this method is widely applicable across biological systems to achieve a greater understanding of how cells

  8. Production of Phytotoxic Metabolite Using Biphasic Fermentation System from Strain C1136 of Lasiodiplodia pseudotheobromae, a Potential Bioherbicidal Agent

    Directory of Open Access Journals (Sweden)

    Charles Oluwaseun ADETUNJI

    2017-09-01

    Full Text Available Formulation of effective and environmental friendly bioherbicides depends on the type of fermentation medium used for the production of phytotoxic metabolites. The effect of biomass, colony forming unit and the phytotoxic metabolite produced from the biphasic fermentation was carried out, while the phytotoxic metabolite was tested in vivo and in-vitro on Echinochola crus-galli and dicotyledonous Chromolaena odorata. The mutant strain of Lasiodiplodia pseudotheobromae C1136 (Lp90 produced the highest amount of conidia and the largest necrotic area on the two tested weeds when compared to its wild strain in the different biphasic media combinations. The study revealed that the biphasic system containing PDB + rice produced the highest bioherbicidal activities. Therefore, the phytotoxic metabolites from strain C1136 are suggested for large scale production of bioherbicides for the management of weeds in conventional farming to improve yield and enhance food security.

  9. Strain and culture medium optimization for production enhancement of prodiginines from marine-derived Streptomyces sp. GQQ-10

    Science.gov (United States)

    Li, Xueping; Zhang, Guojian; Zhu, Tianjiao; Li, Dehai; Gu, Qianqun

    2012-09-01

    A mutant (GQQ-M6) of a Sponge-Derived streptomyces sp. GQQ-10 obtained by UV-induced mutation was used for producing prodiginines (PGs). Single factor experiments and orthogonal array design (OAD) methods were employed for medium optimization. In the single factor method, the effects of soluble starch, glucose, soybean flour, yeast extract and sodium acetate on PGs production were investigated individually. In the subsequent OAD experiments, the concentrations of these 5 key nutritional components combined with salinity were further adjusted. The mutant strain GQQ-M6 gave a 2.2-fold higher PGs production than that of the parent strain; OAD experiments offered a PGs yield of 61mg L-1, which was 10 times higher than that of the initial GQQ-10 strain under the original cultivation mode.

  10. Key Process Conditions for Production of C4 Dicarboxylic Acids in Bioreactor Batch Cultures of an Engineered Saccharomyces cerevisiae Strain

    NARCIS (Netherlands)

    Zelle, R.M.; De Hulster, E.; Kloezen, W.; Pronk, J.T.; Van Maris, A.J.A.

    2010-01-01

    A recent effort to improve malic acid production by Saccharomyces cerevisiae by means of metabolic engineering resulted in a strain that produced up to 59 g liter(-1) of malate at a yield of 0.42 mol (mol glucose)(-1) in calcium carbonate-buffered shake flask cultures. With shake flasks, process

  11. Debaryomyces hansenii strains differ in their production of flavor compounds in a cheese-surface model

    DEFF Research Database (Denmark)

    Gori, Klaus; Sørensen, Louise Marie; Petersen, Mikael Agerlin

    2012-01-01

    Flavor production among12 strains of Debaryomyces hansenii when grown on a simple cheese model mimicking a cheese surface was investigated by dynamic headspace sampling followed by gas chromatography-mass spectrometry. The present study confirmed that D. hansenii possess the ability to produce...

  12. Comparative Analysis of Culture Conditions for the Optimization of Carotenoid Production in Several Strains of the Picoeukaryote Ostreococcus

    Directory of Open Access Journals (Sweden)

    Jean-Baptiste Guyon

    2018-02-01

    Full Text Available Microalgae are promising sources for the sustainable production of compounds of interest for biotechnologies. Compared to higher plants, microalgae have a faster growth rate and can be grown in industrial photobioreactors. The microalgae biomass contains specific metabolites of high added value for biotechnology such as lipids, polysaccharides or carotenoid pigments. Studying carotenogenesis is important for deciphering the mechanisms of adaptation to stress tolerance as well as for biotechnological production. In recent years, the picoeukaryote Ostreococcus tauri has emerged as a model organism thanks to the development of powerful genetic tools. Several strains of Ostreococcus isolated from different environments have been characterized with respect to light response or iron requirement. We have compared the carotenoid contents and growth rates of strains of Ostreococcus (OTTH595, RCC802 and RCC809 under a wide range of light, salinity and temperature conditions. Carotenoid profiles and productivities varied in a strain-specific and stress-dependent manner. Our results also illustrate that phylogenetically related microalgal strains originating from different ecological niches present specific interests for the production of specific molecules under controlled culture conditions.

  13. Do new production concepts and a new management of employment relations, yield higher employee performance and low job strain?

    NARCIS (Netherlands)

    Kraan, K.; Oeij, P.

    2006-01-01

    In this article old versus new production concepts (NPCs) and employment relation instruments, are studied, separately and in combination, to find out which yield high employee performance and low job strain. Therefore, in 2005, TNO conducted coupled surveys among 149 supervisors and employees. In

  14. Combining Protein and Strain Engineering for the Production of Glyco-Engineered Horseradish Peroxidase C1A in Pichia pastoris

    Directory of Open Access Journals (Sweden)

    Simona Capone

    2015-09-01

    Full Text Available Horseradish peroxidase (HRP, conjugated to antibodies and lectins, is widely used in medical diagnostics. Since recombinant production of the enzyme is difficult, HRP isolated from plant is used for these applications. Production in the yeast Pichia pastoris (P. pastoris, the most promising recombinant production platform to date, causes hyperglycosylation of HRP, which in turn complicates conjugation to antibodies and lectins. In this study we combined protein and strain engineering to obtain an active and stable HRP variant with reduced surface glycosylation. We combined four mutations, each being beneficial for either catalytic activity or thermal stability, and expressed this enzyme variant as well as the unmutated wildtype enzyme in both a P. pastoris benchmark strain and a strain where the native α-1,6-mannosyltransferase (OCH1 was knocked out. Considering productivity in the bioreactor as well as enzyme activity and thermal stability, the mutated HRP variant produced in the P. pastoris benchmark strain turned out to be interesting for medical diagnostics. This variant shows considerable catalytic activity and thermal stability and is less glycosylated, which might allow more controlled and efficient conjugation to antibodies and lectins.

  15. Ninety six well microtiter plate as microbioreactors for production of itaconic acid by six Aspergillus terreus strains

    Science.gov (United States)

    Itaconic acid (IA) is a building block platform chemical that is currently produced industrially from glucose by fermentation with Aspergillus terreus. However, lignocellulosic biomass has the potential to serve as low cost source of sugars for production of IA. Previously, 100 A. terreus strains we...

  16. Transcriptome of Aspergillus flavus aswA (AFLA_085170) deletion strain related to sclerotial development and production of secondary metabolites

    Science.gov (United States)

    Aspergillus flavus produces many secondary metabolites including aflatoxins. Besides conidia, the fungus uses sclerotia as another type of propagule. We obtained transcriptomes from four growth conditions of the aswA mutant, a strain impaired in sclerotial development and production of sclerotium-sp...

  17. Ethanol production from kitchen waste using the flocculating yeast Saccharomyces cerevisiae strain KF-7

    Energy Technology Data Exchange (ETDEWEB)

    Tang, Yue-Qin; Liu, Kai; An, Ming-Zhe; Morimura, Shigeru; Kida, Kenji [Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Kumamoto 860-8555 (Japan); Koike, Yoji [Tokyo Gas Co., Ltd., 1-7-7 Suehiro-cho, Tsurumi-ku, Yokohama City, Kanagawa 230-0045 (Japan); Wu, Xiao-Lei [Department of Energy and Resources Engineering, College of Engineering, Peking University, Beijing 100871 (China)

    2008-11-15

    A process for producing ethanol from kitchen waste was developed in this study. The process consists of freshness preservation of the waste, saccharification of the sugars in the waste, continuous ethanol fermentation of the saccharified liquid, and anaerobic treatment of the saccharification residue and the stillage. Spraying lactic acid bacteria (LCB) on the kitchen waste kept the waste fresh for over 1 week. High glucose recovery (85.5%) from LCB-sprayed waste was achieved after saccharification using Nagase N-40 glucoamylase. The resulting saccharified liquid was used directly for ethanol fermentation, without the addition of any nutrients. High ethanol productivity (24.0 g l{sup -1} h{sup -1}) was obtained when the flocculating yeast strain KF-7 was used in a continuous ethanol fermentation process at a dilution rate of 0.8 h{sup -1}. The saccharification residue was mixed with stillage and treated in a thermophilic anaerobic continuous stirred tank reactor (CSTR); a VTS loading rate of 6 g l{sup -1} d{sup -1} with 72% VTS digestion efficiency was achieved. Using this process, 30.9 g ethanol, and 65.2 l biogas with 50% methane, was produced from 1 kg of kitchen waste containing 118.0 g total sugar. Thus, energy in kitchen waste can be converted to ethanol and methane, which can then be used as fuels, while simultaneously treating kitchen waste. (author)

  18. Production of fructanase by a wild strain of Saccharomyces cerevisiae on tequila agave fructan.

    Science.gov (United States)

    Corona-González, R I; Pelayo-Ortiz, C; Jacques, G; Guatemala, G; Arriola, E; Arias, J A; Toriz, G

    2015-01-01

    A new wild strain of Saccharomyces cerevisiae (CF3) isolated from tequila must was evaluated for production of fructanase on Agave tequilana Weber fructan (FT). Fructanase activity (F) was assessed by a 3(3) factorial design (substrate, temperature and pH). High enzymatic activity (31.1 U/ml) was found at 30 °C, pH 5, using FT (10 g/l) as substrate. The effect of initial substrate concentration on F (FT0, 5.7-66 g/l) was studied and it was found that F was highest (44.8 U/ml) at FT0 25 g/l. A 2(2) factorial experimental design with five central points was utilized to study the effect of stirring and aeration on fructanase activity; stirring exhibited a stronger effect on F. The ratio fructanase to invertase (F/S) was 0.57, which confirms that the enzymes are fructanase. Crude fructanase reached high substrate hydrolysis (48 wt%) in 10 h. It is shown that S. cerevisiae CF3 was able to produce large amounts of fructanase by growing it on fructan from A. tequilana.

  19. Production of Bio polymer (PHB) from Whey by Local Strain of Bacillus cereus

    International Nuclear Information System (INIS)

    Abdel Kareem, H.; Hamed, D.; Omar, S.; Gebreel, H.; Khalaf, M.; El-M-Mahalawy, A.

    2008-01-01

    The local strain Bacillus cereus S 3 , which isolated from the soil attached to the rice root, was employed for PHB production from whey and soya extract as the main carbon and nitrogen sources. Some supplements such as (0.5 g) tryptone and (0.5 g) NaCl were added to 75 ml whey and 25 ml soya extract to optimize the PHB accumulation medium. Different parameters including; initial ph of the medium, working volume, NaCl concentration and inoculum age and size; were carried out under shaking flask conditions (150 rpm) at 30 degree C for 48 h of incubation to enhance the PHB accumulation. The maximum PHB accumulation (2.42 gl -1 ) was achieved at ph 6, 100 ml working volume, (0.5-2%) NaCl, at 60 h and 4 ml inoculum age and size, respectively. An experiment was conducted to investigate the effect of gamma irradiation on the activity of B. cereus S 3 towards PHB accumulation. At dose level 1.5 kGy the maximum PHB accumulation obtained was 3.2 gl -1

  20. Ethanol production from kitchen waste using the flocculating yeast Saccharomyces cerevisiae strain KF-7

    International Nuclear Information System (INIS)

    Tang, Y.-Q.; Koike, Yoji; Liu Kai; An, M.-Z.; Morimura, Shigeru; Wu Xiaolei; Kida, Kenji

    2008-01-01

    A process for producing ethanol from kitchen waste was developed in this study. The process consists of freshness preservation of the waste, saccharification of the sugars in the waste, continuous ethanol fermentation of the saccharified liquid, and anaerobic treatment of the saccharification residue and the stillage. Spraying lactic acid bacteria (LCB) on the kitchen waste kept the waste fresh for over 1 week. High glucose recovery (85.5%) from LCB-sprayed waste was achieved after saccharification using Nagase N-40 glucoamylase. The resulting saccharified liquid was used directly for ethanol fermentation, without the addition of any nutrients. High ethanol productivity (24.0 g l -1 h -1 ) was obtained when the flocculating yeast strain KF-7 was used in a continuous ethanol fermentation process at a dilution rate of 0.8 h -1 . The saccharification residue was mixed with stillage and treated in a thermophilic anaerobic continuous stirred tank reactor (CSTR); a VTS loading rate of 6 g l -1 d -1 with 72% VTS digestion efficiency was achieved. Using this process, 30.9 g ethanol, and 65.2 l biogas with 50% methane, was produced from 1 kg of kitchen waste containing 118.0 g total sugar. Thus, energy in kitchen waste can be converted to ethanol and methane, which can then be used as fuels, while simultaneously treating kitchen waste

  1. In Vitro characterization of Lactococcus lactis strains Isolated from Iranian Traditional Dairy Products as a Potential Probiotic

    OpenAIRE

    Fatemeh Nejati; Tobias Oelschlaeger

    2015-01-01

    Few studies have been reported regarding probiotic properties of Lactococcus lactis strains although they are extensively used as starter cultures in the production of dairy products. In this study 8 wild isolates of Lactococcus lactis were evaluated in vitro with regard to resistance to simulated gastric and intestinal juices, adherence ability to Caco-2 cells and HT29-MTX-E12 cell lines, anti-microbial activity, hydrophobicity and antibiotic susceptibility. The results revealed that all iso...

  2. Novel endophytic yeast Rhodotorula mucilaginosa strain PTD3 I: production of xylitol and ethanol.

    Science.gov (United States)

    Bura, Renata; Vajzovic, Azra; Doty, Sharon L

    2012-07-01

    An endophytic yeast, Rhodotorula mucilaginosa strain PTD3, that was isolated from stems of hybrid poplar was found to be capable of production of xylitol from xylose, of ethanol from glucose, galactose, and mannose, and of arabitol from arabinose. The utilization of 30 g/L of each of the five sugars during fermentation by PTD3 was studied in liquid batch cultures. Glucose-acclimated PTD3 produced enhanced yields of xylitol (67% of theoretical yield) from xylose and of ethanol (84, 86, and 94% of theoretical yield, respectively) from glucose, galactose, and mannose. Additionally, this yeast was capable of metabolizing high concentrations of mixed sugars (150 g/L), with high yields of xylitol (61% of theoretical yield) and ethanol (83% of theoretical yield). A 1:1 glucose:xylose ratio with 30 g/L of each during double sugar fermentation did not affect PTD3's ability to produce high yields of xylitol (65% of theoretical yield) and ethanol (92% of theoretical yield). Surprisingly, the highest yields of xylitol (76% of theoretical yield) and ethanol (100% of theoretical yield) were observed during fermentation of sugars present in the lignocellulosic hydrolysate obtained after steam pretreatment of a mixture of hybrid poplar and Douglas fir. PTD3 demonstrated an exceptional ability to ferment the hydrolysate, overcome hexose repression of xylose utilization with a short lag period of 10 h, and tolerate sugar degradation products. In direct comparison, PTD3 had higher xylitol yields from the mixed sugar hydrolysate compared with the widely studied and used xylitol producer Candida guilliermondii.

  3. Construction of Potent Recombinant Strain Through Intergeneric Protoplast Fusion in Endophytic Fungi for Anticancerous Enzymes Production Using Rice Straw.

    Science.gov (United States)

    El-Gendy, Mervat Morsy Abbas Ahmed; Al-Zahrani, Salha Hassan Mastour; El-Bondkly, Ahmed Mohamed Ahmed

    2017-09-01

    Among all fungal endophytes isolates derived from different ethno-medical plants, the hyper-yield L-asparaginase and L-glutaminase wild strains Trichoderma sp. Gen 9 and Cladosporium sp. Gen 20 using rice straw under solid-state fermentation (SSF) were selected. The selected strains were used as parents for the intergeneric protoplast fusion program to construct recombinant strain for prompt improvement production of these enzymes in one recombinant strain. Among 21 fusants obtained, the recombinant strain AYA 20-1, with 2.11-fold and 2.58-fold increase in L-asparaginase and L-glutaminase activities more than the parental isolates Trichoderma sp. Gen 9 and Cladosporium sp. Gen 20, respectively, was achieved using rice straw under SSF. Both therapeutic enzymes L-asparaginase and L-glutaminase were purified and characterized from the culture supernatant of the recombinant AYA 20-1 strain with molecular weights of 50.6 and 83.2 kDa, respectively. Both enzymes were not metalloenzymes. Whereas thiol group blocking reagents such as p-chloromercurybenzoate and iodoacetamide totally inhibited L-asparaginase activity, which refer to sulfhydryl groups and cysteine residues involved in its catalytic activity, they have no effect toward L-glutaminase activity. Interestingly, potent anticancer, antioxidant, and antimicrobial activities were detected for both enzymes.

  4. Prophage-Encoded Staphylococcal Enterotoxin A: Regulation of Production in Staphylococcus aureus Strains Representing Different Sea Regions

    Directory of Open Access Journals (Sweden)

    Nikoleta Zeaki

    2015-12-01

    Full Text Available The present study investigates the nature of the link between the staphylococcal enterotoxin A (SEA gene and the lifecycle of Siphoviridae bacteriophages, including the origin of strain variation regarding SEA production after prophage induction. Five strains representing three different genetic lines of the sea region were studied under optimal and prophage-induced growth conditions and the Siphoviridae lifecycle was followed through the phage replicative form copies and transcripts of the lysogenic repressor, cro. The role of SOS response on prophage induction was addressed through recA transcription in a recA-disruption mutant. Prophage induction was found to increase the abundance of the phage replicative form, the sea gene copies and transcripts and enhance SEA production. Sequence analysis of the sea regions revealed that observed strain variances were related to strain capacity for prophage induction, rather than sequence differences in the sea region. The impact of SOS response activation on the phage lifecycle was demonstrated by the absence of phage replicative form copies in the recA-disruption mutant after prophage induction. From this study it emerges that all aspects of SEA-producing strain, the Siphoviridae phage and the food environment must be considered when evaluating SEA-related hazards.

  5. Strain typing of acetic acid bacteria responsible for vinegar production by the submerged elaboration method.

    Science.gov (United States)

    Fernández-Pérez, Rocío; Torres, Carmen; Sanz, Susana; Ruiz-Larrea, Fernanda

    2010-12-01

    Strain typing of 103 acetic acid bacteria isolates from vinegars elaborated by the submerged method from ciders, wines and spirit ethanol, was carried on in this study. Two different molecular methods were utilised: pulsed field gel electrophoresis (PFGE) of total DNA digests with a number of restriction enzymes, and enterobacterial repetitive intergenic consensus (ERIC) - PCR analysis. The comparative study of both methods showed that restriction fragment PFGE of SpeI digests of total DNA was a suitable method for strain typing and for determining which strains were present in vinegar fermentations. Results showed that strains of the species Gluconacetobacter europaeus were the most frequent leader strains of fermentations by the submerged method in the studied vinegars, and among them strain R1 was the predominant one. Results showed as well that mixed populations (at least two different strains) occurred in vinegars from cider and wine, whereas unique strains were found in spirit vinegars, which offered the most stressing conditions for bacterial growth. Copyright © 2010 Elsevier Ltd. All rights reserved.

  6. Genome comparison and physiological characterization of eight Streptococcus thermophilus strains isolated from Italian dairy products

    DEFF Research Database (Denmark)

    Vendramin, Veronica; Treu, Laura; Campanaro, Stefano

    2017-01-01

    to identify the core and the variable genes, which vary among strains from 196 to 265. Additionally, correlation between the isolation site and the genetic distance was investigated at genomic level. Results highlight that the phylogenetic reconstruction differs from the geographical strain distribution...

  7. Selection of yeast strains for the production of alcohol from lactoserum

    Energy Technology Data Exchange (ETDEWEB)

    Laham-Guillaume, M; Moulin, G; Galzy, P

    1979-01-01

    Five of 11 yeast strains tested fermented 85 g lactose/L to approximately 5% EtOH. Four of these strains, Candida pseudotropicalis CBS 19384 and IP 513, and Kluyveromyces fragilis CBS 397, and CBS 5795, anaerobically fermented deproteinized whey to EtOH.

  8. Differing effects of 2 active dried yeast (Saccharomyces cerevisiae) strains on ruminal acidosis and methane production in nonlactating dairy cows.

