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Sample records for freudenreichii cctcc m207015

  1. 低成本绿色循环工艺发酵生产丙酸联产维生素B12%Green and inexpensive circulation process for propionic acid and vitamin B12 production by Propionibacterium freudenreichii CCTCC M207015

    Institute of Scientific and Technical Information of China (English)

    陈飞; 张力; 冯小海; 吴波; 徐虹

    2011-01-01

    对提取维生素B12后的费氏丙酸杆菌废菌体进行水解处理,考察以菌体水解液作为N源用于丙酸发酵的可行性.利用正交设计得到了提取维生素B12后的废菌体水解优化条件.基于此,构建利用植物纤维床反应器固定化生产丙酸联产维生素B12的低成本绿色循环工艺.结果表明:在4.5L的发酵体系中,单批次总糖质量浓度为200 g/L,发酵进行了5批次共1192h,丙酸生成总量为2 328.75 g,单批次丙酸质量浓度103.50 g/L,丙酸生产效率达0.43 g/(L·h),干菌质量浓度达到19.52 g/L.将菌体注入微好氧发酵罐中发酵获得112.8 mg/L维生素B12.%Waste propionibacterium cells recovered from vitamin B12 extraction were used as nitrogen source for propionic acid fermentation. The optimizing conditions of bacteria hydrolysis were determined by the orthogonal design. Based on the above results, an immobilized fermentation unit with plant fibrous-bed bioreactor( PFB) combined with membrane separation was constructed and applied in the joint production of propionic acid and vitamin B12. By using hydrolyzed waste cells as alternative nitrogen source, propionic acid and vitamin B12 were produced with the green and inexpensive circulation process. The results showed that propionic acid yield reached a total of 2 328. 75 g with a working volume of 4. 5 L after five batches (1 192 h) in PFB immobilized coupling fermentation (single-batch glucose concentration was 200 g/L), single-batch propionic acid concentration was reached 103. 50 g/L with productivity of 0.43 g/(L·h). With the separation combined with fermentation, cell concentration gradually increased and cell dry weight reached 19. 52 g/L The whole cells were washed into the micro-aerobic bio-reactor to synthesis 112. 8 mg/L vitamin B12.

  2. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of health claims related to a combination of Propionibacterium freudenreichii SI 41 and Propionibacterium freudenreichii SI 26 and increasing numbers of gastro-intestinal microorganisms (ID 941, further assessment) pursuant to Article 13(1) of Regulation (EC) No 1924/2006

    DEFF Research Database (Denmark)

    Tetens, Inge

    of Propionibacterium freudenreichii SI 41and Propionibacterium freudenreichii SI 26 and increasing numbers of gastro-intestinal microorganisms. The food constituent that is the subject of the health claim, a combination of Propionibacterium freudenreichii SI 41and Propionibacterium freudenreichii SI 26...

  3. Emmental Cheese Environment Enhances Propionibacterium freudenreichii Stress Tolerance

    Science.gov (United States)

    Gagnaire, Valérie; Jardin, Julien; Rabah, Houem; Briard-Bion, Valérie; Jan, Gwénaël

    2015-01-01

    Dairy propionibacteria are actinomycetales found in various fermented food products. The main species, Propionibacterium freudenreichii, is generally recognized as safe and used both as probiotic and as cheese starter. Its probiotic efficacy tightly depends on its tolerance towards digestive stresses, which can be largely modulated by the ingested delivery vehicle. Indeed, tolerance of this bacterium is enhanced when it is consumed within a fermented dairy product, compared to a dried probiotic preparation. We investigated both stress tolerance and protein neosynthesis upon growth in i) chemically defined or ii) aqueous phase of Emmental cheeses. Although the same final population level was reached in both media, a slower growth and an enhanced survival of CIRM BIA 1 strain of P. freudenreichii subsp. shermanii was observed in Emmental juice, compared to chemically defined medium. This was accompanied by differences in substrates used and products released as well as overexpression of various early stress adaptation proteins in Emmental juice, compared to chemically defined medium, implied in protein folding, in aspartate catabolism, in biosynthesis of valine, leucine and isoleucine, in pyruvate metabolism in citrate cycle, in the propionate metabolism, as well as in oxidoreductases. All these changes led to a higher digestive stress tolerance after growth in Emmental juice. Mechanisms of stress adaptation were induced in this environment, in accordance with enhanced survival. This opens perspectives for the use of hard and semi-hard cheeses as delivery vehicle for probiotics with enhanced efficacy. PMID:26275229

  4. Propionibacterium freudenreichii Surface Protein SlpB Is Involved in Adhesion to Intestinal HT-29 Cells

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    do Carmo, Fillipe L. R.; Rabah, Houem; Huang, Song; Gaucher, Floriane; Deplanche, Martine; Dutertre, Stéphanie; Jardin, Julien; Le Loir, Yves; Azevedo, Vasco; Jan, Gwénaël

    2017-01-01

    Propionibacterium freudenreichii is a beneficial bacterium traditionally used as a cheese ripening starter and more recently for its probiotic abilities based on the release of beneficial metabolites. In addition to these metabolites (short-chain fatty acids, vitamins, and bifidogenic factor), P. freudenreichii revealed an immunomodulatory effect confirmed in vivo by the ability to protect mice from induced acute colitis. This effect is, however, highly strain-dependent. Local action of metabolites and of immunomodulatory molecules is favored by the ability of probiotics to adhere to the host cells. This property depends on key surface compounds, still poorly characterized in propionibacteria. In the present study, we showed different adhesion rates to cultured human intestinal cells, among strains of P. freudenreichii. The most adhesive one was P. freudenreichii CIRM-BIA 129, which is known to expose surface-layer proteins. We evidenced here the involvement of these proteins in adhesion to cultured human colon cells. We then aimed at deciphering the mechanisms involved in adhesion. Adhesion was inhibited by antibodies raised against SlpB, one of the surface-layer proteins in P. freudenreichii CIRM-BIA 129. Inactivation of the corresponding gene suppressed adhesion, further evidencing the key role of slpB product in cell adhesion. This work confirms the various functions fulfilled by surface-layer proteins, including probiotic/host interactions. It opens new perspectives for the understanding of probiotic determinants in propionibacteria, and for the selection of the most efficient strains within the P. freudenreichii species. PMID:28642747

  5. Efficient transformation system for Propionibacterium freudenreichii based on a novel vector

    NARCIS (Netherlands)

    Jore, J.P.M.; Luijk, N. van; Luiten, R.G.M.; Werf, M.J. van der; Pouwels, P.H.

    2001-01-01

    A 3.6-kb endogenous plasmid was isolated from a Propionibacterium freudenreichii strain and sequenced completely. Based on homologies with plasmids from other bacteria, notably a plasmid from Mycobacterium, a region harboring putative replicative functions was defined. Outside this region two

  6. The probiotic Propionibacterium freudenreichii as a new adjuvant for TRAIL-based therapy in colorectal cancer.

    Science.gov (United States)

    Cousin, Fabien J; Jouan-Lanhouet, Sandrine; Théret, Nathalie; Brenner, Catherine; Jouan, Elodie; Le Moigne-Muller, Gwénaëlle; Dimanche-Boitrel, Marie-Thérèse; Jan, Gwénaël

    2016-02-01

    TNF-Related Apoptosis-Inducing Ligand (TRAIL) is a well-known apoptosis inducer, which activates the extrinsic death pathway. TRAIL is pro-apoptotic on colon cancer cells, while not cytotoxic towards normal healthy cells. However, its clinical use is limited by cell resistance to cell death which occurs in approximately 50% of cancer cells. Short Chain Fatty Acids (SCFA) are also known to specifically induce apoptosis of cancer cells. In accordance, we have shown that food grade dairy propionibacteria induce intrinsic apoptosis of colon cancer cells, via the production and release of SCFA (propionate and acetate) acting on mitochondria. Here, we investigated possible synergistic effect between Propionibacterium freudenreichii and TRAIL. Indeed, we hypothesized that acting on both extrinsic and intrinsic death pathways may exert a synergistic pro-apoptotic effect. Whole transcriptomic analysis demonstrated that propionibacterial supernatant or propionibacterial metabolites (propionate and acetate), in combination with TRAIL, increased pro-apoptotic gene expression (TRAIL-R2/DR5) and decreased anti-apoptotic gene expression (FLIP, XIAP) in HT29 human colon cancer cells. The revealed synergistic pro-apoptotic effect, depending on both death receptors (TRAIL-R1/DR4, TRAIL-R2/DR5) and caspases (caspase-8, -9 and -3) activation, was lethal on cancer cells but not on normal human intestinal epithelial cells (HIEC), and was inhibited by Bcl-2 expression. Finally, milk fermented by P. freudenreichii induced HT29 cells apoptosis and enhanced TRAIL cytotoxic activity, as did P. freudenreichii DMEM culture supernatants or its SCFA metabolites. These results open new perspectives for food grade P. freudenreichii-containing products in order to potentiate TRAIL-based cancer therapy in colorectal cancer.

  7. A unique in vivo experimental approach reveals metabolic adaptation of the probiotic Propionibacterium freudenreichii to the colon environment

    Science.gov (United States)

    2013-01-01

    Background Propionibacterium freudenreichii is a food grade bacterium consumed both in cheeses and in probiotic preparations. Its promising probiotic potential, relying largely on the active release of beneficial metabolites within the gut as well as the expression of key surface proteins involved in immunomodulation, deserves to be explored more deeply. Adaptation to the colon environment is requisite for the active release of propionibacterial beneficial metabolites and constitutes a bottleneck for metabolic activity in vivo. Mechanisms allowing P. freudenreichii to adapt to digestive stresses have been only studied in vitro so far. Our aim was therefore to study P. freudenreichii metabolic adaptation to intra-colonic conditions in situ. Results We maintained a pure culture of the type strain P. freudenreichii CIRM BIA 1, contained in a dialysis bag, within the colon of vigilant piglets during 24 hours. A transcriptomic analysis compared gene expression to identify the metabolic pathways induced by this environment, versus control cultures maintained in spent culture medium. We observed drastic changes in the catabolism of sugars and amino-acids. Glycolysis, the Wood-Werkman cycle and the oxidative phosphorylation pathways were down-regulated but induction of specific carbohydrate catabolisms and alternative pathways were induced to produce NADH, NADPH, ATP and precursors (utilizing of propanediol, gluconate, lactate, purine and pyrimidine and amino-acids). Genes involved in stress response were down-regulated and genes specifically expressed during cell division were induced, suggesting that P. freudenreichii adapted its metabolism to the conditions encountered in the colon. Conclusions This study constitutes the first molecular demonstration of P. freudenreichii activity and physiological adaptation in vivo within the colon. Our data are likely specific to our pig microbiota composition but opens an avenue towards understanding probiotic action within the gut

  8. Hyperconcentrated Sweet Whey, a New Culture Medium That Enhances Propionibacterium freudenreichii Stress Tolerance

    Science.gov (United States)

    Huang, Song; Rabah, Houem; Jardin, Julien; Briard-Bion, Valérie; Parayre, Sandrine; Maillard, Marie-Bernadette; Le Loir, Yves; Schuck, Pierre; Jeantet, Romain

    2016-01-01

    ABSTRACT Propionibacterium freudenreichii is used as a cheese-ripening starter and as a probiotic. Its reported physiological effects at the gut level, including modulation of bifidobacteria, colon epithelial cell proliferation and apoptosis, and intestinal inflammation, rely on active metabolism in situ. Survival and activity are thus key factors determining its efficacy, creating stress adaptation and tolerance bottlenecks for probiotic applications. Growth media and growth conditions determine tolerance acquisition. We investigated the possibility of using sweet whey, a dairy by-product, to sustain P. freudenreichii growth. It was used at different concentrations (dry matter) as a culture medium. Using hyperconcentrated sweet whey led to enhanced multistress tolerance acquisition, overexpression of key stress proteins, and accumulation of intracellular storage molecules and compatible solutes, as well as enhanced survival upon spray drying. A simplified process from growth to spray drying of propionibacteria was developed using sweet whey as a 2-in-1 medium to both culture P. freudenreichii and protect it from heat and osmotic injury without harvesting and washing steps. As spray drying is far cheaper and more energy efficient than freeze-drying, this work opens new perspectives for the sustainable development of new starter and probiotic preparations with enhanced robustness. IMPORTANCE In this study, we demonstrate that sweet whey, a dairy industry by-product, not only allows the growth of probiotic dairy propionibacteria, but also triggers a multitolerance response through osmoadaptation and general stress response. We also show that propionibacteria accumulate compatible solutes under these culture conditions, which might account for the limited loss of viability after spray drying. This work opens new perspectives for more energy-efficient production of dairy starters and probiotics. PMID:27235433

  9. The complete genome of Propionibacterium freudenreichii CIRM-BIA1, a hardy actinobacterium with food and probiotic applications.

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    Hélène Falentin

    Full Text Available BACKGROUND: Propionibacterium freudenreichii is essential as a ripening culture in Swiss-type cheeses and is also considered for its probiotic use. This species exhibits slow growth, low nutritional requirements, and hardiness in many habitats. It belongs to the taxonomic group of dairy propionibacteria, in contrast to the cutaneous species P. acnes. The genome of the type strain, P. freudenreichii subsp. shermanii CIRM-BIA1 (CIP 103027(T, was sequenced with an 11-fold coverage. METHODOLOGY/PRINCIPAL FINDINGS: The circular chromosome of 2.7 Mb of the CIRM-BIA1 strain has a GC-content of 67% and contains 22 different insertion sequences (3.5% of the genome in base pairs. Using a proteomic approach, 490 of the 2439 predicted proteins were confirmed. The annotation revealed the genetic basis for the hardiness of P. freudenreichii, as the bacterium possesses a complete enzymatic arsenal for de novo biosynthesis of aminoacids and vitamins (except panthotenate and biotin as well as sequences involved in metabolism of various carbon sources, immunity against phages, duplicated chaperone genes and, interestingly, genes involved in the management of polyphosphate, glycogen and trehalose storage. The complete biosynthesis pathway for a bifidogenic compound is described, as well as a high number of surface proteins involved in interactions with the host and present in other probiotic bacteria. By comparative genomics, no pathogenicity factors found in P. acnes or in other pathogenic microbial species were identified in P. freudenreichii, which is consistent with the Generally Recognized As Safe and Qualified Presumption of Safety status of P. freudenreichii. Various pathways for formation of cheese flavor compounds were identified: the Wood-Werkman cycle for propionic acid formation, amino acid degradation pathways resulting in the formation of volatile branched chain fatty acids, and esterases involved in the formation of free fatty acids and esters

  10. Milk fermented by Propionibacterium freudenreichii induces apoptosis of HGT-1 human gastric cancer cells.

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    Fabien J Cousin

    Full Text Available BACKGROUND: Gastric cancer is one of the most common cancers in the world. The "economically developed countries" life style, including diet, constitutes a risk factor favoring this cancer. Diet modulation may lower digestive cancer incidence. Among promising food components, dairy propionibacteria were shown to trigger apoptosis of human colon cancer cells, via the release of short-chain fatty acids acetate and propionate. METHODOLOGY/PRINCIPAL FINDINGS: A fermented milk, exclusively fermented by P. freudenreichii, was recently designed. In this work, the pro-apoptotic potential of this new fermented milk was demonstrated on HGT-1 human gastric cancer cells. Fermented milk supernatant induced typical features of apoptosis including chromatin condensation, formation of apoptotic bodies, DNA laddering, cell cycle arrest and emergence of a subG1 population, phosphatidylserine exposure at the plasma membrane outer leaflet, reactive oxygen species accumulation, mitochondrial transmembrane potential disruption, caspase activation and cytochrome c release. Remarkably, this new fermented milk containing P. freudenreichii enhanced the cytotoxicity of camptothecin, a drug used in gastric cancer chemotherapy. CONCLUSIONS/SIGNIFICANCE: Such new probiotic fermented milk may thus be useful as part of a preventive diet designed to prevent gastric cancer and/or as a food supplement to potentiate cancer therapeutic treatments.

  11. A temporal-omic study of Propionibacterium freudenreichii CIRM-BIA1 adaptation strategies in conditions mimicking cheese ripening in the cold.

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    Marion Dalmasso

    Full Text Available Propionibacterium freudenreichii is used as a ripening culture in Swiss cheese manufacture. It grows when cheeses are ripened in a warm room (about 24°C. Cheeses with an acceptable eye formation level are transferred to a cold room (about 4°C, inducing a marked slowdown of propionic fermentation, but P. freudenreichii remains active in the cold. To investigate the P. freudenreichii strategies of adaptation and survival in the cold, we performed the first global gene expression profile for this species. The time-course transcriptomic response of P. freudenreichii CIRM-BIA1(T strain was analyzed at five times of incubation, during growth at 30°C then for 9 days at 4°C, under conditions preventing nutrient starvation. Gene expression was also confirmed by RT-qPCR for 28 genes. In addition, proteomic experiments were carried out and the main metabolites were quantified. Microarray analysis revealed that 565 genes (25% of the protein-coding sequences of P. freudenreichii genome were differentially expressed during transition from 30°C to 4°C (P1. At 4°C, a general slowing down was observed for genes implicated in the cell machinery. On the contrary, P. freudenreichii CIRM-BIA1(T strain over-expressed genes involved in lactate, alanine and serine conversion to pyruvate, in gluconeogenesis, and in glycogen synthesis. Interestingly, the expression of different genes involved in the formation of important cheese flavor compounds, remained unchanged at 4°C. This could explain the contribution of P. freudenreichii to cheese ripening even in the cold. In conclusion, P. freudenreichii remains metabolically active at 4°C and induces pathways to maintain its long-term survival.

  12. Antimicrobial activity of Bacillus subtilis CCTCC M207209 against food-borne pathogens%枯草芽孢杆菌CCTCC M207209对食源性病原菌的抑制作用

    Institute of Scientific and Technical Information of China (English)

    孙卉; 师俊玲

    2009-01-01

    采用杯碟法、对峙培养法和平板对扣法分别检测了枯草芽孢杆菌CCTCC M207209的活菌菌碟、无菌体发酵液、多糖提取液、蛋白质粗提液及其挥发性气体产物对金黄色葡萄球菌、肠球菌、化脓链球菌、大肠杆菌、沙门氏菌等食源性病原菌的抑制作用.结果表明,枯草芽孢杆菌CCTCC M207209的活菌菌碟和发酵液对所有参试病原菌均有显著抑制作用;蛋白质粗提液对除化脓链球菌以外的所有病原菌有显著抑制作用;挥发性气体产物对所有病原菌均无抑制作用;所有抑菌活性物质对金黄色葡萄球菌的抑制作用最强,初步确定其为蛋白类物质.%The antimicrobial effects of the agar plug, cell-free broth, polysaccharide extract, crude protein extract and volatile products of Bacillus subtilis CCTCC M207209 on food-borne pathogens, including Staphylococcus aureus, Enterococcus faecalis, Streptococcus pyogenes, Escherichia coli and Salmonella typhimurium, were investigated by cup-plate method, dual-culture plate method and two-sealed-base-plates method.The cell agar plug and cell-free broth showed significant inhibitory activity against all of the tested pathogens.The crude protein extract showed significant inhibitory activity to most pathogens, except for S.pyogenes.The volatile products of B.subtilis CCTCC M207209 had no inhibitory activity to all pathogens.S.aureus was the most sensitive pathogen to all antimicrobial substances, which was preliminarily identified as proteins.

  13. Cloning and expression of the gene encoding (R)-specific carbonyl reductase from Candida parapsilosis CCTCC M203011

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    The gene which encodes (R)-specific carbonyl reductase (rCR) from Candida parapsilosis CCTCC M203011 was cloned, sequenced and compared with genes from the GenBank. The results indicated that rCR gene was 1011 bp, encoding a protein of 336 amino acids with a molecular weight of 35.9 kDa, and its nucleotide sequence showed 99% similarity to those of other members of the alcohol dehydrogenase superfamily. The rCR gene could express in recombinant strain Escherichia coli JM 109, and the expression plasmid could produce (R)-1-pheny-1,2-ethanediol (100% e.e., 80.14% yield) fromβ-hydroxyacetophenone without any additive to regenerate NAD+ from NADH.

  14. Development of healthy whey drink with Lactobacillus rhamnosus, Bifidobacterium bifidum and Propionibacterium freudenreichii subsp. shermanii

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    T.K. Maity

    2008-12-01

    Full Text Available Whey beverage was prepared by utilizing Lactobacillus rhamnosus NCDO 243, Bifidobacterium bifidum NCDO 2715 and Propionibacterium freudenreichii subsp. shermanii MTCC 1371 in order to make a fermented probiotic healthy drink. The product made with 4 % mixed culture (1:1:1 inoculated (initial count - lactobacilli 6.2 x 107 CFU/mL, bifidobacteria 5.4 x 107 CFU/mL, propionibacteria 3.9 x 107 CFU/mL in deprotienized whey (4.6 % lactose, 0.62 % ash, 0.48 % fat and 0.5 % protein adjusted to pH 6.4 and incubated at 37 °C for 8 h has a good technological and dietetic criteria required for a probiotic product. Total bacterial count, lactobacilli count, bifidobacteria count, propionibacteria count, titratable acidity, β-D galactosidase activity, concentration of lactic acid and sensory properties were monitored during storage period. The whey beverage fermented for 8 h and prepared with 4 % inoculum of mixed culture (1:1:1 met the probiotic criterion by maintaining each type of bacterial population at counts greater than 108 CFU/mL up to 10 days of storage period. The titratable acidity as well as sensory properties did not change appreciably during first 7 days of storage. At the end of 15 days of storage, slight acidification was detected, although the beverage still retained an acceptable flavour.

  15. Evaluation of positive interaction for cell growth between Bifidobacterium adolescentis and Propionibacterium freudenreichii using a co-cultivation system with two microfiltration modules.

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    Kouya, Tomoaki; Ishiyama, Yohei; Tanaka, Takaaki; Taniguchi, Masayuki

    2013-02-01

    Using a co-cultivation system developed previously, positive interaction for cell growth between Bifidobacterium adolescentis and Propionibacterium freudenreichii was evaluated. The total dry cell weight (DCW) of these two strains obtained in the co-cultivation system was 1.5-1.7-fold of the sum of the DCWs obtained in two single cultivations of each bacterium.

  16. The effect of Tween 80 on the polymalic acid and pullulan production by Aureobasidium pullulans CCTCC M2012223.

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    Tu, Guangwei; Wang, Yongkang; Ji, Yunchao; Zou, Xiang

    2015-01-01

    The effect of Tween 80 on the fermentative production of polymalic acid (PMA) and pullulan using Aureobasidium pullulans CCTCC M2012223 was investigated. Tween 80 is beneficial for the biosynthesis of PMA and pullulan, and can regulate the ratio of PMA to pullulan in a dose-dependent manner. After adding 0.05 % Tween 80 to the media, the maximal PMA and pullulan production was 46.45 and 28.8 g/L at 60 h in a 5 L fermenter, with an increase of 75.08 and 27.21 % when compared to the control. Tween 80 could regulate and enhance oxygen uptake rate and carbon dioxide evolution rate in the early phase of fermentation, and change the cell morphology. The transcription levels of mitochondrial dicarboxylate transporter and transmembrane transporter were also dramatically upregulated. The present work will be helpful in deeply understanding the mechanism of Tween 80 on the effect of PMA and pullulan production.

  17. Impact of gene dosage on the production of lipase from Rhizopus chinensis CCTCC M201021 in Pichia pastoris.

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    Sha, Chong; Yu, Xiao-Wei; Li, Fei; Xu, Yan

    2013-02-01

    In this work, the high-level expression of the lipase r27RCL was achieved by optimization of the lipase gene copy number in the host strain Pichia pastoris. The copy number of the lipase gene proRCL from Rhizopus chinensis CCTCC M201021 was quantified by real-time quantitative polymerase chain reaction and a range of Mut(+) P. pastoris strains carrying one, three, five, and six copies of proRCL were obtained. The maximum lipase activity was achieved at 12,500 U/mL by the five-copy recombinant strain after 96 h of methanol induction in the 7-L fermenter. However, the enzyme activity of the six-copy recombinant strain decreased remarkably. By transcription analysis of proRCL, ERO1, and PDI, it suggested that unfolded protein response seemed to be triggered in the highest copy recombinant strain after 24 h. Thus, elaborate optimization of foreign gene dosage was very important for the high-level expression of foreign proteins in P. pastoris.

  18. pH Control Strategy in the Polysiaic Acid Fermentation Production by Escherichia coli CCTCC M208088%大肠杆菌CCTCC M208088发酵生产聚唾液酸的pH控制策略

    Institute of Scientific and Technical Information of China (English)

    刘金龙; 詹晓北; 吴剑荣; 郑志永; 李国顺

    2012-01-01

    研究了不同pH值控制策略对大肠杆菌CCTCC M208088发酵生产聚唾液酸的影响.采用高浓度磷酸盐培养基(20 g/L磷酸氢二钾)缓冲pH值,聚唾液酸产量达到1.9 g/L,但大量磷酸盐残留在发酵液中,影响聚唾液酸的后提取处理.把培养基中磷酸盐的质量浓度降至2.5 g/L,同时流加2 mol/L氢氧化钠溶液控制pH,聚唾液酸产量提升至2.3 g/L,但是NH4+在发酵前期16 h即消耗完毕.进一步采取氨水流加控制pH策略,聚唾液酸产量提升至3.2 g/L,同时菌体浓度大幅增加至12.5 g/L,导致40 g/L初始山梨醇在20 h耗尽.最后,在氨水控制pH的同时,向发酵体系中流加山梨醇,聚唾液酸产量和生产强度分别达到了4.8 g/L和0.16 g/(L·h),比优化前(高浓度磷酸盐发酵)分别提高了152%和188%.%Effect of the different pH control strategies on the PSA production by Escherichia coli CCTCC M208088 was compared in this study. Those strategies including: (l)when 20 g/L K2HPO4 was presented in the medium, the titer of PSA achieved at 1. 9 g/L PSA, however, the high-level residual K2HPO4 in broth bring difficult to the following PSA purification. (2) with 2. 5 g/L K2HPO4 and feeding of 2 mol/L NaOH solution, the titer of PSA was 2. 3 g/L; (3)the PSA titer was increased to 3. 2 g/L by feeding of ammonia water as pH control agent. Furthermore, the feeding of ammonia could accelerated sorbitol consummation; (4) To further promote PSA production, a novel strategy using ammonia water for pH control coupling with sorbitol supplementation at a constant feeding speed was adopted. As a result, PSA production and productivity were increased separately to 4. 8 g/L and 0. 16 g/(L·h), which was 152% and 188% higher than that of the contro(without pH control) .

  19. Characteristic of milk whey culture with Propionibacterium freudenreichii ET-3 and its application to the inflammatory bowel disease therapy.

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    Uchida, M; Mogami, O; Matsueda, K

    2007-06-01

    After the screening of microorganism culture, the culture of Propionibacterium freudenreichii ET-3 in the milk whey (milk whey culture) was found to stimulate the growth of our own Bifidobacteria in the colon but not the growth of other microorganisms. One of the active substances was identified as 1,4-dihydroxy-2-naphthoic acid (DHNA).In healthy volunteers, the ingestion containing milk whey culture significantly increased the population of Bifidobacteria to total fecal bacterium. In the TNBS-induced colitis model of rats, milk whey culture significantly accelerated the healing of the colitis in a dose-dependent manner. It has been reported that DHNA inhibited the lymphocyte infiltration through reduction of MAdCAM-1 in DSS colitis model of mice and that the ingestion of milk whey culture was effective in the treatment of ulcerative colitis in human pilot study. These findings suggest that milk whey culture is a useful prebiotic for the therapy of inflammatory bowel disease.

  20. A novel carbonyl reductase with anti-Prelog stereospecificity from Acetobacter sp. CCTCC M209061: purification and characterization.

    Directory of Open Access Journals (Sweden)

    Xiao-Hong Chen

    Full Text Available A novel carbonyl reductase (AcCR catalyzing the asymmetric reduction of ketones to enantiopure alcohols with anti-Prelog stereoselectivity was found in Acetobacter sp. CCTCC M209061 and enriched 27.5-fold with an overall yield of 0.4% by purification. The enzyme showed a homotetrameric structure with an apparent molecular mass of 104 kDa and each subunit of 27 kDa. The gene sequence of AcCR was cloned and sequenced, and a 762 bp gene fragment was obtained. Either NAD(H or NADP(H can be used as coenzyme. For the reduction of 4'-chloroacetophenone, the Km value for NADH was around 25-fold greater than that for NADPH (0.66 mM vs 0.026 mM, showing that AcCR preferred NADPH over NADH. However, when NADH was used as cofactor, the response of AcCR activity to increasing concentration of 4'-chloroacetophenone was clearly sigmoidal with a Hill coefficient of 3.1, suggesting that the enzyme might possess four substrate-binding sites cooperating with each other The Vmax value for NADH-linked reduction was higher than that for NADPH-linked reduction (0.21 mM/min vs 0.17 mM/min. For the oxidation of isopropanol, the similar enzymological properties of AcCR were found using NAD+ or NADP+ as cofactor. Furthermore, a broad range of ketones such as aryl ketones, α-ketoesters and aliphatic ketones could be enantioselectively reduced into the corresponding chiral alcohols by this enzyme with high activity.

  1. Enhancing the vitamin B12 production and growth of Propionibacterium freudenreichii in tofu wastewater via a light-induced vitamin B12 riboswitch.

    Science.gov (United States)

    Yu, Yue; Zhu, Xuan; Shen, Yubiao; Yao, Huanghong; Wang, Peiheng; Ye, Kun; Wang, Xiaofeng; Gu, Qing

    2015-12-01

    The vitamin B12-dependent riboswitch is a crucial factor that regulates gene transcription to mediate the growth of and vitamin B12 synthesis by Propionibacterium freudenreichii. In this study, the effect of various wavelengths of light on the growth rate and vitamin B12 synthesis was studied. Red, green, and blue light-emitting diodes (LEDs) were selected, and a dark condition was used as the control. The microorganism growth rate was measured using a spectrophotometer and plate counting, while the vitamin B12 content was determined using an HPLC-based method. The optical density at 600 nm (OD600) values indicated that P. freudenreichii grew better under the continuous and discontinuous blue light conditions. Moreover, under the blue light condition, P. freudenreichii tended to have a higher growth rate (0.332 h(-1)) and vitamin B12 synthesis (ca. 10 μg/mL) in tofu wastewater than in dark conditions. HPLC analysis also showed that more methylcobalamin was produced under the blue light conditions than in the other conditions. The cbiB gene transcription results showed that blue light induced the synthesis of this vitamin B12 synthesis enzyme. Moreover, the results of inhibiting the expression of green fluorescent protein indicated that blue light removed the inhibition by the vitamin B12-dependent riboswitch. This method can be used to reduce fermentation time and produce more vitamin B12 in tofu wastewater.

  2. Food-Like Growth Conditions Support Production of Active Vitamin B12 by Propionibacterium freudenreichii 2067 without DMBI, the Lower Ligand Base, or Cobalt Supplementation

    Science.gov (United States)

    Deptula, Paulina; Chamlagain, Bhawani; Edelmann, Minnamari; Sangsuwan, Panchanit; Nyman, Tuula A.; Savijoki, Kirsi; Piironen, Vieno; Varmanen, Pekka

    2017-01-01

    Propionibacterium freudenreichii is a traditional dairy bacterium and a producer of short chain fatty acids (propionic and acetic acids) as well as vitamin B12. In food applications, it is a promising organism for in situ fortification with B12 vitamin since it is generally recognized as safe (GRAS) and it is able to synthesize biologically active form of the vitamin. In the present study, vitamin B12 and pseudovitamin biosynthesis by P. freudenreichii was monitored by UHPLC as a function of growth in food-like conditions using a medium mimicking cheese environment, without cobalt or 5,6-dimethylbenzimidazole (DMBI) supplementation. Parallel growth experiments were performed in industrial-type medium known to support the biosynthesis of vitamin B12. The production of other key metabolites in the two media were determined by HPLC, while the global protein production was compared by gel-based proteomics to assess the effect of growth conditions on the physiological status of the strain and on the synthesis of different forms of vitamin. The results revealed distinct protein and metabolite production, which reflected the growth conditions and the potential of P. freudenreichii for synthesizing nutritionally relevant amounts of active vitamin B12 regardless of the metabolic state of the cells. PMID:28337185

  3. Efficient anti-Prelog enantioselective reduction of acetyltrimethylsilane to (R-1-trimethylsilylethanol by immobilized Candida parapsilosis CCTCC M203011 cells in ionic liquid-based biphasic systems

    Directory of Open Access Journals (Sweden)

    Zhang Bo-Bo

    2012-08-01

    Full Text Available Abstract Background Biocatalytic asymmetric reductions with whole cells can offer high enantioselectivity, environmentally benign processes and energy-effective operations and thus are of great interest. The application of whole cell-mediated bioreduction is often restricted if substrate and product have low water solubility and/or high toxicity to the biocatalyst. Many studies have shown that a biphasic system is often useful in this instance. Hence, we developed efficient biphasic reaction systems with biocompatible water-immiscible ionic liquids (ILs, to improve the biocatalytic anti-Prelog enantioselective reduction of acetyltrimethylsilane (ATMS to (R-1-trimethylsilylethanol {(R-1-TMSE}, which is key synthon for a large number of silicon-containing drugs, using immobilized Candida parapsilosis CCTCC M203011 cells as the biocatalyst. Results It was found that the substrate ATMS and the product 1-TMSE exerted pronounced toxicity to immobilized Candida parapsilosis CCTCC M203011 cells. The biocompatible water-immiscible ILs can be applied as a substrate reservoir and in situ extractant for the product, thus greatly enhancing the efficiency of the biocatalytic process and the operational stability of the cells as compared to the IL-free aqueous system. Various ILs exerted significant but different effects on the bioreduction and the performances of biocatalysts were closely related to the kinds and combination of cation and anion of ILs. Among all the water-immiscible ILs investigated, the best results were observed in 1-butyl-3-methylimidazolium hexafluorophosphate (C4mim·PF6/buffer biphasic system. Furthermore, it was shown that the optimum substrate concentration, volume ratio of buffer to IL, buffer pH, reaction temperature and shaking rate for the bioreduction were 120 mM, 8/1 (v/v, 6.0, 30°C and 180 r/min, respectively. Under these optimized conditions, the initial reaction rate, the maximum yield and the product e.e. were 8.1

  4. Simultaneous removal of Cr(Ⅵ) and phenol in consortium culture of Bacillus sp. and Pseudomonas putida Migula (CCTCC AB92019)

    Institute of Scientific and Technical Information of China (English)

    LIU Yun-guo; PAN Cui; XIA Wen-bin; ZENG Guang-ming; ZHOU Ming; LIU Yuan-yuan; KE Jie; HUANG Chao

    2008-01-01

    The simultaneous removal of Cr(Ⅵ) and phenol in a consortium culture containing Cr(Ⅵ) reducer, Bacillus sp. and phenol degrader, Pseudomonas putida Migula (CCTCC AB92019) was studied. Phenol was used as the sole carbon source. Bacillus sp. utilized metabolites formed from phenol degradation as electron donors and energy source for Cr(Ⅵ) reduction. Optimum Cr(Ⅵ) reduction was observed at a phenol concentration of 150 mg/L and an initial Cr(Ⅵ) concentration of 15 mg/L. Both the Cr(Ⅵ) reduction and phenol degradation were influenced by the cell composition of the culture, but the phenol degradation was not significantly affected by the content of Bacillus sp. The experiments also showed that the amount of phenol degraded was more than that stoichiometrically required for Cr(Ⅵ) reduction.

  5. Metabolic flux analysis of propionic acid biosynthesis with two immobilized cell reactors fermentation by Propionibacterium%丙酸杆菌的两种固定化细胞反应器发酵生产丙酸及其代谢通量分析

    Institute of Scientific and Technical Information of China (English)

    陈飞; 冯小海; 吴波; 李莎; 徐虹

    2011-01-01

    根据构建的费氏丙酸杆菌合成丙酸的生化反应网络,利用代谢通量分析法分析了课题组构建的两种固定化细胞反应器对丙酸发酵的影响.结果表明,固定化细胞发酵可以调节葡萄糖-6-磷酸、磷酸烯醇式丙酮酸和丙酮酸节点处的代谢通量分布,从而最终影响生物量和各种有机酸的合成.与游离发酵相比,两种固定化发酵方式戊糖磷酸途径通最都有显著提高,乙酸和乳酸通量均有减少.与植物纤维床反应器主要靠直接加强丙酸合成途径使丙酸产率提高了242.72%不同,多点式棉纤维床反应器发酵主要是通过琥珀酸合成途径的增强而使丙酸产率提高了56.80%.比较了相关关键酶的比活.酶活变化与代谢通量结果基本一致.%The metabolic network of Propionibacterium freudenreichii CCTCC M207015 for propionic acid biosynthesis, with glucose as the sole carbon source, was built based on the stoichiometric balance equations to illustrate the carbon flux distribution of intermediates and metabolites. The propionic acid production with two immobilized cell reactors fermentation was studied by using the metabolic flux analysis method. The results showed that the flux distribution of nodes of glucose 6-phosphate, phosphoenolpyruvate and pyruvate changed after immobilized cells fermentation regulation. Compared with free cells fermentation, the flux of HMP of the two immobilized cells fermentation significantly increased, also with decrease fluxes of acetic acid and lactic acid. The productivity of propionic acid in plant fibrous-bed reactor fermentation increased 242.72% mainly by directly enhancing propionic acid synthesis pathway, while in the multi-point fibrous-bed reactor fermentation it increased 56.80% through the enhancement of succinic acid biosynthetic pathway, and the relevant key enzyme activity and metabolic flux variation had good consistency.

  6. Optimization of culture conditions to produce high yields of active Acetobacter sp. CCTCC M209061 cells for anti-Prelog reduction of prochiral ketones

    Directory of Open Access Journals (Sweden)

    Chen Xiao-Hong

    2011-11-01

    Full Text Available Abstract Background Chiral alcohols are widely used in the synthesis of chiral pharmaceuticals, flavors and functional materials and appropriate whole-cell biocatalysts offer a highly enantioselective, minimally polluting route to these valuable compounds. The recently isolated strain Acetobacter sp. CCTCC M209061 showed exclusive anti-Prelog stereoselectivity for the reduction of prochiral ketones, but the low biomass has limited its commercialization and industrial applications. To tackle this problem, the effects of medium components and culture conditions on the strain's growth and reduction activity were explored. Results By using a one-at-a-time method and a central composite rotatable design (CCRD, the optimal medium and culture conditions were found to be as follows: glucose 8.26 g/L, fructose 2.50 g/L, soy peptone 83.92 g/L, MnSO4·H2O 0.088 g/L, pH 5.70, 30°C and 10% (v/v inoculum. Under the above-mentioned conditions, the biomass after 30 h cultivation reached 1.10 ± 0.03 g/L, which was 9.5-fold higher than that obtained with basic medium. Also, the reduction activity towards 4'-chloroacetophenone was markedly enhanced to 39.49 ± 0.96 μmol/min/g from 29.34 ± 0.65 μmol/min/g, with the product e.e. being above 99%. Comparable improvements were also seen with the enantioselective bioreduction of 4-(trimethylsilyl-3-butyn-2-one to the key pharmaceutical precursor (R - 4-(trimethylsilyl-3-butyn-2-ol. Conclusions The biomass and reduction activity of Acetobacter sp. CCTCC M209061 can be greatly enhanced through the optimization strategy. This facilitates use of the strain in the anti-Prelog stereoselective reduction of prochiral ketones to enantiopure chiral alcohols as building blocks for many industries.

  7. Efficient enantioselective synthesis of (R)-[3,5-bis(trifluoromethyl)phenyl] ethanol by Leifsonia xyli CCTCC M 2010241 using isopropanol as co-substrate.

    Science.gov (United States)

    Ouyang, Qi; Wang, Pu; Huang, Jin; Cai, Jinbo; He, Junyao

    2013-03-01

    (R)-[3,5-Bis(trifluoromethyl)phenyl] ethanol is a key chiral intermediate for the synthesis of aprepitant. In this paper, an efficient synthetic process for (R)-[3,5- bis(trifluoromethyl)phenyl] ethanol was developed via the asymmetric reduction of 3,5-bis(trifluoromethyl) acetophenone, catalyzed by Leifsonia xyli CCTCC M 2010241 cells using isopropanol as the co-substrate for cofactor recycling. Firstly, the substrate and product solubility and cell membrane permeability of biocatalysts were evaluated with different co-substrate additions into the reaction system, in which isopropanol manifested as the best hydrogen donor of coupled NADH regeneration during the bioreduction of 3,5-bis(trifluoromethyl) acetophenone. Subsequently, the optimization of parameters for the bioreduction were undertaken to improve the effectiveness of the process. The determined efficient reaction system contained 200 mM of 3,5-bis(trifluoromethyl) acetophenone, 20% (v/v) of isopropanol, and 300 g/l of wet cells. The bioreduction was executed at 30°C and 200 rpm for 30 h, and 91.8% of product yield with 99.9% of enantiometric excess (e.e.) was obtained. The established bioreduction reaction system could tolerate higher substrate concentrations of 3,5- bis(trifluoromethyl) acetophenone, and afforded a satisfactory yield and excellent product e.e. for the desired (R)-chiral alcohol, thus providing an alternative to the chemical synthesis of (R)-[3,5-bis(trifluoromethyl)phenyl] ethanol.

  8. D190V点突变提高华根霉Rhizopus chinensis CCTCC M201021脂肪酶的最适温度和热稳定性%Improved optimum temperature and thermostability of the lipase from Rhizopus chinensis CCTCC M201021 by site-directed mutagenesis of D190V

    Institute of Scientific and Technical Information of China (English)

    吴厚军; 喻晓蔚; 沙冲; 徐岩

    2013-01-01

    [目的]对来源于Rhizopus chinensis CCTCC M201021的脂肪酶进行了D190V定点突变,提高该酶的最适温度和热稳定性.[方法]对毕赤酵母表达的突变酶D190V与野生型酶r27RCL进行酶学性质比较.[结果]D190V的最适温度比r27RCL高5℃,65℃下的半衰期提高了一倍,在其他性质方面,突变酶D190V与r27RCL基本相似.[结论]通过结构分析表明,定点突变D190V提高该酶稳定性的主要原因可能在于提高了突变位点所在的α螺旋的稳定性以及增强了稳定蛋白质结构的氢键作用力.

  9. Using a water-immiscible ionic liquid to improve asymmetric reduction of 4-(trimethylsilyl-3-butyn-2-one catalyzed by immobilized Candida parapsilosis CCTCC M203011 cells

    Directory of Open Access Journals (Sweden)

    Smith Thomas J

    2009-10-01

    Full Text Available Abstract Background Whole cells are usually employed for biocatalytic reduction reactions to ensure efficient coenzyme regeneration and to avoid problems with enzyme purification and stability. The efficiency of whole cell-catalyzed bioreduction is frequently restricted by pronounced toxicity of substrate and/or product to the microbial cells and in many instances the use of two-phase reaction systems can solve such problems. Therefore, we developed new, biphasic reaction systems with biocompatible water-immiscible ionic liquids (ILs as alternatives to conventional organic solvents, in order to improve the asymmetric reduction of 4-(trimethylsilyl-3-butyn-2-one (TMSB to (S-4-(trimethylsilyl-3-butyn-2-ol {(S-TMSBOL}, a key intermediate for synthesis of 5-lipoxygenase inhibitors, using immobilized Candida parapsilosis CCTCC M203011 cells as the biocatalyst. Results Various ILs exerted significant but different effects on the bioreduction. Of all the tested water-immiscible ILs, the best results were observed with 1-butyl-3-methylimidazolium hexafluorophosphate (C4MIM·PF6, which exhibited not only good biocompatibility with the cells but also excellent solvent properties for the toxic substrate and product, thus markedly improving the efficiency of the bioreduction and the operational stability of the cells as compared to the IL-free aqueous system. 2-Propanol was shown to be the most suitable co-substrate for coenzyme regeneration, and it was found that the optimum volume ratio of buffer to C4MIM·PF6, substrate concentration, buffer pH, 2-propanol concentration and reaction temperature were 4/1 (v/v, 24 mM, 5.5, 130 mM and 30°C, respectively. Under these optimized conditions, the maximum yield and the product e.e. wer 97.7% and >99%, respectively, which are much higher than the corresponding values previously reported. The efficient whole-cell biocatalytic process was shown to be feasible on a 250-mL scale. Conclusion The whole cell

  10. 1,4-Dihydroxy-2-naphthoic acid from Propionibacterium freudenreichii reduces inflammation in interleukin-10-deficient mice with colitis by suppressing macrophage-derived proinflammatory cytokines.

    Science.gov (United States)

    Okada, Yoshikiyo; Tsuzuki, Yoshikazu; Narimatsu, Kazuyuki; Sato, Hirokazu; Ueda, Toshihide; Hozumi, Hideaki; Sato, Shingo; Hokari, Ryota; Kurihara, Chie; Komoto, Shunsuke; Watanabe, Chikako; Tomita, Kengo; Kawaguchi, Atsushi; Nagao, Shigeaki; Miura, Soichiro

    2013-09-01

    The anti-inflammatory mechanism of prebiotics has recently been shown to have an impact on the host immune system. DHNA from Propionibacterium freudenreichii is known to promote the proliferation of Bifidobacterium and can ameliorate colitis, although its mode of action remains unknown. In this study, we investigated whether DHNA attenuates inflammation in piroxicam-treated IL-10(-/-) mice, particularly focusing on the changes of the host immune mechanism. DHNA was administered to IL-10(-/-) mice with colitis, and the expression of adhesion molecules and mRNA levels of proinflammatory cytokines were determined. DHNA pretreatment attenuated the piroxicam-induced histological changes. The increased F4/80-positive cell infiltration and VCAM-1 expression were decreased by DHNA administration. The increased mRNA levels of proinflammatory cytokines were also suppressed by DHNA. In in vitro experiments, increased mRNA levels of proinflammatory cytokines after endotoxin exposure were decreased significantly by DHNA pretreatment in RAW264.7, a macrophage cell line, and IL-10(-/-) mice BMMs, whereas the expression of VCAM-1 in bEnd.3 cells, a endothelial cell line, was not affected. Taken together, these findings suggest that administration of DHNA is useful for the treatment of colitis in piroxicam-treated IL-10(-/-) mice and that attenuation of colitis by DHNA may partly be a result of its direct action on intestinal macrophages to inhibit proinflammatory cytokine production.

  11. Identification of proteins involved in the anti-inflammatory properties of Propionibacterium freudenreichii by means of a multi-strain study.

    Science.gov (United States)

    Deutsch, Stéphanie-Marie; Mariadassou, Mahendra; Nicolas, Pierre; Parayre, Sandrine; Le Guellec, Rozenn; Chuat, Victoria; Peton, Vincent; Le Maréchal, Caroline; Burati, Julien; Loux, Valentin; Briard-Bion, Valérie; Jardin, Julien; Plé, Coline; Foligné, Benoît; Jan, Gwénaël; Falentin, Hélène

    2017-04-13

    Propionibacterium freudenreichii, a dairy starter, can reach a population of almost 10(9) propionibacteria per gram in Swiss-type cheese at the time of consumption. Also consumed as a probiotic, it displays strain-dependent anti-inflammatory properties mediated by surface proteins that induce IL-10 in leukocytes. We selected 23 strains with varied anti-inflammatory potentials in order to identify the protein(s) involved. After comparative genomic analysis, 12 of these strains were further analysed by surface proteomics, eight of them being further submitted to transcriptomics. The omics data were then correlated to the anti-inflammatory potential evaluated by IL-10 induction. This comparative omics strategy highlighted candidate genes that were further subjected to gene-inactivation validation. This validation confirmed the contribution of surface proteins, including SlpB and SlpE, two proteins with SLH domains known to mediate non-covalent anchorage to the cell-wall. Interestingly, HsdM3, predicted as cytoplasmic and involved in DNA modification, was shown to contribute to anti-inflammatory activity. Finally, we demonstrated that a single protein cannot explain the anti-inflammatory properties of a strain. These properties therefore result from different combinations of surface and cytoplasmic proteins, depending on the strain. Our enhanced understanding of the molecular bases for immunomodulation will enable the relevant screening for bacterial resources with anti-inflammatory properties.

  12. Microbial production of propionic acid with Propionibacterium freudenreichii using an anion exchanger-based in situ product recovery (ISPR) process with direct and indirect contact of cells.

    Science.gov (United States)

    Wang, Peng; Wang, Yunshan; Su, Zhiguo

    2012-02-01

    The recovery of an inhibiting product from a bioreactor soon after its formation is an important issue in industrial bioprocess development. In the present study, the potential of the anion exchanger-based in situ product recovery (ISPR) technique for the biocatalytic production of propionic acid was discussed. The focus of the current work was the selection of a suitable configuration of metabolically active cells for application in propionic acid production. Accumulation of propionic acid in fermentation broth caused feedback inhibition of the growth and biotransformation activity of Propionibacterium freudenreichii CICC 10019. Relevant product inhibition kinetics was discussed, and the results showed that keeping the aqueous propionic acid concentration below 10.02 g L⁻¹ was an essential prerequisite for ISPR process. A batch study, in which three ISPR configuration mode designs were compared, was conducted. The comparison indicated that employing an external direct mode had significant advantages over other modes in terms of increased productivity and product yield, with a corresponding decrease in the number of downstream processing steps, as well as in substrate consumption. The fed-batch culture using an external direct mode for the continuous accumulation of propionic acid resulted in a cumulative propionic acid concentration of 62.5 g L⁻¹, with a corresponding product yield of 0.78 g propionic acid/g glucose.