    Science.gov (United States)

    Chung, Y-H; Walker, N D; McGinn, S M; Beauchemin, K A

    2011-05-01

    Fifteen ruminally cannulated, nonlactating Holstein cows were used to measure the effects of 2 strains of Saccharomyces cerevisiae, fed as active dried yeasts, on ruminal pH and fermentation and enteric methane (CH(4)) emissions. Nonlactating cows were blocked by total duration (h) that their ruminal pH was below 5.8 during a 6-d pre-experimental period. Within each block, cows were randomly assigned to control (no yeast), yeast strain 1 (Levucell SC), or yeast strain 2 (a novel strain selected for enhanced in vitro fiber degradation), with both strains (Lallemand Animal Nutrition, Montréal, QC, Canada) providing 1 × 10(10) cfu/head per day. Cows were fed once daily a total mixed ration consisting of a 50:50 forage to concentrate ratio (dry matter basis). The yeast strains were dosed via the rumen cannula daily at the time of feeding. During the 35-d experiment, ruminal pH was measured continuously for 7 d (d 22 to 28) by using an indwelling system, and CH(4) gas was measured for 4 d (d 32 to 35) using the sulfur hexafluoride tracer gas technique (with halters and yokes). Rumen contents were sampled on 2 d (d 22 and 26) at 0, 3, and 6h after feeding. Dry matter intake, body weight, and apparent total-tract digestibility of nutrients were not affected by yeast feeding. Strain 2 decreased the average daily minimum (5.35 vs. 5.65 or 5.66), mean (5.98 vs. 6.24 or 6.34), and maximum ruminal pH (6.71 vs. 6.86 or 6.86), and prolonged the time that ruminal pH was below 5.8 (7.5 vs. 3.3 or 1.0 h/d) compared with the control or strain 1, respectively. The molar percentage of acetate was lower and that of propionate was greater in the ruminal fluid of cows receiving strain 2 compared with cows receiving no yeast or strain 1. Enteric CH(4) production adjusted for intake of dry matter or gross energy, however, did not differ between either yeast strain compared with the control but it tended to be reduced by 10% when strain 2 was compared with strain 1. The study shows that

  9. Do mechanical strain and TNF-α interact to amplify pro-inflammatory cytokine production in human annulus fibrosus cells?

    Science.gov (United States)

    Likhitpanichkul, Morakot; Torre, Olivia M; Gruen, Jadry; Walter, Benjamin A; Hecht, Andrew C; Iatridis, James C

    2016-05-03

    During intervertebral disc (IVD) injury and degeneration, annulus fibrosus (AF) cells experience large mechanical strains in a pro-inflammatory milieu. We hypothesized that TNF-α, an initiator of IVD inflammation, modifies AF cell mechanobiology via cytoskeletal changes, and interacts with mechanical strain to enhance pro-inflammatory cytokine production. Human AF cells (N=5, Thompson grades 2-4) were stretched uniaxially on collagen-I coated chambers to 0%, 5% (physiological) or 15% (pathologic) strains at 0.5Hz for 24h under hypoxic conditions with or without TNF-α (10ng/mL). AF cells were treated with anti-TNF-α and anti-IL-6. ELISA assessed IL-1β, IL-6, and IL-8 production and immunocytochemistry measured F-actin, vinculin and α-tubulin in AF cells. TNF-α significantly increased AF cell pro-inflammatory cytokine production compared to basal conditions (IL-1β:2.0±1.4-84.0±77.3, IL-6:10.6±9.9-280.9±214.1, IL-8:23.9±26.0-5125.1±4170.8pg/ml for basal and TNF-α treatment, respectively) as expected, but mechanical strain did not. Pathologic strain in combination with TNF-α increased IL-1β, and IL-8 but not IL-6 production of AF cells. TNF-α treatment altered F-actin and α-tubulin in AF cells, suggestive of altered cytoskeletal stiffness. Anti-TNF-α (infliximab) significantly inhibited pro-inflammatory cytokine production while anti-IL-6 (atlizumab) did not. In conclusion, TNF-α altered AF cell mechanobiology with cytoskeletal remodeling that potentially sensitized AF cells to mechanical strain and increased TNF-α-induced pro-inflammatory cytokine production. Results suggest an interaction between TNF-α and mechanical strain and future mechanistic studies are required to validate these observations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Strain improvement and optimization for β-glucosidase production in Aspergillus niger by low-energy N+ implantation

    International Nuclear Information System (INIS)

    Diao Jinshan; Wang Li; Chen Zhen; Liu Hui; Nie Guangjun; Zheng Zhiming

    2010-01-01

    Low-energy N + implantation was employed to mutate Aspergillus niger Au to enhance productivity of β-glucosidase. Effects of N + on strains, survival and mutation rate were studied. After several rounds of implantation, activity of β-glucosidase of the final mutant Au 0847 reached 13.75 U/mL, which is higher by 106.8% than that of original strain Au, and its heritability was stabilized. Activity of β-glucosidase of Au 0847 reached 30.53 U/mL after further fermentation condition optimization. (authors)

  11. Optimization of the cultivation conditions for mushroom production with European wild strains of Agaricus subrufescens and Brazilian cultivars.

    Science.gov (United States)

    Llarena-Hernández, Carlos R; Largeteau, Michèle L; Ferrer, Nathalie; Regnault-Roger, Catherine; Savoie, Jean-Michel

    2014-01-15

    The almond mushroom Agaricus subrufescens (formerly Agaricus blazei or Agaricus brasiliensis) is cultivated at commercial level in Brazil and some Asian countries on local substrates and casing mixtures. Despite its tropical origin, A. subrufescens might be a seasonal option for mushroom growers in western countries, where some wild strains have been isolated. For this purpose, cultivation conditions were developed starting from the substrate and casing mixture commonly used for commercial production of the button mushroom Agaricus bisporus in France. The commercial compost, based on wheat straw and horse manure, used for A. bisporus and the casing mixture (peat and limestone) supplemented with fine sand proved efficient to grow A. subrufescens. Increasing the depth of the casing layer improved significantly the yield and time to fruiting. Daily variations in temperature did not markedly modify the yield. Significantly higher mushroom biomass was obtained with three wild European strains compared with three Brazilian cultivars. The very productive wild strain CA438-A gave mushrooms of size and dry matter content comparable to those of a cultivar. Commercial production of A. subrufescens can be developed in western countries on the wheat straw-based substrate commonly used for A. bisporus in these regions, by a simple modification of the casing mixture and maintaining the incubation temperature throughout the crop, which is expected to save energy during summer. Good yields were obtained cultivating European strains under optimised parameters. © 2013 Society of Chemical Industry.

  12. Improving the productivity of S-adenosyl-l-methionine by metabolic engineering in an industrial Saccharomyces cerevisiae strain.

    Science.gov (United States)

    Zhao, Weijun; Hang, Baojian; Zhu, Xiangcheng; Wang, Ri; Shen, Minjie; Huang, Lei; Xu, Zhinan

    2016-10-20

    S-Adenosyl-l-methionine (SAM) is an important metabolite having prominent roles in treating various diseases. In order to improve the production of SAM, the regulation of three metabolic pathways involved in SAM biosynthesis were investigated in an industrial yeast strain ZJU001. GLC3 encoded glycogen-branching enzyme (GBE), SPE2 encoded SAM decarboxylase, as well as ERG4 and ERG6 encoded key enzymes in ergosterol biosynthesis, were knocked out in ZJU001 accordingly. The results indicated that blocking of either glycogen pathway or SAM decarboxylation pathway could improve the SAM accumulation significantly in ZJU001, while single disruption of either ERG4 or ERG6 gene had no obvious effect on SAM production. Moreover, the double mutant ZJU001-GS with deletion of both GLC3 and SPE2 genes was also constructed, which showed further improvement of SAM accumulation. Finally, SAM2 was overexpressed in ZJU001-GS to give the best SAM-producing recombinant strain ZJU001-GS-SAM2, in which 12.47g/L SAM was produced by following our developed pseudo-exponential fed-batch cultivation strategy, about 81.0% increase comparing to its parent strain ZJU001. The present work laid a solid base for large-scale SAM production with the industrial Saccharomyces cerevisiae strain. Copyright © 2016 Elsevier B.V. All rights reserved.

  13. Investigation of lactic acid bacterial strains for meat fermentation and the product's antioxidant and angiotensin-I-converting-enzyme inhibitory activities.

    Science.gov (United States)

    Takeda, Shiro; Matsufuji, Hisashi; Nakade, Koji; Takenoyama, Shin-Ichi; Ahhmed, Abdulatef; Sakata, Ryoichi; Kawahara, Satoshi; Muguruma, Michio

    2017-03-01

    In the lactic acid bacteria (LAB) strains screened from our LAB collection, Lactobacillus (L.) sakei strain no. 23 and L. curvatus strain no. 28 degraded meat protein and tolerated salt and nitrite in vitro. Fermented sausages inoculated strains no. 23 and no. 28 showed not only favorable increases in viable LAB counts and reduced pH, but also the degradation of meat protein. The sausages fermented with these strains showed significantly higher antioxidant activity than those without LAB or fermented by each LAB type strain. Angiotensin-I-converting-enzyme (ACE) inhibitory activity was also significantly higher in the sausages fermented with strain no. 23 than in those fermented with the type strain. Higher ACE inhibitory activity was also observed in the sausages fermented with strain no. 28, but did not differ significantly from those with the type strain. An analysis of the proteolysis and degradation products formed by each LAB in sausages suggested that those bioactivities yielded fermentation products such as peptides. Therefore, LAB starters that can adequately ferment meat, such as strains no. 23 and no. 28, should contribute to the production of bioactive compounds in meat products. © 2016 Japanese Society of Animal Science.

  14. Improved ethanol production at high temperature by consolidated bioprocessing using Saccharomyces cerevisiae strain engineered with artificial zinc finger protein.

    Science.gov (United States)

    Khatun, M Mahfuza; Yu, Xinshui; Kondo, Akihiko; Bai, Fengwu; Zhao, Xinqing

    2017-12-01

    In this work, the consolidated bioprocessing (CBP) yeast Saccharomyces cerevisiae MNII/cocδBEC3 was transformed by an artificial zinc finger protein (AZFP) library to improve its thermal tolerance, and the strain MNII-AZFP with superior growth at 42°C was selected. Improved degradation of acid swollen cellulose by 45.9% led to an increase in ethanol production, when compared to the control strain. Moreover, the fermentation of Jerusalem artichoke stalk (JAS) by MNII-AZFP was shortened by 12h at 42°C with a concomitant improvement in ethanol production. Comparative transcriptomics analysis suggested that the AZFP in the mutant exerted beneficial effect by modulating the expression of multiple functional genes. These results provide a feasible strategy for efficient ethanol production from JAS and other cellulosic biomass through CBP based-fermentation at elevated temperatures. Copyright © 2017 Elsevier Ltd. All rights reserved.

  15. Bioreactor studies on the production of bacitracin by mutant strain of bacillus licheniformis BLS-NTTG 4

    International Nuclear Information System (INIS)

    Sabeen, M.; Arif, R.; Saleem, M.; Ghouri, S.M.; Baig, S.; Syed, Q.

    2005-01-01

    The present study deals with the production of antibiotic bacitracin by Bacillus licheniformis using submerged fermentation technique in stirred fermenter. Altogether 15 samples were isolated from local habitat such as Government College Lahore. Of all the culture tested BLS 13 gave maximum titer (81 plus minus li. Micro /ml) in the fermentation broth. To develop the process for the production of antibiotic bacitracin agro industrial wastes used as a sole source of carbon in different fermentation medias (M1-M10). The culture of Bacillus licheniformis BLS 13 was improved by UV irradiation by exposing the cells of BLS-UV 16 mutated for 60 minutes gave the maximum production i.e. 129.1 plus minus 0.7 i. Micro/ml. The parental strain (i.e. BLS 13 was also improved by using chemical mutagen NTTG which enhance the antibiotic activity, and the mutated strain number i.e. BLS-NTTG 4 gave the maximum production 143.0 plus minus 1.0 i. Micro/ml. In stirred fermenter chemically mutated strain was used to check the productivity of bacitracin in stirred fermenter was increased up to 156 plus minus 1 i. micro /ml. (author)

  16. Enhanced production of xylanase from locally isolated fungal strain using agro-industrial residues under solid-state fermentation.

    Science.gov (United States)

    Abdullah, Roheena; Nisar, Kinza; Aslam, Aafia; Iqtedar, Mehwish; Naz, Shagufta

    2015-01-01

    This study is related to the isolation of fungal strain for xylanase production using agro-industrial residues. Forty fungal strains with xylanolytic potential were isolated by using xylan agar plates and quantitatively screened in solid-state fermentation. Of all the tested isolates, the strain showing highest ability to produce xylanase was assigned the code Aspergillus niger LCBT-14. For the enhanced production of the enzyme, five different fermentation media were evaluated. Out of all media, M4 containing wheat bran gave maximum enzyme production. Effect of different variables including incubation time, temperature, pH, carbon and nitrogen sources has been investigated. The optimum enzyme production was obtained after 72 h at 30°C and pH 4. Glucose as a carbon source while ammonium sulphate and yeast extract as nitrogen sources gave maximum xylanase production (946 U/mL/min). This study was successful in producing xylanase by A. niger LCBT-14 economically by utilising cheap indigenous substrate.

  17. UTILIZATION OF PINE NEEDLES AS BED MATERIAL IN SOLID STATE FERMENTATION FOR PRODUCTION OF LACTIC ACID BY LACTOBACILLUS STRAINS

    Directory of Open Access Journals (Sweden)

    Manoj Kumar Ghosh

    Full Text Available Pine needles, which are abundantly found as underexploited biomass in coniferous forests, are responsible for fire hazards and air pollution. Utilization of pine needles as bed material in lactic acid production with solid state fermentation (SSF has been studied here. This investigation compared lactic acid production by pure strains of Lactobacilli, (1 L. delbrueckii (NCIM2025; (2 L. pentosus (NCIM 2912; (3 Lactobacillus sp. (NCIM 2734; (4 Lactobacillus sp. (NCIM2084; and a co-culture of the first two strains. The studies required 6 g per flask powdered dry pine needles as bed material, 2 g/L (inoculum, liquid production media based on pure glucose or whey substituted glucose, at 60, 80, and 120 g/L sugar levels, 37 oC, and an initial pH of 6.5. Co-culture attained a maximum lactic acid concentration of 45.10 g/L, followed by that of strain-1, 43.87 g/L and strain-4, 26.15 g/L, in 80 g/L pure glucose media. With 120g/L total sugar in whey-substituted media, the co-culture attained maximum lactic acid production of 44.88 g/L followed by that of strain-1, 43.67 g/L. The present experimental studies indicated better compatibility of pine needle bed with co-culture in solid state fermentation of lactic acid, which may prove to be an eco-friendly technology for utilization of biomass as well as minimizing fires in coniferous forests.

  18. The gene bap, involved in biofilm production, is present in Staphylococcus spp. strains from nosocomial infections.

    Science.gov (United States)

    Potter, Amina; Ceotto, Hilana; Giambiagi-Demarval, Marcia; dos Santos, Kátia Regina Netto; Nes, Ingolf F; Bastos, Maria do Carmo de Freire

    2009-06-01

    This study analyzed ten strains of coagulase-negative staphylococci (CNS) involved in nosocomial infections in three Brazilian hospitals. Their antibiotic susceptibility profile showed that most strains exhibited multiple antibiotic resistance and possessed the mecA gene. The ability of these strains to adhere to polystyrene microtiter plates was also tested and nine of them proved to be biofilm producers at least in one of the three conditions tested: growth in TSB, in TSB supplemented with NaCl, or in TSB supplemented with glucose. The presence of the bap gene, which codes for the biofilm-associated protein (Bap), was investigated in all ten strains by PCR. AU strains were bop-positive and DNA sequencing experiments confirmed that the fragments amplified were indeed part of a bap gene. The presence of the icaA gene, one of the genes involved in polysaccharide intercellular adhesin (PIA) formation, was also detected by PCR in eight of the ten strains tested. The two icaA-negative strains were either weak biofilm producer or no biofilm producer, although they were bop-positive. To our knowledge, this is the first report demonstrating the presence of the bap gene in nosocomial isolates of CNS, being also the first report on the presence of this gene in Staphylococcus haemolyticus and S. cohnii.

  19. Leuconostoc strains isolated from dairy products: Response against food stress conditions.

    Science.gov (United States)

    D'Angelo, Luisa; Cicotello, Joaquín; Zago, Miriam; Guglielmotti, Daniela; Quiberoni, Andrea; Suárez, Viviana

    2017-09-01

    A systematic study about the intrinsic resistance of 29 strains (26 autochthonous and 3 commercial ones), belonging to Leuconostoc genus, against diverse stress factors (thermal, acidic, alkaline, osmotic and oxidative) commonly present at industrial or conservation processes were evaluated. Exhaustive result processing was made by applying one-way ANOVA, Student's test (t), multivariate analysis by Principal Component Analysis (PCA) and Matrix Hierarchical Cluster Analysis. In addition, heat adaptation on 4 strains carefully selected based on previous data analysis was assayed. The strains revealed wide diversity of resistance to stress factors and, in general, a clear relationship between resistance and Leuconostoc species was established. In this sense, the highest resistance was shown by Leuconostoc lactis followed by Leuconostoc mesenteroides strains, while Leuconostoc pseudomesenteroides and Leuconostoc citreum strains revealed the lowest resistance to the stress factors applied. Heat adaptation improved thermal cell survival and resulted in a cross-resistance against the acidic factor. However, all adapted cells showed diminished their oxidative resistance. According to our knowledge, this is the first study regarding response of Leuconostoc strains against technological stress factors and could establish the basis for the selection of "more robust" strains and propose the possibility of improving their performance during industrial processes. Copyright © 2017 Elsevier Ltd. All rights reserved.

  20. Application of Chinese Jun-Cao technique for the production of Brazilian Ganoderma lucidum strains

    Directory of Open Access Journals (Sweden)

    Leonardo do Nascimento Rolim

    2014-06-01

    Full Text Available Ganoderma lucidum is a medicinal mushroom traditionally used in China against a wide range of diseases such as cancer and also for its prevention. In this work, commercial Chinese strains G. lucidum were compared to wild Brazilian strains aiming to determine the cultivation potential through the use of Jun-Cao. Six formulations were tested and the strains presented good response to the applied method. In general, the mixture between the grass and wood was well suited for the basidiomycetes, contributing to the preparation of substrates that generated better results in Jun Cao.

  1. Production of ethanol from cassava pulp via fermentation with a surface-engineered yeast strain displaying glucoamylase

    Energy Technology Data Exchange (ETDEWEB)

    Kosugi, Akihiko; Murata, Yoshinori; Arai, Takamitsu; Mori, Yutaka [Post-harvest Science and Technology Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki 305-8686 (Japan); Kondo, Akihiko [Department of Chemical Science and Engineering, Faculty of Engineering, Kobe University, Nada-ku, Kobe, 657-8501 (Japan); Ueda, Mitsuyoshi [Department of Applied Biochemistry, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Sakyo-ku, Kyoto 606-8502 (Japan); Vaithanomsat, Pilanee; Thanapase, Warunee [Nanotechnology and Biotechnology Division, Kasetsart Agricultural and Agro-Industrial Product Improvement Institute (KAPI), Kasetsart University, 50 Chatuchak, Ladyao, Bangkok 10900 (Thailand)

    2009-05-15

    Cassava (Manihot esculenta Crantz) pulp, produced in large amounts as a by-product of starch manufacturing, is a major biomass resource in Southeast Asian countries. It contains abundant starch (approximately 60%) and cellulose fiber (approximately 20%). To effectively utilize the cassava pulp, an attempt was made to convert its components to ethanol using a sake-brewing yeast displaying glucoamylase on the cell surface. Saccharomyces cerevisiae Kyokai no. 7 (strain K7) displaying Rhizopus oryzae glucoamylase, designated strain K7G, was constructed using the C-terminal-half region of {alpha}-agglutinin. A sample of cassava pulp was pretreated with a hydrothermal reaction (140 C for 1 h), followed by treatment with a Trichoderma reesei cellulase to hydrolyze the cellulose in the sample. The K7G strain fermented starch and glucose in pretreated samples without addition of amylolytic enzymes, and produced ethanol in 91% and 80% of theoretical yield from 5% and 10% cassava pulp, respectively. (author)

  2. Pectinase production by fungal strains in solid-state fermentation using agro-industrial bioproduct

    Directory of Open Access Journals (Sweden)

    Natalia Martin

    2004-09-01

    Full Text Available Pectin lyase and polygalacturonase production by newly isolated fungal strains was carried out in solid-state fermentation. Moniliella SB9 and Penicillium sp EGC5 produced polygalcturonase (PG and pectin lyase (PL on mixture of orange bagasse, sugar cane bagasse and wheat bran as substrate. PG and PL produced by Moniliella presented optimum activity at pH 4.5 and 10.0 and at 55 and 45°C, respectively, while these enzymes from Penicillium sp presented optimum activity at pH 4.5-5.0 and 9.0 and 40°C, respectively.A produção de pectina liase e poligalacturonase por linhagens de fungos filamentosos isoladas, foi estudada através de fermentação em estado sólido utilizando subprodutos agro-industriais. Os fungos Moniliella sp SB9 e Penicillium sp EGC5 produziram consideráveis quantidades de PG e PL em substrato composto por mistura de bagaço de laranja, bagaço de cana de açúcar e farelo de trigo (1:1:1. As enzimas PG e PL, produzidas por Moniliella sp, apresentaram atividades ótimas em pH de 4,5 e 10,0 e em temperaturas de 55°C e 45°C, respectivamente. As mesmas enzimas, produzidas por Penicillium sp apresentaram atividades ótimas em pH 4,5-5,9 e 9,0 e 40°C, respectivamente.