  13. Identification of proteins involved in the anti-inflammatory properties of Propionibacterium freudenreichii by means of a multi-strain study

    Science.gov (United States)

    Deutsch, Stéphanie-Marie; Mariadassou, Mahendra; Nicolas, Pierre; Parayre, Sandrine; Le Guellec, Rozenn; Chuat, Victoria; Peton, Vincent; Le Maréchal, Caroline; Burati, Julien; Loux, Valentin; Briard-Bion, Valérie; Jardin, Julien; Plé, Coline; Foligné, Benoît; Jan, Gwénaël; Falentin, Hélène

    2017-01-01

    Propionibacterium freudenreichii, a dairy starter, can reach a population of almost 109 propionibacteria per gram in Swiss-type cheese at the time of consumption. Also consumed as a probiotic, it displays strain-dependent anti-inflammatory properties mediated by surface proteins that induce IL-10 in leukocytes. We selected 23 strains with varied anti-inflammatory potentials in order to identify the protein(s) involved. After comparative genomic analysis, 12 of these strains were further analysed by surface proteomics, eight of them being further submitted to transcriptomics. The omics data were then correlated to the anti-inflammatory potential evaluated by IL-10 induction. This comparative omics strategy highlighted candidate genes that were further subjected to gene-inactivation validation. This validation confirmed the contribution of surface proteins, including SlpB and SlpE, two proteins with SLH domains known to mediate non-covalent anchorage to the cell-wall. Interestingly, HsdM3, predicted as cytoplasmic and involved in DNA modification, was shown to contribute to anti-inflammatory activity. Finally, we demonstrated that a single protein cannot explain the anti-inflammatory properties of a strain. These properties therefore result from different combinations of surface and cytoplasmic proteins, depending on the strain. Our enhanced understanding of the molecular bases for immunomodulation will enable the relevant screening for bacterial resources with anti-inflammatory properties. PMID:28406170

  14. Exopolysaccharides from co-cultures of Weissella confusa 11GU-1 and Propionibacterium freudenreichii JS15 act synergistically on wheat dough and bread texture.

    Science.gov (United States)

    Tinzl-Malang, Saskia Katharina; Rast, Peter; Grattepanche, Franck; Sych, Janice; Lacroix, Christophe

    2015-12-02

    The storage of bread is limited by both physical (staling) and microbial (mainly fungal) spoilage. Exopolysaccharides (EPS) produced by lactic acid bacteria (LAB) and organic acids from propionibacteria (PAB) have been used to enhance texture and extend shelf-life of bakery products. In this study the functionality of EPS of Weissella confusa A3/2-1 (dextran), W. confusa F3/2-2 (dextran and levan), W. confusa 11GU-1 (dextran and ropy capsular polysaccharide) was evaluated in wheat bread. Two strains of Propionibacterium freudenreichii (Pf), shown to produce a heteropolysaccharide (Pf JS15) or a β-glucan (Pf DF30), were tested in single and mixed cultures with W. confusa (Wc). The EPS fermentates were prepared by batch fermentation of cereal- or malt-based medium using sucrose (Wc) or lactic acid (Pf) as carbon source. Incorporation of EPS from single culture fermentates and 1:1 Weissella-Propionibacterium fermentate mixtures revealed strong positive effects of dextran and ropy capsular polysaccharide produced by Wc 11GU-1 on bread staling retardation, with synergistic effects of EPS mixture from Wc 11GU-1 and Pf JS15. A co-fermentation of Wc 11GU-1 and Pf JS15 was developed to produce EPS together with antifungal organic acid mixture (acetate and propionate) in a single step process. The addition of 15% (w/w flour base) co-culture, yielding EPS, acetate and propionate concentrations of 1.5, 0.5 and 1g/kg dough, respectively, resulted in improved bread texture, increased loaf volume and decreased crumb firming during storage for 3days compared with control breads and breads supplemented with equivalent levels of chemical organic acids. Our data showed that EPS could compensate for the negative effects of chemical acetate and propionate in a concentration range exerting antifungal effects. The natural bioingredient produced by Wc 11GU-1 and Pf JS15 has potential for applications as antifungal, texture-building and anti-staling agent in breads, consistent with

  15. pH对费氏丙酸杆菌细菌素抑菌性的影响%Effect of pH on Antimicrobial Activity of Propionibacterium freudenreichii Bacteriocins

    Institute of Scientific and Technical Information of China (English)

    郑丽雪; 郭晨; 谢建松; 唐亚进; 王立梅; 齐斌

    2016-01-01

    以实验室保存的一株费氏丙酸杆菌CS1420(Propionibacterium freudenreichii CS1420)为试验菌株。首先考察了不同初始pH下对其发酵产细菌素的影响。结果表明:以大肠杆菌ATCC25922为指示菌,当培养基初始pH为6.0时,细菌素抑菌效果最好;以Saccharomyces cerevisiae 2-10515为指示菌,当培养基初始pH为5.5时,细菌素抑菌效果最好。然后用丙酸调节细菌素粗提物的pH进行抑菌试验,结果表明,pH对粗提得到的细菌素抑菌性影响很大,当用丙酸将细菌素溶液的pH调至5.5时,细菌素的抑菌活性有显著提高。%Propionibacterium freudenreichii CS1420 was as experimental strains. Firstly,the effects of different pH on fermentation of Propionibacterium freudenreichii Bacteriocins were investigated. The result showed:the antimicrobial activity were best by using Eoli ATCC25922 as indicator strains when the medium initial pH was 6.0,the antimicrobial activity were best by using Saccharomyces cerevisiae 2-10515 as indicator strains when the medium initial pH was 5.5. Then,bacteriocins crude extract pH were adjusted and then bacteriostatic exper-iments were carried out. The result showed:the effect of pH on crude extract bacteriocins was tremendous. The antimicrobial activity of bacteriocins improved significantly when crude extract bacteriocins solution pH were adjusted 5.5 with propionic acid.

  16. Effects of probiotic Lactobacillus rhamnosus GG and Propionibacterium freudenreichii ssp. shermanii JS supplementation on intestinal and systemic markers of inflammation in ApoE*3Leiden mice consuming a high-fat diet.

    Science.gov (United States)

    Oksaharju, Anna; Kooistra, Teake; Kleemann, Robert; van Duyvenvoorde, Wim; Miettinen, Minja; Lappalainen, Jani; Lindstedt, Ken A; Kovanen, Petri T; Korpela, Riitta; Kekkonen, Riina A

    2013-07-14

    A high-fat diet disturbs the composition and function of the gut microbiota and generates local gut-associated and also systemic responses. Intestinal mast cells, for their part, secrete mediators which play a role in the orchestration of physiological and immunological functions of the intestine. Probiotic bacteria, again, help to maintain the homeostasis of the gut microbiota by protecting the gut epithelium and regulating the local immune system. In the present study, we explored the effects of two probiotic bacteria, Lactobacillus rhamnosus GG (GG) and Propionibacterium freudenreichii spp. shermanii JS (PJS), on high fat-fed ApoE*3Leiden mice by estimating the mast cell numbers and the immunoreactivity of TNF-α and IL-10 in the intestine, as well as plasma levels of several markers of inflammation and parameters of lipid metabolism. We found that mice that received GG and PJS exhibited significantly lower numbers of intestinal mast cells compared with control mice. PJS lowered intestinal immunoreactivity of TNF-α, while GG increased intestinal IL-10. PJS was also observed to lower the plasma levels of markers of inflammation including vascular cell adhesion molecule 1, and also the amount of gonadal adipose tissue. GG lowered alanine aminotransferase, a marker of hepatocellular activation. Collectively, these data demonstrate that probiotic GG and PJS tend to down-regulate both intestinal and systemic pro-inflammatory changes induced by a high-fat diet in this humanised mouse model.

  17. Co-expression of formate dehydrogenase form Candida boidinii and (R)-specific carbonyl reductase form Candida parapsilosis CCTCC M203011 in Escherichia coli%(R)-专一性羰基还原酶与甲酸脱氢酶基因在大肠杆菌中的共表达

    Institute of Scientific and Technical Information of China (English)

    孙莹; 张荣珍; 徐岩

    2008-01-01

    [目的]通过研究-专一性拨基还原酶和甲酸脱氢酶基因在大肠杆菌中的共表达,解决较高底物浓度下不对称转化反应的辅酶限制性问题.[方法]分别以近平滑假丝酵母(candida parapsitosis CCTCC M203011)和博伊丁假丝酵母(Candida boidinii)基因组为模板,采用PCR方法扩增得到(R伏)-专一性拨基还原酶基因(rcr)和甲酸脱氢酶基因(fdh),克隆到共表达载体pETDuetTM-1中进行表达.共表达质粒 pETDuet-rcr-fdh转化稀有密码子优化型菌株E.coli Rosetta,获得重组菌E.coli Rosetta/pETDuet-rcr-fdh.结果 在30℃ 条件下,经1mmol/L IPTG诱导表达8h后,SDS-PAGE结果表明-专一性拨基还原酶和甲酸脱氢酶均有明显的表达,其相对分子质量分别为37kDa和40kDa.以高浓度(6glL)2一经基苯乙酮为底物时,0.19重组菌细胞催化产生(R)-苯基乙二醇,产物光学纯度为100% e.e.,产率为85.9%.与无甲酸脱氢酶参与辅酶再生循环的重组菌E.coli Rosetta/pETDuet-rrc;相比,产物光学纯度和产率分别提高了1.3和2.7倍.讨论 该重组菌的构建为基因工程法生物合成(R)-苯基乙二醇的工业应用奠定了基础.

  18. Understanding the role of clay minerals in the chromium(VI) bioremoval by Pseudomonas aeruginosa CCTCC AB93066 under growth condition: microscopic, spectroscopic and kinetic analysis.

    Science.gov (United States)

    Kang, Chunxi; Wu, Pingxiao; Li, Yuewu; Ruan, Bo; Li, Liping; Tran, Lytuong; Zhu, Nengwu; Dang, Zhi

    2015-11-01

    Laboratory batch experiments were conducted to investigate the role of clay minerals, e.g., kaolinite and vermiculite, in microbial Cr(VI) reduction by Pseudomonas aeruginosa under growth condition in glucose-amended mediums as a method for treating Cr(VI)-contaminated subsurface environment such as soil. Our results indicated that glucose could acted as an essential electron donor, and clay minerals significantly enhanced microbial Cr(VI) reduction rates by improving the consumption rate of glucose and stimulating the growth and propagation of P. aeruginosa. Cr(VI) bioreduction by both free cells and clay minerals-amended cells followed the pseudo-first-order kinetic model, with the latter one fitting better. The mass balance analyses and X-ray photoelectron spectroscopy analysis found that Cr(VI) was reduced to Cr(III) and the adsorption of total chromium on clay minerals-bacteria complex was small, implying that Cr(VI) bioremoval was not mainly due to the adsorption of Cr(VI) onto cells or clay minerals or clay minerals-cells complex but mainly due to the Cr(VI) reduction capacity of P. aeruginosa under the experimental conditions studied (e.g., pH 7). Atomic force microscopy revealed that the addition of clay minerals (e.g. vermiculite) decreased the surface roughness of Cr(VI)-laden cells and changed the cell morphology and dimension. Fourier transform infrared spectroscopy revealed that organic matters such as aliphatic species and/or proteins played an important role in the combination of cells and clay minerals. Scanning electron microscopy confirmed the attachment of cells on the surface of clay minerals, indicating that clay minerals could provide a microenvironment to protect cells from Cr(VI) toxicity and serve as growth-supporting materials. These findings manifested the underlying influence of clay minerals on microbial reduction of Cr(VI) and gave an understanding of the interaction between pollutants, the environment and the biota.

  19. Markedly improving asymmetric oxidation of 1-(4-methoxyphenyl) ethanol with Acetobacter sp. CCTCC M209061 cells by adding deep eutectic solvent in a two-phase system

    OpenAIRE

    Wei, Ping; Liang, Jing; Cheng, Jing; Zong, Min-Hua; Lou, Wen-Yong

    2016-01-01

    Background Enantiopure (S)-1-(4-methoxyphenyl) ethanol {(S)-MOPE} can be employed as an important synthon for the synthesis of cycloalkyl [b] indoles with the treatment function for general allergic response. To date, the biocatalytic resolution of racemic MOPE through asymmetric oxidation in the biphasic system has remained largely unexplored. Additionally, deep eutectic solvents (DESs), as a new class of promising green solvents, have recently gained increasing attention in biocatalysis for...

  20. Fermented dairy products in the context of digestive cancer : the case of Propionibacterium freudenreichii, a cheese starter bacterium with pro-apoptotic properties

    OpenAIRE

    2013-01-01

    Background and objectives.[br/] Dairy propionibacteria are isolated from various ecological niches, including soil, grass, silage, rumen, raw milk and dairy plants, showing great adaptability and robustness. Used as ripening starters in Swiss type cheeses, these food grade bacteria are described as nutraceutical producers and they release into the external medium short chain fatty acids (SCFA), folic acid, cobalamin and the bifidogenic 1,4- dihydroxy-2-naphthoic acid (DHNA). These compounds ...

  1. Dairy Propionibacterium extends the mean lifespan of Caenorhabditis elegans via activation of the innate immune system.

    Science.gov (United States)

    Kwon, Gayeung; Lee, Jiyun; Lim, Young-Hee

    2016-08-17

    Dairy Propionibacterium freudenreichii is a candidate non-lactic acid probiotic. However, little information is available on the effect of P. freudenreichii on lifespan extension in humans. The aim of this study was to evaluate the effects of P. freudenreichii on lifespan extension and to elucidate the mechanism of P. freudenreichii-dependent lifespan extension in Caenorhabditis elegans. The results showed that P. freudenreichii significantly (p OP50, a standard food for the worm. Analysis of age-related biomarkers showed that P. freudenreichii retards ageing. Moreover, P. freudenreichii increased resistance against a human pathogen, Salmonella typhimurium, through the activation of skn-1, which is involved in pathogen resistance in C. elegans. Furthermore, P. freudenreichii-fed daf-16, jnk-1, skn-1 or daf-7 loss-of-function mutants showed an extended mean lifespan compared with E. coli OP50-fed worms. However, the increase in lifespan was not observed in pmk-1, sek-1, mek-1, dbl-1, daf-12 or daf-2 mutants, which suggests potential roles for these genes in P. freudenreichii-induced longevity in C. elegans. In conclusion, P. freudenreichii extends the lifespan of C. elegans via the p38 MAPK pathway involved in stress response and the TGF-β pathways associated with anti-inflammation processes in the immune system.

  2. 醋酸杆菌醇脱氢酶粗酶液的特性研究%Properties of Alcohol Dehydrogenase(ADH)Preparation from Acetobacter sp.CCTCC M209061

    Institute of Scientific and Technical Information of China (English)

    肖仔君; 黄国清; 钟瑞敏

    2010-01-01

    乙醇脱氢酶(ADH)是醋酸杆菌发酵生产醋酸的关键酶.以硫酸铵为沉淀剂,采用盐析法对醋酸杆菌中乙醇脱氢酶进行初步的分离纯化,并研究其酶学特性.结果表明:ADH比活力从粗酶液的0.201 U/mg提高到0.460 U/mg,纯化倍数为2.289;其最适作用pH值为7.5~8.0,pH值为7.8时酶活力达到最大,pH值为7.0时酶较为稳定;最适作用温度为35℃,温度为30℃~40℃时酶活力较为稳定,温度超过45℃后酶活力急剧下降.通过对乙醇底物浓度对ADH活力影响的研究,ADH对乙醇的米氏常数Km为2.59×10-2mol/L.

  3. Effects of probiotic Lactobacillus rhamnosus GG and Propionibacterium freudenreichii ssp. shermanii JS supplementation on intestinal and systemic markers of inflammation in ApoE*3Leiden mice consuming a high-fat diet

    NARCIS (Netherlands)

    Oksaharju, A.; Kooistra, T.; Kleemann, R.; Duyvenvoorde, W. van; Miettinen, M.; Lappalainen, J.; Lindstedt, K.A.; Kovanen, P.T.; Korpela, R.; Kekkonen, R.A.

    2013-01-01

    A high-fat diet disturbs the composition and function of the gut microbiota and generates local gut-associated and also systemic responses. Intestinal mast cells, for their part, secrete mediators which play a role in the orchestration of physiological and immunological functions of the intestine.

  4. Effects of probiotic Lactobacillus rhamnosus GG and Propionibacterium freudenreichii ssp. shermanii JS supplementation on intestinal and systemic markers of inflammation in ApoE*3Leiden mice consuming a high-fat diet

    NARCIS (Netherlands)

    Oksaharju, A.; Kooistra, T.; Kleemann, R.; Duyvenvoorde, W. van; Miettinen, M.; Lappalainen, J.; Lindstedt, K.A.; Kovanen, P.T.; Korpela, R.; Kekkonen, R.A.

    2013-01-01

    A high-fat diet disturbs the composition and function of the gut microbiota and generates local gut-associated and also systemic responses. Intestinal mast cells, for their part, secrete mediators which play a role in the orchestration of physiological and immunological functions of the intestine. P

  5. Clinical trial: multispecies probiotic supplementation alleviates the symptoms of IBS and stabilises intestinal microbiota

    NARCIS (Netherlands)

    Kajander, K.; Myllyluoma, E.; Rajlic-Stojanovic, M.; Kyronpalo, S.S.; Rasmussen, M.; Jarvenpaa, S.S.; Zoetendal, E.G.; Vos, de W.M.; Vapaatalo, H.; Korpela, R.

    2008-01-01

    Aim To investigate the effects of multispecies probiotic supplementation (Lactobacillus rhamnosus GG, L. rhamnosus Lc705, Propionibacterium freudenreichii ssp. shermanii JS and Bifidobacterium animalis ssp. lactis Bb12) on abdominal symptoms, quality of life, intestinal microbiota and inflammatory m

  6. Lactobacillus crispatus M206119 exacerbates murine DSS-colitis by interfering with inflammatory responses

    Institute of Scientific and Technical Information of China (English)

    Fu-Xi Zhou; Lu Chen; Xiao-Wei Liu; Chun-Hui Ouyang; Xiao-Ping Wu; Xue-Hong Wang; Chun-Lian Wang

    2012-01-01

    AIM:To investigate the role of Lactobacillus crispatus (L.crispatus) strain China Center for Type Culture Collection (CCTCC) M206119 in intestinal inflammation.METHODS:Forty 8-wk-old Balb/c mice (20 ± 2 g)were divided into four groups of 10 mice each.Three groups that had received dextran sulfate sodium (DSS)were administered normal saline,sulfasalazine or CCTCC M206119 strain,and the fourth group received none of these.We assessed the severity of colitis using a disease activity index,measured the colon length and weight,collected stools and mesenteric lymph nodes for bacterial microflora analysis.One centimeter of the proximal colon,middle colon and distal colon were collected and fixed in 10% buffered formalin,dehydrated in ethanol,and embedded in paraffin.Interleukin (IL)-1β,IL-6 and tumor necrosis factor (TNF)-α expression was detected using reverse transcription polymerase chain reaction.Protective factors zonula occludens (ZO)-1 and β-defensin 2 were detected by immunoblotting.The features of CCTCC M206119 strain were identified based on morphology,biochemical profile,and 16S RNA sequencing.RESULTS:DSS-colitis animals treated with CCTCC M206119 had markedly more severe disease,with greater weight loss,diarrhea,fecal bleeding,and shortened colon length.In addition,the CCTCC-M206119-treated group had comparatively higher histological scores and more neutrophil infiltration than the controls.Expression of protective factors ZO-1 and β-defensin 2 was downregulated due to destruction of the mucosal barrier after CCTCC M206119 strain treatment.An in vitro assay demonstrated that CCTCC M206119 strain increased the nuclear translocation of nuclear factor-κB in epithelial cells.Intestinal proinflammatory or anti-inflammatory cytokine responses were evaluated.Proinflammatory colonic cytokine (IL-1β,IL-6and TNF-α) levels were clearly increased in CCTCC-M206119-treated animals,whereas anti-inflammatory colonic cytokine (IL-10) level was lowered compared with

  7. Modification of digestive system microbiome of lactating dairy cows by feeding Bovamine: effect on ruminal fermentation

    Science.gov (United States)

    We evaluated the immune modulatory effects as well as effects on productivity of Bovamine® (Lactobacillus acidophilus strain NP51 and Probionibacterium freudenreichii) on the digestive system microbiome of dairy cattle during late lactation (average DIM = 202). To unveil the underlying mechanisms, ...

  8. Modification of immune responses and digestive system microbiota of lactating dairy cows by feeding Bovamine(R)

    Science.gov (United States)

    We evaluated the immune modulatory effects as well as effects on productivity of Bovamine® (Lactobacillus acidophilus strain NP51 and Probionibacterium freudenreichii) fed to Holstein and Jersey dairy cows during late lactation (average DIM = 202.44 days on wk-0). Cows were randomized to treatment g...

  9. Probiotic Bovamine fine-tunes the ruminal microbiome for enhanced fermentation and immunity

    Science.gov (United States)

    We evaluated the effect of Bovamine, Lactobacillus acidophilus strain NP51 and Propionibacterium freudenreichii strain strain NP24, feeding on the microbial composition of the digestive system microbiota of dairy cattle during late lactation (average DIM = 202 days on time 0). To examine the underly...

  10. Dicty_cDB: Contig-U12852-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available id:none) Nocardia farcinica IFM 10152 DN... 54 4e-06 AM944375_1( AM944375 |pid:none) Propionibacterium fre...udenreichii s... 54 4e-06 AE016825_167( AE016825 |pid:none) Chromobacterium violace

  11. Dicty_cDB: Contig-U16081-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available 1291 |pid:none) Cyanothece sp. PCC 7424, comple... 81 4e-37 AJ535197_1( AJ535197 |pid:none) Propionibacterium freud...eumoniae 342, comp... 75 4e-32 AJ535196_1( AJ535196 |pid:none) Propionibacterium freudenreichii s... 72 4e-3

  12. Dicty_cDB: Contig-U11181-1 [Dicty_cDB

    Lifescience Database Archive (English)

    Full Text Available id:none) Pyrobaculum aerophilum str. IM2,... 94 2e-17 AM944373_1( AM944373 |pid:n...one) Propionibacterium freudenreichii s... 93 3e-17 CP001114_2205( CP001114 |pid:none) Deinococcus deserti V

  13. Great interspecies and intraspecies diversity of dairy propionibacteria in the production of cheese aroma compounds.

    Science.gov (United States)

    Yee, Alyson L; Maillard, Marie-Bernadette; Roland, Nathalie; Chuat, Victoria; Leclerc, Aurélie; Pogačić, Tomislav; Valence, Florence; Thierry, Anne

    2014-11-17

    Flavor is an important sensory property of fermented food products, including cheese, and largely results from the production of aroma compounds by microorganisms. Propionibacterium freudenreichii is the most widely used species of dairy propionibacteria; it has been implicated in the production of a wide variety of aroma compounds through multiple metabolic pathways and is associated with the flavor of Swiss cheese. However, the ability of other dairy propionibacteria to produce aroma compounds has not been characterized. This study sought to elucidate the effect of interspecies and intraspecies diversity of dairy propionibacteria on the production of aroma compounds in a cheese context. A total of 76 strains of Propionibacterium freudenreichii, Propionibacterium jensenii, Propionibacterium thoenii, and Propionibacterium acidipropionici were grown for 15 days in pure culture in a rich medium derived from cheese curd. In addition, one strain each of two phylogenetically related non-dairy propionibacteria, Propionibacterium cyclohexanicum and Propionibacterium microaerophilum were included. Aroma compounds were analyzed using headspace trap-gas chromatography-mass spectrometry (GC-MS). An analysis of variance performed on GC-MS data showed that the abundance of 36 out of the 45 aroma compounds detected showed significant differences between the cultures. A principal component analysis (PCA) was performed for these 36 compounds. The first two axes of the PCA, accounting for 60% of the variability between cultures, separated P. freudenreichii strains from P. acidipropionici strains and also differentiated P. freudenreichii strains from each other. P. freudenreichii strains were associated with greater concentrations of a variety of compounds, including free fatty acids from lipolysis, ethyl esters derived from these acids, and branched-chain acids and alcohols from amino acid catabolism. P. acidipropionici strains produced less of these compounds but more sulfur

  14. Metabolic engineering of Torulopsis glabrata for malate production.

    Science.gov (United States)

    Chen, Xiulai; Xu, Guoqiang; Xu, Nan; Zou, Wei; Zhu, Pan; Liu, Liming; Chen, Jian

    2013-09-01

    The yeast Torulopsis glabrata CCTCC M202019, which is used for industrial pyruvate production, was chosen to explore the suitability of engineering this multi-vitamin auxotrophic yeast for increased malate production. Various metabolic engineering strategies were used to manipulate carbon flux from pyruvate to malate: (i) overexpression of pyruvate carboxylase and malate dehydrogenase; (ii) identification of the bottleneck in malate production by model iNX804; (iii) simultaneous overexpression of genes RoPYC, RoMDH and SpMAE1. Using these strategies, 8.5gL(-1) malate was accumulated in the engineered strain T.G-PMS, which was about 10-fold greater than that of the control strain T.G-26. The results presented here suggest that T. glabrata CCTCC M202019 is a promising candidate for industrial malate production.