  3. Comparison of hydrogen-production capability of four different Enterobacteriaceae strains under growing and non-growing conditions

    Energy Technology Data Exchange (ETDEWEB)

    Seol, Eunhee; Kim, Seohyoung; Raj, S. Mohan; Park, Sunghoon [Department of Chemical and Biochemical Engineering, Pusan National University, Busan 609-735 (Korea)

    2008-10-15

    Non-growing cells can function as whole-cell biocatalysts for hydrogen (H{sub 2}) production, a process that has recently drawn much attention. In order to evaluate their potential as whole-cell biocatalysts, we compared the H{sub 2}-production capability of four Enterobacteriaceae strains (Citrobacter amalonaticus Y19, Escherichia coli K-12 MG1655, Escherichia coli DJT135, and Enterobacter aerogenes) under growing and non-growing conditions. We evaluated their H{sub 2}-production activity at varying temperatures (25-45 C) and pH conditions (6.0-8.0) using glucose or formate as the carbon source. Under growing conditions with 10 mM glucose as a substrate, E. aerogenes exhibited the highest H{sub 2}-production activity (17.0 {+-} 0.2 {mu}mol H{sub 2} mg cell{sup -1} h{sup -1}) among the four strains, but the final H{sub 2} yield was similar (1.7-1.8 mol H{sub 2} mol{sup -1} glucose) in all four strains. H{sub 2} production in the four strains proceeded through a formate-dependent pathway that involved the formate hydrogen lyase (FHL) complex. Under non-growing conditions with 20 mM formate as a substrate, we obtained high H{sub 2}-production activities, in the range of 95.5-195.2 {mu}mol H{sub 2} mg cell{sup -1} h{sup -1}, with E. coli DJT135 exhibiting the highest activity (195.2 {mu}mol H{sub 2} mg{sup -1} h{sup -1}) at pH 6.0 and 45 C. In contrast, using glucose as the carbon substrate in non-growing cell experiments greatly reduced the H{sub 2}-production activity to 6.1-7.7 {mu}mol H{sub 2} mg cell{sup -1} h{sup -1}. This study indicated that formate is a better substrate than glucose for H{sub 2} production by non-growing cells, and that the H{sub 2}-production performance among the strains did not vary significantly, with the exception of E. coli K-12 MG1655. (author)

  4. Isolation and Selection of Microalgal Strains from Natural Water Sources in Viet Nam with Potential for Edible Oil Production.

    Science.gov (United States)

    Thao, Tran Yen; Linh, Dinh Thi Nhat; Si, Vo Chi; Carter, Taylor W; Hill, Russell T

    2017-06-23

    Industrial vegetable oil production in Viet Nam depends on oil seeds and crude plant oils that are currently more than 90% imported. As the first step in investigating the feasibility of using microalgae to provide Viet Nam with a domestic source of oil for food and edible oil industries, fifty lipid-producing microalgae were isolated and characterized. The microalgae were isolated from water sources ranging from freshwater to brackish and marine waters from a wide geographic distribution in Viet Nam. Initial analyses showed that 20 of the 50 strains had good growth rates, produced high biomass and had high lipid content, ranging up to 50% of dry weight biomass. 18S rRNA gene sequence analyses of the 50 strains showed a great diversity in this assemblage of microalgae, comprising at least 38 species and representatives of 25 genera : Chlamydomonas , Poterioochromonas , Scenedesmus , Desmodesmus , Chlorella , Bracteacoccus , Monoraphidium , Selenastrum , Acutodesmus , Mychonastes , Ankistrodesmus , Kirchneriella , Raphidocelis , Dictyosphaerium , Coelastrella , Schizochlamydella , Oocystidium , Nannochloris , Auxenochlorella , Chlorosarcinopsis , Stichococcus , Picochlorum , Prasinoderma , Chlorococcum , and Marvania. Some of the species are closely related to well-known lipid producers such as Chlorella sorokiniana , but some other strains are not closely related to the strains found in public sequence databases and likely represent new species. Analysis of oil quality showed that fatty acid profiles of the microalgal strains were very diverse and strain-dependent. Fatty acids in the microalgal oils comprised saturated fatty acids (SFAs), poly-unsaturated fatty acids (PUFAs), and mono-unsaturated fatty acids (MUFAs). The main SFA was palmitic acid. MUFAs and PUFAs were dominated by oleic acid, and linoleic and linolenic acids, respectively. Some strains were especially rich in the essential fatty acid α-linolenic acid (ALA), which comprised more than 20% of the

  5. Nitric oxide (NO) production in mammalian non-tumorigenic epithelial cells of the small intestine and macrophages induced by individual strains of lactobacilli and bifidobacteria

    DEFF Research Database (Denmark)

    Pipenbaher, Natasa; Møller, Peter Lange; Dolinsek, Jan

    2009-01-01

    and absence of interferon gamma (INF-¿). Production of NO in intestinal epithelium was stimulated by individual strains of lactobacilli without INF-¿ priming. While none of the tested bifidobacteria were capable of inducing NO production, most constitutively secreted NO. Most tested strains induced...

  6. Selection of yeast starter culture strains for the production of marula fruit wines and distillates.

    Science.gov (United States)

    Fundira, M; Blom, M; Pretorius, I S; van Rensburg, P

    2002-03-13

    Juice of the Sclerocarya birrea subsp. caffra (marula) fruit was fermented by indigenous microflora and different commercial Saccharomyces cerevisiae yeast strains at different temperatures, namely, 15 and 30 degrees C. Volatile acids, esters, and higher alcohols were quantified in the wine and distillates, and the results were interpreted using a multivariate analysis of variance and an average linkage cluster analysis. Significant differences between 15 and 30 degrees C and also among yeasts with respect to volatile compounds were observed. Yeast strains VIN7 and FC consistently produced wines and final distillates significantly different from the other strains. A panel of tasters and marula and brandy producers was asked to select wines and distillates that had an acceptable and typical marula "nose". They were also asked to detect the differences among wines and distillates fermented with the same yeast strain at different temperatures.

  7. Recent advances in microbial production of mannitol: utilization of low-cost substrates, strain development and regulation strategies.

    Science.gov (United States)

    Zhang, Min; Gu, Lei; Cheng, Chao; Ma, Jiangfeng; Xin, Fengxue; Liu, Junli; Wu, Hao; Jiang, Min

    2018-02-26

    Mannitol has been widely used in fine chemicals, pharmaceutical industries, as well as functional foods due to its excellent characteristics, such as antioxidant protecting, regulation of osmotic pressure and non-metabolizable feature. Mannitol can be naturally produced by microorganisms. Compared with chemical manufacturing, microbial production of mannitol provides high yield and convenience in products separation; however the fermentative process has not been widely adopted yet. A major obstacle to microbial production of mannitol under industrial-scale lies in the low economical efficiency, owing to the high cost of fermentation medium, leakage of fructose, low mannitol productivity. In this review, recent advances in improving the economical efficiency of microbial production of mannitol were reviewed, including utilization of low-cost substrates, strain development for high mannitol yield and process regulation strategies for high productivity.

  8. Preliminary Evaluation of Probiotic Properties of Lactobacillus Strains Isolated from Sardinian Dairy Products

    OpenAIRE

    Pisano, Maria Barbara; Viale, Silvia; Conti, Stefania; Fadda, Maria Elisabetta; Deplano, Maura; Melis, Maria Paola; Deiana, Monica; Cosentino, Sofia

    2014-01-01

    Twenty-three Lactobacillus strains of dairy origin were evaluated for some functional properties relevant to their use as probiotics. A preliminary subtractive screening based on the abilities to inhibit the growth of microbial pathogens and hydrolyze conjugated bile salts was applied, and six strains were selected for further characterization including survival under gastrointestinal environmental conditions, adhesion to gut epithelial tissue, enzymatic activity, and some safety properties. ...

  9. The dynamics of toxic microcystis strains and microcystin production in two hypertrofic South African reservoirs

    OpenAIRE

    Barnard, Sandra; Conradie, Karin Ronel

    2012-01-01

    The South African impoundments of Hartbeespoort and Roodeplaat experience excessive blooms of Microcystis species each year. Microcystins, produced primarily by strains of cyanobacteria belonging to the genera Microcystis, Anabaena and Planktothrix, are harmful cyanobacterial hepatotoxins. These bloom-forming cyanobacteria form toxic and non-toxic strains that co-occur and are visually indistinguishable, but can be identified and quantified molecularly. We described the relationships between ...

  10. Lactic Acid Production from Pretreated Hydrolysates of Corn Stover by a Newly Developed Bacillus coagulans Strain

    OpenAIRE

    Jiang, Ting; Qiao, Hui; Zheng, Zhaojuan; Chu, Qiulu; Li, Xin; Yong, Qiang; Ouyang, Jia

    2016-01-01

    An inhibitor-tolerance strain, Bacillus coagulans GKN316, was developed through atmospheric and room temperature plasma (ARTP) mutation and evolution experiment in condensed dilute-acid hydrolysate (CDH) of corn stover. The fermentabilities of other hydrolysates with B. coagulans GKN316 and the parental strain B. coagulans NL01 were assessed. When using condensed acid-catalyzed steam-exploded hydrolysate (CASEH), condensed acid-catalyzed liquid hot water hydrolysate (CALH) and condensed acid-...

  11. Lactose-mediated carbon catabolite repression of putrescine production in dairy Lactococcus lactis is strain dependent.

    Science.gov (United States)

    del Rio, Beatriz; Ladero, Victor; Redruello, Begoña; Linares, Daniel M; Fernández, Maria; Martín, Maria Cruz; Alvarez, Miguel A

    2015-06-01

    Lactococcus lactis is the lactic acid bacterial (LAB) species most widely used as a primary starter in the dairy industry. However, several strains of L. lactis produce the biogenic amine putrescine via the agmatine deiminase (AGDI) pathway. We previously reported the putrescine biosynthesis pathway in L. lactis subsp. cremoris GE2-14 to be regulated by carbon catabolic repression (CCR) via glucose but not lactose (Linares et al., 2013). The present study shows that both these sugars repress putrescine biosynthesis in L. lactis subsp. lactis T3/33, a strain isolated from a Spanish artisanal cheese. Furthermore, we demonstrated that both glucose and lactose repressed the transcriptional activity of the aguBDAC catabolic genes of the AGDI route. Finally, a screening performed in putrescine-producing dairy L. lactis strains determined that putrescine biosynthesis was repressed by lactose in all the L. lactis subsp. lactis strains tested, but in only one L. lactis subsp. cremoris strain. Given the obvious importance of the lactose-repression in cheese putrescine accumulation, it is advisable to consider the diversity of L. lactis in this sense and characterize consequently the starter cultures to select the safest strains. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Flux Balance Analysis Inspired Bioprocess Upgrading for Lycopene Production by a Metabolically Engineered Strain of Yarrowia lipolytica

    Directory of Open Access Journals (Sweden)

    Komi Nambou

    2015-12-01

    Full Text Available Genome-scale metabolic models embody a significant advantage of systems biology since their applications as metabolic flux simulation models enable predictions for the production of industrially-interesting metabolites. The biotechnological production of lycopene from Yarrowia lipolytica is an emerging scope that has not been fully scrutinized, especially for what concerns cultivation conditions of newly generated engineered strains. In this study, by combining flux balance analysis (FBA and Plackett-Burman design, we screened chemicals for lycopene production from a metabolically engineered strain of Y. lipolytica. Lycopene concentrations of 126 and 242 mg/L were achieved correspondingly from the FBA-independent and the FBA-assisted designed media in fed-batch cultivation mode. Transcriptional studies revealed upregulations of heterologous genes in media designed according to FBA, thus implying the efficiency of model predictions. Our study will potentially support upgraded lycopene and other terpenoids production from existing or prospect bioengineered strains of Y. lipolytica and/or closely related yeast species.

  13. Molecular characterization of an unauthorized genetically modified Bacillus subtilis production strain identified in a vitamin B2 feed additive.

    Science.gov (United States)

    Paracchini, Valentina; Petrillo, Mauro; Reiting, Ralf; Angers-Loustau, Alexandre; Wahler, Daniela; Stolz, Andrea; Schönig, Birgit; Matthies, Anastasia; Bendiek, Joachim; Meinel, Dominik M; Pecoraro, Sven; Busch, Ulrich; Patak, Alex; Kreysa, Joachim; Grohmann, Lutz

    2017-09-01

    Many food and feed additives result from fermentation of genetically modified (GM) microorganisms. For vitamin B2 (riboflavin), GM Bacillus subtilis production strains have been developed and are often used. The presence of neither the GM strain nor its recombinant DNA is allowed for fermentation products placed on the EU market as food or feed additive. A vitamin B 2 product (80% feed grade) imported from China was analysed. Viable B. subtilis cells were identified and DNAs of two bacterial isolates (LHL and LGL) were subjected to three whole genome sequencing (WGS) runs with different devices (MiSeq, 454 or HiSeq system). WGS data revealed the integration of a chloramphenicol resistance gene, the deletion of the endogenous riboflavin (rib) operon and presence of four putative plasmids harbouring rib operons. Event- and construct-specific real-time PCR methods for detection of the GM strain and its putative plasmids in food and feed products have been developed. Copyright © 2017 Elsevier Ltd. All rights reserved.

  14. Screening and Evaluation of Some Green Algal Strains (Chlorophyceae Isolated from Freshwater and Soda Lakes for Biofuel Production

    Directory of Open Access Journals (Sweden)

    Ramganesh Selvarajan

    2015-07-01

    Full Text Available Microalgae are photosynthetic microorganisms that can produce lipids, proteins and carbohydrates in large amounts and within short periods of time and these can be processed into both biofuels and other useful commercial products. Due to this reason microalgae are considered as a potential source of renewable energy; and one of the most important decisions in obtaining oil from microalgae is the choice of species. In this study, the potential of Chlorophyceae species isolated from freshwater and soda lakes in Hungary and Romania (Central Europe were characterized and evaluated by determining their biomass accumulation, lipid productivity, fatty acid profiles, and biodiesel properties besides protein and carbohydrate productivity. Out of nine strains tested, three accumulated more than 40% dry weight of protein, four accumulated more than 30% dry weight of carbohydrate and the strain Chlorella vulgaris LC8 accumulated high lipid content (42.1% ± 2.6% with a favorable C16-C18 fatty acid profile (77.4% as well as suitable biodiesel properties of high cetane number (57.3, low viscosity (4.7 mm2/s, lower iodine number (75.18 g I2/100 g, relative cloud point (8.8 °C and negative cold filter plugging point (−6.5 °C. Hence the new strain, Chlorella vulgaris LC8 has potential as a feedstock for the production of excellent quality biodiesel.

  15. GABA production and structure of gadB/gadC genes in Lactobacillus and Bifidobacterium strains from human microbiota.

    Science.gov (United States)

    Yunes, R A; Poluektova, E U; Dyachkova, M S; Klimina, K M; Kovtun, A S; Averina, O V; Orlova, V S; Danilenko, V N

    2016-12-01

    Gamma-amino butyric acid (GABA) is an active biogenic substance synthesized in plants, fungi, vertebrate animals and bacteria. Lactic acid bacteria are considered the main producers of GABA among bacteria. GABA-producing lactobacilli are isolated from food products such as cheese, yogurt, sourdough, etc. and are the source of bioactive properties assigned to those foods. The ability of human-derived lactobacilli and bifidobacteria to synthesize GABA remains poorly characterized. In this paper, we screened our collection of 135 human-derived Lactobacillus and Bifidobacterium strains for their ability to produce GABA from its precursor monosodium glutamate. Fifty eight strains were able to produce GABA. The most efficient GABA-producers were Bifidobacterium strains (up to 6 g/L). Time profiles of cell growth and GABA production as well as the influence of pyridoxal phosphate on GABA production were studied for L. plantarum 90sk, L. brevis 15f, B. adolescentis 150 and B. angulatum GT102. DNA of these strains was sequenced; the gadB and gadC genes were identified. The presence of these genes was analyzed in 14 metagenomes of healthy individuals. The genes were found in the following genera of bacteria: Bacteroidetes (Bacteroides, Parabacteroides, Alistipes, Odoribacter, Prevotella), Proteobacterium (Esherichia), Firmicutes (Enterococcus), Actinobacteria (Bifidobacterium). These data indicate that gad genes as well as the ability to produce GABA are widely distributed among lactobacilli and bifidobacteria (mainly in L. plantarum, L. brevis, B. adolescentis, B. angulatum, B. dentium) and other gut-derived bacterial species. Perhaps, GABA is involved in the interaction of gut microbiota with the macroorganism and the ability to synthesize GABA may be an important feature in the selection of bacterial strains - psychobiotics. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. Do New Production Concepts and a new Management of Employment Relations, yield higher Employee Performance and lower Job Strain?

    Directory of Open Access Journals (Sweden)

    Karolus Kraan

    2007-05-01

    Full Text Available In this article old versus new production concepts (NPCs and employment relation instruments, are studied, separately and in combination, to find out which yield high employee performance and low job strain. Therefore, in 2005, TNO conducted coupled surveys among 149 supervisors and employees. In the past decades, in reaction to dysfunctions of Tayloristic and professional bureaucratic production concepts and employment relations, several new forms of employment relations and NPCs, appeared. Examples are the Socio-technical NPC and customized employment relations. In this study both this NPC and customized employment relations - i.c. customized performance targets - demonstrate positive associations with employee performance. According to Socio-technical theory the design of employment relations is relatively unimportant, as human resources are mobilised primarily by the production concept. Our results for this NPC show the legitimacy of this assumption, because its high employee performance is irrespective of the employment relation instruments. On the contrary, in the other NPCs and in professional bureaucracies, the (employment relation instruments of respectively an increased period needed for learning the job, and customized performance targets can compensate for the lower employee performance in these production concepts. The results do not show increased job strain, due to new production concepts, or new employment relations. production concepts, employment relations, labour productivity, socio-technical theory

  17. Impact of the Botrytis cinerea strain and metabolism on (-)-geosmin production by Penicillium expansum in grape juice.

    Science.gov (United States)

    La Guerche, Stéphane; De Senneville, Laure; Blancard, Dominique; Darriet, Philippe

    2007-10-01

    Geosmin, an off-flavour of some rotten grapes, has been implicated in wine defects. Botrytis cinerea and Penicillium expansum were the most common among the numerous microorganisms isolated from rotten grapes. P. expansum produces geosmin on model media but not healthy grape juice. However, geosmin synthesis by P. expansum was demonstrated in grape juice and on crushed grapes that had been pre-cultured with certain B. cinerea strains. 34 out of 156 B. cinerea strains ([bot +] phenotype) isolated from the centre of grape bunches were able to induce high geosmin production, up to 494 ng/l, by P. expansum in grape juice. A study of the impact of grape juice composition on geosmin synthesis by P. expansum revealed the importance of nitrogen composition, particularly amino-acid deficiency. Metabolism of amino acids by B. cinerea was shown to be favourable to geosmin synthesis by P. expansum. However, the amino-acid and ammonium concentrations in grape juices pre-cultured with B. cinerea [bot -] and [bot +] strains were very similar implying that other factors are involved as well. Indeed, an ethanol-precipitable fraction, probably a polysaccharide, synthesized by B. cinerea [bot -], but not [bot +] strains, inhibited geosmin production by P. expansum.

  18. Optimization of liquid-state fermentation conditions for the glyphosate degradation enzyme production of strain Aspergillus oryzae by ultraviolet mutagenesis.

    Science.gov (United States)

    Fu, Gui-Ming; Li, Ru-Yi; Li, Kai-Min; Hu, Ming; Yuan, Xiao-Qiang; Li, Bin; Wang, Feng-Xue; Liu, Cheng-Mei; Wan, Yin

    2016-11-16

    This study aimed to obtain strains with high glyphosate-degrading ability and improve the ability of glyphosate degradation enzyme by the optimization of fermentation conditions. Spore from Aspergillus oryzae A-F02 was subjected to ultraviolet mutagenesis. Single-factor experiment and response surface methodology were used to optimize glyphosate degradation enzyme production from mutant strain by liquid-state fermentation. Four mutant strains were obtained and named as FUJX 001, FUJX 002, FUJX 003, and FUJX 004, in which FUJX 001 gave the highest total enzyme activity. Starch concentration at 0.56%, GP concentration at 1,370 mg/l, initial pH at 6.8, and temperature at 30°C were the optimum conditions for the improved glyphosate degradation endoenzyme production of A. oryzae FUJX 001. Under these conditions, the experimental endoenzyme activity was 784.15 U/100 ml fermentation liquor. The result (784.15 U/100 ml fermentation liquor) was approximately 14-fold higher than that of the original strain. The result highlights the potential of glyphosate degradation enzyme to degrade glyphosate.

  19. Potential of Zimbabwean commercial probiotic products and strains of Lactobacillus plantarum as prophylaxis and therapy against diarrhoea caused by Escherichia coli in children.

    Science.gov (United States)

    Chingwaru, Walter; Vidmar, Jerneja

    2017-01-01

    To evaluate the potential of commercial fermented products sold in the country, and strains of Lactobacillus plantarum (L. plantarum) as prophylaxis and therapy against diarrhoea in children. The antimicrobial potential of cultures of lactobacilli enriched from 4 Zimbabwean commercial food/beverage products: Dairibord Lacto sour milk (DLSM), Probrand sour milk (PSM), Kefalos Vuka cheese (KVC) and Chibuku opaque beer (COB); and four strains of L. plantarum obtained from Balkan traditional cheeses against clinical strains of Escherichia coli (E. coli) was assayed using the well diffusion method. Three commercial paediatric antidiarrhoeal drug products: Biogaia (BG), Prolife (PL) and Probio Junior (PJ) and a mutant strain of E. coli [strain 11105 (ATCC) - a vitamin B-12 auxotroph and penicillin G acylase-producing strain] were used as controls. An agar diffusion assay and a competitive exclusion assay were carried out on Mueller Hinton agar. Crude cultures of putative lactobacillus strains obtained from Zimbabwean dairy products (Probrand sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer) had significantly higher antimicrobial activities against clinical strains of E. coli than strains of L. plantarum isolated from Balkan cheeses (CLP1, CLP2 or CLP3) and crude microbial cultures from commercial paediatric probiotic products (BG, PJ and PL) of a culture of Lactobacillus rhamnosus LGG (P sour milk, Kefalos Vuka vuka cheese and Chibuku opaque beer), and three strains of L. plantarum from Balkan cheeses (CLP1, CLP2 or CLP3) exhibited high antibacterial activities that can be harnessed to control paediatric diarrhoea that is caused by pathogenic strains of E. coli. Studies to characterise the probiotic potential of the live cultures in the products and the new strains of L. plantarum are underway. Copyright © 2017 Hainan Medical University. Production and hosting by Elsevier B.V. All rights reserved.

  20. A new highly productive Propionibacterium acidipropionici FL-48 strain with increased resistance to propionic acid and the scaling up of its production for industrial bioreactors

    Directory of Open Access Journals (Sweden)

    M. A. Kartashov

    2016-09-01

    Full Text Available Propionic acid bacteria, including Propionibacterium acidipropionici, are widely used in the chemical industry to produce propionic acid and also for food and feed preservation. However, the efficiency of the industrial production of these bacteria is limited by their sensitivity to high concentrations of propionic acid excreted into the cultivation medium. Therefore, the development of new biotechnological processes and strains able to overcome this limitation and to improve the profitability of the microbiological production remains  a relevant problem. A new P. acidipropionici FL-48 strain characterized by an increased resistance to 10 g/L of propionic acid (the number of viable cells after 24-h cultivation reached 1.05 × 106 was developed by a two-step induced mutagenesis using UV and diethyl sulphate from the P. acidipropionici VKPM B-5723 strain. The mutant strain exceeded the parental strain in the biomass accumulation rate and the amount of produced propionic and acetic acids by 35%, 20%, and 16%, respectively. The stability of such important characteristics as the biomass accumulation rate and the viability on media containing heightened concentrations of propionic acid was confirmed by three sequential monoclonal subculturings on a medium supplemented with 10 g/L of propionic acid. The optimization of the cultivation technology made it possible to determine the optimum seed inoculum dose (10% of the fermentation medium volume and the best pH level for the active growth stage (6.1 ± 0.1. The scaling up of the fermentation to a 100-L bioreactor under observance of optimum cultivation conditions demonstrated a high biomass growth rate with a sufficient reproducability; after 20 h of fermentation, the number of viable cells in the culture broth exceeded 1 × 1010 CFU/mL. The new strain could be interesting as the component of silage and haylage biopreservatives and also could be used as an efficient producer of propionic acid.

  1. SCREENING OF SELECTED OLEAGINOUS YEASTS FOR LIPID PRODUCTION FROM GLYCEROL AND SOME FACTORS WHICH AFFECT LIPID PRODUCTION BY YARROWIA LIPOLYTICA STRAINS

    Directory of Open Access Journals (Sweden)

    Salinee Sriwongchai

    2013-04-01

    Full Text Available The ability of eight yeast strains to utilize glycerol as a sole carbon source and accumulate lipids in a chemically defined medium was screened. Among the yeasts, Yarrowia lipolytica strains DSM 70561 and JDC 335 grew to high cell densities on glycerol. These strains were further tested for lipid accumulation under varying nutritional conditions in Erlenmeyer flasks. The results showed that strains DSM 70561 and JDC 335 accumulated lipids up to 37.1 % and 54.4 % of total cell dry weight, respectively, when the defined medium was supplemented with 1 g/L urea and 2 g/L yeast extract. The lipids accumulated by the two yeasts contained a high proportion of C16:0, C18:1, C18:2 and C18:0 fatty acids. The results suggest that Y. lipolytica strains DSM 70561 and JDC 335 have the potential for converting crude glycerol into fatty acids which can in turn be utilized as substrate for biodiesel production.

  2. Biosurfactant production from marine hydrocarbon-degrading consortia and pure bacterial strains using crude oil as carbon source

    OpenAIRE

    Antoniou, Eleftheria; Fodelianakis, Stilianos; Korkakaki, Emmanouela; Kalogerakis, Nicolas

    2015-01-01

    Biosurfactants (BS) are green amphiphilic molecules produced by microorganisms during biodegradation, increasing the bioavailability of organic pollutants. In this work, the BS production yield of marine hydrocarbon degraders isolated from Elefsina bay in Eastern Mediterranean Sea has been investigated. The drop collapse test was used as a preliminary screening test to confirm biosurfactant producing strains or mixed consortia. The community structure of the best consortia based on the drop c...

  3. Antibiotic resistance patterns of Escherichia coli strains isolated from surface water and groundwater samples in a pig production area

    OpenAIRE

    Roger Neto Schneider; André Nadvorny; Verônica Schmidt

    2009-01-01

    The use of antibiotics, so excessive and indiscriminate in intensive animal production, has triggered an increase in the number of resistant microorganisms which can be transported to aquatic environments. The aim of this study was to determine the profile of the antimicrobial resistance of samples of Escherichia coli isolated from groundwater and surface water in a region of pig breeding. Through the test of antimicrobial susceptibility, we analyzed 205 strains of E. coli. A high rate of res...

  4. Production of Phytotoxic Metabolite Using Biphasic Fermentation System from Strain C1136 of Lasiodiplodia pseudotheobromae, a Potential Bioherbicidal Agent

    OpenAIRE

    Charles Oluwaseun ADETUNJI; Julius Kola OLOKE; Gandham PRASAD; Moses ABALAKA; Emenike Onyebum IROKANULO

    2017-01-01

    Formulation of effective and environmental friendly bioherbicides depends on the type of fermentation medium used for the production of phytotoxic metabolites. The effect of biomass, colony forming unit and the phytotoxic metabolite produced from the biphasic fermentation was carried out, while the phytotoxic metabolite was tested in vivo and in-vitro on Echinochola crus-galli and dicotyledonous Chromolaena odorata. The mutant strain of Lasiodiplodia pseudotheobromae C1136 (Lp90) produced th...

  5. Laccase Production from a Temperature and pH Tolerant Fungal Strain of Trametes hirsuta (MTCC 11397

    Directory of Open Access Journals (Sweden)

    Kusum Dhakar

    2013-01-01

    Full Text Available Laccase production by a temperature and pH tolerant fungal strain (GBPI-CDF-03 isolated from a glacial site in Indian Himalayan Region (IHR has been investigated. The fungus developed white cottony mass on potato dextrose agar and revealed thread-like mycelium under microscope. ITS region analysis of fungus showed its 100% similarity with Trametes hirsuta. The fungus tolerated temperature from 4 to 48°C ± 2 (25°C opt. and pH 3–13 (5–7 opt.. Molecular weight of laccase was determined approximately 45 kDa by native PAGE. Amplification of laccase gene fragment (corresponding to the copper-binding conserved domain contained 200 bp. The optimum pH for laccase production, at optimum growth temperature, was determined between 5.5 and 7.5. In optimization experiments, fructose and ammonium sulfate were found to be the best carbon and nitrogen sources, respectively, for enhancing the laccase production. Production of laccase was favored by high carbon/nitrogen ratio. Addition of CuSO4 (up to 1.0 mM induced laccase production up to 2-fold, in case of 0.4 mM concentration. Addition of organic solvents also induced the production of laccase; acetone showed the highest (2-fold induction. The study has implications in bioprospecting of ecologically resilient microbial strains.

  6. Antimicrobial Susceptibility and Biofilm Production by Salmonella sp. Strains Isolated from Frozen Poultry Carcasses

    Directory of Open Access Journals (Sweden)

    MJ Sereno

    Full Text Available ABSTRACT The objectives of this study were to evaluate the antimicrobial resistance and the biofilm-producing ability of Salmonella sp. strains isolated from frozen poultry carcasses. Antimicrobial susceptibility was tested by the disk-diffusion method. Biofilm-producing ability was determined in 96-well polystyrene microplates stained with crystal violet at 1%. Out of the 22 strains tested, all were multiresistant, that is, resistant to more than three antimicrobial classes, and 72.7% were able to form biofilms. The highest resistance rates obtained were against sulfonamides, tetracycline, and quinolones. On the other hand, 100% of the strains were sensitive to chloramphenicol. According to the rate of biofilm formation, 3 (13.6% and 13 (59.1% strains were classified as moderate and weak biofilm-producers, respectively, and 27.3% did not form biofilms. Biofilms increase the tolerance of microorganisms to stress, reducing their sensitivity to disinfectants and antimicrobials; favor equipment corrosion; and act as substrates for the adhesion of bacteria with lower biofilm-producing capacity. The results of the present study stress the importance of cleaning procedures in food processing plants and highlight the public health risks related to the emergence of multiresistant strains.

  7. Potential of functional strains, isolated from traditional Maasai milk, as starters for the production of fermented milks.

    Science.gov (United States)

    Patrignani, Francesca; Lanciotti, Rosalba; Mathara, Julius Maina; Guerzoni, Maria Elisabetta; Holzapfel, Wilhelm H

    2006-03-01

    The purpose of this research was the evaluation of technological features and of the ability of functional LAB strains with desirable sensory characteristics, to produce fermented milk. Eight strains of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus paracasei and Lactococcus lactis, isolated from Maasai traditional fermented milk in Kenya and previously tested for their probiotic properties, were selected for this investigation. Technological features such as growth kinetics in fresh heat-treated whole milk medium and survival in the final product during storage at 4 degrees C, were studied. The strains Lb. acidophilus BFE 6,059, Lb. paracasei BFE 5,264 and Lc. lactis BFE 6,049 showed the best potential and were thus selected for use as starter cultures in further trials with the objective to improve their technological performance and to optimise the sensory features of fermented milk obtained. The effects of fat (F), non-fat milk solids (S) and fermentation temperature (T), modulated according to a Central Composite Design, on fermentation rates and viability losses during refrigerated storage of the chosen starters, and on product texture parameters, were studied. From the data analysis, it was possible to select optimum conditions for enhancing positive sensory traits of final products and for improving the survival of these potentially probiotic cultures.

  8. Improved production, purification, and characterization of biosurfactants produced by Serratia marcescens SM3 and its isogenic SMRG-5 strain.

    Science.gov (United States)

    Rosas-Galván, Nashbly Sarela; Martínez-Morales, Fernando; Marquina-Bahena, Silvia; Tinoco-Valencia, Raunel; Serrano-Carreón, Leobardo; Bertrand, Brandt; León-Rodríguez, Renato; Guzmán-Aparicio, Josefina; Alvaréz-Berber, Laura; Trejo-Hernández, María R

    2018-02-19

    In this study, the biosurfactants (Bs) production of two Serratia marcescens strains (SM3 and its isogenic SMRG-5 strain) was improved and the tenso-active agents were purified and characterized. A 2 3 factorial design was used to evaluate the effect of nitrogen and carbon sources on the surface tension (ST) reduction and emulsion index (EI 24 ) of the produced Bs. Optimum Bs production by SM3 was achieved at high concentrations of carbon and nitrogen, reducing ST to 26.5 ± 0.28 dynes/cm, with an EI 24 of 79.9 ± 0.2%. Meanwhile, the best results for SMRG-5 were obtained at low concentrations, reducing the ST to 25.2 ± 0.2 dynes/cm, with an EI 24 of 89.7 ± 0.28%. The optimal conditions for Bs production were scaled up in a 2-L reactor, yielding 4.8 and 5.2 g/L for SM3 and SMRG-5, respectively. Gas Chromatography-Mass Spectrometry (GC-MS) analysis revealed the presence of two different lipopeptides (hidrofobic fractions: octadecanoic and hexadecanoic acid for SM3 and SMRG5, respectively). Both strains were capable of benzo [a] pyrene removal (59% after 72 H of culture). © 2018 International Union of Biochemistry and Molecular Biology, Inc.

  9. Robust cellulosic ethanol production from SPORL-pretreated lodgepole pine using an adapted strain Saccharomyces cerevisiae without detoxification.

    Science.gov (United States)

    Tian, S; Luo, X L; Yang, X S; Zhu, J Y

    2010-11-01

    This study reports an ethanol yield of 270L/ton wood from lodgepole pine pretreated with sulfite pretreatment to overcome recalcitrance of lignocellulose (SPORL) using an adapted strain, Saccharomyces cerevisiae Y5, without detoxification. The enzymatic hydrolysate produced from pretreated cellulosic solids substrate was combined with pretreatment hydrolysate before fermentation. Detoxification of the pretreatment hydrolysate using overliming or XAD-4 resin before being combined with enzymatic hydrolysate improved ethanol productivity in the first 4h of fermentation and overall fermentation efficiency. However, detoxification did not improve final ethanol yield because of sugar losses. The Y5 strain showed excellent ethanol productivities of 2.0 and 0.8g/L/h averaged over a period of 4 and 24h, respectively, in the undetoxified run. The furan metabolization rates of the Y5 strain were significantly higher for the undetoxified run than those for the detoxidfied runs, suggesting it can tolerate even higher furan concentrations than those studied. Preliminary mass and energy balances were conducted. SPORL produced an excellent monomeric sugar recovery value of about 85% theoretical and a net energy output of 4.05GJ/ton wood with an ethanol energy production efficiency of 178% before distillation.

  10. Cellulase and xylanase productions by isolated Amazon Bacillus strains using soybean industrial residue based solid-state cultivation

    Directory of Open Access Journals (Sweden)

    Heck Júlio X.

    2002-01-01

    Full Text Available In Brazil, a large amount of a fibrous residue is generated as result of soybean (Glycine max protein production. This material, which is rich in hemicellulose and cellulose, can be used in solid state cultivations for the production of valuable metabolites and enzymes. In this work, we studied the bioconversion of this residue by bacteria strains isolated from water and soil collected in the Amazon region. Five strains among 87 isolated bacteria selected for their ability to produce either celullases or xylanases were cultivated on the aforementioned residue. From strain BL62, identified as Bacillus subtilis, it was obtained a preparation showing the highest specific cellulase activity, 1.08 UI/mg protein within 24 hours of growth. Concerning xylanase, the isolate BL53, also identified as Bacillus subtilis, showed the highest specific activity for this enzyme, 5.19 UI/mg protein within 72 hours of cultivation. It has also been observed the production of proteases that were associated with the loss of cellulase and xylanase activities. These results indicated that the selected microorganisms, and the cultivation process, have great biotechnological potential.

  11. In Vitro characterization of Lactococcus lactis strains Isolated from Iranian Traditional Dairy Products as a Potential Probiotic

    Directory of Open Access Journals (Sweden)

    Fatemeh Nejati

    2015-12-01

    Full Text Available Few studies have been reported regarding probiotic properties of Lactococcus lactis strains although they are extensively used as starter cultures in the production of dairy products. In this study 8 wild isolates of Lactococcus lactis were evaluated in vitro with regard to resistance to simulated gastric and intestinal juices, adherence ability to Caco-2 cells and HT29-MTX-E12 cell lines, anti-microbial activity, hydrophobicity and antibiotic susceptibility. The results revealed that all isolates had better survival after exposure to simulated gastrointestinal tract stresses in comparison to control probiotic Lactobacillus rhamnosus GG. Regarding adherence efficiency, almost all isolates exhibited similar adherence with control. Three isolates showed antibacterial activity against Gram-positive pathogens (Staphylococcus aureus and Listeria monocytogenes through spot-agar method. Almost all isolates (seven out of eight showed similar hydrophobicity to control probiotic. Regarding to antibiotic resistance, all isolates were susceptible to gentamicin, ampicillin, ciprofloxacin, erythromycin, tetracycline, penicillin, kanamycin and nitrofurantoin. Although, further investigations are necessary, it was concluded that strains derived from raw milk and home-made dairy products could be a remarkable reservoir for identification of new potential probiotic strains.

  12. Bakery by-products based feeds borne-Saccharomyces cerevisiae strains with probiotic and antimycotoxin effects plus antibiotic resistance properties for use in animal production.

    Science.gov (United States)

    Poloni, Valeria; Salvato, Lauranne; Pereyra, Carina; Oliveira, Aguida; Rosa, Carlos; Cavaglieri, Lilia; Keller, Kelly Moura

    2017-09-01

    The aim of this study was to select S. cerevisiae strains able to exert probiotic and antimycotoxin effects plus antibiotics resistance properties for use in animal production. S. cerevisiae LL74 and S. cerevisiae LL83 were isolated from bakery by-products intended for use in animal feed and examined for phenotypic characteristics and nutritional profile. Resistance to antibiotic, tolerance to gastrointestinal conditions, autoaggregation and coaggregation assay, antagonism to animal pathogens and aflatoxin B 1 binding were studied. S. cerevisiae LL74 and S. cerevisiae LL83 showed resistance to all the antibiotics assayed (ampicillin, streptomycin, neomycin, norfloxacin, penicillin G, sulfonamide and trimethoprim). The analysis showed that exposure time to acid pH had a significant impact onto the viable cell counts onto both yeast strains. Presence of bile 0.5% increased significantly the growth of the both yeast strains. Moreover, they were able to tolerate the simulated gastrointestinal conditions assayed. In general, the coaggregation was positive whereas the autoaggregation capacity was not observed. Both strains were able to adsorb AFB 1 . In conclusion, selected S. cerevisiae LL74 and S. cerevisiae LL83 have potential application to be used as a biological method in animal feed as antibiotic therapy replacement in, reducing the adverse effects of AFB 1 and giving probiotic properties. Copyright © 2017 Elsevier Ltd. All rights reserved.

  13. Screening and selection of Lactobacillus strains for use as adjunct cultures in production of semi-hard cheese.

    Science.gov (United States)

    Antonsson, Martin; Ardö, Ylva; Nilsson, Bengt Frans; Molin, Göran

    2002-08-01

    Thirty-three Lactobacillus strains were tested as adjuncts in a cheese model system. Eighteen strains originated from cheese (nine Lactobacillus spp. and nine Lb. paracasei/casei) and 15 from human intestinal mucosa (11 Lb. rhamnosus; three Lb. paracasei; one Lb. plantarum). Model cheeses weighing 120 g were made of cheese grains from full-scale production of washed curd semi-hard cheese (Herrgård). The model system was reproducible and similar to full-scale production with respect to moisture, salt content, pH and microbial flora. The model cheeses were sampled for aerobic and anaerobic plate count and viable counts of Lactobacillus and Lactococcus. The presence of adjuncts in the model cheeses was confirmed by typing isolates with Randomly Amplified Polymorphic DNA (RAPD). The sensory properties of model cheeses were described. In a first trial 23 of the 33 adjuncts were re-isolated from the corresponding model cheeses after 9 or 13 weeks. Adjuncts of Lb. paracasei were re-isolated more frequently than adjuncts of Lb. rhamnosus. Nine strains were selected, on the basis of their ability to grow and be a dominating part of the microflora of model cheese with interesting sensory properties. These strains were further studied together with two commercial cultures. The sensory influences on model cheeses of six of the adjuncts were confirmed, and flavour scores were in the range of 2.9-7.1 for model cheeses with different adjuncts while the control had a flavour score of 5.6 (0-10 scale). Survival and growth of seven out of the nine strains correlated with the results of the first trial. Growth and influence on flavour of four adjunct cultures were confirmed in experimental cheese manufactured in a 400-1 open vat.