  15. Effect of Trichoderma harzianum on maize rhizosphere microbiome and biocontrol of Fusarium Stalk rot.

    Science.gov (United States)

    Saravanakumar, Kandasamy; Li, Yaqian; Yu, Chuanjin; Wang, Qiang-Qiang; Wang, Meng; Sun, Jianan; Gao, Jin-Xin; Chen, Jie

    2017-05-11

    Fusarium stalk rot (FSR) caused by Fusarium graminearum (FG) significantly affects the productivity of maize grain crops. Application of agrochemicals to control the disease is harmful to environment. In this regard, use of biocontrol agent (BCA) is an alternative to agrochemicals. Although Trichoderma species are known as BCA, the selection of host-pathogen specific Trichoderma is essential for the successful field application. Hence, we screened a total of 100 Trichoderma isolates against FG, selected Trichoderma harzianum (CCTCC-RW0024) for greenhouse experiments and studied its effect on changes of maize rhizosphere microbiome and biocontrol of FSR. The strain CCTCC-RW0024 displayed high antagonistic activity (96.30%), disease reduction (86.66%), biocontrol-related enzyme and gene expression. The root colonization of the strain was confirmed by eGFP tagging and qRT-PCR analysis. Pyrosequencing revealed that exogenous inoculation of the strain in maize rhizosphere increased the plant growth promoting acidobacteria (18.4%), decreased 66% of FG, and also increased the plant growth. In addition, metabolites of this strain could interact with pathogenicity related transcriptional cofactor FgSWi6, thereby contributing to its inhibition. It is concluded that T. harzianum strain CCTCC-RW0024 is a potential BCA against FSR.

  16. Characterization of Feruloyl Esterases Produced by the Four Lactobacillus Species: L. amylovorus, L. acidophilus, L. farciminis and L. fermentum, Isolated from Ensiled Corn Stover

    Directory of Open Access Journals (Sweden)

    Zhenshang Xu

    2017-06-01

    Full Text Available Lactic acid bacteria (LAB play important roles in silage fermentation, which depends on the production of sufficient organic acids to inhibit the growth of undesirable microorganisms. However, LAB are not able to degrade cellulose and hemicellulose. Bacteria and fibrolytic enzymes are usually used as inoculants to improve the silage quality and digestibility. In the present study, we isolated four Lactobacillus strains (L. amylovorus CGMCC 11056, L. acidophilus CCTCC AB2010208, L. farciminis CCTCC AB2016237 and L. fermentum CCTCC AB2010204 with feruloyl esterase (FAE activities from ensiled corn stover (CS by a plate screening assay. The genes encoding FAEs were cloned and hetero-expressed in Escherichia coli. The optimal temperature and pH of these purified enzymes ranged from 45 to 50°C and from 7.0 to 8.0, respectively. They could hydrolyze hydroxycinnamoyl esters in a substrate-specific manner when methyl ferulate, methyl caffeate, methyl ρ-coumarate and methyl sinapinate were used as substrates. Moreover, these four FAEs were able to hydrolyze CS to release hydroxycinnamic acids. Furthermore, these strains could degrade hydroxycinnamic esters, and L. amylovorus CGMCC 11056 was the most efficient strain among these four isolates. These results provided a new target for the development of inoculants to improve silage quality and digestibility.

  17. Production of wheat bread without preservatives using sourdough starters.

    Science.gov (United States)

    Denkova, Rositsa; Ilieva, Svetla; Denkova, Zapryana; Georgieva, Ljubka; Yordanova, Mariya; Nikolova, Dilyana; Evstatieva, Yana

    2014-09-03

    In order for the beneficial effects of sourdough application in breadmaking to take place a proper selection of lactic acid bacteria species and strains, an appropriate technology and effective control of the purity and activity of the selected cultures. Four symbiotic starters for sourdough for the production of bread were developed and probated in a production laboratory using the selected strains Lactobacillus brevis LBRZ7, L. buchneri LBRZ6, L. plantarum X2, L. paracasei RN5, L. sanfranciscensis R and L. fermentum LBRH10 and the probiotic strain Propionibacterium freudenreichii ssp. shermanii NBIMCC 327. The starter sourdoughs that include Propionibacterium freudenreichii ssp. shermanii NBIMCC 327 had greater antimicrobial activity against saprophytic microorganisms: Bacillus subtilis, B. mesentericus, Aspergillus niger, Penicillium sp. and Rhizopus sp., but none of them inhibited the growth of bakery yeasts Saccharomyces cerevisiae. It was established that in order to prevent bacterial spoilage 10% of the selected starter sourdoughs had to be added in the breadmaking process, while for prevention of mold spoilage the necessary amount of starter sourdough had to be between 15% and 20%.The application of the developed starters for the production of wheat bread guarantees longer shelf life and no adverse alterations in the features of the final bread.

  18. Clarification of interactions among microorganisms and development of co-culture system; Biseibutsukan sokosayo no kaiseki to kongo baiyo shisutemu no kaihatsu

    Energy Technology Data Exchange (ETDEWEB)

    Taniguchi, Masayuki [Niigata University, Niigata (Japan). Dept. of Materials Science and Technology

    1999-03-10

    Co-culture systems containing two microorganisms for production of useful substances are described. The co-culture of Bifidobacterium longum and Propionibacterium freudenreichii, where lactic acid produced once from lactose by B. longum is converted to acetic and propionic acids by P. freudenreichii, was carried out. Thought the sequential conversion of lactose using the two microorganisms, the culture supernatant containing a mixture of acetic and propionic acids without lactic acid is produced. The antimicrobial activity of the mixture is higher than that obtained in the cultivation of B. longum alone. We developed a novel co-culture system composed of two fermentors and two micro filtration modules for efficient ethanol production from a mixture of glucose and xylose by co-culture of Pichia stipitis and Saccharomyces cerevisiae. The proposed co-culture system allowed regulation of the dissolved oxygen concentration at a level suitable for an individual yeast in each fermentor, as well as the successful exchange of culture medium between two fermentors. When P. stipitis and S. cerevisiae are cultivated individually under different oxygen supply conditions in the new co-culture system, the yield and productivity of ethanol from a glucose and xylose mixture are higher than in single culture of P. stipitis alone. By clarifying the interactions among microorganisms, new bioprocesses in which similar performance to co-culture systems is expressed even using a single microorganism, are expected to be developed for improvement of biochemical reaction systems. (author)

  19. A methodological approach to screen diverse cheese-related bacteria for their ability to produce aroma compounds.

    Science.gov (United States)

    Pogačić, Tomislav; Maillard, Marie-Bernadette; Leclerc, Aurélie; Hervé, Christophe; Chuat, Victoria; Yee, Alyson L; Valence, Florence; Thierry, Anne

    2015-04-01

    Microorganisms play an important role in the development of cheese flavor. The aim of this study was to develop an approach to facilitate screening of various cheese-related bacteria for their ability to produce aroma compounds. We combined i) curd-based slurry medium incubated under conditions mimicking cheese manufacturing and ripening, ii) powerful method of extraction of volatiles, headspace trap, coupled to gas chromatography-mass spectrometry (HS-trap-GC-MS), and iii) metabolomics-based method of data processing using the XCMS package of R software and multivariate analysis. This approach was applied to eleven species: five lactic acid bacteria (Leuconostoc lactis, Lactobacillus sakei, Lactobacillus paracasei, Lactobacillus fermentum, and Lactobacillus helveticus), four actinobacteria (Brachybacterium articum, Brachybacterium tyrofermentans, Brevibacterium aurantiacum, and Microbacterium gubbeenense), Propionibacterium freudenreichii, and Hafnia alvei. All the strains grew, with maximal populations ranging from 7.4 to 9.2 log (CFU/mL). In total, 52 volatile aroma compounds were identified, of which 49 varied significantly in abundance between bacteria. Principal component analysis of volatile profiles differentiated species by their ability to produce ethyl esters (associated with Brachybacteria), sulfur compounds and branched-chain alcohols (H. alvei), branched-chain acids (H. alvei, P. freudenreichii and L. paracasei), diacetyl and related carbonyl compounds (M. gubbeenense and L. paracasei), among others.

  20. Quick Estimates of Peak Overpressure from Two Simultaneous Blast Waves

    Science.gov (United States)

    1977-12-01

    C. S. Knauth, Nuclear Blast Standard (1 kT), Air Force Weapons Laboratory, Krtland A-r Force Base, New Mexico , AFWL-TR-73-55, (Revised) 1975. 6 C. E...Needham, Private Communication, Air Force Weapons Laboratory, Kirtland Air Force Base, New Mexico , November 1977. 36i If 1I * I I| ~~’.0 DISTRIBUTION...for Ops. & Plans Defense Communications Agency ATTN: Dep. Dir. for Nuc. Chem. Matters ATTN: Code 930 ATTN: MOCA -ADL ATTN: CCTC/C672, F. Moore Deputy

  1. 基于代谢通量分析的SAM和GSH联产溶氧控制策略%The Dissolved Oxygen Control Strategy Based on Metabolic Flux Analysis for Fermentative Co-Production of S-Adenosylmethionine and Glutathione

    Institute of Scientific and Technical Information of China (English)

    王大慧; 王玉磊; 卫功元; 邵娜; 周捷

    2013-01-01

    In order to improve the co-production of SAM and GSH under batch cultivation, the effect of dissolved oxygen (DO) level on fermentative co-production of S-adenosylmethionine (SAM) and glutathione (GSH) in a 5 L stirred fermentor with Candida utilis CCTCC M 209298 was investigated. The metabolic fluxes of intermediates and energy during batch fermentative co-production of SAM and GSH under different agitation rates were calculated by using metabolic flux analysis method. Based on the flux distribution of main intermediates, an optimum DO control strategy of a constant agitation rate of 350 r⋅min−1 maintained during the first 9 h of fermentation followed with a DO level not less than 30% after 9 h was proposed as an efficient approach for high co-production of SAM and GSH. Under this DO control mode, the product of 520.7 mg⋅L−1 for batch co-production of SAM and GSH with C.utilis CCTCC M 209298 was achieved. Furthermore, the reason for the improvement of the co-production of SAM and GSH under the proposed strategy was quantitatively interpreted by the fermentative kinetics, metabolic flux analysis and respiratory quotient, respectively.%  为了提高S-腺苷甲硫氨酸(SAM)和谷胱甘肽(GSH)的联产量,考察了溶氧条件对Candida utilis CCTCC M 209298发酵联产SAM和GSH的影响.采用代谢通量分析方法对不同转速下SAM和GSH联产发酵过程中的物质和能量代谢进行分析,确定了较优的溶氧控制策略:前9 h的发酵恒定转速350 r⋅min−1,9 h后控制溶氧不低于30%.在此策略下,C. utilis CCTCC M 209298分批发酵生产SAM和GSH的联产量为520.7 mg⋅L−1.最后分别从发酵动力学、代谢通量分析以及呼吸商变化规律对该溶氧控制策略促进SAM和GSH联合高产的原因进行了定量解释.

  2. Synthesis, Characterization and Biological Activities of Novel Water-soluble Metalloporphyrins

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The synthesis, characterization, and biological activities of a series of metal porphyrins were described. The complexes 3a~3e were prepared from the reaction of compound 2 with M(OAc)2 in CHCl3, and the treatment of 3a~3f with pyridine gave corresponding complexes 4a~4f. These new compounds were identified by absorption spectroscopies, 1H NMR and elemental analysis. The results of biological activity testing revealed that 4a and 4c had stronger inhibiting action for Escherichia coli (CCTCC AB91115).

  3. Antifungal Activity of Selected Lactic Acid Bacteria and Propionic Acid Bacteria against Dairy-Associated Spoilage Fungi

    DEFF Research Database (Denmark)

    Aunsbjerg, Stina Dissing

    for prolonging shelf-life of food without the addition of specific preservatives. Increased interest in the use of these bacteria for biopreservation has led to identification of a range of potent strains, and in addition, isolation and identification of various antifungal metabolites produced by these cultures......Bacterial cultures of lactic and propionic acid bacteria are widely used in fermented products including dairy products. Spoilage fungi may constitute a quality and safety issue in these products. The antifungal properties of some lactic and propionic acid bacteria make them potential candidates...... defined interaction medium (CDIM) was developed allowing growth of protective Lb. paracasei and P. freudenreichii subsp. shermaniii as well as the spoilage fungi, Penicillium spp., Rhodotorula mucilaginosa and Debaryomyces hansenii isolated from fermented dairy products. Lb. paracasei and P...

  4. Development of a chemically defined medium for studying foodborne bacterial-fungal interactions

    DEFF Research Database (Denmark)

    Aunsbjerg, Stina Dissing; Honoré, Anders Hans; Vogensen, Finn Kvist

    2015-01-01

    There is a growing interest for using natural preservatives in the food and dairy industries including the application of bacterial cultures to inhibit fungal spoilage. Several antifungal metabolites from bacteria have been identified, but their relative importance has been difficult to establish....... In dynamic systems such as fermented milk products, the complexity of the food matrix affects detection, identification and quantification of antifungal metabolites, and thereby the understanding of the bacterial-fungal interactions. To ease the identification and quantification of bacterial metabolites (as...... judged by ultra-performance liquid chromatography/mass spectrometry) a chemically defined interaction medium (CDIM) was developed. The medium supported growth of antifungal cultures such as Lactobacillus paracasei and Propionibacterium freudenreichii, as well as spoilage moulds and yeasts isolated from...

  5. Antifungal Activity of Selected Lactic Acid Bacteria and Propionic Acid Bacteria against Dairy-Associated Spoilage Fungi

    DEFF Research Database (Denmark)

    Aunsbjerg, Stina Dissing

    Bacterial cultures of lactic and propionic acid bacteria are widely used in fermented products including dairy products. Spoilage fungi may constitute a quality and safety issue in these products. The antifungal properties of some lactic and propionic acid bacteria make them potential candidates...... diacetyl and lactic acid, 6 antifungal hydroxy acids were identified. Of these, 3 have previously been reported from antifungal lactic acid bacteria, whereas the other 3 hydroxy acids have not previously been reported produced by antifungal lactic acid bacteria....... factors thereby facilitating development of new protective cultures. The aim of the current thesis was to determine and study metabolites involved in antifungal activity of protective Lactobacillus paracasei and Propionibacterium freudenreichii subsp. shermanii. This involved development of a suitable...

  6. Exometabolomic Profiling of Bacterial Cultures

    DEFF Research Database (Denmark)

    Honoré, Anders Hans

    was required to maintain a diacetyl concentration sufficient for the antifungal effect. Over time, the concentration of diacetyl decreased and mold developed similar to an acidified un‐inoculated medium. Besides diacetyl, production of lactic acid and other 2‐hydroxy acids contributed weakly to the antifungal......) and Propionibacterium freudenreichii subsp. shermanii (PAB A). The strategy was to investigate the effect of the composition of exometabolome of the co‐culture on mold growth. A biological model system was developed in order to have a simplified system for studying the growth of bacteria and the subsequent effect...... on mold growth represented by two strains of Penicillium (manuscript III). Characterization of mold growth was performed by a spectral clustering algorithm on data from multispectral imaging (manuscript VI). Untargeted analysis of the exometabolome was performed on liquid chromatography/mass spectrometry...

  7. Eyeblink conditioning in unmedicated schizophrenia patients: a positron emission tomography study.

    Science.gov (United States)

    Parker, Krystal L; Andreasen, Nancy C; Liu, Dawei; Freeman, John H; O'Leary, Daniel S

    2013-12-30

    Previous studies suggest that patients with schizophrenia exhibit dysfunctions in a widely distributed circuit-the cortico-cerebellar-thalamic-cortical circuit, or CCTCC-and that this may explain the multiple cognitive deficits observed in the disorder. This study uses positron emission tomography (PET) with O(15) H₂O to measure regional cerebral blood flow (rCBF) in response to a classic test of cerebellar function, the associative learning that occurs during eyeblink conditioning, in a sample of 20 unmedicated schizophrenia patients and 20 closely matched healthy controls. The PET paradigm examined three phases of acquisition and extinction (early, middle and late). The patients displayed impaired behavioral performance during both acquisition and extinction. The imaging data indicate that, compared to the control subjects, the patients displayed decreases in rCBF in all three components of the CCTCC during both acquisition and extinction. Specifically, patients had less rCBF in the middle and medial frontal lobes, anterior cerebellar lobules I/V and VI, as well as the thalamus during acquisition and although similar areas were found in the frontal lobe, ipsilateral cerebellar lobule IX showed consistently less activity in patients during extinction. Thus this study provides additional support for the hypothesis that patients with schizophrenia have a cognitive dysmetria--an inability to smoothly coordinate many different types of mental activity--that affects even a very basic cognitive task that taps into associative learning.

  8. Isolation of brefeldin A from Eupenicillium brefeldianum broth using macroporous resin adsorption chromatography.

    Science.gov (United States)

    Wang, Ya-Jun; Wu, Ye-Fei; Xue, Feng; Wu, Zhi-Xian; Xue, Ya-Ping; Zheng, Yu-Guo; Shen, Yin-Chu

    2012-05-01

    Brefeldin A (BFA) is a macrolide lactone antibiotic, possessing antitumor, antiviral, antifungal activities. In this work, a separation strategy involving one-step macroporous resin adsorption chromatography combined with crystallization was established for BFA purification from Eupenicillium brefeldianum CCTCC M 208113 fermentation broth. Among six macroporous resin adsorbents tested, the non-polar resin HZ830 had the best adsorption and desorption performance. The static equilibrium adsorption data fitted well with the Freundlich equation, and the adsorption kinetic followed the pseudo-second order model. Through experimental optimization of column adsorption and desorption, BFA in purity of 90.4% (w/w), 92.1% (w/w) yield was obtained by a one-step macroporous resin adsorption chromatography, using a stepwise elution protocol. Furthermore, high purity (>99%, w/w) of BFA crystals were prepared from E. brefeldianum CCTCC M 208113 fermentation broth in an overall recovery of 67.0% (w/w), using a combination of adsorption chromatography packed with non-polar macroporous adsorbent HZ830 and crystallization in acetone. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. Production of a single cyclic type of fructooligosaccharide structure by inulin-degrading Paenibacillus sp. LX16 newly isolated from Jerusalem artichoke root.

    Science.gov (United States)

    Yao, Zhihua; Guo, Jiqiang; Tang, Wenzhu; Sun, Zhen; Hou, Yingmin; Li, Xianzhen

    2016-05-01

    A novel inulin-degrading bacterium was isolated from a soil sample collected on Jerusalem artichoke roots. It is a Gram-positive, aerobic, motile and central endospore-forming straight rod, and exhibits phenotypic properties being consistent with its classification in the genus Paenibacillus. The predominant cellular fatty acids were anteiso-C15:0, C16:0 and anteiso-C17:0. This strain represents a novel species of the genus Paenibacillus on the basis of phenotypic data together with phylogenetic analysis, and it is here designated as LX16 and deposited in China centre for type collection, China (= CCTCC 2015256). Strain LX16 could produce a cyclofructooligosaccharide fructanotransferase catalysing the formation of one type of fructooligosaccharide (FOS) from inulin. The FOS was identified as a cyclofructooligosaccharide with a degree of polymerization of 6. Such homology in inulin degradation products may be beneficial for the functional FOS production.

  10. Copper sulfate improves pullulan production by bioconversion using whole cells of Aureobasidium pullulans as the catalyst.

    Science.gov (United States)

    Wang, Dahui; Ju, Xiaomin; Zhang, Gaochuan; Wang, Donghua; Wei, Gongyuan

    2016-10-01

    The effects of mineral salts on pullulan production by bioconversion using whole cells of Aureobasidium pullulans CCTCC M 2012259 as the catalyst were investigated. Copper sulfate (CuSO4) improved pullulan production by 36.2% and 42.3% when added at the optimum concentration of 0.2mg/L to the bioconversion broth or seed medium, respectively, as compared with controls without CuSO4 addition. Pullulan production was further enhanced when CuSO4 was added to both seed medium and bioconversion broth simultaneously. In order to probe the mechanism of CuSO4 improvement, cell viability, membrane integrity, intracellular adenosine triphosphate (ATP) levels and the activities of key enzymes involved in pullulan biosynthesis were determined. As a result, CuSO4 increased the activities of key biosynthetic enzymes, maintained intracellular ATP at a higher level, and accelerated the rate of pullulan secretion, all of which contributed to improved pullulan production by bioconversion.

  11. Roseovarius antarcticus sp. nov., isolated from a decayed whale bone.

    Science.gov (United States)

    Deng, Sangsang; Jiang, Fan; Chang, Xulu; Qu, Zhihao; Ren, Lvzhi; Zhang, Yumin; Kan, Wenjing; Da, Xuyang; Qiu, Xia; Kim, Myongchol; Fang, Chengxiang; Peng, Fang

    2015-07-01

    A pale yellow, ovoid- to rod-shaped and budding bacterium, designated strain M-S13-148(T), was isolated from a decayed bone of whale from the eastern coast of King George Island, South Shetlands, Antarctica. Strain M-S13-148(T) exhibited motility, aerobic growth and was Gram-stain-negative. Strain M-S13-148(T) was positive for catalase and oxidase. Growth was observed at pH 6.0-9.0, at 4-42 °C and with 0-14% (w/v) NaCl. The novel strain contained diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine and an unknown phospholipid as the major polar lipids. The dominant cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c), (58.8%) and C16 : 0 (11.7%). The respiratory quinone was Q-10 and the DNA G + C content was 60.9 mol%. Neighbour-joining, maximum-likelihood and minimum-evolution phylogenetic trees, based on 16S rRNA gene sequences, indicated that strain M-S13-148(T) belonged to the genus Roseovarius and was most closely related to Roseovarius nanhaiticus CCTCC AB 208317(T) (93.72% 16S rRNA gene sequence similarity). The 16S rRNA gene sequence similarity with respect to members of the genus Roseovarius ranged from 91.81 to 93.94%. On the basis of phenotypic, molecular and chemotaxonomic characteristics, strain M-S13-148 is considered to represent a novel species of the genus Roseovarius, for which the name Roseovarius antarcticus sp. nov., is proposed. The type strain is M-S13-148(T) ( = CCTCC AB2014072(T) = LMG 28420(T)).

  12. 植物乳杆菌与费氏丙酸菌转化生成甘露醇的比较%Comparison of D-Mannitol Formation from D-fructose by Biotransformation with Lactobacilli or Propionibacteria

    Institute of Scientific and Technical Information of China (English)

    胡奇杰; 戚民; 姚兆敏

    2011-01-01

    对1株植物乳杆菌(Lactobacillus plantarum HSC235)和1株费氏丙酸菌(Propionibacterium freudenreichii HZP-35)生物转化果糖生成甘露醇能力进行了比较研究。在生长细胞、静息细胞以及无完整细胞粗酶提取物3种不同转化反应体系中,植物乳杆菌HSC235和费氏丙酸菌HZP-35都表现出甘露醇脱氢酶活性,而生长细胞甘露醇产量和得率较高,分别为38g/L和27%,静息细胞和粗酶提取物反应体系甘露醇的产量及得率则相关无几,表明甘露醇的转化过程与细胞生长紧密%D-Mannitol production from fructose by lactic acid bacteria (LAB, Lactobacillus plantarum HSC 235) and propionate acid bacteria (PAB, Propionibacterium freudenreichii HZP-35) was investigated. Fructose can be con- verted into mannitol and sorbitol. Three kinds of reaction systems of growing cells, resting cells, and free cell extract were compared in mannitol formation using fructose as substrate. The resuhs showed that the growing ceils reaction process gave out a higher mannitol yield at 38 g/L, the Yp/s at 27% , while the last two reaction processes had a similar low mannitol yield. This suggested that mannitol formation was coincided with the cell growth of LAB and PAB. Initial fructose concentration hadn't appear effect on mannitol formation between 50-150 g/L where the mannitol Yp/ s was at 27% , but high up to 200 g/L fructose had a inhibition on the conversion process. Although LAB and PAB can grow together as in many food, but the mixed cultivation of the two bacteria would not produce more mannitol compared with any single pure strain fermentation. In the ability of mannitol production, LAB showed a high level to PAB.