  14. Biosurfactant production from marine hydrocarbon-degrading consortia and pure bacterial strains using crude oil as carbon source

    Directory of Open Access Journals (Sweden)

    Eleftheria eAntoniou

    2015-04-01

    Full Text Available Biosurfactants (BS are green amphiphilic molecules produced by microorganisms during biodegradation, increasing the bioavailability of organic pollutants. In this work, the BS production yield of marine hydrocarbon degraders isolated from Elefsina bay in Eastern Mediterranean Sea has been investigated. The drop collapse test was used as a preliminary screening test to confirm biosurfactant producing strains or mixed consortia. The community structure of the best consortia based on the drop collapse test was determined by 16S-rDNA pyrotag screening. Subsequently, the effect of incubation time, temperature, substrate and supplementation with inorganic nutrients, on biosurfactant production, was examined. Two types of BS - lipid mixtures were extracted from the culture broth; the low molecular weight BS Rhamnolipids and Sophorolipids. Crude extracts were purified by silica gel column chromatography and then identified by thin layer chromatography (TLC and Fourier transform infrared spectroscopy (FT-IR. Results indicate that biosurfactant production yield remains constant and low while it is independent of the total culture biomass, carbon source, and temperature. A constant BS concentration in a culture broth with continuous degradation of crude oil implies that the BS producing microbes generate no more than the required amount of biosurfactants that enables biodegradation of the crude oil. Isolated pure strains were found to have higher specific production yields than the complex microbial marine community-consortia. The heavy oil fraction of crude oil has emerged as a promising substrate for BS production (by marine BS producers with fewer impurities in the final product. Furthermore, a particular strain isolated from sediments, Paracoccus marcusii, may be an optimal choice for bioremediation purposes as its biomass remains trapped in the hydrocarbon phase, not suffering from potential dilution effects by sea currents.

  15. Biosurfactant production from marine hydrocarbon-degrading consortia and pure bacterial strains using crude oil as carbon source.

    Science.gov (United States)

    Antoniou, Eleftheria; Fodelianakis, Stilianos; Korkakaki, Emmanouela; Kalogerakis, Nicolas

    2015-01-01

    Biosurfactants (BSs) are "green" amphiphilic molecules produced by microorganisms during biodegradation, increasing the bioavailability of organic pollutants. In this work, the BS production yield of marine hydrocarbon degraders isolated from Elefsina bay in Eastern Mediterranean Sea has been investigated. The drop collapse test was used as a preliminary screening test to confirm BS producing strains or mixed consortia. The community structure of the best consortia based on the drop collapse test was determined by 16S-rDNA pyrotag screening. Subsequently, the effect of incubation time, temperature, substrate and supplementation with inorganic nutrients, on BS production, was examined. Two types of BS - lipid mixtures were extracted from the culture broth; the low molecular weight BS Rhamnolipids and Sophorolipids. Crude extracts were purified by silica gel column chromatography and then identified by thin layer chromatography and Fourier transform infrared spectroscopy. Results indicate that BS production yield remains constant and low while it is independent of the total culture biomass, carbon source, and temperature. A constant BS concentration in a culture broth with continuous degradation of crude oil (CO) implies that the BS producing microbes generate no more than the required amount of BSs that enables biodegradation of the CO. Isolated pure strains were found to have higher specific production yields than the complex microbial marine community-consortia. The heavy oil fraction of CO has emerged as a promising substrate for BS production (by marine BS producers) with fewer impurities in the final product. Furthermore, a particular strain isolated from sediments, Paracoccus marcusii, may be an optimal choice for bioremediation purposes as its biomass remains trapped in the hydrocarbon phase, not suffering from potential dilution effects by sea currents.

  16. Biosurfactant production from marine hydrocarbon-degrading consortia and pure bacterial strains using crude oil as carbon source

    Science.gov (United States)

    Antoniou, Eleftheria; Fodelianakis, Stilianos; Korkakaki, Emmanouela; Kalogerakis, Nicolas

    2015-01-01

    Biosurfactants (BSs) are “green” amphiphilic molecules produced by microorganisms during biodegradation, increasing the bioavailability of organic pollutants. In this work, the BS production yield of marine hydrocarbon degraders isolated from Elefsina bay in Eastern Mediterranean Sea has been investigated. The drop collapse test was used as a preliminary screening test to confirm BS producing strains or mixed consortia. The community structure of the best consortia based on the drop collapse test was determined by 16S-rDNA pyrotag screening. Subsequently, the effect of incubation time, temperature, substrate and supplementation with inorganic nutrients, on BS production, was examined. Two types of BS – lipid mixtures were extracted from the culture broth; the low molecular weight BS Rhamnolipids and Sophorolipids. Crude extracts were purified by silica gel column chromatography and then identified by thin layer chromatography and Fourier transform infrared spectroscopy. Results indicate that BS production yield remains constant and low while it is independent of the total culture biomass, carbon source, and temperature. A constant BS concentration in a culture broth with continuous degradation of crude oil (CO) implies that the BS producing microbes generate no more than the required amount of BSs that enables biodegradation of the CO. Isolated pure strains were found to have higher specific production yields than the complex microbial marine community-consortia. The heavy oil fraction of CO has emerged as a promising substrate for BS production (by marine BS producers) with fewer impurities in the final product. Furthermore, a particular strain isolated from sediments, Paracoccus marcusii, may be an optimal choice for bioremediation purposes as its biomass remains trapped in the hydrocarbon phase, not suffering from potential dilution effects by sea currents. PMID:25904907

  17. Genome Sequence of Enterococcus faecium Strain ICIS 96 Demonstrating Intermicrobial Antagonism Associated with Bacteriocin Production.

    Science.gov (United States)

    Pashkova, Tatiana M; Vasilchenko, Alexey S; Khlopko, Yuriy A; Kochkina, Elena E; Kartashova, Olga L; Sycheva, Maria V

    2018-03-08

    We report here the complete genome sequence of Enterococcus faecium strain ICIS 96, which was isolated from the feces of a horse. Bacteriological characterization of strain ICIS 96 revealed the absence of pathogenicity factors, while its spectrum of antagonistic activity was found to be broad, having activities associated with both Gram-positive and Gram-negative bacteria. Analysis of the E. faecium ICIS 96 genome revealed five genes associated with antimicrobial activity (enterocin [ent] A, ent B, lactobin A/cerein 7b, and ent L50 A/B). No genes that correlate with human pathogenicity were identified. Copyright © 2018 Pashkova et al.

  18. Advances in metabolic pathway and strain engineering paving the way for sustainable production of chemical building blocks

    DEFF Research Database (Denmark)

    Chen, Yun; Nielsen, Jens

    2013-01-01

    Bio-based production of chemical building blocks from renewable resources is an attractive alternative to petroleum-based platform chemicals. Metabolic pathway and strain engineering is the key element in constructing robust microbial chemical factories within the constraints of cost effective...... production. Here we discuss how the development of computational algorithms, novel modules and methods, omics-based techniques combined with modeling refinement are enabling reduction in development time and thus advance the field of industrial biotechnology. We further discuss how recent technological...

  19. Production, extraction and characterization of exopolysaccharides produced by the native Leuconostoc pseudomesenteroides R2 strain

    Directory of Open Access Journals (Sweden)

    Elinalva M. Paulo

    2012-06-01

    Full Text Available The genus Leuconostoc belongs to a group of lactic acid bacteria usually isolated from fermented vegetables, which includes species involved in the production of exopolysaccharides (EPS. These biopolymers possess considerable commercial potential. Because of the wide variety of industrial applications of EPS, this study aimed to produce and characterize the native exopolysaccharide strain Leuconostoc pseudomesenteroides R2, which was isolated from cabbage collected in a semi-arid region of Bahia. We employed the following conditions for the production of EPS: 10.7% sucrose, pH 8.2, without agitation and incubation at 28ºC for 30 hours. The fermentation broth was treated with ethanol and generated two types of polysaccharide substances (EPS I and EPS II. The identification of EPS I and EPS II was conducted using FT-IR, ¹H, 13C and DEPT-135 NMR spectra. The two substances were identified as linear dextran α polysaccharides (1 → 6 which indicated different characteristics with respect to thermal analysis and density of free packaging, viscosity and time of solubilization. Both dextrans are of low density, possess high thermal stability and exhibited the behavior characteristic of pseudoplastic polymers.O gênero Leuconostoc pertence a um grupo de bactérias lácticas normalmente isoladas de vegetais fermentados, que inclui espécies envolvidas na produção de exopolissacarídeos (EPS. Esses biopolímeros possuem potencial comercial considerável. Devido à grande variedade de aplicações industriais, de EPS, o presente estudo teve como objetivo produzir e caracterizar o nativo exopolissacarídeo cepa Leuconostoc pseudomesenteroides R2, que foi isolado de repolho coletado em uma região semi-árida da Bahia. Utilizamos as seguintes condições para a produção de EPS: 10,7% de sacarose, pH 8,2, sem agitação e incubação a 28º C por 30 horas. O caldo fermentado foi tratado com etanol, gerando dois tipos de substâncias de polissacar

  20. Statistical optimization of process parameters for inulinase production from Tithonia weed by Arthrobacter mysorens strain no.1.

    Science.gov (United States)

    Kamble, Prajakta P; Kore, Maheshkumar V; Patil, Sushama A; Jadhav, Jyoti P; Attar, Yasmin C

    2018-06-01

    Tithonia rotundifolia is an easily available and abundant inulin rich weed reported to be competitive and allelopathic. This weed inulin is hydrolyzed by inulinase into fructose. Response surface methodology was employed to optimize culture conditions for the inulinase production from Arthrobacter mysorens strain no.1 isolated from rhizospheric area of Tithonia weed. Initially, Plackett- Burman design was used for screening 11 nutritional parameters for inulinase production including inulin containing weeds as cost effective substrate. The experiment shows that amongst the 11 parameters studied, K 2 HPO 4 , Inulin, Agave sisalana extract and Tithonia rotundifolia were the most significant variables for inulinase production. Quantitative effects of these 4 factors were further investigated using Box Behnken design. The medium having 0.27% K 2 HPO 4 , 2.54% Inulin, 6.57% Agave sisalana extract and 7.27% Tithonia rotundifolia extract were found to be optimum for maximum inulinase production. The optimization strategies used showed 2.12 fold increase in inulinase yield (1669.45 EU/ml) compared to non-optimized medium (787 EU/ml). Fructose produced by the action of inulinase was further confirmed by spectrophotometer, osazone, HPTLC and FTIR methods. Thus Tithonia rotundifolia can be used as an eco-friendly, economically feasible and promising alternative substrate for commercial inulinase production yielding fructose from Arthrobacter mysorens strain no.1. Copyright © 2018 Elsevier B.V. All rights reserved.

  1. Hydrogen production and metal-dye bioremoval by a Nostoc linckia strain isolated from textile mill oxidation pond.

    Science.gov (United States)

    Mona, Sharma; Kaushik, Anubha; Kaushik, C P

    2011-02-01

    Biohydrogen production by Nostoc linckia HA-46, isolated from a textile-industry oxidation-pond was studied by varying light/dark period, pH, temperature and ratio of carbon-dioxide and argon in the gas-mixture. Hydrogen production rates were maximum under 18 h of light and 6 h of darkness, pH 8.0, 31°C, a CO(2):Ar ratio 2:10. Hydrogen production of the strain acclimatized to 20 mg/L of chromium/cobalt and 100 mg/L of Reactive red 198/crystal violet dye studied in N-supplemented/deficient medium was 6-10% higher in the presence of 1.5 g/L of NaNO(3). Rates of hydrogen production in the presence of dyes/metals by the strain (93-105 μmol/h/mg Chlorophyll) were significantly higher than in medium without metals/dyes serving as control (91.3 μmol/h/mg Chlorophyll). About 58-60% of the two metals and 35-73% of dyes were removed by cyanobacterium. Optimal conditions of temperature, pH and metals/dyes concentration for achieving high hydrogen production and wastewater treatment were found practically applicable as similar conditions are found in the effluent of regional textile-mills. Copyright © 2010 Elsevier Ltd. All rights reserved.

  2. Extracellular polysaccharide production by a novel osmotolerant marine strain of Alteromonas macleodii and its application towards biomineralization of silver.

    Directory of Open Access Journals (Sweden)

    Ananya Mehta

    Full Text Available The present study demonstrates exopolysaccharide production by an osmotolerant marine isolate and also describes further application of the purified polysaccharide for production of colloidal suspension of silver nanoparticles with narrow size distribution. Phylogenetic analysis based on 16S r RNA gene sequencing revealed close affinity of the isolate to Alteromonas macleodii. Unlike earlier reports, where glucose was used as the carbon source, lactose was found to be the most suitable substrate for polysaccharide production. The strain was capable of producing 23.4 gl(-1 exopolysaccharide with a productivity of 7.8 gl(-1 day(-1 when 15% (w/v lactose was used as carbon source. Furthermore, the purified polysaccharide was able to produce spherical shaped silver nanoparticles of around 70 nm size as characterized by Uv-vis spectroscopy, Dynamic light scattering and Transmission electron microscopy. These observations suggested possible commercial potential of the isolated strain for production of a polysaccharide which has the capability of synthesizing biocompatible metal nanoparticle.

  3. Preliminary Study and Improve the Production of Metabolites with Antifungal Activity by A Bacillus Sp Strain IBA 33

    Directory of Open Access Journals (Sweden)

    María Antonieta Gordillo

    2009-01-01

    Full Text Available Bacillus sp strain IBA 33 metabolites, isolated from decaying lemon fruits, were evaluated for the control of pathogenic and non-pathogenic fungi (Penicillium digitatum, Geotrichum candidum, Penicillium expansum, Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, and Fusarium moniliforme. These metabolites were recovered from Landy medium (LM without aminoacids. In order to optimize metabolites production the LM was modified by adding different concentrations and sources of amino acids and carbohydrates at different culture conditions.Bacillus sp strain IBA 33 metabolites efficacy to control fungi were evaluated with in vitro and in vivo assays. A. flavus growth inhibition was 52% with the metabolites of Bacillus sp strain IBA 33 recovered from LM (MBLM in vitro assays. MBLM supplemented with 0.5% glutamic acid, inhibited the growth of P. digitatum, G. candidum, A. clavatus, A. niger and F. moniliforme by 65%, 88.44%, 84%, 34% and 92% respectively. The highest inhibition of P. expansum was 45% with MBLM supplemented with 0.5% aspartic acid. Similar results were obtained in vivo assays. These results showed that Bacillus sp strain IBA 33 metabolites specificity against fungi depended on the composition of the LM.

  4. Preliminary Study and Improve the Production of Metabolites with Antifungal Activity by A Bacillus Sp Strain IBA 33

    Directory of Open Access Journals (Sweden)

    María Antonieta Gordillo

    2009-04-01

    Full Text Available Bacillus sp strain IBA 33 metabolites, isolated from decaying lemon fruits, were evaluated for the control of pathogenic and non-pathogenic fungi (Penicillium digitatum, Geotrichum candidum, Penicillium expansum, Aspergillus clavatus, Aspergillus flavus, Aspergillus niger, and Fusarium moniliforme. These metabolites were recovered from Landy medium (LM without aminoacids. In order to optimize metabolites production the LM was modified by adding different concentrations and sources of amino acids and carbohydrates at different culture conditions. Bacillus sp strain IBA 33 metabolites efficacy to control fungi were evaluated with in vitro and in vivo assays. A. flavus growth inhibition was 52% with the metabolites of Bacillus sp strain IBA 33 recovered from LM (MBLM in vitro assays. MBLM supplemented with 0.5% glutamic acid, inhibited the growth of P. digitatum, G. candidum, A. clavatus, A. niger and F. moniliforme by 65%, 88.44%, 84%, 34% and 92% respectively. The highest inhibition of P. expansum was 45% with MBLM supplemented with 0.5% aspartic acid. Similar results were obtained in vivo assays. These results showed that Bacillus sp strain IBA 33 metabolites specificity against fungi depended on the composition of the LM.

  5. The ACEII recombinant Trichoderma reesei QM9414 strains with enhanced xylanase production and its applications in production of xylitol from tree barks.

    Science.gov (United States)

    Xiong, Lili; Kameshwar, Ayyappa Kumar Sista; Chen, Xi; Guo, Zhiyun; Mao, Canquan; Chen, Sanfeng; Qin, Wensheng

    2016-12-28

    ACEII transcription factor plays a significant role in regulating the expression of cellulase and hemicellulase encoding genes. Apart from ACEII, transcription factors such as XYR1, CRE1, HAP2/3/5 complex and ACEI function in a coordinated pattern for regulating the gene expression of cellulases and hemicellulases. Studies have demonstrated that ACEII gene deletion results in decreased total cellulase and xylanase activities with reduced transcript levels of lignocellulolytic enzymes. In this study, we have successfully transformed the ACEII transcription factor encoding gene in Trichoderma reesei to significantly improve its degrading abilities. Transformation experiments on parental strain T. reesei QM9414 has resulted in five genetically engineered strains T/Ace2-2, T/Ace2-5, T/Ace2-8, T/Ace5-4 and T/Ace10-1. Among which, T/Ace2-2 has exhibited significant increase in enzyme activity by twofolds, when compared to parental strain. The T/Ace2-2 was cultured on growth substrates containing 2% bark supplemented with (a) sugar free + MA medium (b) glucose + MA medium and (c) xylose + MA medium. The bark degradation efficiency of genetically modified T/Ace2-2 strain was assessed by analyzing the xylitol production yield using HPAEC. By 6th day, about 10.52 g/l of xylitol was produced through enzymatic conversion of bark (2% bark + MA + xylose) by the T/Ace2-2 strain and by 7th day the conversion rate was found to be 0.21 g/g. Obtained results confirmed that bark growth medium supplemented with D-xylose has profoundly increased the conversion rate of bark by T/Ace2-2 strain when compared to sugar free and glucose supplemented growth media. Results obtained from scanning electron microscopy has endorsed our current results. Bark samples inoculated with T/Ace2-2 strain has showed large number of degraded cells with clearly visible cavities and fractures, by exposing the microfibrillar interwoven complex. We propose a cost effective and ecofriendly method for

  6. A multi-phase approach to select new wine yeast strains with enhanced fermentative fitness and glutathione production.

    Science.gov (United States)

    Bonciani, Tommaso; De Vero, Luciana; Mezzetti, Francesco; Fay, Justin C; Giudici, Paolo

    2018-03-01

    The genetic improvement of winemaking yeasts is a virtually infinite process, as the design of new strains must always cope with varied and ever-evolving production contexts. Good wine yeasts must feature both good primary traits, which are related to the overall fermentative fitness of the strain, and secondary traits, which provide accessory features augmenting its technological value. In this context, the superiority of "blind," genetic improvement techniques, as those based on the direct selection of the desired phenotype without prior knowledge of the genotype, was widely proven. Blind techniques such as adaptive evolution strategies were implemented for the enhancement of many traits of interest in the winemaking field. However, these strategies usually focus on single traits: this possibly leads to genetic tradeoff phenomena, where the selection of enhanced secondary traits might lead to sub-optimal primary fermentation traits. To circumvent this phenomenon, we applied a multi-step and strongly directed genetic improvement strategy aimed at combining a strong fermentative aptitude (primary trait) with an enhanced production of glutathione (secondary trait). We exploited the random genetic recombination associated to a library of 69 monosporic clones of strain UMCC 855 (Saccharomyces cerevisiae) to search for new candidates possessing both traits. This was achieved by consecutively applying three directional selective criteria: molybdate resistance (1), fermentative aptitude (2), and glutathione production (3). The strategy brought to the selection of strain 21T2-D58, which produces a high concentration of glutathione, comparable to that of other glutathione high-producers, still with a much greater fermentative aptitude.

  7. Citric acid production and citrate synthase genes in distinct strains of ...

    African Journals Online (AJOL)

    Citric acid is an important organic acid, multifunctional with a wide array of uses. The objectives of this study were the isolation and selection strains of the genus Aspergillus, investigating the solubilization of phosphate of these isolates, verifying the expression rate of genes involved in the identification of isolates, and ...

  8. Survival of diverse bacillus thuringiensis strains in gypsy moth (Lepidotera: Lymantriidae) is correlated with urease production

    Science.gov (United States)

    Bacillus thuringiensis is an entomopathogenic bacterium that can kill a variety of pest insects, but seldom causes epizootics because it replicates poorly in insects. By attempting to repeatedly pass lepidopteran-active B. thuringiensis strains through gypsy moth larvae, we found that only those str...

  9. New hybrids between Saccharomyces sensu stricto yeast species found among wine and cider production strains

    DEFF Research Database (Denmark)

    Masneuf, I; Hansen, J.; Groth, C

    1998-01-01

    Two yeast isolates, a wine-making yeast first identified as a Mel(+) strain (ex. S. uvarum) and a cider-making yeast, were characterized for their nuclear and mitochondrial genomes, Electrophoretic karyotyping analyses, restriction fragment length polymorphism maps of PCR-amplified MET2 gene...

  10. Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion.

    Science.gov (United States)

    Hatvani, Lóránt; Manczinger, László; Kredics, László; Szekeres, András; Antal, Zsuzsanna; Vágvölgyi, Csaba

    2006-01-01

    The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T. atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 microg/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 microg/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T. atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.

  11. Comparisons of five Saccharomyces cerevisiae strains for ethanol production from SPORL pretreated lodgepole pine

    Science.gov (United States)

    The performances of 5 yeast strains under three levels of toxicity were evaluated using hydrolysates from lodgepole pine pretreated by Sulfite Pretreatment to Overcome the Recalcitrance of Lignocelluloses (SPORL). The highest level of toxicity was represented by the whole pretreated biomass slurry, ...

  12. Microbial succession of Debaryomyces hansenii strains during the production of Danish surfaced-ripened cheeses

    DEFF Research Database (Denmark)

    Petersen, Karen Mee; Westall, Signe; Jespersen, Lene

    2002-01-01

    to be the dominant yeast species throughout the ripening period, whereas other yeast species such as Trichosporon spp., Rhodotorula spp., and Candida spp. were found in minor concentrations during early stages of cheese ripening. Mitochondrial DNA RFLP was used to show that several strains of D. hansenii were...

  13. Saccharomyces cerevisiae strains tor second-generation ethanol production : from academie exploration to industrial implementation

    NARCIS (Netherlands)

    Jansen, Mickel L.A.; Bracher, J.M.; Papapetridis, I.; Verhoeven, M.D.; de Bruijn, J.A.; de Waal, P.; van Maris, A.J.A.; Klaassen, P; Pronk, J.T.

    2017-01-01

    The recent start-up of several full-scale ‘second generation’ ethanol plants marks a major milestone in the development of Saccharomyces cerevisiae strains for fermentation of lignocellulosic hydrolysates of agricultural residues and energy crops. After a discussion of the challenges that these

  14. Comparisons of five Saccharomyces cerevisiae strains for ethanol production from SPORL pretreated lodgepole pine

    Science.gov (United States)

    Haifeng Zhou; Tianqing Lan; Bruce S. Dien; Ronald E. Hector; J.Y. Zhu

    2014-01-01

    The performances of five yeast strains under three levels of toxicity were evaluated using hydrolysates from lodgepole pine pretreated by Sulfite Pretreatment to Overcome the Recalcitrance of Lignocelluloses (SPORL). The highest level of toxicity was represented by the whole pretreated biomass slurry, while intermediate toxicity was represented by the...

  15. Biodiesel Production from Selected Microalgae Strains and Determination of its Properties and Combustion Specific Characteristics

    Directory of Open Access Journals (Sweden)

    N. Kokkinos

    2015-11-01

    Full Text Available Biofuels are gaining importance as significant substitutes for the depleting fossil fuels. Recent focus is on microalgae as the third generation feedstock. In the present research work, two indigenous fresh water and two marine Chlorophyte strains have been cultivated successfully under laboratory conditions using commercial fertilizer (Nutrileaf 30-10-10, initial concentration=70 g/m3 as nutrient source. Gas chromatographic analysis data showed that microalgae biodiesel obtained from Chlorophyte strains biomass were composed of fatty acid methyl esters. The produced microalgae biodiesel achieved a range of 2.2 - 10.6 % total lipid content and an unsaturated FAME content between 49 mol% and 59 mol%. The iodine value, the cetane number, the cold filter plugging point, the oxidative stability as well as combustion specific characteristics of the final biodiesels were determined based on the compositions of the four microalgae strains. The calculated biodiesel properties compared then with the corresponding properties of biodiesel from known vegetable oils, from other algae strains and with the specifications in the EU (EN 14214 and US (ASTM D6751 standards. The derived biodiesels from indigenous Chlorophyte algae were significantly comparable in quality with other biodiesels.

  16. Metabolic characterization and transformation of the non-dairy Lactococcus lactis strain KF147, for production of ethanol from xylose

    DEFF Research Database (Denmark)

    Petersen, Kia Vest; Liu, Jianming; Chen, Jun

    2017-01-01

    producing ethanol as the sole fermentation product with a high yield corresponding to 83% of the theoretical maximum. The results clearly indicate the great potential of using the more metabolically diverse non-dairy L. lactis strains for bio-production based on xylose containing feedstocks.......The non-dairy lactic acid bacterium Lactococcus lactis KF147 can utilize xylose as the sole energy source. To assess whether KF147 could serve as a platform organism for converting second generation sugars into useful chemicals, we characterized growth and product formation for KF147 when grown...... the arcA gene encoding the arginine deiminase. The fermentation product profile suggested two routes for xylose degradation, the phosphoketolase pathway and the pentose phosphate pathway. Inactivation of the phosphoketolase pathway redirected the entire flux through the pentose phosphate pathway whereas...

  17. Simultaneous production of 2,3-butanediol, ethanol and hydrogen with a Klebsiella sp. strain isolated from sewage sludge.

    Science.gov (United States)

    Wu, Ken-Jer; Saratale, Ganesh D; Lo, Yung-Chung; Chen, Wen-Ming; Tseng, Ze-Jing; Chang, Ming-Ching; Tsai, Ben-Ching; Su, Ay; Chang, Jo-Shu

    2008-11-01

    A Klebsiella sp. HE1 strain isolated from hydrogen-producing sewage sludge was examined for its ability to produce H2 and other valuable soluble metabolites (e.g., ethanol and 2,3-butanediol) from sucrose-based medium. The effect of pH and carbon substrate concentration on the production of soluble and gaseous products was investigated. The major soluble metabolite produced from Klebsiella sp. HE1 was 2,3-butanediol, accounting for over 42-58% of soluble microbial products (SMP) and its production efficiency enhanced after increasing the initial culture pH to 7.3 (without pH control). The HE1 strain also produced ethanol (contributing to 29-42% of total SMP) and a small amount of lactic acid and acetic acid. The gaseous products consisted of H2 (25-36%) and CO2 (64-75%). The optimal cumulative hydrogen production (2.7 l) and hydrogen yield (0.92mol H2 mol sucrose(-1)) were obtained at an initial sucrose concentration of 30g CODl(-1) (i.e., 26.7gl(-1)), which also led to the highest production rate for H2 (3.26mmol h(-1)l(-1)), ethanol (6.75mmol h(-1)l(-1)) and 2,3-butanediol (7.14mmol h(-1)l(-1)). The highest yield for H2, ethanol and 2,3-butanediol was 0.92, 0.81 and 0.59molmol-sucrose(-1), respectively. As for the overall energy production performance, the highest energy generation rate was 27.7kJ h(-1)l(-1) and the best energy yield was 2.45kJmolsucrose(-1), which was obtained at a sucrose concentration of 30 and 20g CODl(-1), respectively.

  18. Optimization of endoglucanase production from thermophilic strain of Bacillus licheniformis RT-17 and its application for saccharification of sugarcane bagasse

    International Nuclear Information System (INIS)

    Tariq, R.; Qadir, F.; Ahmed, A.; Shariq, M.; Zafar, U.; Khan, S.A.

    2018-01-01

    Thermostable cellulases are required for a variety of commercial processes. Bacillus is a house of thermostable proteins. Screening of indigenously isolated strains of bacteria revealed the promising production of cellulase by a strain, RT-17, at 50 degree C. The strain was identified on the basis of biochemical and molecular characteristics as B. licheniformis. The factors affecting cellulase production from B. licheniformis RT-17 were evaluated for their significant effect using Plackett Burman Design and were optimized by employing Box-Behnken Design. The model predicted 9.808 IU/ml of endoglucanase (EG) under optimum conditions of 50 degree C; 10% inoculum size; pH 5; and 1% peptone in fermentation medium. Practically, a titer of 9.128 IU/ml was obtained, showed the validity of the model. The enzyme preparation from B. licheniformis RT-17 was applied in combination with xylanase and pectinase preparations from indigenous yeasts for the hydrolysis of sugarcane bagasse (SCB). A higher degree of synergy (7.1 folds) was observed when yeast pectinase was used with bacterial cellulase for the hydrolysis of alkali treated SCB. Whereas, the degree of synergy was lower when bacterial cellulase was mixed with yeast xylanase. The study revealed the possibility of utilization of combination of yeast and bacterial enzymes for biomass saccharification. (author)

  19. Production performance, use of nest box, and external appearance of two strains of laying hens kept in conventional and enriched cages.

    Science.gov (United States)

    Onbaşılar, E E; Ünal, N; Erdem, E; Kocakaya, A; Yaranoğlu, B

    2015-04-01

    The aim of this study was to investigate the differences in production performance, use of nest box, and external appearance of 2 strains of laying hens kept in conventional and enriched cages. Lohmann Brown Classic (LB, n=532) and Lohmann LSL Classic (LW, n=532) hens were housed from 16 to 73 wk in either conventional cages or enriched cages. Enriched cages had a nesting area, scratch pad, perch, and nail shortener. Body weight (BW), hen-day egg production, egg weight, feed intake, feed conversion ratio (FCR), cracked and dirty eggs, use of nest box for lay, and external appearance were determined. Laying period influenced the hen-day egg production, egg weight, feed intake, and feed conversion ratio. Cage type affected the hen-day egg production and feed conversion ratio, while strain affected the egg weight, feed intake, and feed conversion ratio. Laying period×cage type and laying period×strain interactions affected egg production, egg weight, and feed conversion ratio. Both strains preferred to lay in the nest box. Percentages of cracked and dirty eggs of LW hens in enriched cages were higher than that in conventional cages. Most of the dirty eggs laid by both strains were found outside of the nest box. The LW hens laid more dirty eggs than the LB hens. Cage type and cage type×strain interaction were important for total feather score. Final claw length was affected by cage type, strain, and cage type×strain interaction. This study suggests that cage type, strain, and also cage type×strain and period×strain interactions should be considered when alternative housing systems are used. © 2015 Poultry Science Association Inc.

  20. Engineering of a Bacillus subtilis strain with adjustable levels of intracellular biotin for secretory production of functional streptavidin.

    Science.gov (United States)

    Wu, Sau-Ching; Wong, Sui-Lam

    2002-03-01

    Streptavidin is a biotin-binding protein which has been widely used in many in vitro and in vivo applications. Because of the ease of protein recovery and availability of protease-deficient strains, the Bacillus subtilis expression-secretion system is an attractive system for streptavidin production. However, attempts to produce streptavidin using B. subtilis face the problem that cells overproducing large amounts of streptavidin suffer poor growth, presumably because of biotin deficiency. This problem cannot be solved by supplementing biotin to the culture medium, as this will saturate the biotin binding sites in streptavidin. We addressed this dilemma by engineering a B. subtilis strain (WB800BIO) which overproduces intracellular biotin. The strategy involves replacing the natural regulatory region of the B. subtilis chromosomal biotin biosynthetic operon (bioWAFDBIorf2) with an engineered one consisting of the B. subtilis groE promoter and gluconate operator. Biotin production in WB800BIO is induced by gluconate, and the level of biotin produced can be adjusted by varying the gluconate dosage. A level of gluconate was selected to allow enhanced intracellular production of biotin without getting it released into the culture medium. WB800BIO, when used as a host for streptavidin production, grows healthily in a biotin-limited medium and produces large amounts (35 to 50 mg/liter) of streptavidin, with over 80% of its biotin binding sites available for future applications.

  1. Direct ethanol production from lignocellulosic sugars and sugarcane bagasse by a recombinant Trichoderma reesei strain HJ48.

    Science.gov (United States)

    Huang, Jun; Chen, Dong; Wei, Yutuo; Wang, Qingyan; Li, Zhenchong; Chen, Ying; Huang, Ribo

    2014-01-01

    Trichoderma reesei can be considered as a candidate for consolidated bioprocessing (CBP) microorganism. However, its ethanol yield needs to be improved significantly. Here the ethanol production of T. reesei CICC 40360 was improved by genome shuffling while simultaneously enhancing the ethanol resistance. The initial mutant population was generated by nitrosoguanidine treatment of the spores, and an improved population producing more than fivefold ethanol than wild type was obtained by genome shuffling. The results show that the shuffled strain HJ48 can efficiently convert lignocellulosic sugars to ethanol under aerobic conditions. Furthermore, it was able to produce ethanol directly from sugarcane bagasse, demonstrating that the shuffled strain HJ48 is a suitable microorganism for consolidated bioprocessing.

  2. Production, Structural Elucidation, and In Vitro Antitumor Activity of Trehalose Lipid Biosurfactant from Nocardia farcinica Strain.

    Science.gov (United States)

    Christova, Nelly; Lang, Siegmund; Wray, Victor; Kaloyanov, Kaloyan; Konstantinov, Spiro; Stoineva, Ivanka

    2015-04-01

    The objective of this study was to isolate and identify the chemical structure of a biosurfactant produced by Nocardia farcinica strain BN26 isolated from soil, and evaluate its in vitro antitumor activity on a panel of human cancer cell lines. Strain BN26 was found to produce glycolipid biosurfactant on n-hexadecane as the sole carbon source. The biosurfactant was purified using medium-pressure liquid chromatography and characterized as trehalose lipid tetraester (THL) by nuclear magnetic resonance spectroscopy and mass spectrometry. Subsequently, the cytotoxic effects of THL on cancer cell lines BV-173, KE-37 (SKW-3), HL-60, HL-60/DOX, and JMSU-1 were evaluated by MTT assay. It was shown that THL exerted concentration-dependent antiproliferative activity against the human tumor cell lines and mediated cell death by the induction of partial oligonucleosomal DNA fragmentation. These findings suggest that THL could be of potential to apply in biomedicine as a therapeutic agent.

  3. Systematic mutagenesis method for enhanced production of bacitracin by Bacillus licheniformis mutant strain UV-MN-HN-6

    Directory of Open Access Journals (Sweden)

    Muhammad Nauman Aftab

    2012-03-01

    Full Text Available The purpose of the current study was intended to obtain the enhanced production of bacitracin by Bacillus licheniformis through random mutagenesis and optimization of various parameters. Several isolates of Bacillus licheniformis were isolated from local habitat and isolate designated as GP-35 produced maximum bacitracin production (14±0.72 IU ml-1. Bacitracin production of Bacillus licheniformis GP-35 was increased to 23±0.69 IU ml-1 after treatment with ultraviolet (UV radiations. Similarly, treatment of vegetative cells of GP-35 with chemicals like N-methyl N'-nitro N-nitroso guanidine (MNNG and Nitrous acid (HNO2 increased the bacitracin production to a level of 31±1.35 IU ml-1 and 27±0.89 IU ml-1 respectively. Treatment of isolate GP-35 with combined effect of UV and chemical treatment yield significantly higher titers of bacitracin with maximum bacitracin production of 41.6±0.92 IU ml-1. Production of bacitracin was further enhanced (59.1±1.35 IU ml-1 by optimization of different parameters like phosphate sources, organic acids as well as temperature and pH. An increase of 4.22 fold in the production of bacitracin after mutagenesis and optimization of various parameters was achieved in comparison to wild type. Mutant strain was highly stable and produced consistent yield of bacitracin even after 15 generations. On the basis of kinetic variables, notably Yp/s (IU/g substrate, Yp/x (IU/g cells, Yx/s (g/g, Yp/s, mutant strain B. licheniformis UV-MN-HN-6 was found to be a hyperproducer of bacitracin.

  4. Beta Glucan Production from Two Strains of Agrobacterium sp in Medium Containing of Molases and Uracil Combine

    Directory of Open Access Journals (Sweden)

    KUSMIATI

    2007-04-01

    Full Text Available Production of β-glucan by Agrobacterium sp is influenced by the composition of nutrition in the fermentation media. Molases has been used successfully by others in the fermentation media of S. cerevisiae to increase the yield of -glucan, and similarly, uracil has been used in the fermentation media of Agrobacterium sp to increase the yield of -glucan. Investigations to increase the yield of -glucan by two strains of Agrobacterium sp, i.e. A1.5 (reference and B4.4 (local strain, have been carried out by addition of various combination of molases and uracil into fermentation media, i.e. 5%(v/v molase-0,05%(b/v uracil; 5% molase-0,025% uracil; 10% molase-0,05% uracil; and 10% molase-0,025% uracil. The β-1,3-glucan and β-1,2-glucan fractions were separated by extraction method. Beta-glucan concentration was determined as the glucose monomer using the phenol-sulphate spectrophotometric method at 490 nm. The protein content was determined by a modified Lowry-spectrophotometric method at 750 nm. The results showed that all combination of molases and uracil in the fermentation media of Agrobacterium sp A1.5 and B4.4 strains have increased both the dry-weight yield of β-glucan (crude and the β¬glucan content, with the highest was in a medium containing 10% molases-0,025% uracil combination. In the above medium, the A1.5 strain produced the highest β-glucan (7,5% with the lowest protein content ( 8,4% in the β-1,3-glucan fraction, while the β-glucan content in the β-1,2-glucan fraction were all lower than in the control media, while the protein content were all higher than in the control media. In the above media, the B4.4 strain produced the highest β-glucan, 7,2% in the β-1,3-glucan fraction, and 13,1% in β-1,2-glucan fraction, while the lowest protein content ( 8,4% was in the β-1,3-glucan fraction. In conclusion, fermentation media of Agrobacterium sp A1.5 strain or B4.4 strain containing molase and uracil combination have increased both

  5. Rep-PCR typing of Staphylococcus spp. strains in meat paste production line and identification of their origin

    Directory of Open Access Journals (Sweden)

    Ivan Manga

    2015-05-01

    Full Text Available A meat paste production line and its microbial parameters have been evaluated in single Czech company. The raw meat paste samples before heat treatment were tested positively for the presence of three staphylococci species: Staphylococcus aureus, Staphylococcus haemolyticus and Staphylococcus epidermidis. Subsequent microbial analysis of meat paste components and ingredients (fresh meat, water, spices, equipment identified only the spices used as positive for S. aureus (coriander, cinnamon, badian, mustard – (10 - 40 cfu/g and S. haemolyticus strains (juniper, ginger. The collection of sixteen collected strains (S. aureus (n = 4, S. haemolyticus (n = 4, S. epidermidis (n = 8 has been typed with the rep-PCR method utilising (GTG5 primer. Analysis of the fingerprints using the unweighted pair-group method using arithmetic averages (UPGMA clustering method revealed presence of eleven strain clusters with similarity lower than 90%: two fingerprint clusters of S. aureus, three individual clusters characteristic for S. haemolyticus and six different S. epidermidis specific clusters. The S. aureus strains from different types of spice were identical, resp. very similar. Molecular tracking composed from the rep-PCR analysis of acquired isolates and comparison among all collected fingerprints confirmed the spices to be the source of both S. aureus and S. haemolyticus strains identified in raw meat paste. The additional rep-PCR analysis of the S. epidermidis collection confirmed usability and performance of this method. The antibiotic susceptibility to fourteen individual antibiotics has been examined among the collected staphylococci strains. The predominant erythromycin resistance (68.8% was followed with the resistance to amoxicillin/clavulanic acid (56.2%. Other resistances observed were less frequent (clindamycin – 12.5%, oxacillin – 6.3%, tetracycline – 6.3%, sulphamethoxazole-trimethoprim – 6.3%, chloramphenicol – 6.3%, novobiocin – 6

  6. Patulin and secondary metabolite production by marine-derived Penicillium strains

    DEFF Research Database (Denmark)

    Vansteelandt, Marieke; Kerzaon, Isabelle; Blanchet, Elodie

    2012-01-01

    )–mass spectrometry (MS)/MS. Each strain was grown on six different culture media to enhance the number of observable metabolites.Thirty-two secondary metabolites were detected in crude extracts with twenty first observations for studied species. Patulin, a major mycotoxin, was classically detected in extracts...... of these fungi in shellfish farming areas.Patulin induced acute neurotoxicity on Diptera larvae, indicating the interest of this bioassay as an additional tool for detection of this major mycotoxin in crude extracts....

  7. Penicillin production in industrial strain Penicillium chrysogenum P2niaD18 is not dependent on the copy number of biosynthesis genes.

    Science.gov (United States)

    Ziemons, Sandra; Koutsantas, Katerina; Becker, Kordula; Dahlmann, Tim; Kück, Ulrich

    2017-02-16

    Multi-copy gene integration into microbial genomes is a conventional tool for obtaining improved gene expression. For Penicillium chrysogenum, the fungal producer of the beta-lactam antibiotic penicillin, many production strains carry multiple copies of the penicillin biosynthesis gene cluster. This discovery led to the generally accepted view that high penicillin titers are the result of multiple copies of penicillin genes. Here we investigated strain P2niaD18, a production line that carries only two copies of the penicillin gene cluster. We performed pulsed-field gel electrophoresis (PFGE), quantitative qRT-PCR, and penicillin bioassays to investigate production, deletion and overexpression strains generated in the P. chrysogenum P2niaD18 background, in order to determine the copy number of the penicillin biosynthesis gene cluster, and study the expression of one penicillin biosynthesis gene, and the penicillin titer. Analysis of production and recombinant strain showed that the enhanced penicillin titer did not depend on the copy number of the penicillin gene cluster. Our assumption was strengthened by results with a penicillin null strain lacking pcbC encoding isopenicillin N synthase. Reintroduction of one or two copies of the cluster into the pcbC deletion strain restored transcriptional high expression of the pcbC gene, but recombinant strains showed no significantly different penicillin titer compared to parental strains. Here we present a molecular genetic analysis of production and recombinant strains in the P2niaD18 background carrying different copy numbers of the penicillin biosynthesis gene cluster. Our analysis shows that the enhanced penicillin titer does not strictly depend on the copy number of the cluster. Based on these overall findings, we hypothesize that instead, complex regulatory mechanisms are prominently implicated in increased penicillin biosynthesis in production strains.