  13. Recrystallized S-layer protein of a probiotic Propionibacterium: structural and nanomechanical changes upon temperature or pH shifts probed by solid-state NMR and AFM.

    Science.gov (United States)

    de sa Peixoto, Paulo; Roiland, Claire; Thomas, Daniel; Briard-Bion, Valérie; Le Guellec, Rozenn; Parayre, Sandrine; Deutsch, Stéphanie-Marie; Jan, Gwénaël; Guyomarc'h, Fanny

    2015-01-01

    Surface protein layers (S layers) are common constituents of the bacterial cell wall and originate from the assembly of strain-dependent surface layer proteins (Slps). These proteins are thought to play important roles in the bacteria's biology and to have very promising technological applications as biomaterials or as part of cell-host cross-talk in probiotic mechanism. The SlpA from Propionibacterium freudenreichii PFCIRM 118 strain was isolated and recrystallized to investigate organization and assembly of the protein using atomic force microscopy and solid-state (1)H and (13)C-nuclear magnetic resonance. SlpA was found to form hexagonal p1 monolayer lattices where the protein exhibited high proportions of disordered regions and of bound water. The lattice structure was maintained, but softened, upon mild heating or acidification, probably in relation with the increasing mobilities of the disordered protein regions. These results gave structural insights on the mobile protein regions exposed by S layer films, upon physiologically relevant changes of their environmental conditions.

  14. Comparison of 16S ribosomal RNA genes in Clavibacter michiganensis subspecies with other coryneform bacteria.

    Science.gov (United States)

    Li, X; De Boer, S H

    1995-10-01

    Nearly complete sequences (97-99%) of the 16S rRNA genes were determined for type strains of Clavibacter michiganensis subsp. michiganensis, Clavibacter michiganensis subsp. insidiosus, Clavibacter michiganensis subsp. sepedonicus, and Clavibacter michiganensis subsp. nebraskensis. The four subspecies had less than 1% dissimilarity in their 16S rRNA genes. Comparative studies indicated that the C. michiganensis subsp. shared relatively high homology with the 16S rRNA gene of Clavibacter xyli. Further comparison with representatives of other Gram-positive coryneform and related bacteria with high G+C% values showed that this group of bacteria was subdivided into three clusters. One cluster consisted of the Clavibacter michiganensis subsp., Clavibacter xyli, Arthrobacter globiformis, Arthrobacter simplex, and Frankia sp.; another cluster consisted of members of the corynebacteria-mycobacteria-nocardia (CMN) group of Mycobacteriaceae including Tsukamurella paurometabolum; and Propionibacterium freudenreichii alone formed a unique cluster, which was remote from other coryneform bacteria analyzed. The three clusters may reflect a systematic rank higher than the genus level among these bacteria.

  15. Reduced-fat Cheddar and Swiss-type cheeses harboring exopolysaccharide-producing probiotic Lactobacillus mucosae DPC 6426.

    Science.gov (United States)

    Ryan, P M; Burdíková, Z; Beresford, T; Auty, M A E; Fitzgerald, G F; Ross, R P; Sheehan, J J; Stanton, C

    2015-12-01

    Exopolysaccharide-producing Lactobacillus mucosae DPC 6426 was previously shown to have promising hypocholesterolemic activity in the atherosclerosis-prone apolipoprotein-E-deficient (apoE(-/-)) murine model. The aim of this study was to investigate the suitability of reduced-fat Cheddar and Swiss-type cheeses as functional (carrier) foods for delivery of this probiotic strain. All cheeses were manufactured at pilot-scale (500-L vats) in triplicate, with standard commercially available starters: for Cheddar, Lactococcus lactis; and for Swiss-type cheese, Streptococcus thermophilus, Lactobacillus helveticus, and Propionibacterium freudenreichii. Lactobacillus mucosae DPC 6426 was used as an adjunct culture during cheese manufacture, at a level of ~10(6) cfu·mL(-1) cheese milk (subsequently present in the cheese curd at>10(7) cfu·g(-1)). The adjunct strain remained viable at >5×10(7) cfu·g(-1) in both Swiss-type and Cheddar cheeses following ripening for 6 mo. Sensory analysis revealed that the presence of the adjunct culture imparted a more appealing appearance in Swiss-type cheese, but had no significant effect on the sensory characteristics of Cheddar cheeses. Moreover, the adjunct culture had no significant effect on cheese composition, proteolysis, pH, or instrumentally quantified textural characteristics of Cheddar cheeses. These data indicate that low-fat Swiss-type and Cheddar cheeses represent suitable food matrices for the delivery of the hypocholesterolemic Lactobacillus mucosae DPC 6426 in an industrial setting.

  16. Interaction of Campylobacter spp. and human probiotics in chicken intestinal mucus.

    Science.gov (United States)

    Ganan, M; Martinez-Rodriguez, A J; Carrascosa, A V; Vesterlund, S; Salminen, S; Satokari, R

    2013-03-01

    Campylobacter is the most common cause of bacterial food-borne diarrhoeal disease throughout the world. The principal risk of human contamination is handling and consumption of contaminated poultry meat. To colonize poultry, Campylobacter adheres to and persists in the mucus layer that covers the intestinal epithelium. Inhibiting adhesion to the mucus could prevent colonization of the intestine. The aim of this study was to investigate in vitro the protective effect of defined commercial human probiotic strains on the adhesion of Campylobacter spp. to chicken intestinal mucus, in a search for alternatives to antibiotics to control this food-borne pathogen. The probiotic strains Lactobacillus rhamnosus GG and Propionibacterium freudenreichii ssp. shermanii JS and a starter culture strain Lactococcus lactis ssp. lactis adhered well to chicken intestinal mucus and were able to reduce the binding of Campylobacter spp. when the mucus was colonized with the probiotic strain before contacting the pathogen. Human-intended probiotics could be useful as prophylactics in poultry feeding for controlling Campylobacter spp. colonization. © 2012 Blackwell Verlag GmbH.

  17. Probiotic intervention has strain-specific anti-inflammatory effects in healthy adults

    Institute of Scientific and Technical Information of China (English)

    Riina A Kekkonen; Riitta Korpela; Netta Lummela; Heli Karjalainen; Sinikka Latvala; Soile Tynkkynen; Salme Jarvenpaa; Hannu Kautiainen; Ilkka Julkunen; Heikki Vapaatalo

    2008-01-01

    AIM: To evaluate the effects of three potentially anti-inflammatory probiotic bacteria from three different genera on immune variables in healthy adults in a clinical setting based on previous in vitro characterization of cytokine responses.METHODS: A total of 62 volunteers participated in this randomized, double-blind and placebo-controlled parallel group intervention study. The volunteers were randomized to receive a milk-based drink containing either Lactobadllus rhamnosus GG (LGG), Bifidobacterium animalis ssp. lactis Bbl2 (Bbl2), or Propionibacterium freudenreichii ssp. shermanii JS (PJS) or a placebo drink for 3 wk. Venous blood and saliva samples were taken at baseline and on d 1, 7 and 21. Fecal samples were collected at baseline and at the end of intervention.RESULTS: The serum hsCRP expressed as the median AUCo-21 (minus baseline) was 0.018 mg/L in the placebo group, -0.240 mg/L in the LGG group, 0.090 mg/L in the Bbl2 group and -0.085 mg/L in the PJS group (P = 0.014). In vitro production of TNF-a from in vitro cultured peripheral blood mononuclear cells (PBMC) was significantly lower in subjects receiving LGG vs placebo. IL-2 production from PBMC in the Bbl2 group was significantly lower compared with the other groups.CONCLUSION: In conclusion, probiotic bacteria have strain-specific anti-inflammatory effects in healthy adults.

  18. Survey of bacterial proteins released in cheese: a proteomic approach.

    Science.gov (United States)

    Gagnaire, Valérie; Piot, Michel; Camier, Bénédicte; Vissers, Johannes P C; Jan, Gwénaël; Léonil, Joëlle

    2004-07-15

    During the ripening of Emmental cheese, the bacterial ecosystem confers its organoleptic characteristics to the evolving curd both by the action of the living cells, and through the release of numerous proteins, including various types of enzymes into the cheese when the cells lyse. In Emmental cheese these proteins can be released from thermophilic lactic acid bacteria used as starters like Lactobacillus helveticus, Lb delbruecki subsp. lactis and Streptococcus salivarius subsp. thermophilus and ripening bacteria such as Propionibacterium freudenreichii. The aim of this study was to obtain a proteomic view of the different groups of proteins within the cheese using proteomic tools to create a reference map. A methodology was therefore developed to reduce the complexity of cheese matrix prior to 2D-PAGE analysis. The aqueous phase of cheese was prefractionated by size exclusion chromatography, bacterial and milk proteins were separated and subsequently characterised by mass spectrometry, prior to peptide mass fingerprint and sequence homology database search. Five functional groups of proteins were identified involved in: (i) proteolysis, (ii) glycolysis, (iii) stress response, (iv) DNA and RNA repair and (v) oxidoreduction. The results revealed stress responses triggered by thermophilic lactic acid bacteria and Propionibacterium strains at the end of ripening. Information was also obtained regarding the origin and nature of the peptidases released into the cheese, thus providing a greater understanding of casein degradation mechanisms during ripening. Different peptidases arose from St thermophilus and Lb helveticus, suggesting that streptococci are involved in peptide degradation in addition to the proteolytic activity of lactobacilli.

  19. A riboswitch sensor to determine vitamin B12 in fermented foods.

    Science.gov (United States)

    Zhu, Xuan; Wang, Xiaofeng; Zhang, Chen; Wang, Xiaoqi; Gu, Qing

    2015-05-15

    We describe a sensitive and selective method for determination of vitamin B12 content in fermented foods using riboswitch sensor. A riboswitch amplicon from Propionibacterium freudenreichii was cloned in p519NGFP vector in Escherichia coli BL21 (DE3). The expression of green fluorescence protein was revers correlated to the concentrations of adenosylcobalamin. Adenosylcobalamin directly binds to riboswitch region leading to conformational changes in the secondary structure of mRNA, thus inhibiting expression. After various examinations, a standard curve was obtained from 10 to 1000 ng/mL of cyanocobalamin. The limit of determination is 10 ng/mL. The inter-assay coefficients of variation were 7.5% for the range of 10-1000 ng/mL. The recovery of this method was 92.3%. This method has no or less responses to nucleic acid, pseudovitamin B12, vitamin B12 bound to intrinsic factor and haptocorrin. The riboswitch sensor results were similar with HPLC, but they were Ca. 24% lower than the microbiological assay results.

  20. Effect of a multispecies probiotic supplement on quantity of irritable bowel syndrome-related intestinal microbial phylotypes

    Directory of Open Access Journals (Sweden)

    Lyra Anna

    2010-09-01

    Full Text Available Abstract Background Probiotics can alleviate the symptoms of irritable bowel syndrome (IBS, possibly by stabilizing the intestinal microbiota. Our aim was to determine whether IBS-associated bacterial alterations were reduced during multispecies probiotic intervention consisting of Lactobacillus rhamnosus GG, L. rhamnosus Lc705, Propionibacterium freudenreichii ssp. shermanii JS and Bifidobacterium breve Bb99. The intervention has previously been shown to successfully alleviate gastrointestinal symptoms of IBS. Methods The faecal microbiotas of 42 IBS subjects participating in a placebo-controlled double-blind multispecies probiotic intervention were analysed using quantitative real-time polymerase chain reaction (qPCR. Eight bacterial targets within the gastrointestinal microbiota with a putative IBS association were measured. Results A phylotype with 94% similarity to Ruminococcus torques remained abundant in the placebo group, but was decreased in the probiotic group during the intervention (P = 0.02 at 6 months. In addition, the clostridial phylotype, Clostridium thermosuccinogenes 85%, was stably elevated during the intervention (P = 0.00 and P = 0.02 at 3 and 6 months, respectively. The bacterial alterations detected were in accordance with previously discovered alleviation of symptoms. Conclusions The probiotic supplement was thus shown to exert specific alterations in the IBS-associated microbiota towards the bacterial 16S rDNA phylotype quantities described previously for subjects free of IBS. These changes may have value as non-invasive biomarkers in probiotic intervention studies.

  1. Bacillus composti sp. nov. and Bacillus thermophilus sp. nov., two thermophilic, Fe(III)-reducing bacteria isolated from compost.

    Science.gov (United States)

    Yang, Guiqin; Chen, Ming; Yu, Zhen; Lu, Qin; Zhou, Shungui

    2013-08-01

    Two novel thermophilic bacteria, designated SgZ-9(T) and SgZ-10(T), were isolated from compost. Cells of the two strains were catalase-positive, endospore-forming and Gram-staining-positive rods. Strain SgZ-9(T) was oxidase-positive and non-motile, and strain SgZ-10(T) was oxidase-negative and motile. The highest 16S rRNA gene sequence similarity for both strains SgZ-9(T) and SgZ-10(T) was observed with Bacillus fortis (97.5 % and 96.9 %, respectively). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain SgZ-9(T) formed a cluster with B. fortis R-6514(T) and Bacillus fordii R-7190(T), and SgZ-10(T) formed a cluster with Bacillus farraginis R-6540(T). The DNA-DNA pairing studies showed that SgZ-9(T) displayed 41.6 % and 30.7 % relatedness to the type strains of B. fortis and B. fordii, respectively. The 16S rRNA gene sequence similarity between strains SgZ-9(T) and SgZ-10(T) was 97.2 %, and the level of DNA-DNA relatedness between them was 39.2 %. The DNA G+C content of SgZ-9(T) and SgZ-10(T) was 45.3 and 47.9 mol%, respectively. Chemotaxonomic analysis revealed that both strains contained the menaquinone 7 (MK-7) as the predominant respiratory quinone. The major cellular fatty acids (>5 %) were iso-C15 : 0, anteiso-C15 : 0, anteiso-C17 : 0, iso-C16 : 0 and iso-C17 : 0 in SgZ-9(T) and iso-C15 : 0, anteiso-C15 : 0, iso-C17 : 0, anteiso-C17 : 0 and iso-C16 : 0 in SgZ-10(T). Based on the phenotypic characteristics, chemotaxonomic features, DNA-DNA hybridization with the nearest phylogenetic neighbours and phylogenetic analysis based on the 16S rRNA gene sequences, the two strains were determined to be two distinct novel species in the genus Bacillus, and the names proposed are Bacillus composti sp. nov. SgZ-9(T) ( = CCTCC AB2012109(T) = KACC 16872(T)) and Bacillus thermophilus sp. nov. SgZ-10(T) (CCTCC AB2012110(T) = KACC 16873(T)).

  2. Zhongshania antarctica gen. nov., sp. nov. and Zhongshania guokunii sp. nov., gammaproteobacteria respectively isolated from coastal attached (fast) ice and surface seawater of the Antarctic.

    Science.gov (United States)

    Li, Hui-Juan; Zhang, Xi-Ying; Chen, Chun-Xiao; Zhang, Yan-Jiao; Gao, Zhao-Ming; Yu, Yong; Chen, Xiu-Lan; Chen, Bo; Zhang, Yu-Zhong

    2011-09-01

    Two Gram-negative, motile, aerobic, catalase- and oxidase-positive, rod-shaped strains, designated ZS5-23(T) and ZS6-22(T), were respectively isolated from Antarctic coastal attached (fast) ice and surface seawater samples. Both strains could grow at 4-35 °C (optimum 30 °C) and in the absence of NaCl. Analyses of 16S rRNA gene sequences revealed that strains ZS5-23(T) and ZS6-22(T) were closely related to each other (99.0 % sequence similarity) and belonged to the class Gammaproteobacteria, with their closest relatives being Spongiibacter and Melitea species (93.1-94.3 % sequence similarity). The predominant cellular fatty acids in both strains were C₁₇:₁ω8c, C₁₇:₀ and summed feature 3 (C₁₆:₁ω7c and/or iso-C₁₅:₀ 2-OH). Genomic DNA G+C contents of strains ZS5-23(T) and ZS6-22(T) were 51.5 and 51.8 mol%, respectively. The DNA-DNA relatedness between strains ZS5-23(T) and ZS6-22(T) was 50.9 %. Strains ZS5-23(T) and ZS6-22(T) could be differentiated from each other and from Spongiibacter and Melitea species by differences in a number of phenotypic properties. Based on the data presented, strains ZS5-23(T) and ZS6-22(T) represent two novel species in a new genus in the class Gammaproteobacteria, for which the names Zhongshania antarctica gen. nov., sp. nov. (the type species) and Zhongshania guokunii sp. nov. are proposed. The type strain of Zhongshania antarctica is ZS5-23(T) ( = KACC 14066(T)  = CCTCC AB 209246(T)) and that of Zhongshania guokunii is ZS6-22(T) ( = KACC 14532(T)  = CCTCC AB 209247(T)).

  3. Prebióticos, probióticos e simbióticos na prevenção e tratamento das doenças alérgicas Prebiotics, probiotics and symbiotics on prevention and treatment of allergic diseases

    Directory of Open Access Journals (Sweden)

    Fabíola Suano Souza

    2010-03-01

    Full Text Available OBJETIVO: Avaliar o papel dos probióticos, prebióticos e simbióticos no equilíbrio do sistema imunológico do lactente, bem como seu efeito preventivo no desenvolvimento de doenças alérgicas na criança. FONTE DE DADOS: A partir do levantamento de todos os ensaios clínicos duplo-cegos e randômicos em seres humanos, publicados nos últimos cinco anos na base de dados Medline e que contivessem unitermos relacionados a prebióticos (oligossacarídeos, probióticos e simbióticos versus hipersensibilidade, analisou-se seu papel quanto à utilização em doenças alérgicas. SÍNTESE DE DADOS: Foram incluídos nesta revisão três trabalhos com prebióticos, os quais utilizaram a mistura GOS:FOS (9:1 em fórmulas infantis em lactentes nos primeiros meses de vida; 24 trabalhos com probióticos, sendo os micro-organismos utilizados na suplementação L. rhamnosus GG, B. lactis, L. casei, L. paracasei, L. reuteri, L. acidophilus, B. longum, B. breve e P. freudenreichii sp., e dois estudos com simbióticos. CONCLUSÕES: Apesar das evidências de benefícios da suplementação precoce de probióticos com algumas cepas específicas, prebióticos e simbióticos na prevenção da dermatite atópica, em crianças de alto risco para alergias, e do uso de probióticos no tratamento das dermatites atópicas moderadas e graves mediadas por IgE, há necessidade de ampliar os estudos quanto ao tempo de observação dos indivíduos suplementados, quanto à segurança e aos efeitos em longo prazoOBJECTIVE: To review current evidence about the effects of probiotics, prebiotics and symbiotics on the immune development as well as on the prevention of allergic diseases in children. DATA SOURCES: Randomized, double-blind clinical trials in humans published in the last five years, in the Medline database, containing the following keywords: prebiotics (oligosaccharides, probiotics, symbiotics and hypersensitivity. DATA SYNTHESIS: For this review three papers with

  4. Effect of applying bacterial inoculants containing different types of bacteria to corn silage on the performance of dairy cattle.

    Science.gov (United States)

    Arriola, K G; Kim, S C; Staples, C R; Adesogan, A T

    2011-08-01

    This study examined the effect of applying different bacterial inoculants to corn silage at the time of ensiling on the performance of lactating dairy cows. Corn plants were harvested at 35% dry matter (DM), chopped, and ensiled in 2.4-m-wide bags after application of (1) no inoculant (CON); (2) Biotal Plus II (B2) containing Pediococcus pentosaceus and Propionibacteria freudenreichii; (3) Buchneri 40788 (BUC) containing Lactobacillus buchneri; or (4) Buchneri 500 (B500) containing Pediococcus pentosaceus and L. buchneri. All inoculants were supplied by Lallemand Animal Nutrition (Milwaukee, WI). Each of the 4 silages was included in separate total mixed rations consisting of 44% corn silage, 50% concentrate, and 6% alfalfa hay (DM basis). Fifty-two lactating Holstein cows were stratified according to milk production and parity and randomly assigned at 22 d in milk to the 4 dietary treatments. Cows were fed for ad libitum consumption and milked twice daily for 49 d. Dietary treatment did not affect intakes (kg/d) of DM (20.0), crude protein (CP; 3.7), neutral detergent fiber (NDF; 5.7), or acid detergent fiber (ADF; 3.6), or digestibility (%) of DM (73.9) or CP (72.4). However, NDF digestibility was lower in cows fed B2 compared with those fed other diets (45.3 vs. 53.0%). Consequently, cows fed B2 had lower digestible NDF intake (kg/d) than those fed other diets (2.5 vs. 3.0 kg/d). Dietary treatment did not affect milk yield (32.3 kg/d), efficiency of milk production (1.61), concentrations of milk fat (3.18%) and protein (2.79%), or yields of milk fat (1.03 kg/d) and protein (1.26 kg/d). Inoculant application to corn silage did not affect milk yield or feed intake of cows.

  5. Role of vitamin B12 on methylmalonyl-CoA mutase activity

    Institute of Scientific and Technical Information of China (English)

    Tóshiko TAKAHASHI-I(N)IGUEZ; Enrique GARC(I)A-HERNANDEZ; Roberto ARREGU(I)N-ESPINOSA; Maria Elena FLORES

    2012-01-01

    Vitamin B12 is an organometallic compound with important metabolic derivatives that act as cofactors of certain enzymes,which have been grouped into three subfamilies depending on their cofactors.Among them,methylmalonyl-CoA mutase (MCM) has been extensively studied.This enzyme catalyzes the reversible isomerization of L-methylmalonyl-CoA to succinyI-CoA using adenosylcobalamin (AdoCbl) as a cofactor participating in the generation of radicals that allow isomerization of the substrate.The crystal structure of MCM determined in Propionibacterium freudenreichii var.shermanii has helped to elucidate the role of this cofactor AdoCbl in the reaction to specify the mechanism by which radicals are generated from the coenzyme and to clarify the interactions between the enzyme,coenzyme,and substrate.The existence of human methylmalonic acidemia (MMA) due to the presence of mutations in MCM shows the importance of its role in metabolism.The recent crystallization of the human MCM has shown that despite being similar to the bacterial protein,there are significant differences in the structural organization of the two proteins.Recent studies have identified the involvement of an accessory protein called MMAA,which interacts with MCM to prevent MCM's inactivation or acts as a chaperone to promote regeneration of inactivated enzyme.The interdisciplinary studies using this protein as a model in different organisms have helped to elucidate the mechanism of action of this isomerase,the impact of mutations at a functional level and their repercussion in the development and progression of MMA in humans.It is still necessary to study the mechanisms involved in more detail using new methods.

  6. Identification and quantification of Bifidobacterium species isolated from food with genus-specific 16S rRNA-targeted probes by colony hybridization and PCR.