  8. Comparison of SHF and SSF processes from steam-exploded wheat straw for ethanol production by xylose-fermenting and robust glucose-fermenting Saccharomyces cerevisiae strains

    DEFF Research Database (Denmark)

    Tomas Pejo, Elia; Oliva, Jose M.; Ballesteros, Mercedes

    2008-01-01

    In this study, bioethanol production from steam-exploded wheat straw using different process configurations was evaluated using two Saccharomyces cerevisiae strains, F12 and Red Star. The strain F12 has been engineerically modified to allow xylose consumption as cereal straw contain considerable ...

  9. Citric acid production and citrate synthase genes in distinct strains of ...

    African Journals Online (AJOL)

    SAM

    2014-05-28

    May 28, 2014 ... synthase in lactic acid production by A. niger and with the ... A number of microorganisms, including both bacteria and fungi, possess the capacity ..... citric acid production by solid-state fermentation from cassava bagasse and ...

  10. Advances in metabolic pathway and strain engineering paving the way for sustainable production of chemical building blocks.

    Science.gov (United States)

    Chen, Yun; Nielsen, Jens

    2013-12-01

    Bio-based production of chemical building blocks from renewable resources is an attractive alternative to petroleum-based platform chemicals. Metabolic pathway and strain engineering is the key element in constructing robust microbial chemical factories within the constraints of cost effective production. Here we discuss how the development of computational algorithms, novel modules and methods, omics-based techniques combined with modeling refinement are enabling reduction in development time and thus advance the field of industrial biotechnology. We further discuss how recent technological developments contribute to the development of novel cell factories for the production of the building block chemicals: adipic acid, succinic acid and 3-hydroxypropionic acid. Copyright © 2013 Elsevier Ltd. All rights reserved.

  11. Application of statistical experimental design for optimisation of bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium.

    Science.gov (United States)

    Ben Khedher, Saoussen; Jaoua, Samir; Zouari, Nabil

    2013-01-01

    In order to overproduce bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L(-1) starch, 30 g L(-1) soya bean and 9 g L(-1) sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch) when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch). Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis bioinsecticides at industrial scale which is of great importance in practical point of view.

  12. Application of statistical experimental design for optimisation of bioinsecticides production by sporeless Bacillus thuringiensis strain on cheap medium

    Directory of Open Access Journals (Sweden)

    Saoussen Ben Khedher

    2013-09-01

    Full Text Available In order to overproduce bioinsecticides production by a sporeless Bacillus thuringiensis strain, an optimal composition of a cheap medium was defined using a response surface methodology. In a first step, a Plackett-Burman design used to evaluate the effects of eight medium components on delta-endotoxin production showed that starch, soya bean and sodium chloride exhibited significant effects on bioinsecticides production. In a second step, these parameters were selected for further optimisation by central composite design. The obtained results revealed that the optimum culture medium for delta-endotoxin production consists of 30 g L-1 starch, 30 g L-1 soya bean and 9g L-1 sodium chloride. When compared to the basal production medium, an improvement in delta-endotoxin production up to 50% was noted. Moreover, relative toxin yield of sporeless Bacillus thuringiensis S22 was improved markedly by using optimised cheap medium (148.5 mg delta-endotoxins per g starch when compared to the yield obtained in the basal medium (94.46 mg delta-endotoxins per g starch. Therefore, the use of optimised culture cheap medium appeared to be a good alternative for a low cost production of sporeless Bacillus thuringiensis bioinsecticides at industrial scale which is of great importance in practical point of view.

  13. Co-fermentation using Recombinant Saccharomyces cerevisiae Yeast Strains Hyper-secreting Different Cellulases for the Production of Cellulosic Bioethanol.

    Science.gov (United States)

    Lee, Cho-Ryong; Sung, Bong Hyun; Lim, Kwang-Mook; Kim, Mi-Jin; Sohn, Min Jeong; Bae, Jung-Hoon; Sohn, Jung-Hoon

    2017-06-30

    To realize the economical production of ethanol and other bio-based chemicals from lignocellulosic biomass by consolidated bioprocessing (CBP), various cellulases from different sources were tested to improve the level of cellulase secretion in the yeast Saccharomyces cerevisiae by screening an optimal translational fusion partner (TFP) as both a secretion signal and fusion partner. Among them, four indispensable cellulases for cellulose hydrolysis, including Chaetomium thermophilum cellobiohydrolase (CtCBH1), Chrysosporium lucknowense cellobiohydrolase (ClCBH2), Trichoderma reesei endoglucanase (TrEGL2), and Saccharomycopsis fibuligera β-glucosidase (SfBGL1), were identified to be highly secreted in active form in yeast. Despite variability in the enzyme levels produced, each recombinant yeast could secrete approximately 0.6-2.0 g/L of cellulases into the fermentation broth. The synergistic effect of the mixed culture of the four strains expressing the essential cellulases with the insoluble substrate Avicel and several types of cellulosic biomass was demonstrated to be effective. Co-fermentation of these yeast strains produced approximately 14 g/L ethanol from the pre-treated rice straw containing 35 g/L glucan with 3-fold higher productivity than that of wild type yeast using a reduced amount of commercial cellulases. This process will contribute to the cost-effective production of bioenergy such as bioethanol and biochemicals from cellulosic biomass.

  14. Production and some properties of crude alkaline proteases of indigenous Central Amazonian rhizobia strains

    Directory of Open Access Journals (Sweden)

    Arlem Nascimento de Oliveira

    2010-10-01

    Full Text Available Two rhizobia strains isolated from soils of the Central Amazonian floodplain produced appreciable quantities of crude alkaline protease extracts with inexpensive carbon and nitrogen sources. These protease crude extracts were optimally active at pH 9.0-11.0. The optimum temperatures were 35 ºC for Rhizobium sp. strain R-986 and 55 ºC for Bradyrhizobium sp. strain R-993. Protease activities in the crude extracts were enhanced in the presence of 5 mM metal ions, such as Na+, Ca2+, Mg2+ and Mn2+. Rhizobia proteases were strongly inhibited by PMSF, a serine-protease inhibitor. The enzymes were active in the presence of surfactants (SDS and Triton X-100 and stable in oxidizing (H2O2 and reducing agents (β-mercaptoethanol, and organic solvents (acetone, hexane, methanol, 1-propanol and toluene.Duas estirpes de rizóbia isoladas de solos de várzea da Amazônia Central produziram grandes quantidades de proteases alcalinas extracelulares, usando fontes baratas de carbono e nitrogênio. Os extratos brutos de proteases foram ativos em pH 9,0-11,0. As temperaturas ótimas foram de 35 ºC para a enzima do Rhizobium R-986 e de 55 ºC para a do Bradyrhizobium R-993. As atividades proteolíticas aumentaram na presença de 5 mM dos íons Na+, Ca2+ , Mg2+ e Mn2+ . As proteases secretadas pelos rizóbios foram fortemente inibidas por PMSF, um inibidor de serina protease. As enzimas foram ativas na presença de surfactantes (SDS e Triton X-100, e estáveis na presença de agentes oxidantes (H2O2 e redutores (β-mercaptoetanol e solventes orgânicos (acetona, hexano, metanol, 1-propanol e tolueno.

  15. Production of xylan-degrading enzymes by a Trichoderma harzianum strain

    Directory of Open Access Journals (Sweden)

    Cacais André O.Guerreiro

    2001-01-01

    Full Text Available Trichoderma harzianum strain 4 produced extracellular xylan-degrading enzymes, namely beta-xylanase, beta-xylosidase and alpha-arabinofuranosidase, when grown in liquid medium cultures containing oat spelt xylan as inducer. Cellulase activity was not detected. The pattern of xylan-degrading enzymes induction was influenced by the form of xylan present in the medium. They were detected in different incubation periods. Electrophoretic separation of the proteins from liquid culture filtrates by SDS-PAGE showed a variety of bands with high and low molecular weights.

  16. Novel starters for old processes: use of Saccharomyces cerevisiae strains isolated from artisanal sourdough for craft beer production at a brewery scale.

    Science.gov (United States)

    Marongiu, Antonella; Zara, Giacomo; Legras, Jean-Luc; Del Caro, Alessandra; Mascia, Ilaria; Fadda, Costantino; Budroni, Marilena

    2015-01-01

    The deliberate inoculation of yeast strains isolated from food matrices such as wine or bread, could allow the transfer of novel properties to beer. In this work, the feasibility of the use of baker's yeast strains as starters for craft beer production has been evaluated at laboratory and brewery scale. Nine out of 12 Saccharomyces cerevisiae strains isolated from artisanal sourdoughs metabolized 2 % maltose, glucose and trehalose and showed growth rates and cell populations higher than those of the brewer's strain Safbrew-S33. Analysis of allelic variation at 12 microsatellite loci clustered seven baker's strains and Safbrew-S33 in the main group of bread isolates. Chemical analyses of beers produced at a brewery scale showed significant differences among the beers produced with the baker's strain S38 or Safbrew-S33, while no significant differences were observed when S38 or the brewer's strain Safbrew-F2 was used for re-fermentation. The sensory profile of beers obtained with S38 or the brewer's yeasts did not show significant differences, thus suggesting that baker's strains of S. cerevisiae could represent a reservoir of biodiversity for the selection of starter strains for craft beer production.

  17. Mycotoxin production capability of Penicillium roqueforti in strains isolated from mould-ripened traditional Turkish civil cheese.

    Science.gov (United States)

    Cakmakci, Songul; Gurses, Mustafa; Hayaloglu, A Adnan; Cetin, Bulent; Sekerci, Pinar; Dagdemir, Elif

    2015-01-01

    Mould-ripened civil is a traditional cheese produced mainly in eastern Turkey. The cheese is produced with a mixture of civil and whey curd cheeses (lor). This mixture is pressed into goat skins or plastic bags and is ripened for more than three months. Naturally occurring moulds grow on the surface and inside of the cheese during ripening. In this research, 140 Penicillium roqueforti strains were isolated from 41 samples of mould-ripened civil cheese collected from Erzurum and around towns in eastern Turkey. All strains were capable of mycotoxin production and were analysed using an HPLC method. It was established that all the strains (albeit at very low levels) produced roquefortine C, penicillic acid, mycophenolic acid and patulin. The amounts of toxins were in the ranges 0.4-47.0, 0.2-43.6, 0.1-23.1 and 0.1-2.3 mg kg(-1), respectively. Patulin levels of the samples were lower than the others. The lowest level and highest total mycotoxin levels were determined as 1.2 and 70.1 mg kg(-1) respectively. The results of this preliminary study may help in the choice of secondary cultures for mould-ripened civil cheese and other mould-ripened cheeses.

  18. Production and Purification of a Novel Xanthan Lyase from a Xanthan-Degrading Microbacterium sp. Strain XT11

    Directory of Open Access Journals (Sweden)

    Fan Yang

    2014-01-01

    Full Text Available A xanthan lyase was produced and purified from the culture supernatant of an excellent xanthan-modifying strain Microbacterium sp. XT11. Xanthan lyase was induced by xanthan but was inhibited by its structural monomer glucose. Its production by strain XT11 is much higher than that by all other reported strains. The purified xanthan lyase has a molecular mass of 110 kDa and a specific activity of 28.2 U/mg that was much higher than that of both Paenibacillus and Bacillus lyases. It was specific on the pyruvated mannosyl residue in the intact xanthan molecule, but about 50% lyase activity remained when xanthan was partially depyruvated. Xanthan lyase was optimally active at pH 6.0–6.5 and 40°C and alkali-tolerant at a high pH value of 11.0. The metal ions including K+, Ca2+, Na+, Mg2+, Mn2+, and Li+ strongly stimulated xanthan lyase activity but ions Zn2+ and Cu2+ were its inhibitor. Xanthan lyase should be a novel enzyme different from the other xanthan lyases ever reported.

  19. Taxonomic identification and lipid production of two Chilean Chlorella-like strains isolated from a marine and an estuarine coastal environment

    Science.gov (United States)

    González, Mariela A.; Pröschold, Thomas; Palacios, Yussi; Aguayo, Paula; Inostroza, Ingrid; Gómez, Patricia I.

    2013-01-01

    The genus Chlorella was the first microalga to be massively cultured as food, feed and as a source of nutraceuticals. More recently, some species have been suggested as candidates for biodiesel production. One of the most difficult tasks in studying the systematics of green coccoids is the identification of species assigned to the genus Chlorella. In the context of several projects carried out by our research group we isolated two Chlorella-like strains from a marine and an estuarine coastal environment in Chile (Coliumo strain and Baker strain, respectively). The main objectives of this research were to identify these Chilean strains—at the species level—and determine and compare their lipid production when cultured under identical conditions. Cell size and shape, autospore number and sizes, and chloroplast and pyrenoid ultrastructure were considered as taxonomic descriptors, and 18S rDNA sequences and internal transcribed spacer ITS-1 + ITS-2 sequences and secondary structure were adopted as phylogenetic tools. The combined use of these morphological, ultrastructural and molecular attributes revealed that only the Baker strain belongs to the genus Chlorella (C. vulgaris), while the Coliumo strain corresponds to the recently amended genus Chloroidium (C. saccharophilum). Lipid characterization of the biomass obtained from these strains showed that Chlorella vulgaris (Baker strain) appears to be suitable as a raw material for biodiesel production, while Chloroidium saccharophilum (Coliumo strain) would be more appropriate for animal nutrition.

  20. Influence of Nitrogen Source on 2,4-diacetylphloroglucinol Production by the Biocontrol Strain Pf-5

    OpenAIRE

    M, Hultberg; B, Alsanius

    2008-01-01

    The production of 2,4-diacetylphloroglucinol (DAPG) by the biocontrol agent Pseudomonas fluorescens Pf-5 was studied in nutrient-solution based media with varying nitrogen content. No production of DAPG was observed when organic nitrogen was omitted from the media, regardless of the inorganic nitrogen source used. Furthermore, a micromolar concentration range of organic nitrogen was insufficient to sustain production. When a millimolar concentration range of organic nitrogen was used, DAPG pr...

  1. Geographical markers for Saccharomyces cerevisiae strains with similar technological origins domesticated for rice-based ethnic fermented beverages production in North East India.

    Science.gov (United States)

    Jeyaram, Kumaraswamy; Tamang, Jyoti Prakash; Capece, Angela; Romano, Patrizia

    2011-11-01

    Autochthonous strains of Saccharomyces cerevisiae from traditional starters used for the production of rice-based ethnic fermented beverage in North East India were examined for their genetic polymorphism using mitochondrial DNA-RFLP and electrophoretic karyotyping. Mitochondrial DNA-RFLP analysis of S. cerevisiae strains with similar technological origins from hamei starter of Manipur and marcha starter of Sikkim revealed widely separated clusters based on their geographical origin. Electrophoretic karyotyping showed high polymorphism amongst the hamei strains within similar mitochondrial DNA-RFLP cluster and one unique karyotype of marcha strain was widely distributed in the Sikkim-Himalayan region. We conceptualized the possibility of separate domestication events for hamei strains in Manipur (located in the Indo-Burma biodiversity hotspot) and marcha strains in Sikkim (located in Himalayan biodiversity hotspot), as a consequence of less homogeneity in the genomic structure between these two groups, their clear separation being based on geographical origin, but not on technological origin and low strain level diversity within each group. The molecular markers developed based on HinfI-mtDNA-RFLP profile and the chromosomal doublets in chromosome VIII position of Sikkim-Himalayan strains could be effectively used as geographical markers for authenticating the above starter strains and differentiating them from other commercial strains.

  2. Enhanced cellulase recovery without β-glucosidase supplementation for cellulosic ethanol production using an engineered strain and surfactant.

    Science.gov (United States)

    Huang, Renliang; Guo, Hong; Su, Rongxin; Qi, Wei; He, Zhimin

    2017-03-01

    Recycling cellulases by substrate adsorption is a promising strategy for reducing the enzyme cost of cellulosic ethanol production. However, β-glucosidase has no carbohydrate-binding module (CBM). Thus, additional enzymes are required in each cycle to achieve a high ethanol yield. In this study, we report a new method of recycling cellulases without β-glucosidase supplementation using lignocellulosic substrate, an engineered strain expressing β-glucosidase and Tween 80. The cellulases and Tween 80 were added to an aqueous suspension of diluted sulfuric acid/ammonia-treated corncobs in a simultaneous saccharification and fermentation (SSF) process for ethanol production. Subsequently, the addition of fresh pretreated corncobs to the fermentation liquor and remaining solid residue provided substrates with absorbed cellulases for the next SSF cycle. This method provided excellent ethanol production in three successive SSF cycles without requiring the addition of new cellulases. For a 10% (w/v) solid loading, a cellulase dosage of 30 filter paper units (FPU)/g cellulose, 0.5% Tween 80, and 2 g/L of the engineered strain, approximately 90% of the initial ethanol concentration from the first SSF process was obtained in the next two SSF processes, with a total ethanol production of 306.27 g/kg corncobs and an enzyme productivity of 0.044 g/FPU. Tween 80 played an important role in enhancing cellulase recovery. This new enzyme recycling method is more efficient and practical than other reported methods. Biotechnol. Bioeng. 2017;114: 543-551. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  3. Comparative evaluation of two Trichoderma harzianum strains for major secondary metabolite production and antifungal activity.

    Science.gov (United States)

    Ahluwalia, Vivek; Kumar, Jitendra; Rana, Virendra S; Sati, Om P; Walia, S

    2015-01-01

    This investigation was undertaken to identify the major secondary metabolite, produced by two Trichoderma harzianum strains (T-4 and T-5) with their antifungal activity against phytopathogenic fungi using poison food technique. The ethyl acetate extract was subjected to column chromatography using n-hexane, ethyl acetate and methanol gradually. Chromatographic separation of ethyl acetate extract of T. harzianum (T-4) resulted in the isolation and identification of palmitic acid (1), 1,8-dihydroxy-3-methylanthraquinone (2), 6-pentyl-2H-pyran-2-one (3), 2(5H)-furanone (4), stigmasterol (5) and β-sitosterol (6), while T. harzianum (T-5) gave palmitic acid (1), 1-hydroxy-3-methylanthraquinone (7), δ-decanolactone (8), 6-pentyl-2H-pyran-2-one (3), ergosterol (9), harzianopyridone (10) and 6-methyl-1,3,8-trihydroxyanthraquinone (11) as major metabolites. Among compounds screened for antifungal activity, compound 10 was found to be most active (EC50 35.9-50.2 μg mL(-1)). In conclusion, the present investigation provided significant information about antifungal activity and compounds isolated from two different strains of T. harzianum obtained from two different Himalayan locations.

  4. Selection, application and monitoring of Lactobacillus paracasei strains as adjunct cultures in the production of Gouda-type cheeses.

    Science.gov (United States)

    Van Hoorde, Koenraad; Van Leuven, Isabelle; Dirinck, Patrick; Heyndrickx, Marc; Coudijzer, Kathleen; Vandamme, Peter; Huys, Geert

    2010-12-15

    Raw milk cheeses have more intense flavours than cheeses made from pasteurized milk and harbour strains with potential adjunct properties. Two Lactobacillus paracasei strains, R-40926 and R-40937, were selected as potential adjunct cultures from a total of 734 isolates from good quality artisan raw milk Gouda-type cheeses on the basis of their prevalence in different cheese types and/or over several production batches, safety and technological parameters. Conventional culturing, isolation and identification and a combined PCR-DGGE approach using total cheese DNA extracts and DNA extracts obtained from culturable fractions were employed to monitor viability of the introduced adjuncts and their effect on the cheese microbiota. The control cheese made without adjuncts was dominated by members of the starter, i.e. Lactococcus lactis and Leuconostoc pseudomesenteroides. In the cheeses containing either R-40926 or R-40937, the respective adjuncts increased in number as ripening progressed indicating that both strains are well adapted to the cheese environment and can survive in a competitive environment in the presence of a commercial starter culture. Principal component analysis of cheese volatiles determined by steam distillation-extraction and gas chromatography-mass spectrometry could differentiate cheeses made with different concentrations of adjunct R-40926 from the control cheese, and these differences could be correlated to the proteolytic and lipolytic properties of this strain. Collectively, results from microbiological and metabolic analyses indicate that the screening procedure followed throughout this study was successful in delivering potential adjunct candidates to enrich or extend the flavour palette of artisan Gouda-type cheeses under more controlled conditions. Copyright © 2010 Elsevier B.V. All rights reserved.

  5. Statistical optimization of thermo-alkali stable xylanase production from Bacillus tequilensis strain ARMATI

    Directory of Open Access Journals (Sweden)

    Ameer Khusro

    2016-07-01

    Conclusions: The cellulase-free xylanase showed an alkali-tolerant and thermo-stable property with potentially applicable nature at industrial scale. This statistical approach established a major contribution in enzyme production from the isolate by optimizing independent factors and represents a first reference on the enhanced production of thermo-alkali stable cellulase-free xylanase from B. tequilensis.