    Science.gov (United States)

    Kaufmann, P; Pfefferkorn, A; Teuber, M; Meile, L

    1997-01-01

    A Bifidobacterium genus-specific target sequence in the V9 variable region of the 16S rRNA has been elaborated and was used to develop a hybridization probe. The specificity of this probe, named lm3 (5'-CGGGTGCTI*CCCACTTTCATG-3'), was used to identify all known type strains and distinguish them from other bacteria. All of the 30 type strains of Bifidobacterium which are available at the German culture collection Deutsche Sammlung von Mikroorganismen und Zellkulturen, 6 commercially available production strains, and 34 closely related relevant strains (as negative controls) were tested. All tested bifidobacteria showed distinct positive signals by colony hybridization, whereas all negative controls showed no distinct dots except Gardnerella vaginalis DSM4944 and Propionibacterium freudenreichii subsp. shermanii DSM4902, which gave slight signals. Furthermore, we established a method for isolation and identification of bifidobacteria from food by using a PCR assay without prior isolation of DNA but breaking the cells with proteinase K. By this method, all Bifidobacterium strains lead to a DNA product of the expected size. We also established a quick assay to quantitatively measure Bifidobacterium counts in food and feces by dilution plating and colony hybridization. We were able to demonstrate that 2.1 x 10(6) to 2.3 x 10(7) colonies/g of sour milk containing bifidobacteria hybridized with the specific nucleotide probe. With these two methods, genus-specific colony hybridization and genus-specific PCR, it is now possible to readily and accurately detect any bifidobacteria in food and fecal samples and to discriminate between them and members of other genera. PMID:9097423

  7. Probiotic Lactobacillus rhamnosus downregulates FCER1 and HRH4 expression in human mast cells

    Institute of Scientific and Technical Information of China (English)

    Anna Oksaharju; Matti Kankainen; Riina A Kekkonen; Ken A Lindstedt; Petri T Kovanen; Riitta Korpela; Minja Miettinen

    2011-01-01

    AIM: To investigate the effects of four probiotic bacteria and their combination on human mast cell gene expression using microarray analysis.METHODS: Human peripheral-blood-derived mast cells were stimulated with Lactobacillus rhamnosus (L.rhamnosus ) GG (LGG(R)), L. rhamnosus Lc705 (Lc705),Propionibacterium freudenreichii ssp. shermanii JS (PJS) and Bifidobacterium animalis ssp. lactis Bb12 (Bb12) and their combination for 3 or 24 h, and were subjected to global microarray analysis using an Affymetrix GeneChip(R) Human Genome U133 Plus 2.0 Array. The gene expression differences between unstimulated and bacteria-stimulated samples were further analyzed with GOrilla Gene Enrichment Analysis and Visualization Tool and MeV Multiexperiment Viewer-tool.RESULTS: LGG and Lc705 were observed to suppress genes that encoded allergy-related high-affinity IgE receptor subunits α and γ (FCER1A and FCER1G, respectively)and histamine H4 receptor. LGG, Lc705 and the combination of four probiotics had the strongest effect on the expression of genes involved in mast cell immune system regulation, and on several genes that encoded proteins with a pro-inflammatory impact, such as interleukin (IL)-8 and tumour necrosis factor alpha. Also genes that encoded proteins with anti-inflammatory functions, such as IL-10, were upregulated. CONCLUSION: Certain probiotic bacteria might diminish mast cell allergy-related activation by downregulation of the expression of high-affinity IgE and histamine receptor genes, and by inducing a pro-inflammatory response.

  8. Composition of Asarum heterotropoides var. mandshuricum radix oil from different extraction methods and activities against human body odor-producing bacteria

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    A.S.M. Tanbirul Haque

    2016-10-01

    Full Text Available In this study, oils from Asarum heterotropoides were extracted by traditional solvent extraction and supercritical CO2 (SC-CO2 extraction methods and their antioxidant activities along with antimicrobial and inhibitory activities against five human body odor-producing bacteria (Staphylococcus epidermidis, Propionibacterium freudenreichii, Micrococcus luteus, Corynebacterium jeikeium, and Corynebacterium xerosis were evaluated. The oil was found to contain 15 components, among which the most abundant component was methyl eugenol (37.6%, which was identified at every condition studied in different extraction methods. The oil extracted with n-hexane and ethanol mixture exhibited a strong antioxidant activity (92% ± 2% and the highest ABTS and 2,2-diphenyl-1-picrylhydrazyl scavenging activities (89% ± 0.2%. The highest amounts of total phenolic content and total flavonoid content were 23.1±0.4 mg/g and 4.9±0.1 mg/g, respectively, in the traditional method. In the SC-CO2 method performed at 200 bar/50°C using ethanol as an entrainer, the highest inhibition zone was recorded against all the aforementioned bacteria. In particular, strong antibacterial activity (38±2 mm was found against M. luteus. The minimum inhibitory concentration (MIC for the oil against bacteria ranged from 10.1±0.1 μg/mL to 46±2 μg/mL. The lowest MIC was found against M. luteus. Methyl eugenol was found to be one of the major compounds working against human body odor-producing bacteria.

  9. Quinone-dependent D-lactate dehydrogenase Dld (Cg1027 is essential for growth of Corynebacterium glutamicum on D-lactate

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    Oikawa Tadao

    2010-12-01

    Full Text Available Abstract Background Corynebacterium glutamicum is able to grow with lactate as sole or combined carbon and energy source. Quinone-dependent L-lactate dehydrogenase LldD is known to be essential for utilization of L-lactate by C. glutamicum. D-lactate also serves as sole carbon source for C. glutamicum ATCC 13032. Results Here, the gene cg1027 was shown to encode the quinone-dependent D-lactate dehydrogenase (Dld by enzymatic analysis of the protein purified from recombinant E. coli. The absorption spectrum of purified Dld indicated the presence of FAD as bound cofactor. Inactivation of dld resulted in the loss of the ability to grow with D-lactate, which could be restored by plasmid-borne expression of dld. Heterologous expression of dld from C. glutamicum ATCC 13032 in C. efficiens enabled this species to grow with D-lactate as sole carbon source. Homologs of dld of C. glutamicum ATCC 13032 are not encoded in the sequenced genomes of other corynebacteria and mycobacteria. However, the dld locus of C. glutamicum ATCC 13032 shares 2367 bp of 2372 bp identical nucleotides with the dld locus of Propionibacterium freudenreichii subsp. shermanii, a bacterium used in Swiss-type cheese making. Both loci are flanked by insertion sequences of the same family suggesting a possible event of horizontal gene transfer. Conclusions Cg1067 encodes quinone-dependent D-lactate dehydrogenase Dld of Corynebacterium glutamicum. Dld is essential for growth with D-lactate as sole carbon source. The genomic region of dld likely has been acquired by horizontal gene transfer.

  10. Streptomyces caeruleatus sp. nov., with dark blue diffusible pigment.

    Science.gov (United States)

    Zhu, Hong-hui; Guo, Jun; Yao, Qing; Yang, Song-zhen; Deng, Ming-rong; Li, Tai-hui

    2011-03-01

    An actinomycete, designated strain GIMN4.002(T), was isolated from a tomato rhizosphere soil sample in Guangzhou, China. The strain produces white aerial mycelium and dark blue diffusible pigment on Gause's synthetic agar, and microscopic observation revealed that it produces looped chains of spiny spores. Morphological and chemotaxonomic characteristics of the strain are typical of the genus Streptomyces. Melanin was produced and antibacterial activity was detected against Gram-positive micro-organisms, such as Bacillus subtilis, Micrococcus luteus and Staphylococcus aureus. The 16S rRNA gene sequence of strain GIMN4.002(T) had highest similarity (99.4  %) to Streptomyces lincolnensis B91; however, DNA-DNA relatedness between strain GIMN4.002(T) and S. lincolnensis NBRC 13054(T) was only 32.17  %. Further, the morphological, physiological and biochemical characteristics of strain GIMN4.002(T) are distinct from S. lincolnensis and other species of the genus Streptomyces with which this strain has high 16S rRNA gene sequence similarity (98-99  %). On the basis of the physiological and molecular properties observed, it is proposed that strain GIMN4.002(T) represents a novel species of the genus Streptomyces, for which the name Streptomyces caeruleatus sp. nov. is proposed, with GIMN4.002(T) (=CCTCC M 208213(T) =NRRL B-24802(T)) as the type strain.

  11. Actinotalea ferrariae sp. nov., isolated from an iron mine, and emended description of the genus Actinotalea.

    Science.gov (United States)

    Li, Yanzhi; Chen, Fang; Dong, Kun; Wang, Gejiao

    2013-09-01

    A Gram-stain-positive, aerobic, non-motile, rod-shaped bacterium, designated strain CF5-4(T), was isolated from iron mining powder. 16S rRNA gene sequence analysis grouped strain CF5-4(T) in a single cluster with Actinotalea fermentans DSM 3133(T) (97.6% similarity). The major fatty acids (>5%) of strain CF5-4(T) were anteiso-C(15:0), anteiso-C(15:1) A, C(16:0), iso-C(16:0), iso-C(15:0) and anteiso-C(17:0). The predominant respiratory quinone was MK-10(H₄) and the genomic DNA G+C content was 74.7 mol%. The major polar lipids were diphosphatidylglycerol and one unidentified phosphoglycolipid. The peptidoglycan type of strain CF5-4(T) was A4β, containing L-Orn-D-Ser-D-Asp. The cell-wall sugars were rhamnose, fucose, mannose and galactose. The results of DNA-DNA hybridization in combination with the comparison of phenotypic and phylogenetic characteristics among strain CF5-4(T) and related micro-organisms revealed that the isolate represents a novel species of the genus Actinotalea, for which the name Actinotalea ferrariae sp. nov. is proposed. The type strain is CF5-4(T) ( =KCTC 29134(T) =CCTCC AB2012198(T)).

  12. Arenimonas metalli sp. nov., isolated from an iron mine.

    Science.gov (United States)

    Chen, Fang; Shi, Zunji; Wang, Gejiao

    2012-08-01

    A Gram-staining-negative, aerobic, rod-shaped bacterium (CF5-1(T)) was isolated from Hongshan Iron Mine, Daye City, Hubei province, China. The major cellular fatty acids (>10%) were iso-C(16:0), iso-C(15:0), C(16:1)ω7c alcohol and iso-C(17:1)ω9c. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The major respiratory quinone was Q-8. The genomic DNA G+C content was 70.5 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain CF5-1(T) was most closely related to Arenimonas malthae (95.3% gene sequence similarity), Arenimonas oryziterrae (94.7%), Arenimonas donghaensis (94.6%) and Arenimonas composti (94.5%). A taxonomic study using a polyphasic approach showed that strain CF5-1(T) represents a novel species of the genus Arenimonas, for which the name Arenimonas metalli sp. nov. is proposed. The type strain is CF5-1(T) ( = CGMCC 1.10787(T) = KCTC 23460(T) = CCTCC AB 2010449(T)).

  13. Development of a freeze-dried fungal wettable powder preparation able to biodegrade chlorpyrifos on vegetables.

    Directory of Open Access Journals (Sweden)

    Jie Liu

    Full Text Available Continuous use of the pesticide chlorpyrifos has resulted in harmful contaminations in environment and species. Based on a chlorpyrifos-degrading fungus Cladosporium cladosporioides strain Hu-01 (collection number: CCTCC M 20711, a fungal wettable powder preparation was developed aiming to efficiently remove chlorpyrifos residues from vegetables. The formula was determined to be 11.0% of carboxymethyl cellulose-Na, 9.0% of polyethylene glycol 6000, 5.0% of primary alcohol ethoxylate, 2.5% of glycine, 5.0% of fucose, 27.5% of kaolin and 40% of freeze dried fungi by response surface methodology (RSM. The results of quality inspection indicated that the fungal preparation could reach manufacturing standards. Finally, the degradation of chlorpyrifos by this fungal preparation was determined on pre-harvest cabbage. Compared to the controls without fungal preparation, the degradation of chlorpyrifos on cabbages, which was sprayed with the fungal preparation, was up to 91% after 7 d. These results suggested this freeze-dried fungal wettable powder may possess potential for biodegradation of chlorpyrifos residues on vegetables and provide a potential strategy for food and environment safety against pesticide residues.

  14. L-Tryptophan Production in Escherichia coli Improved by Weakening the Pta-AckA Pathway.

    Science.gov (United States)

    Liu, Lina; Duan, Xuguo; Wu, Jing

    2016-01-01

    Acetate accumulation during the fermentation process of Escherichia coli FB-04, an L-tryptophan production strain, is detrimental to L-tryptophan production. In an initial attempt to reduce acetate formation, the phosphate acetyltransferase gene (pta) from E. coli FB-04 was deleted, forming strain FB-04(Δpta). Unfortunately, FB-04(Δpta) exhibited a growth defect. Therefore, pta was replaced with a pta variant (pta1) from E. coli CCTCC M 2016009, forming strain FB-04(pta1). Pta1 exhibits lower catalytic capacity and substrate affinity than Pta because of a single amino acid substitution (Pro69Leu). FB-04(pta1) lacked the growth defect of FB-04(Δpta) and showed improved fermentation performance. Strain FB-04(pta1) showed a 91% increase in L-tryptophan yield in flask fermentation experiments, while acetate production decreased by 35%, compared with its parent FB-04. Throughout the fed-batch fermentation process, acetate accumulation by FB-04(pta1) was slower than that by FB-04. The final L-tryptophan titer of FB-04(pta1) reached 44.0 g/L, representing a 15% increase over that of FB-04. Metabolomics analysis showed that the pta1 genomic substitution slightly decreased carbon flux through glycolysis and significantly increased carbon fluxes through the pentose phosphate and common aromatic pathways. These results indicate that this strategy enhances L-tryptophan production and decreases acetate accumulation during the L-tryptophan fermentation process.

  15. Resting-state cerebellar-cerebral networks are differently affected in first-episode, drug-naive schizophrenia patients and unaffected siblings.

    Science.gov (United States)

    Guo, Wenbin; Liu, Feng; Chen, Jindong; Wu, Renrong; Zhang, Zhikun; Yu, Miaoyu; Xiao, Changqing; Zhao, Jingping

    2015-11-26

    Dysconnectivity hypothesis posits that schizophrenia is a disorder with dysconnectivity of the cortico-cerebellar-thalamic-cortical circuit (CCTCC). However, it remains unclear to the changes of the cerebral connectivity with the cerebellum in schizophrenia patients and unaffected siblings. Forty-nine patients with first-episode, drug-naive schizophrenia patients, 46 unaffected siblings of schizophrenia patients and 46 healthy controls participated in the study. Seed-based resting-state functional connectivity approach was employed to analyze the data. Compared with the controls, the patients and the siblings share increased default-mode network (DMN) seed - right Crus II connectivity. The patients have decreased right dorsal attention network (DAN) seed - bilateral cerebellum 4,5 connectivity relative to the controls. By contrast, the siblings exhibit increased FC between the right DAN seed and the right cerebellum 6 and right cerebellum 4,5 compared to the controls. No other abnormal connectivities (executive control network and salience network) are observed in the patients/siblings relative to the controls. There are no correlations between abnormal cerebellar-cerebral connectivities and clinical variables. Cerebellar-cerebral connectivity of brain networks within the cerebellum are differently affected in first-episode, drug-naive schizophrenia patients and unaffected siblings. Increased DMN connectivity with the cerebellum may serve as potential endophenotype for schizophrenia.

  16. Mutational analysis of the hyc-operon determining the relationship between hydrogenase-3 and NADH pathway in Enterobacter aerogenes.

    Science.gov (United States)

    Pi, Jian; Jawed, Muhammad; Wang, Jun; Xu, Li; Yan, Yunjun

    2016-01-01

    In this study, the hydrogenase-3 gene cluster (hycDEFGH) was isolated and identified from Enterobacter aerogenes CCTCC AB91102. All gene products were highly homologous to the reported bacterial hydrogenase-3 (Hyd-3) proteins. The genes hycE, hycF, hycG encoding the subunits of hydrogenase-3 were targeted for genetic knockout to inhibit the FHL hydrogen production pathway via the Red recombination system, generating three mutant strains AB91102-E (ΔhycE), AB91102-F (ΔhycF) and AB91102-G (ΔhycG). Deletion of the three genes affected the integrity of hydrogenase-3. The hydrogen production experiments with the mutant strains showed that no hydrogen was detected compared with the wild type (0.886 mol/mol glucose), demonstrating that knocking out any of the three genes could inhibit NADH hydrogen production pathway. Meanwhile, the metabolites of the mutant strains were significantly changed in comparison with the wild type, indicating corresponding changes in metabolic flux by mutation. Additionally, the activity of NADH-mediated hydrogenase was found to be nil in the mutant strains. The chemostat experiments showed that the NADH/NAD(+) ratio of the mutant strains increased nearly 1.4-fold compared with the wild type. The NADH-mediated hydrogenase activity and NADH/NAD(+) ratio analysis both suggested that NADH pathway required the involvement of the electron transport chain of hydrogenase-3.

  17. Preparative separation of echinocandin B from Aspergillus nidulans broth using macroporous resin adsorption chromatography.

    Science.gov (United States)

    Zou, Shu-Ping; Liu, Miao; Wang, Qiu-Liang; Xiong, Yan; Niu, Kun; Zheng, Yu-Guo; Shen, Yin-Chu

    2015-01-26

    Echinocandin B (ECB), an echinocandin type of lipopeptide antibiotic produced by Aspergillus nidulans, is a precursor for the synthesis of novel anti-fungal drug - anidulafungin. In this work, a separation strategy involving one-step macroporous resin adsorption chromatography was established for ECB purification from Aspergillus nidulans CCTCC M 2010275 fermentation broth. Among nine macroporous resin adsorbents tested, the non-polar resin HP-20 had the best adsorption and desorption performance. The static equilibrium adsorption data fitted well with the Langmuir equation, and the adsorption kinetic followed the pseudo-second order model. The separation parameters of ECB from broth were optimised by dynamic adsorption/desorption experiments with the column packed with HP-20 resin. Under optimal conditions, the purity increased by 3.8-fold from 23.2% in broth to 88.5% in eluent with 87.1% recovery yield by a one-step treatment. Our study provided a one-step and effective method for large-scale production of ECB, and offered references for separating other echinocandins from broth. Copyright © 2014 Elsevier B.V. All rights reserved.

  18. Bacillus thermophilum sp. nov., isolated from a microbial fuel cell.

    Science.gov (United States)

    Tang, Jia; Yang, Guiqin; Wen, Junlin; Yu, Zhen; Zhou, Shungui; Liu, Zhi

    2014-09-01

    A novel thermophilic, Gram-staining positive bacterium, designated DX-2(T), was isolated from the anode biofilm of a microbial fuel cell. Cells of the strain were oxidase positive, catalase positive, facultative anaerobic, motile rods. The isolate grew at 30-60 °C (optimum 50 °C) and pH 5-9 (optimum pH 8-8.5). The pairwise 16S rRNA gene sequence similarities showed that strain DX-2(T) was most closely related to Bacillus fumarioli LMG 17489(T) (96.2 %), B. firmus JCM 2512(T) (96.0 %) and B. foraminis DSM 19613(T) (95.7 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain DX-2(T) formed a cluster with B. smithii (95.5 %) and B. infernus (94.9 %). The genomic G+C content of DX-2(T) was 43.7 mol%. The predominant respiratory quinone was MK-7. The polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and unknown phospholipids. The major cellular fatty acid was iso-C16:0. Based on its phenotypic characteristics, chemotaxonomic features, and results of phylogenetic analysis, the strain was identified to represent a distinct novel species in the genus Bacillus, and the name proposed is B. thermophilum sp. nov. The type strain is DX-2(T) (=CCTCC AB2012194(T) = KCTC 33128(T)).

  19. Two-stage pH Control Mode in Batch Fermentation of a Novel Bioflocculant from Corynebacterium Glutamicum

    Institute of Scientific and Technical Information of China (English)

    HE Ning; WU Xiao-jie; DENG Xu; LU Ying-hua; LI Qing-biao

    2004-01-01

    The effect of pH of the fermentation medium on cell growth and the production of a novel bioflocculant (named REA-11 ) by Corynebacterium glutamicum CCTCC M201005 were investigated. The maximum biomass (2.23 g/L) and flocculating activity (142.2 U/mL) were simultaneously obtained at the 14th hour when the pH value of the culture medium was maintained at 7.0 during the whole fermentation process. The production of REA-11 kept on a trend of increase till the later phase of fermentation process, which resulted in the ultimate flocculating activity of the culture broth to enhance to nearly 100 U/mL at pH 6.0. A twostage pH control mode was adopted in REA-11 production in which the pH value of the culture medium was controlled at 7.0 during the first 14 h, then decreased to 6.0 that was maintained until the end of the fermentation process. With the two-stage pH control mode, the maximum flocculating activity reached 178.8 U/mL which was 30% higher than that obtained under the condition of pH 7.0 and the biomass enhanced about 15%. Compared with the fermentation process without pH control, REA-11 production and cell growth via the two-stage pH control mode increased 80% and 25%, respectively.

  20. Enhancement of lipase r27RCL production in Pichia pastoris by regulating gene dosage and co-expression with chaperone protein disulfide isomerase.

    Science.gov (United States)

    Sha, Chong; Yu, Xiao-Wei; Lin, Nai-Xin; Zhang, Meng; Xu, Yan

    2013-12-10

    Pichia pastoris has been successfully used in the production of many secreted and intracellular recombinant proteins, but there is still a large room of improvement for this expression system. Two factors drastically influence the lipase r27RCL production from Rhizopus chinensis CCTCC M201021, which are gene dosage and protein folding in the endoplasmic reticulum (ER). Regarding the effect of gene dosage, the enzyme activity for recombinant strain with three copies lipase gene was 1.95-fold higher than that for recombinant strain with only one copy lipase gene. In addition, the lipase production was further improved by co-expression with chaperone PDI involved in the disulfide bond formation in the ER. Overall, the maximum enzyme activity reached 355U/mL by the recombinant strain with one copy chaperone gene PDI plus five copies lipase gene proRCL in shaking flasks, which was 2.74-fold higher than that for the control strain with only one copy lipase gene. Overall, co-expression with PDI vastly increased the capacity for processing proteins of ER in P. pastoris.

  1. Efficient secretion of lipase r27RCL in Pichia pastoris by enhancing the disulfide bond formation pathway in the endoplasmic reticulum.

    Science.gov (United States)

    Sha, Chong; Yu, Xiao-Wei; Zhang, Meng; Xu, Yan

    2013-11-01

    The lipase r27RCL from Rhizopus chinensis CCTCC M201021 was heterologously expressed in Pichia pastoris GS115 by simultaneous co-expression with two secretion factors ERO1p and PDI involved in the endoplasmic reticulum (ER). Compared to the expression of the lipase alone (12,500 U/ml), co-expression with these two proteins resulted in the production of larger total quantities of enzymes. The largest increase was seen when the combined ERO1p/PDI system was co-expressed, resulting in approximately 30 % higher enzyme yields (16,200 U/ml) than in the absence of co-expressed secretion factors. The extracellular protein concentration of the recombinant strain Co XY RCL-5 reached 9.39 g/l in the 7-l fermentor. Simultaneously, the fermentation time was also shortened by about 8 h compared to that of the control. The substrate-specific consumption rate (Qs) and the product-specific production rate (Qp) were both investigated in this research. In conclusion, the space-time yield was improved by co-expression with ERO1p and PDI. This is a potential strategy for high level expression of other heterologous proteins in P. pastoris.

  2. Metabolic network modeling and metabolic flux analysis of production of novel bioflocculant REA-11%新型生物絮凝剂REA-11的代谢模型建立与代谢网络分析

    Institute of Scientific and Technical Information of China (English)

    何宁; 李寅; 陈坚

    2005-01-01

    利用代谢通量分析方法,对谷氨酸棒杆菌Corynebacterium glutamicum CCTCC M201005分批发酵不同阶段和不同溶氧浓度下的代谢网络模型进行了计算,考察了代谢节点对絮凝剂REA-11合成的影响,并对C. glutamicum生长代谢过程中能量和还原力的产生及消耗问题进行分析.结果发现,磷酸戊糖途径(PP)通量在整个发酵过程中始终维持在较高的水平;REA-11合成通量随溶氧浓度的增加而降低,菌体合成通量则随溶氧水平的增加而增加;ATP通量的增加可以促进菌体生长,而与REA-11的合成呈负相关.

  3. Rhizobium yantingense sp. nov., a mineral-weathering bacterium.

    Science.gov (United States)

    Chen, Wei; Sheng, Xia-Fang; He, Lin-Yan; Huang, Zhi

    2015-02-01

    A Gram-stain-negative, rod-shaped bacterial strain, H66(T), was isolated from the surfaces of weathered rock (purple siltstone) found in Yanting, Sichuan Province, PR China. Cells of strain H66(T) were motile with peritrichous flagella. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain H66(T) belongs to the genus Rhizobium. It is closely related to Rhizobium huautlense SO2(T) (98.1 %), Rhizobium alkalisoli CCBAU 01393(T) (98.0 %) and Rhizobium cellulosilyticum ALA10B2(T) (98.0 %). Analysis of the housekeeping genes, recA, glnII and atpD, showed low levels of sequence similarity (Rhizobium. The predominant components of the cellular fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C16 : 0. The G+C content of strain H66(T) was 60.3 mol%. Strain H66(T) is suggested to be a novel species of the genus Rhizobium based on the low levels of DNA-DNA relatedness (ranging from 14.3 % to 40.0 %) with type strains of species of the genus Rhizobium and on its unique phenotypic characteristics. The namehttp://dx.doi.org/10.1601/nm.1279Rhizobium yantingense sp. nov. is proposed for this novel species. The type strain is H66(T) ( = CCTCC AB 2014007(T) = LMG 28229(T)).

  4. Rhizobium phenanthrenilyticum sp. nov., a novel phenanthrene-degrading bacterium isolated from a petroleum residue treatment system.