  6. Screening of phospholipase A activity and its production by new actinomycete strains cultivated by solid-state fermentation

    Directory of Open Access Journals (Sweden)

    Priscila Sutto-Ortiz

    2017-07-01

    Full Text Available Novel microbial phospholipases A (PLAs can be found in actinomycetes which have been poorly explored as producers of this activity. To investigate microbial PLA production, efficient methods are necessary such as high-throughput screening (HTS assays for direct search of PLAs in microbial cultures and cultivation conditions to promote this activity. About 200 strains isolated with selected media for actinomycetes and mostly belonging to Streptomyces (73% and Micromonospora (10% genus were first screened on agar-plates containing the fluorophore rhodamine 6G and egg yolk phosphatidylcholine (PC to detect strains producing phospholipase activity. Then, a colorimetric HTS assay for general PLA activity detection (cHTS-PLA using enriched PC (≈60% as substrate and cresol red as indicator was developed and applied; this cHTS-PLA assay was validated with known PLAs. For the first time, actinomycete strains were cultivated by solid-state fermentation (SSF using PC as inductor and sugar-cane bagasse as support to produce high PLA activity (from 207 to 2,591 mU/g of support. Phospholipase activity of the enzymatic extracts from SSF was determined using the implemented cHTS-PLA assay and the PC hydrolysis products obtained, were analyzed by TLC showing the presence of lyso-PC. Three actinomycete strains of the Streptomyces genus that stood out for high accumulation of lyso-PC, were selected and analyzed with the specific substrate 1,2-α-eleostearoyl-sn-glycero-3-phosphocholine (EEPC in order to confirm the presence of PLA activity in their enzymatic extracts. Overall, the results obtained pave the way toward the HTS of PLA activity in crude microbial enzymatic extracts at a larger scale. The cHTS-PLA assay developed here can be also proposed as a routine assay for PLA activity determination during enzyme purification,directed evolution or mutagenesis approaches. In addition, the production of PLA activity by actinomycetes using SSF allow find and

  7. Malt sprout, an underused beer by-product with promising potential for the growth and dehydration of lactobacilli strains.

    Science.gov (United States)

    Cejas, Luján; Romano, Nelson; Moretti, Ana; Mobili, Pablo; Golowczyc, Marina; Gómez-Zavaglia, Andrea

    2017-12-01

    Malt sprout (MS), a by-product of the malt industry obtained by removing rootlets and sprouts from the seed of germinated barley ( Hordeum vulgare L.), was used as culture, dehydration and storage medium of three strains of lactobacilli: Lactobacillus salivarius CM-CIDCA 1231B and CM-CIDCA 1232Y and Lactobacillus plantarum CIDCA 83114. The three strains were grown in MS and MS supplemented with 20% w/v fructo-oligosaccharides (MS FOS). Bacterial growth was determined by registering the decrease of pH and by plate counting. Comparable results with those of microorganisms grown in MRS (controls) were observed in terms of lag times, ΔpH and acidification rates. Furthermore, during fermentation, a significant increase of DP6 (FOS with degree of polymerization 6) was observed at expenses of inulin and DP7, probably indicating their hydrolysis. A concomitant decrease of DP3, sucrose and monosaccharides was also observed, as result of their bacterial consumption during growth. The presence of FOS in the fermented media protected microorganisms during freeze-drying and storage, as no decrease of culturability was observed after 60 days at 4 °C (> 10 8 CFU/mL). Using MS appears as an innovative strategy for the production of lactobacilli at large scale, supporting their use for the elaboration of functional foods containing prebiotics and probiotics.

  8. High-level ethanol production from starch by a flocculent Saccharomyces cerevisiae strain displaying cell-surface glucoamylase

    Energy Technology Data Exchange (ETDEWEB)

    Kondo, A.; Shigechi, H.; Abe, M.; Uyama, K. [Dept. of Chemical Science and Engineering, Kobe Univ., Nadaku, Kobe (Japan); Matsumoto, T.; Fukuda, H. [Div. of Molecular Science, Kobe Univ., Nadaku, Kobe (Japan); Takahashi, S.; Ueda, M.; Tanaka, A. [Dept. of Synthetic Chemistry and Biological Chemistry, Kyoto Univ., Sakyoku, Kyoto (Japan); Kishimoto, M. [Dept. of Biotechnology, Osaka Univ., Osaka (Japan)

    2002-07-01

    A Strain of host yeast YF207, which is a tryptophan auxotroph and shows strong flocculation ability, was obtained from Saccharomyces diastaticus ATCC60712 and S. cerevisiae W303-1B by tetrad analysis. The plasmid pGA11, which is a multicopy plasmid for cell-surface expression of the Rhyzopus oryzae glucoamylase/{alpha}-agglutinin fusion protein, was then introduced into this flocculent yeast strain (YF207/pGA11). Yeast YF207/pGA11 grew rapidly under aerobic condition (dissolved oxygen 2.0 ppm), using soluble starch. The harvested cells were used for batch fermentation of soluble starch to ethanol under anaerobic condition and showed high ethanol production rates (0.71 g h{sup -1} I{sup -1}) without a time lag, because glucoamylase was immobilized on the yeast cell surface. During repeated utilization of cells for fermentation, YF207/pGA11 maintained high ethanol production rates over 300 h. Moreover, in fed-batch fermentation with YF207/pGA11 for approximately 120 h, the ethanol concentration reached up to 50 g I{sup -1}. In conclusion, flocculent yeast cells displaying cell-surface glucoamylase are considered to be very effective for the direct fermentation of soluble starch to ethanol. (orig.)

  9. Approach toward enhancement of halophilic protease production by Halobacterium sp. strain LBU50301 using statistical design response surface methodology.

    Science.gov (United States)

    Chuprom, Julalak; Bovornreungroj, Preeyanuch; Ahmad, Mehraj; Kantachote, Duangporn; Dueramae, Sawitree

    2016-06-01

    A new potent halophilic protease producer, Halobacterium sp. strain LBU50301 was isolated from salt-fermented fish samples ( budu ) and identified by phenotypic analysis, and 16S rDNA gene sequencing. Thereafter, sequential statistical strategy was used to optimize halophilic protease production from Halobacterium sp. strain LBU50301 by shake-flask fermentation. The classical one-factor-at-a-time (OFAT) approach determined gelatin was the best nitrogen source. Based on Plackett - Burman (PB) experimental design; gelatin, MgSO 4 ·7H 2 O, NaCl and pH significantly influenced the halophilic protease production. Central composite design (CCD) determined the optimum level of medium components. Subsequently, an 8.78-fold increase in corresponding halophilic protease yield (156.22 U/mL) was obtained, compared with that produced in the original medium (17.80 U/mL). Validation experiments proved the adequacy and accuracy of model, and the results showed the predicted value agreed well with the experimental values. An overall 13-fold increase in halophilic protease yield was achieved using a 3 L laboratory fermenter and optimized medium (231.33 U/mL).

  10. Approach toward enhancement of halophilic protease production by Halobacterium sp. strain LBU50301 using statistical design response surface methodology

    Directory of Open Access Journals (Sweden)

    Julalak Chuprom

    2016-06-01

    Full Text Available A new potent halophilic protease producer, Halobacterium sp. strain LBU50301 was isolated from salt-fermented fish samples (budu and identified by phenotypic analysis, and 16S rDNA gene sequencing. Thereafter, sequential statistical strategy was used to optimize halophilic protease production from Halobacterium sp. strain LBU50301 by shake-flask fermentation. The classical one-factor-at-a-time (OFAT approach determined gelatin was the best nitrogen source. Based on Plackett–Burman (PB experimental design; gelatin, MgSO4·7H2O, NaCl and pH significantly influenced the halophilic protease production. Central composite design (CCD determined the optimum level of medium components. Subsequently, an 8.78-fold increase in corresponding halophilic protease yield (156.22 U/mL was obtained, compared with that produced in the original medium (17.80 U/mL. Validation experiments proved the adequacy and accuracy of model, and the results showed the predicted value agreed well with the experimental values. An overall 13-fold increase in halophilic protease yield was achieved using a 3 L laboratory fermenter and optimized medium (231.33 U/mL.

  11. Optimization of laccase production by two strains of Ganoderma lucidum using phenolic and metallic inducers

    Directory of Open Access Journals (Sweden)

    Francisco Kuhar

    Full Text Available Ganoderma lucidum (Curtis P. Karst is a white rot fungus that is able to degrade the lignin component in wood. The ability of two strains of this species to produce the ligninolytic enzyme laccase was assessed. After the evaluation of induction with heavy metals and phenolic compounds, it was found that among the tested substances, copper and ferulic acid are the best laccase inducers. It was also observed that the two types of inducers (phenolic and metallic produce different electrophoretic patterns of laccase activity. Optimized concentrations of inducers were obtained through a factorial design and the thermal stability of optimized supernatants was studied at a wide range of acidic pH. We found that the enzyme is more thermostable at higher pH values.

  12. EVALUATION OF STRAIN-STRESS STATE OF THE RAILS IN THE PRODUCTION

    Directory of Open Access Journals (Sweden)

    V. V. Muravev

    2017-01-01

    Full Text Available High values of residual stresses is one of the most common reason of breaking lots of metal constructions, including rails. These stresses can reach values of flow limit, especially in the area of faults. Estimation of residual stresses values allows to get information about technical condition of the rail and also allow to avoid abnormal situations So, the aim of the research is creating the model of stress-strain state of the rail, which was hardened in its top and bottom, and to compare modeling results with experimental measurements of stresses and discrepancy of the housing.For creating the model and making evaluations by finite element method we used a program COMSOL. Forces on the top and bottom of the rail cause tension stresses, forces on the web of the rail cause tensile stresses. We compared calculated values of stresses with discrepancy of the housing. The discrepancy of the housing is informative characteristic for estimating the residual stresses according to standards. For experimental measurements we used an acoustic structuroscope SEMA. This structuroscope uses the acoustoelastic phenomenon for measurements. We made measurements of the five rails.According to the calculation results of the model, critical discrepancy of the housing in 2 mm corresponded to the following values of maximum stresses: –54 MPa in the top of the rail, 86 MPa in the web and –62 MPa in the bottom of the rail. Experimental measurements are the following: from –48 MPa to – 64 MPa in the top of the rail, 54 MPa to 93 MPa in the web of the rail, and –59 MPA to –74 MPa in the bottom of the rail. Absolute error was ±5 MPa.Thus we created the model, which allowed to analyze strain-stress state and compare real values of stresses with discrepancy of the housing. Results of the modeling showed coincidence with structure of distribution of residual stresses in five probes of rails. 

  13. Proteolysis in tempeh-type products obtained with Rhizopus and Aspergillus strains from grass pea (Lathyrus sativus) seeds.

    Science.gov (United States)

    Starzyńska-Janiszewska, Anna; Stodolak, Bożena; Wikiera, Agnieszka

    2015-01-01

    Tempeh is a food product obtained from legumes by means of solid-state fermentation with Rhizopus sp. Our previous research proved that mixed-culture inoculum may also be successfully applied. The objective of present research was to study the proteolytic activity of R. microsporus var. chinensis and A. oryzae during tempeh-type fermentation of grass pea seeds, and the effect of inoculum composition on the protein level and in vitro protein bioavailability in products. Fermentation substrate were soaked and cooked grass pea seeds. Material was mixed with single- or mixed-culture inoculum, and incubated in perforated plastic bags at 30°C for 32 hrs. In the products, the proteolytic activity (pH 3, 5 and 7), glucosamine, total protein and free amino acids levels, as well as protein in vitro bioavailability and degree of protein hydrolysis were obtained. The significant correlation was found between glucosamine content and proteolytic activity in grass pea seeds fermented with Rhizopus or Aspergillus. The activities of Rhizopus proteases were higher than Aspergillus ones, which corresponded with the degree of seed protein hydrolysis. Both strains showed the highest activity of protease at pH 3. Tempeh made with pure culture of Rhizopus had 37% protein of 69% in-vitro bioavailability. Mixed-culture fermentation improved nutritional parameters of products only when the dose of Aspergillus spores in the inoculum was equal and lower than that of Rhizopus. This process resulted in higher in-vitro bioavailability of protein, slightly more efficient protein hydrolysis and higher level of free amino acids, as compared to standard tempeh. The activity of A. oryzae in tempeh-type fermentation is beneficial as long as it does not dominate the activity and/or growth of Rhizopus strain.

  14. The early use of yellow fever virus strain 17D for vaccine production in Brazil - a review

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    Paulo Roberto Post

    2001-08-01

    Full Text Available The use of yellow fever (YF virus 17D strain for vaccine production adapted in Brazil since its introduction in 1937 was reviewed. This was possible due to the availability of official records of vaccine production. The retrieved data highlight the simultaneous use of several serially passaged 17D substrain viruses for both inocula and vaccine preparation that allowed uninterrupted production. Substitution of these substrain viruses became possible with the experience gained during quality control and human vaccination. Post-vaccinal complications in humans and the failure of some viruses in quality control tests (neurovirulence for monkeys indicated that variables needed to be reduced during vaccine production, leading to the development of the seed lot system. The 17DD substrain, still used today, was the most frequently used substrain and the most reliable in terms of safety and efficacy. For this reason, it is possible to derive an infectious cDNA clone of this substrain combined with production in cell culture that could be used to direct the expression of heterologous antigens and lead to the development of new live vaccines.

  15. Strain Screening from Traditional Fermented Soybean Foods and Induction of Nattokinase Production in Bacillus subtilis MX-6.

    Science.gov (United States)

    Man, Li-Li; Xiang, Dian-Jun; Zhang, Chun-Lan

    2018-02-06

    The plasminogen-free fibrin plate assay method was used to isolate Bacillus subtilis MX-6, a strain with high production of nattokinase from Chinese douchi. The presence of aprN, a gene-encoding nattokinase, was verified with PCR method. The predicted amino acid sequence was aligned with homologous sequences, and a phylogenetic tree was constructed. Nattokinase was sublimated with ammonium sulfate, using a DEAE-Sepharose Fast Flow column, a CM-Sepharose Fast Flow column and a Sephadex G-75 gel filtration column. SDS-PAGE analysis indicated that the molecular weight of the purified nattokinase from Bacillus subtilis MX-6 was about 28 kDa. Fermentation of Bacillus subtilis MX-6 nattokinase showed that nattokinase production was maximized after 72 h; the diameter of clear zone reached 21.60 mm on the plasminogen-free fibrin plate. Nattokinase production by Bacillus subtilis MX-6 increased significantly after supplementation with supernatant I, supernatant II and soy peptone but decreased substantially after the addition of amino acids. This result indicated that the nattokinase production by B. subtilis MX-6 might be induced by soybean polypeptides. The addition of MgSO 4 and CaCl 2 increased B. subtilis MX-6 nattokinase production.

  16. Exposure of clinical MRSA heterogeneous strains to β-lactams redirects metabolism to optimize energy production through the TCA cycle.

    Directory of Open Access Journals (Sweden)

    Mignon A Keaton

    adaptation that HeR-MRSA clinical strains undergo when exposed to β-lactam pressure, indicating that the energy production is redirected to supply the cell wall synthesis/metabolism, which in turn contributes to the survival response in the presence of β-lactam antibiotics.

  17. Exposure of clinical MRSA heterogeneous strains to β-lactams redirects metabolism to optimize energy production through the TCA cycle.

    Science.gov (United States)

    Keaton, Mignon A; Rosato, Roberto R; Plata, Konrad B; Singh, Christopher R; Rosato, Adriana E

    2013-01-01

    R-MRSA clinical strains undergo when exposed to β-lactam pressure, indicating that the energy production is redirected to supply the cell wall synthesis/metabolism, which in turn contributes to the survival response in the presence of β-lactam antibiotics.

  18. Isolation and partial characterization of a new strain of Klebsiella pneumoniae capable of high 1,3 propanediol production from glycerol

    Directory of Open Access Journals (Sweden)

    B. Sen

    2015-04-01

    Full Text Available Glycerol is a promising feedstock for microbial cultivation and production of 1,3 propanediol (1,3 PDO. Here we report a newly isolated bacterial strain BA11 from soil, capable of fermenting glycerol to 1,3 PDO, and has been identified to be a strain of Klebsiella pneumoniae. Strain BA11 was fast growing showing peak 1,3 PDO production in 6 h of cultivation with productivity of 1.2 g/L-h without the addition of Vitamin B12. Based on the optimum glycerol utilization (75% and 1,3 PDO production (8.3 g/L and yield (0.56 mol/mol glycerol utilized, the most appropriate glycerol concentration for cultivation was 20 g/L. The strain BA11 could tolerate the pH range of 6 to 8.5 as no inhibitory effects were seen on growth as well as 1,3 PDO production. Strain BA11 was most active and could produce high 1,3 PDO in the incubation temperature range of 25 to 40 oC. The production of 1,3 PDO was maximum (9.3 g/L under aerobic condition with 95.8% glycerol utilization. Addition of glucose to the glycerol fermentation led to increased cell mass but no improvement in the 1,3 PDO production.

  19. Development of a rapid SNP-typing assay to differentiate Bifidobacterium animalis ssp. lactis strains used in probiotic-supplemented dairy products.

    Science.gov (United States)

    Lomonaco, Sara; Furumoto, Emily J; Loquasto, Joseph R; Morra, Patrizia; Grassi, Ausilia; Roberts, Robert F

    2015-02-01

    Identification at the genus, species, and strain levels is desirable when a probiotic microorganism is added to foods. Strains of Bifidobacterium animalis ssp. lactis (BAL) are commonly used worldwide in dairy products supplemented with probiotic strains. However, strain discrimination is difficult because of the high degree of genome identity (99.975%) between different genomes of this subspecies. Typing of monomorphic species can be carried out efficiently by targeting informative single nucleotide polymorphisms (SNP). Findings from a previous study analyzing both reference and commercial strains of BAL identified SNP that could be used to discriminate common strains into 8 groups. This paper describes development of a minisequencing assay based on the primer extension reaction (PER) targeting multiple SNP that can allow strain differentiation of BAL. Based on previous data, 6 informative SNP were selected for further testing, and a multiplex preliminary PCR was optimized to amplify the DNA regions containing the selected SNP. Extension primers (EP) annealing immediately adjacent to the selected SNP were developed and tested in simplex and multiplex PER to evaluate their performance. Twenty-five strains belonging to 9 distinct genomic clusters of B. animalis ssp. lactis were selected and analyzed using the developed minisequencing assay, simultaneously targeting the 6 selected SNP. Fragment analysis was subsequently carried out in duplicate and demonstrated that the assay yielded 8 specific profiles separating the most commonly used commercial strains. This novel multiplex PER approach provides a simple, rapid, flexible SNP-based subtyping method for proper characterization and identification of commercial probiotic strains of BAL from fermented dairy products. To assess the usefulness of this method, DNA was extracted from yogurt manufactured with and without the addition of B. animalis ssp. lactis BB-12. Extracted DNA was then subjected to the minisequencing

  20. Bacillus coagulans MA-13: a promising thermophilic and cellulolytic strain for the production of lactic acid from lignocellulosic hydrolysate.

    Science.gov (United States)

    Aulitto, Martina; Fusco, Salvatore; Bartolucci, Simonetta; Franzén, Carl Johan; Contursi, Patrizia

    2017-01-01

    The transition from a petroleum-based economy towards more sustainable bioprocesses for the production of fuels and chemicals (circular economy) is necessary to alleviate the impact of anthropic activities on the global ecosystem. Lignocellulosic biomass-derived sugars are suitable alternative feedstocks that can be fermented or biochemically converted to value-added products. An example is lactic acid, which is an essential chemical for the production of polylactic acid, a biodegradable bioplastic. However, lactic acid is still mainly produced by Lactobacillus species via fermentation of starch-containing materials, the use of which competes with the supply of food and feed. A thermophilic and cellulolytic lactic acid producer was isolated from bean processing waste and was identified as a new strain of Bacillus coagulans , named MA-13. This bacterium fermented lignocellulose-derived sugars to lactic acid at 55 °C and pH 5.5. Moreover, it was found to be a robust strain able to tolerate high concentrations of hydrolysate obtained from wheat straw pre-treated by acid-catalysed (pre-)hydrolysis and steam explosion, especially when cultivated in controlled bioreactor conditions. Indeed, unlike what was observed in microscale cultivations (complete growth inhibition at hydrolysate concentrations above 50%), B. coagulans MA-13 was able to grow and ferment in 95% hydrolysate-containing bioreactor fermentations. This bacterium was also found to secrete soluble thermophilic cellulases, which could be produced at low temperature (37 °C), still retaining an optimal operational activity at 50 °C. The above-mentioned features make B. coagulans MA-13 an appealing starting point for future development of a consolidated bioprocess for production of lactic acid from lignocellulosic biomass, after further strain development by genetic and evolutionary engineering. Its optimal temperature and pH of growth match with the operational conditions of fungal enzymes hitherto

  1. Characterisation of the thermostable protease AprX in strains of Pseudomonas fluorescens and impact on the shelf-life of dairy products: preliminary results

    Directory of Open Access Journals (Sweden)

    Nadia Andrea Andreani

    2016-12-01

    Full Text Available Bacterial proteases are involved in food spoilage and shelf-life reduction. Among the bacterial pro