    Science.gov (United States)

    Wen, Ya; Zhang, Juan; Yan, Qiuxiang; Li, Shunpeng; Hong, Qing

    2011-01-01

    Strain F11(T), a phenanthrene-degrading bacterium, was isolated from a petroleum residue treatment system, and classified under the genus Rhizobium based on the similarity analysis of its 16S rRNA and recA gene sequences. Strain F11(T) falls into the same phylogenetic clade with Rhizobium oryzae Alt 505(T) (96.8% 16S rRNA gene sequence similarity) and Rhizobium pseudoryzae J34A-127(T) (96.2%). Major cellular fatty acids of strain F11(T) are C(16:0) (6.24%) and summed feature 8 (C(18:1ω7c) and/or C(18:1ω6c), 76.59%), which are also the major fatty acids of R. oryzae Alt 505(T) and R. pseudoryzae J34A-127(T). The DNA G+C content of strain F11(T) was 59.3±0.4 mol%. Based on the phylogenetic analysis as well as biochemical and physiological characteristics, strain F11(T) could be separated from all recognized Rhizobium species. Strain F11(T) (=DSM 21882(T) =CCTCC AB 209029(T)) was considered to be representative of a novel species of Rhizobium, for which the name Rhizobium phenanthrenilyticum sp. nov. is proposed.

  5. Purification and characterization of a novel carbonyl reductase with high stereo-selectivity

    Institute of Scientific and Technical Information of China (English)

    YANG Ming; XU Yan; MU Xiaoqing; XIAO Rong

    2007-01-01

    A novel NADPH-dependent carbonyl reductase was separated from Candida parapsilosis CCTCC 203011.The enzyme gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE),which was purified through ammonium sulfate,Diethylamino Ethanol (DEAE) sepharose Fast flow (FF),phenyl-sepharose FF and blue sepharose FF chromatography from cell-free extract.The molecular mass of the enzyme was about 30 kDa.The optimum pH and temperature for reduction were 4.5℃ and 35℃,respectively.The Cu2+ had strong restrictive effect on enzyme activity.In addition,the carbonyl reductase was an enzyme with high substrate specificity and stereo-selectivity,and showed high asymmetric reduction activity towards α-hydroxyacetophenone and ethyl 4-chloro acetoacetate.For the asymmetric reduction of α-hydroxyacetophenone and ethyl 4-chloro acetoacetate,(S)-1-phenyl-1,2-ethanediol and (R)-ethyl 4-chloro-3-hydroxybutanoate were produced by the purified enzyme,with the 100% and 94.3% e.e.value,respectively.Therefore,the enzyme could be one of the effective biocatalysts for asymmetric synthesis of chiral alcohols.The amino acid sequences of one peptide from the purified enzyme were analyzed by LC-MASS-MASS,and the carbonyl reductase showed some identity to the hypothetical protein CaO 19.10414 reported.

  6. Synthetic activity enhancement of membrane-bound lipase from Rhizopus chinensis by pretreatment with isooctane.

    Science.gov (United States)

    Wang, Dong; Xu, Yan; Teng, Yun

    2007-05-01

    The cell-bound lipase from Rhizopus chinensis CCTCC M201021 with high catalysis ability for ester synthesis was located as a membrane-bound lipase by the treatments of Yatalase firstly. In order to improve its synthetic activity in non-aqueous phase, the pretreatments of this enzyme with various organic solvents were investigated. The pretreatment with isooctane improved evidently the lipase synthetic activity, resulting in about 139% in relative synthetic activity and 115% in activity recovery. The morphological changes of mycelia caused by organic solvent pretreatments could influence the exposure of the membrane-bound enzyme from mycelia and the exhibition of the lipase activity. The pretreatment conditions with isooctane and acetone were further investigated, and the optimum effect was obtained by the isooctane pretreatment at 4 degrees C for 1 h, resulting in 156% in relative synthetic activity and 126% in activity recovery. When the pretreated lipases were employed as catalysts for the esterification production of ethyl hexanoate in heptane, higher initial reaction rate and higher final molar conversion were obtained using the lipase pretreated with isooctane, compared with the untreated lyophilized one. This result suggested that the pretreatment of the membrane-bound lipase with isooctane could be an effective method to substitute the lyophilization for preparing biocatalysts used in non-aqueous phase reactions.

  7. Acinetobacter plantarum sp. nov. isolated from wheat seedlings plant.

    Science.gov (United States)

    Du, Juan; Singh, Hina; Yu, Hongshan; Jin, Feng-Xie; Yi, Tae-Hoo

    2016-07-01

    Strain THG-SQM11(T), a Gram-negative, aerobic, non-motile, coccus-shaped bacterium, was isolated from wheat seedlings plant in P. R. China. Strain THG-SQM11(T) was closely related to members of the genus Acinetobacter and showed the highest 16S rRNA sequence similarities with Acinetobacter junii (97.9 %) and Acinetobacter kookii (96.1 %). DNA-DNA hybridization showed 41.3 ± 2.4 % DNA reassociation with A. junii KCTC 12416(T). Chemotaxonomic data revealed that strain THG-SQM11(T) possesses ubiquinone-9 as the predominant respiratory quinone, C18:1 ω9c, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), and C16:0 as the major fatty acids. The major polar lipids were found to be diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylcholine. The DNA G+C content was 41.7 mol %. These data, together with phenotypic characterization, suggest that the isolate represents a novel species, for which the name Acinetobacter plantarum sp. nov. is proposed, with THG-SQM11(T) as the type strain (=CCTCC AB 2015123(T) =KCTC 42611(T)).

  8. Lysinibacillus tabacifolii sp. nov., a novel endophytic bacterium isolated from Nicotiana tabacum leaves.

    Science.gov (United States)

    Duan, Yan-Qing; He, Song-Tao; Li, Qing-Qing; Wang, Ming-Feng; Wang, Wen-Yuan; Zhe, Wei; Cao, Yong-Hong; Mo, Ming-He; Zhai, Yu-Long; Li, Wen-Jun

    2013-06-01

    A Gram-positive, catalase- and oxidase-positive, strictly aerobic, endospore-forming rod bacterium, designated K3514(T), was isolated from the leaves of Nicotiana tabacum. The strain was able to grow at temperatures of 8-40°C, pH 5.0-10.0 and NaCl concentrations of 0-7%. The predominant quinones (>30%) of this strain were MK-7(H2) and MK-7. Phylogenetic analysis of 16S rRNA gene sequence showed that strain K3514(T) was affiliated to the genus Lysinibacillus, with its closest relatives being Lysinibacillus mangiferihumi (98.3% sequence similarity), Lysinibacillus sphaericus (97.9% sequence similarity), Lysinibacillus fusiformis (97.4% sequence similarity), and Lysinibacillus xylanilyticus (97.3% sequence similarity). However, low levels of DNA-DNA relatedness values suggested that the isolate was distinct from the other closest Lysinibacillus species. Additionally, based on analysis of morphological, physiological, and biochemical characteristics, the isolate could be differentiated from the closest known relatives. Therefore, based on polyphasic taxonomic data, the novel isolate likely represents a novel species, for which the name Lysinibacillus tabacifolii sp. nov. and the type strain K3514(T) (=KCTC 33042(T) =CCTCC AB 2012050(T)) are proposed.

  9. Isolation and Characterization of a New Subspecies of Aeromonas salmonicida (Aeromonas salmonicida subsp.Flounderacida subsp.Nov.) from Stone Flounder(Kareius bicoloratus L.)

    Institute of Scientific and Technical Information of China (English)

    Zhang Xiaojun(张晓君); Zhan Wenbin; Chen Cuizhen; Fang Hai

    2004-01-01

    Biological properties were studied to appropriate pathogenic bacteria which were isolated from di-seased (or dead) stone flounder (Kareius bicoloratus L.) which expressed bacterial septicaemia, including morphological characteristics, colony characteristics, physiological and biochemical characteristics and serum homology of isolates, the results showed that the isolates belonged to a new subspecies of A.salmonicida. In addition, the representative strains have been re-checked and detected the mol% G+C ratio of the DNA by China Center for Type Culture Collection (CCTCC), the examined strains were also regarded as a new subspecies of A.salmonicida, and designated as Aeromonas salmonicida subsp. flounderacida subsp.nov.by its isolated fish (Kareius bicoloratus). Molecular identification of analysis of the nucleotide sequence of the 16S rRNA gene were applied, the results showed high similarity (99%) with the 16S rRNA gene of Aeromonas salmonicida from GenBank database. Cluster analysis of phylogenetic tree revealed that the representative strain formed separately bootstrap-supported cluster.

  10. 产细菌纤维素菌株Gluconacetobacter intermedius的分离与发酵条件优化%Construction and Optimization of Two-Dimensional Electrophoresis for Proteome Analysis of Rhizopus chinensis

    Institute of Scientific and Technical Information of China (English)

    郑礼月; 王栋; 徐岩

    2015-01-01

    华根霉(Rhizopus chinensis CCTCC M201021)是从我国传统微生物载体——大曲中分离筛选得到的一株丝状真菌,具有重要的工业应用前景.为建立一种适于华根霉胞内蛋白质的双向电泳体系以进行蛋白质组学研究,作者对华根霉胞内蛋白质的提取方法及双向电泳流程相关参数进行了考察和优化.确定采用液氮研磨与高速珠磨相结合的方法对华根霉进行细胞破碎,用TCA/丙酮对提取的蛋白质进行纯化,采用被动水化的上样方式上样,24 cm、pH 4~7的线性IPG胶条,上样量为1 200 μg蛋白质,考马斯亮蓝G-250胶体染色法,最终得到了背景清晰、分辨率高的双向电泳图谱.

  11. Paenibacillus guangzhouensis sp. nov., an Fe(III)- and humus-reducing bacterium from a forest soil.

    Science.gov (United States)

    Li, Jibing; Lu, Qin; Liu, Ting; Zhou, Shungui; Yang, Guiqin; Zhao, Yong

    2014-11-01

    A Gram-reaction-variable, rod-shaped, motile, facultatively aerobic and endospore-forming bacterium, designated strain GSS02(T), was isolated from a forest soil. Strain GSS02(T) was capable of reducing humic substances and Fe(III) oxides. Strain GSS02(T) grew optimally at 35 °C, at pH 78 and in the presence of 1% NaCl. The predominant menaquinone was MK-7. The major cellular fatty acids were anteiso-C(15:0) and iso-C(16:0) and the polar lipid profile contained mainly phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol, with moderate amounts of two unknown aminophospholipids and a minor amount of one unknown lipid. The DNA G+C content was 53.4 mol%. Comparative 16S rRNA gene sequence analysis showed that strain GSS02(T) was related most closely to Paenibacillus terrigena JCM 21741(T) (98.1% similarity). Mean DNA-DNA relatedness between strain GSS02(T) and P. terrigena JCM 21741(T) was 58.8 ± 0.5%. The phylogenetic, chemotaxonomic and phenotypic results clearly demonstrated that strain GSS02(T) belongs to the genus Paenibacillus and represents a novel species, for which the name Paenibacillus guangzhouensis sp. nov. is proposed. The type strain is GSS02(T) ( =KCTC 33171(T) =CCTCC AB 2013236(T)).

  12. 华根霉合成活性脂肪酶液态发酵菌体形态调控的研究%Studies on morphologic regulation of Rhizopus chinensis in submerged fermentation of lipase with synthetic activity

    Institute of Scientific and Technical Information of China (English)

    周艺博; 王栋; 徐岩; 朱增亮; 李鸣

    2011-01-01

    Membrane-bound lipase with high activity produced by Rhizopus chinensis (CCTCC 2010021) could catalyze the synthesis of ester in non-aqueous phase and had potential applications in the production of flavor and fragrance compounds, biodiesel, and so on. Similar to other filamentous fungi, Rhizopus chinensis would form different morphologies in submerged fermentation, which influenced the production of membrane-bound lipase with synthetic activity obviously. Based on mycelial morphology and synthetic activity of Rhizopus chinensis lipase, the influences of fermentation conditions on the mycelial morphologies and lipase synthetic activity were investigated. The results showed that the medium volume, rotation speed and baffled flask remarkably influenced the Rhizopus chinensis morphology and the higher lipase synthetic activity could be obtained by clump mycelia. The optimized fermentation conditions for clump mycelia were as follows: medium volume 20 mL/250 flask, 200 r/min, initial pH 5. 5, inoculum 5 × 107 spores/mL. Under the optimized fermentation conditions, clump mycelia were formed and synthetic activity of Rhizopus chinensis lipase reached up to 1080 U/g.%华根霉(CCTCC M201021)膜结合脂肪酶在非水相中具有突出的催化酯合成的能力,在生物香料、生物柴油生产等工业应用中具有良好的前景.与许多丝状真茵形态影响其初级及次级代谢产物的生产相类似,华根霉在液态发酵中也会形成不同的茵体形态,显著影响合成活性膜结合脂肪酶的发酵水平.本研究以华根霉菌体形态及脂肪酶合成活性为指标,考察了不同发酵条件对华根霉液态培养茵体形态的影响以及与脂肪酶合成活性的关系.研究结果表明,发酵摇瓶装液量、摇床转速及摇瓶中挡板的设置对华根霉茵体形态影响显著,块状茵体可以得到较高的脂肪酶活性.确定在装液量20 mL/250 mL发酵摇瓶,摇床转速200 r/min,发酵液初始pH 5.5,接种量5

  13. Themoanaerobacterium calidifontis sp. nov., a novel anaerobic, thermophilic, ethanol-producing bacterium from hot springs in China.

    Science.gov (United States)

    Shang, Shu-mei; Qian, Long; Zhang, Xu; Li, Kun-zhi; Chagan, Irbis

    2013-06-01

    A novel thermophilic Gram staining positive strain Rx1 was isolated from hot springs in Baoshan of Yunnan Province, China. The strain was characterized as a hemicellulose-decomposing obligate anaerobe bacterium that is rod-shaped (diameter: 0.5-0.7 μm; length: 2.0-6.7 μm), spore-forming, and motile. Its growth temperature range is 38-68 °C (optimum 50-55 °C) and pH range is 4.5-8.0 (optimum 7.0). The maximum tolerance concentration of NaCl was 3 %. Rx1 converted thiosulfate to elemental sulfur and reduced sulfite to hydrogen sulfide. The bacterium grew by utilizing xylan and starch, as well as a wide range of monosaccharide and polysaccharides, including glucose and xylose. The main products of fermentation were ethanol, lactate, acetate, CO2, and H2. The maximum xylanase activity in the culture supernatant after 30 h of incubation at 55 °C was 16.2 U/ml. Rx1 DNA G + C content was 36 mol %. 16S rRNA gene sequence analysis indicated that strain Rx1 belonged to the genus Thermoanaerobacterium of the family 'Thermoanaerobacteriaceae' (Firmicutes), with Thermoanaerobacterium aciditolerans 761-119 (99.2 % 16S rRNA gene sequence similarity) being its closest relative. DNA-DNA hybridization between Rx1 and T. aciditolerans 761-119 showed 36 % relatedness. Based on its physiological and biochemical tests and DNA-DNA hybridization analyses, the isolate is considered to represent a novel species in the genus Thermoanaerobacterium, for which the name Thermoanaerobacterium calidifontis sp. nov. is proposed, with the type strain is Rx1 (=JCM 18270 = CCTCC M 2011109).

  14. Identification of "Bacillus cellulasensis" strain NIO-1130(T) as a member of Bacillus altitudinis and emendation of the latter.

    Science.gov (United States)

    Liu, Yang; Lai, Qiliang; Shao, Zongze

    2016-10-01

    In the study by Mawlankar et al. in Arch Microbiol 198:83-89 (2016), the phylogenetic position of strain "Bacillus cellulasensis" NIO-1130(T) based on 16S rRNA and gyrB genes was inconsistent. Therefore, the aim of this study is to re-determine its taxonomic status using diverse genotypic approaches including single gene analysis, multilocus sequence analysis, and genomic analyses. The reconstructed phylogenetic trees based on 16S rRNA gene and six concatenated genes showed that "B. cellulasensis" NIO-1130(T) (=NCIM 5461(T) = CCTCC AB 2011126(T)) revealed the closest genetic relationship with type strain Bacillus altitudinis 41KF2b(T), with 98.6-100 % similarities of 16S rRNA gene, gyrB, pycA, pyrE, mutL, aroE, trpB, and six concatenated housekeeping genes. The high similarities for gene(s) sequences between "B. cellulasensis" NIO-1130(T) and B. altitudinis 41KF2b(T) indicated that they should be conspecific. The DNA G+C content for strain NIO-1130(T) was determined to be 41.3 mol% and identical to that of B. altitudinis 41KF2b(T). Moreover, 88.4 % of digital DNA-DNA hybridization and 98.7 % of average nucleotide identity values between two strains were much higher than the standard criteria for delineation of bacterial species, suggesting that they belonged to the same species. Therefore, the data from the combined genotypic analyses suggest that "Bacillus cellulasensis" should be classified as a member of Bacillus altitudinis.

  15. Bacillus mesophilus sp. nov., an alginate-degrading bacterium isolated from a soil sample collected from an abandoned marine solar saltern.

    Science.gov (United States)

    Zhou, Yan-Xia; Liu, Guo-Hong; Liu, Bo; Chen, Guan-Jun; Du, Zong-Jun

    2016-07-01

    A novel Gram-stain positive, endospore-forming bacterium, designated SA4(T), was isolated from a soil sample collected from an abandoned marine solar saltern at Wendeng, Shandong Province, PR China. Cells were observed to be rod shaped, alginase positive, catalase positive and motile. The strain was found to grow at temperatures ranging from 15 to 40 °C (optimum 35 °C), and pH 5.0-11.0 (optimum pH 8.0) with 0-7.0 % (w/v) NaCl concentration (optimum NaCl 3.0 %). Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain SA4(T) belongs to the genus Bacillus and exhibits 16S rRNA gene sequence similarities of 96.6, 96.5, 96.3 and 96.2 % with Bacillus horikoshii DSM 8719(T), Bacillus acidicola 105-2(T), Bacillus shackletonii LMG 18435(T) and Bacillus pocheonensis Gsoil 420(T), respectively. The menaquinone was identified as MK-7 and the major polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The major fatty acids detected were anteiso-C15:0 (22.3 %), iso-C15:0 (22.6 %), iso-C16:0 (14.8 %) and iso-C14:0 (14.7 %). The DNA G+C content was determined to be 42.4 mol %. Phenotypic, chemotaxonomic and genotypic properties clearly indicated that isolate SA4(T) represents a novel species within the genus Bacillus, for which the name Bacillus mesophius sp. nov. is proposed. The type strain is SA4(T) (=DSM 101000(T)=CCTCC AB 2015209(T)).

  16. Nonomuraea flavida sp. nov., a novel species of soil actinomycete isolated from Aconitum napellus rhizosphere.

    Science.gov (United States)

    Chen, Shaofeng; Shi, Jindi; Li, Dan; Wu, Yingying; Huang, Yaojian

    2015-11-01

    A novel actinomycete strain, YN-5-1T, isolated from the rhizosphere soil of a medicinal plant, Aconitum napellus, was characterized by a polyphasic approach to determine its taxonomic position. The strain showed highest 16S rRNA gene sequence similarities of 97.3, 97.2 and 97.1 % to Nonomuraea turkmeniaca DSM 43926T, Nonomuraea ferruginea DSM 43553T and Nonomuraea candida DSM 45086T, respectively. A wide range of genotypic and phenotypic characteristics, as well as levels of DNA-DNA relatedness between strain YN-5-1T and N. turkmeniaca DSM 43926T (57.46 %), N. ferruginea DSM 43553T (53.50 %) and N. candida DSM 45086T (48.80 %), distinguished the novel isolate from its closest phylogenetic neighbours. The morphological characteristics of strain YN-5-1T were typical of the genus Nonomuraea. Chemotaxonomic characteristics, such as diagnostic diamino acid of the peptidoglycan, whole-cell sugars, phospholipid type, major menaquinone and major fatty acids, further supported the assignment of strain YN-5-1T to the genus Nonomuraea. The G+C content of the genomic DNA was 72.1 mol%. Based on the above data, strain YN-5-1T is considered to represent a novel species of the genus Nonomuraea, for which the name Nonomuraea flavida sp. nov. is proposed. The type strain is YN-5-1T ( = CCTCC AB 2012909T = KCTC 29143T).

  17. Novosphingobium lotistagni sp. nov., isolated from a lotus pond.

    Science.gov (United States)

    Ngo, Hien T T; Trinh, Huan; Kim, Jung-Hee; Yang, Jung-Eun; Won, Kyung-Hwa; Kim, Ju-Han; Kook, MooChang; Yi, Tae-Hoo

    2016-11-01

    A Gram-staining-negative, aerobic, non-motile, rod-shaped and yellow-pigmented bacterium, designated strain THG-DN6.20T, was isolated from a lotus pond near Donghaksa temple in Daejeon, Republic of Korea. According to 16S rRNA gene sequence comparisons, strain THG-DN6.20T was found to be most closely related to Novosphingobium rosa IFO 15208T (97.6 % sequence similarity), Novosphingobium sediminicola HU1-AH51T (97.5 %) and Novosphingobium barchaimii LL02T (96.9 %). The DNA-DNA relatedness between strain THG-DN6.20T and its phylogenetically closest neighbours was below 60.0 %. The respiratory quinone and polyamine detected in strain THG-DN6.20T were ubiquinone Q-10 and spermidine, respectively. The DNA G+C content was 63.1 mol%. The major polar lipids were found to be phosphatidylethanolamine, diphosphatidylglycerol, sphingoglycolipid and phosphatidylcholine. The major fatty acids were identified as C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C14 : 0 2-OH. These data supported the affiliation of strain THG-DN6.20T to the genus Novosphingobium. Strain THG-DN6.20T could be distinguished from related species of the genus Novosphingobium by physiological and biochemical characteristics. Therefore, the novel isolate represents a novel species, for which the name Novosphingobium lotistagni sp. nov. is proposed, with THG-DN6.20T as the type strain (=KACC 18541T=CCTCC AB 2015354T).

  18. Niabella hibiscisoli sp. nov., isolated from soil of a Rose of Sharon garden.

    Science.gov (United States)

    Ngo, Hien T T; Trinh, Huan; Yan, Zheng-Fei; Moya, Gabriela; Kook, MooChang; Yi, Tae-Hoo

    2016-10-21

    A Gram-stain-negative, strictly aerobic, non-motile, rod-shaped and yellow-pigmented bacterium, designated strain THG-DN5.5T, was isolated from soil of a Rose of Sharon garden in Daejeon, South Korea. According to 16S rRNA gene sequence comparisons, strain THG-DN5.5T was found to be most closely related to Niabella yanshanensis CCBAU 05354T (97.7 % sequence similarity), Niabella ginsengisoli GR10-1T (97.0 %), Niabella terrae ICM 1-15T (96.0 %), Niabella soli DSM 19437T (95.7 %), and Niabella aquatica RP-2T (95.6 %). The DNA-DNA relatedness between strain THG-DN5.5T and its phylogenetically closest neighbours was below 50.0 %. The DNA G+C content was 43.1 mol%. The major polar lipid was found to be phosphati¬dylethanolamine. The major fatty acids were identified as C16:0, iso-C15:0, iso-C15:1 G, and iso-C17:0 3OH. MK-7 was the only menaquinone present. These data supported the affiliation of strain THG-DN5.5T to the genus Niabella. Strain THG-DN5.5T was distinguished from related Niabella species by physiological and biochemical tests. In this study, the novel isolate represents a novel species, for which the name Niabella hibiscisoli sp. nov. is proposed, with THG-DN5.5T as the type strain (= KACC 18857T = CCTCC AB 2016086T).

  19. Abyssicoccus albus gen. nov., sp. nov., a novel member of the family Staphylococcaceae isolated from marine sediment of the Indian Ocean.

    Science.gov (United States)

    Jiang, Zhao; Yuan, Chang-Guo; Xiao, Min; Tian, Xin-Peng; Khan, Inam-Ullah; Kim, Chang-Jin; Zhi, Xiao-Yang; Li, Wen-Jun

    2016-08-01

    A Gram-stain positive, aerobic, non-motile, asporogenous, coccoid shaped bacterium, designated YIM M12140(T), was isolated from a marine sediment sample collected from the Indian Ocean. Phylogenetic analysis showed that strain YIM M12140(T) forms a separate clade within the family Staphylococcaceae. Strain YIM M12140(T) shares high 16S rRNA gene sequence similarity with Macrococcus brunensis DSM 19358(T) (92.9 %). The isolate was found to grow at 0-10 % (w/v) NaCl (optimum, 2-3 %), pH 6.0-10.0 (optimum, pH 8.0) and temperature 5-40 °C (optimum, 28 °C). The polar lipids were identified as diphosphatidylglycerol, phosphatidylglycerol, one unidentified aminophospholipid and two unidentified polar lipids. The major cellular fatty acids of the strain were identified as anteiso-C15:0, -C17:0, iso-C16:0, anteiso-C19:0 and C20:0. The respiratory menaquinones were found to be MK-6 (94 %) and MK-7 (6 %). The cell wall amino acids were found to contain Lys, Ala, Glu, Gly, Asp, Ser and Thr. Whole cell sugars were identified as mannose, ribose, rhamnose, glucose, galactose and xylose. The G+C content of the genomic DNA of strain YIM M12140(T) was determined to be 42.4 mol %. Based on phenotypic, chemotaxonomic data and phylogenetic analysis, it is proposed that strain YIM M12140(T) represents a novel species of a new genus in the family Staphylococcaceae, for which the name Abyssicoccus albus gen. nov., sp. nov. is proposed. The type strain is YIM M12140(T) (= DSM 29158(T) = CCTCC AB 2014213(T)).

  20. Roseomonas eburnea sp. nov., isolated from activated sludge.

    Science.gov (United States)

    Wang, Chenghong; Deng, Shikai; Liu, Xin; Yao, Li; Shi, Chao; Jiang, Jin; Kwon, Soon-Wo; He, Jian; Li, Jiayou

    2016-01-01

    A Gram-stain-negative, aerobic, short rod-shaped, non-endospore-forming, ivory-pigmented and non-motile bacterium, designated strain BUT-5T, was isolated from activated sludge of an herbicides-manufacturing wastewater treatment facility in Jiangsu Province, China. The major fatty acids (>5 % of total fatty acids) were C16 : 0, C18 : 1 2-OH and summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c). The predominant respiratory quinone was ubiquinone Q-10. The polar lipids profile of strain BUT-5T included diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine and two unknown aminolipids. The DNA G+C content was 67.6 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain BUT-5T showed the highest sequence similarities to Roseomonas soli 5N26T (97.5 % 16S rRNA gene sequence similarity), followed by Roseomonas lacus TH-G33T (97.3 %) and Roseomonas terrae DS-48T (97.1 %). Strain BUT-5T showed low DNA-DNA relatedness with Roseomonas soli KACC 16376T (41 %), Roseomonas lacus KACC 11678T (46 %) and Roseomonas terrae KACC 12677T (42 %), respectively. On the basis of phenotypic and genotypic properties, as well as chemotaxonomic data, strain BUT-5T represents a novel species of the genus Roseomonas, for which the name Roseomonas eburnea sp. nov. is proposed. The type strain is BUT-5T ( = CCTCC AB2013276T = KACC 17166T).

  1. Chryseobacterium chengduensis sp. nov. isolated from the air of captive giant panda enclosures in Chengdu, China.

    Science.gov (United States)

    Wen, Cai-Fang; Xi, Li-Xin; Zhao, Shan; Hao, Zhong-Xiang; Luo, Lu; Liao, Hong; Chen, Zhen-Rong; She, Rong; Han, Guo-Quan; Cao, San-Jie; Wu, Rui; Yan, Qi-Gui; Hou, Rong

    2016-08-01

    A Gram-negative, aerobic, non-motile, rod-shaped bacterial strain, designated 25-1(T), was isolated from the air inside giant panda enclosures at the Chengdu Research Base of Giant Panda Breeding, China. Strain 25-1(T) grew optimally at pH 7.0-8.0, at 28-30 °C and in the presence of NaCl concentrations from 0.0% to 0.5 %. 16S rRNA gene sequence analysis indicated that strain 25-1(T) belongs to the genus Chryseobacterium within the family Flavobacteriaceae and is related most closely to C. carnis G81(T) (96.4% similarity), C. lathyri RBA2-6(T) (95.8% similarity), and C. zeae JM1085(T) (95.8% similarity). Its genomic DNA G+C molar composition was 36.2%. The major cellular fatty acids were iso-C15:0 (44.0%), iso-C17:0 3OH (19.8%) and C16:1 ω7c/16:1 ω6c (12.7%). The only isoprenoid quinone was menaquinone 6 (MK-6). The major polar lipids were phosphatidylethanolamine, two unidentified amino lipids and two unidentified lipids. The DNA-DNA relatedness between strain 25-1(T) and C. lathyri RBA2-6(T) was 38%. Phenotypic, genotypic, and phylogenetic characteristics indicated that strain 25-1(T) is a novel member of the genus Chryseobacterium, for which the name C. chengduensis sp. nov. is proposed. The type strain is 25-1(T) (CCTCC AB2015133(T)=DSM 100396(T)).

  2. Halalkalicoccus paucihalophilus sp. nov., a halophilic archaeon from Lop Nur region in Xinjiang, northwest of China.

    Science.gov (United States)

    Liu, Bing-Bing; Tang, Shu-Kun; Zhang, Yong-Guang; Lu, Xin-Hua; Li, Li; Cheng, Juan; Zhang, Yuan-Ming; Zhang, Li-Li; Li, Wen-Jun

    2013-05-01

    Two extremely halophilic archaea, designated YIM 93701(T) and YIM 93664, were isolated from Lop Nur region in Xinjiang Province, northwest of China. The cells of the two strains were observed to be cocci, non-motile and Gram-negative. The organisms were determined to be aerobic and required at least 6 % NaCl for growth (optimum 20-25 % and maximum 35 %). Growth was found to occur in the ranges of 16-50 °C (optimum 37 °C) and pH 6.0-8.5 (optimum 6.5-7.5). Cells did not lyse in distilled water. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the two strains belongs to the genus Halalkalicoccus and possessed 99.3 and 99.5 % similarities with their closest phylogenetic relative Halalkalicoccus tibetensis JCM 11890(T). Major polar lipids of the two strains were determined to be phosphatidylglycerol(PG),phosphatidylglycerol phosphate methyl ester (PGP-Me), phosphatidylglycerol sulfate (PGS) and three unidentified glycolipids. The DNA G+C contents were determined to be 60.0-60.4 mol%. The DNA hybridization between the two strains was 92.0 %. In addition, the hybridizations of both strains to H. tibetensis were 49 and 52 %, respectively, and to Halalkalicoccus jeotali were 38 and 33 %, respectively. On the basis of physiological, biochemical tests and phylogenetic differentiations, strains YIM 93701(T) and YIM 93664 were classified as the same species which represent a novel species in the genus Halalkalicoccus, for which the name Halalkalicoccus paucihalophilus sp. nov. is proposed. The type strain is YIM 93701(T) (=JCM 17505(T) = CCTCC 2012803(T)).

  3. Transfer of Bacillus mucilaginosus and Bacillus edaphicus to the genus Paenibacillus as Paenibacillus mucilaginosus comb. nov. and Paenibacillus edaphicus comb. nov.

    Science.gov (United States)

    Hu, Xiu-Fang; Li, Shi-Xiao; Wu, Jin-Guang; Wang, Jian-Feng; Fang, Qiong-Lou; Chen, Ji-Shuang

    2010-01-01

    Bacillus mucilaginosus and Bacillus edaphicus were reclassified based on their 16S rRNA and gyrB gene sequences, DNA-DNA hybridization, fatty acid methyl esters and other taxonomic characteristics. Phylogenetic analysis based on 16S rRNA and gyrB gene sequences indicated that strains of B. mucilaginosus and B. edaphicus were members of the genus Paenibacillus, with over 90.4 % and 70.3 % sequence similarity, respectively. Their DNA G+C contents were 54.5-56.8 mol%. The DNA-DNA relatedness values of B. edaphicus VKPM B-7517(T) with B. mucilaginosus KNP414 and B. mucilaginosus CGMCC 1.236 were 89.2 % and 88.7 %, respectively. The major isoprenoid quinone of B. mucilaginosus and B. edaphicus was MK-7 (94.1-95.7 %). The peptidoglycan type was A1gamma (meso-diaminopimelic acid) and the major polar lipids were phosphatidylglycerol and diphosphatidylglycerol. The major fatty acids were anteiso-C(15 : 0), C(16 : 1)omega11c and C(16 : 0). Phenotypic features and fatty acid profiles supported the similarity of B. mucilaginosus and B. edaphicus to Paenibacillus validus CCTCC 95016(T) and confirmed their relationship with members of the genus Paenibacillus. Therefore, it is proposed that Bacillus mucilaginosus and Bacillus edaphicus be transferred to the genus Paenibacillus as Paenibacillus mucilaginosus comb. nov. (type strain HSCC 1605(T)=VKPM B-7519(T)=VKM B-1480D(T)=CIP 105815(T)=KCTC 3870(T)) and Paenibacillus edaphicus comb. nov. (type strain VKPM B-7517(T)=DSM 12974(T)=CIP 105814(T)), respectively.

  4. Rhizobium flavum sp. nov., a triazophos-degrading bacterium isolated from soil under the long-term application of triazophos.

    Science.gov (United States)

    Gu, Tao; Sun, Li Na; Zhang, Jun; Sui, Xin Hua; Li, Shun Peng

    2014-06-01

    A Gram-stain-negative, non-motile, pale yellow, rod-shaped bacterial strain, YW14(T), was isolated from soil and its taxonomic position was investigated by a polyphasic study. Strain YW14(T) did not form nodules on three different legumes, and the nodD and nifH genes were not detected by PCR. Strain YW14(T) contained Q-10 as the predominant ubiquinone. The major cellular fatty acid was C(18 : 1)ω7c. Phylogenetic analyses based on 16S rRNA gene sequences and seven housekeeping gene sequences (recA, atpD, glnII, gyrB, rpoB, dnaK and thrC) showed that strain YW14(T) belonged to the genus Rhizobium. Strain YW14(T) showed 16S rRNA gene sequence similarity of 93.4-97.3% to the type strains of recognized species of the genus Rhizobium. DNA-DNA relatedness between strain YW14(T) and the type strains of Rhizobium sullae IS123(T) and Rhizobium yanglingense CCBAU 71623(T) was 19.6-25.7%, indicating that strain YW14(T) was distinct from them genetically. Strain YW14(T) could also be differentiated from these phylogenetically related species of the genus Rhizobium by various phenotypic properties. On the basis of phenotypic properties, phylogenetic distinctiveness and genetic data, strain YW14(T) is considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium flavum sp. nov. is proposed. The type strain is YW14(T) ( = KACC 17222(T) = CCTCC AB2013042(T)).

  5. Rhizobium smilacinae sp. nov., an endophytic bacterium isolated from the leaf of Smilacina japonica.

    Science.gov (United States)

    Zhang, Lei; Shi, Xu; Si, Meiru; Li, Changfu; Zhu, Lingfang; Zhao, Liang; Shen, Xihui; Wang, Yao

    2014-10-01

    During a study of endophytic bacteria from traditional Chinese medicinal plants, a bacterial strain, designated PTYR-5(T), was isolated from the leaf of Smilacina japonica A. Gray collected from Taibai Mountain in Shaanxi Province, north-west China. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain PTYR-5(T) is a member of the genus Rhizobium, exhibiting the highest sequence similarities to R. cellulosilyticum LMG 23642(T) (97.2%), R. huautlense LMG 18254(T) (97.2%) and R. alkalisoli CCBAU 01393(T) (97.1%). The levels of 16S rRNA gene sequence similarity with respect to other Rhizobium species with validly published names were less than 97.0%. Phylogenies of the housekeeping genes atpD, recA and glnII confirmed its distinct position, showing low similarity with respect to those of recognized Rhizobium species (no more than 94.1, 90.0 and 88.0% similarity, respectively). The DNA-DNA relatedness values of strain PTYR-5(T) with R. cellulosilyticum LMG 23642(T), R. huautlense LMG 18254(T) and R. alkalisoli CCBAU 01393(T) were 33.6, 21.4 and 29.5 %, respectively. Based on phenotypic, phylogenetic and genotypic data, strain PTYR-5(T) is considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium smilacinae sp. nov. is proposed. The type strain is PTYR-5(T) (=CCTCC AB 2013016(T)=KCTC 32300(T)=LMG 27604(T)).

  6. Asticcacaulis endophyticus sp. nov., a prosthecate bacterium isolated from the root of Geum aleppicum.

    Science.gov (United States)

    Zhu, Lingfang; Long, Mingxiu; Si, Meiru; Wei, Linfang; Li, Changfu; Zhao, Liang; Shen, Xihui; Wang, Yao; Zhang, Lei

    2014-12-01

    A strictly aerobic, light-yellow-coloured, stalked bacterium, designated strain ZFGT-14(T), was isolated from the root of Geum aleppicum Jacq. collected from Taibai Mountain in Shaanxi province, north-west China, and was subjected to a taxonomic study using a polyphasic approach. This novel isolate grew at 7-33 °C (optimum 25-28 °C) and pH 6.0-10.0 (optimum pH 7.0-8.0). Flexirubin-type pigments were not produced. Cells were Gram-stain-negative, rod-shaped and motile with a single polar flagellum. The predominant respiratory quinone was Q-10. The major cellular fatty acids were summed feature 8 (comprising C18 : 1ω7c/C18 : 1ω6c), C16 : 0, C19 : 0 cyclo ω8c and summed feature 3 (comprising C16 : 1ω7c and/or C16 : 1ω6c) and the major polar lipids were phosphatidylglycerol and glycolipids. The DNA G+C content was 57.8 mol%. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain ZFGT-14(T) was most closely related to the genus Asticcacaulis and had low sequence similarity (95.0-95.9 %) with all species with validly published names within the genus Asticcacaulis. Based on the phenotypic, phylogenetic and genotypic data, strain ZFGT-14(T) is considered to represent a novel species of the genus Asticcacaulis, for which the name Asticcacaulis endophyticus sp. nov. is proposed. The type strain is ZFGT-14(T) ( = CCTCC AB 2013012(T) = KCTC 32296(T) = LMG 27605(T)).

  7. Surface display of monkey metallothionein {alpha} tandem repeats and EGFP fusion protein on Pseudomonas putida X4 for biosorption and detection of cadmium

    Energy Technology Data Exchange (ETDEWEB)

    He, Xiaochuan; Chen, Wenli; Huang, Qiaoyun [Huazhong Agricultural Univ., Wuhan (China). State Key Lab. of Agricultural Microbiology

    2012-09-15

    Monkey metallothionein {alpha} domain tandem repeats (4mMT{alpha}), which exhibit high cadmium affinity, have been displayed for the first time on the surface of a bacterium using ice nucleation protein N-domain (inaXN) protein from the Xanthomonas campestris pv (ACCC - 10049) as an anchoring motif. The shuttle vector pIME, which codes for INAXN-4mMT{alpha}-EGFP fusion, was constructed and used to target 4mMT{alpha} and EGFP on the surface of Pseudomonas putida X4 (CCTCC - 209319). The surface location of the INAXN-4mMT{alpha}-EGFP fusion was further verified by western blot analysis and immunofluorescence microscopy. The growth of X4 showed resistance to cadmium presence. The presence of surface-exposed 4mMT{alpha} on the engineered strains was four times higher than that of the wild-type X4. The Cd{sup 2+} accumulation by X4/pIME was not only four times greater than that of the original host bacterial cells but was also remarkably unaffected by the presence of Cu{sup 2+} and Zn{sup 2+}. Moreover, the surface-engineered strains could effectively bind Cd{sup 2+} under a wide range of pH levels, from 4 to 7. P. putida X4/pIME with surface-expressed 4mMT{alpha}-EGFP had twice the cadmium binding capacity as well as 1.4 times the fluorescence as the cytoplasmic 4mMTa-EGFP. These results suggest that P. putida X4 expressing 4mMT{alpha}-EGFP with the INAXN anchor motif on the surface would be a useful tool for the remediation and biodetection of environmental cadmium contaminants. (orig.)

  8. Lysobacter novalis sp. nov., isolated from fallow farmland soil.

    Science.gov (United States)

    Singh, Hina; Du, Juan; Won, Kyung-Hwa; Yang, Jung-Eun; Akter, Shahina; Kim, Ki-Young; Yi, Tae-Hoo

    2015-09-01

    A novel bacterial strain, designated THG-PC7(T), was isolated from fallow farmland soil in Yongin, South Korea. Cells of strain THG-PC7(T) were Gram-stain-negative, dark yellow, aerobic, rod-shaped and had gliding motility. Strain THG-PC7(T) grew optimally at 25-35 °C, at pH 7 and in the absence of NaCl. Comparative 16S rRNA gene sequence analysis identified strain THG-PC7(T) as belonging to the genus Lysobacter, exhibiting highest sequence similarity with Lysobacter ximonensis KCTC 22336(T) (98.7%) followed by Lysobacter niastensis KACC 11588(T) (95.7%). In DNA-DNA hybridization tests, DNA relatedness between strain THG-PC7(T) and its closest phylogenetic neighbour L. ximonensis was below 25%. The DNA G+C content of the novel isolate was determined to be 62.5 mol%. Flexirubin-type pigments were found to be present. The major cellular fatty acids were determined to be iso-C15 : 0, iso-C16 : 0, anteiso-C15 : 0 and iso-C17 : 1ω9c. The major respiratory quinone was identified as ubiquonone-8 (Q8). The predominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified aminophospolipid. On the basis of results from DNA-DNA hybridization and the polyphasic data, strain THG-PC7(T) represents a novel species of the genus Lysobacter, for which the name Lysobacter novalis sp. nov. is proposed. The type strain is THG-PC7(T)( = KACC 18276(T) = CCTCC AB 2014319(T)).

  9. Streptomyces indoligenes sp. nov., isolated from rhizosphere soil of Populus euphratica.

    Science.gov (United States)

    Luo, Xiaoxia; Sun, Yong; Xie, Sinan; Wan, Chuanxing; Zhang, Lili

    2016-06-01

    A novel actinobacterium, designated TRM 43006T, was isolated from the rhizosphere soil of Populus euphratica in Xinjiang Province, north-west China. Phylogenetic and phenotypic analysis demonstrated that strain TRM 43006T belongs to the genus Streptomyces. The strain was aerobic and Gram-stain-positive; the aerial mycelium branched monopodially, forming chains of arthrospores. The spores were oval to cylindrical with smooth surfaces. The whole-cell sugar pattern of strain TRM 43006T consisted of xylose, mannitol, galactose and ribose. The menaquinones were MK-9(H6), MK-9(H8) and MK-9(H10). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol, phosphatidylinositol mannosides and four unknown phospholipids. Major fatty acids were iso-C16 : 0, iso-C16 : 1, iso-C14 : 0 and anteiso-C15 : 0. The G+C content of the genomic DNA was 69.0 mol%. Comparative 16S rRNA gene sequence analysis indicated that strain TRM 43006T was phylogenetically most closely related to Streptomyces roseolilacinus NBRC 12815T (98.6 % similarity) and Streptomycessudanensis SD 504T (98.3 %); however, DNA-DNA hybridization studies between S. roseolilacinus NBRC 12815T, S. sudanensis SD 504T and TRM 43006T showed only 30.28 and 30.65  % relatedness, respectively. Based on the evidence from this polyphasic study, strain TRM 43006T represents a novel species of the genus Streptomyces, for which the name Streptomyces indoligenes sp. nov. is proposed. The type strain is TRM 43006T (=KCTC 39611T=CCTCC AA 2015010T).

  10. Streptomyces alfalfae sp. nov. and comparisons with its closest taxa Streptomyces silaceus, Streptomyces flavofungini and Streptomyces intermedius.

    Science.gov (United States)

    She, Wenqing; Sun, Zhongfeng; Yi, Lei; Zhao, Shumiao; Liang, Yunxiang

    2016-01-01

    A novel streptomycete strain, designated XY25T, was isolated from the rhizosphere soil in an alfalfa field in Jingyang, Shanxi, China. The isolate showed optimal growth at 37 °C, and was capable of growing at pH 6-10 and in the presence of 0-6 % (w/v) NaCl. Mycelia of strain XY25T appeared spiral and developed into white spore chains with long-rod spores and a smooth surface. The 16S rRNA gene sequence of XY25T was determined and was found to be highly similar to those of species of the genus Streptomyces including Streptomyces silaceus DSM 41861T (99.11 % 16S rRNA gene sequence similarity), Streptomyces flavofungini DSM 40366T (98.49 %) and Streptomyces intermedius DSM 40372T (98.43 %), all of which were used for further characterization. Each of the four streptomycetes showed distinctive patterns of carbon usage and fatty acids composition. Analysis of cellular components of strain XY25T revealed ll-diaminopimelic acid as diagnostic diamino acid and xylose as the major sugar, whereas polar lipids were determined as phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylinositol, an unknown phospholipid, two unknown phosphatidylinositol mannosides and several unknown lipids. Menaquinones were dominated by MK-9(H6) and MK-9(H8), and the main fatty acids were anteiso-C15 : 0, iso-C16 : 0 and anteiso-C17 : 0. DNA-DNA hybridization studies indicated that strain XY25T showed relatedness values of 35.2-40.42 % with the closest related species. Based on these results, strain XY25T represents a novel species of the genus Streptomyces, for which the name Streptomyces alfalfae sp. nov. is proposed. The type strain is XY25T ( = KCTC 39571T = CCTCC AA2015019T).

  11. Streptomyces bohaiensis sp. nov., a novel actinomycete isolated from Scomberomorus niphonius in the Bohai Sea.

    Science.gov (United States)

    Pan, Hua-Qi; Cheng, Juan; Zhang, Dao-Feng; Yu, Su-Ya; Khieu, Thi-Nhan; Son, Chu Ky; Jiang, Zhao; Hu, Jiang-Chun; Li, Wen-Jun

    2015-04-01

    A novel actinomycete strain, designated 11A07(T), was isolated from young Scomberomorus niphonius in the Bohai Sea. Basic local alignment search tool analyses showed that this isolate had the highest 16S rRNA gene sequence similarity of 97.41% with Streptomyces rimosus subsp. paromomycinus DSM 41429(T). Phylogenetic tree revealed that strain 11A07(T) formed a distinct lineage clustered with Streptomyces panacagri Gsoil 519(T), Streptomyces sodiiphilus YIM 80305(T) and Streptomyces albus subsp. albus NRRL B-2365(T) having similarities of 97.30%, 97.10% and 96.83%, respectively. Multilocus sequence analysis further demonstrated that the new isolate was different from the selected representatives of Streptomyces as a separate phylogenetic line. Strain 11A07(T) produced straight or rectiflexibile spore chains with smooth surface, white aerial mycelia and brown diffusible pigments on international streptomyces project 2 medium. Maximum tolerated NaCl concentration for growth was 11.0%. Whole-cell sugars were mannose, ribose, glucose, galactose and xylose. The predominant menaquinones were MK-9(H2), MK-9(H4) and MK-9 (H6). The fatty-acid profile contained iso-C16:0, C18:0 10-methyl (tuberculostearic acid) and anteiso-C17:0 as the major compositions. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol, phosphatidylinositol mannoside and an unknown phospholipid. The G+C content of the genomic DNA was 71.4 mol%. These morphological, phenotypic and chemotaxonomic properties showed that strain 11A07(T) could be readily distinguished from the most closely related members of the genus Streptomyces. Thus, based on the polyphasic taxonomic data, strain 11A07(T) (=JCM 19630(T)=CCTCC AA 2013020(T)=KCTC 29263(T)) represents a novel species within the genus Streptomyces, for which the name Streptomyces bohaiensis sp. nov. is proposed.

  12. Marisediminicola antarctica gen. nov., sp. nov., an actinobacterium isolated from the Antarctic.

    Science.gov (United States)

    Li, Hui-Rong; Yu, Yong; Luo, Wei; Zeng, Yin-Xin

    2010-11-01

    Strain ZS314(T) was isolated from a sandy intertidal sediment sample collected from the coastal area off the Chinese Antarctic Zhongshan Station, east Antarctica (6 9° 22' 13″ S 76 ° 21' 41″ E). The cells were Gram-positive, motile, short rods. The temperature range for growth was 0-26 °C and the pH for growth ranged from 5 to 10, with optimum growth occurring within the temperature range 18-23 °C and pH range 6.0-8.0. Growth occurred in the presence of 0-6 % (w/v) NaCl, with optimum growth occurring in the presence of 2-4 % (w/v) NaCl. Strain ZS314(T) had MK-10 as the major menaquinone and anteiso-C(15 : 0), iso-C(16 : 0) and anteiso-C(17 : 0) as major fatty acids. The cell-wall peptidoglycan type was B2β with ornithine as the diagnostic diamino acid. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The genomic DNA G+C content was approximately 67 mol%. Phylogenetic analysis based on 16S rRNA gene sequence similarity showed that strain ZS314(T) represents a new lineage in the family Microbacteriaceae. On the basis of the phylogenetic analyses and phenotypic characteristics, a new genus, namely Marisediminicola gen. nov., is proposed, harbouring the novel species Marisediminicola antarctica sp. nov. with the type strain ZS314(T) (=DSM 22350(T) =CCTCC AB 209077(T)).

  13. The mechanism of improved intracellular organic selenium and glutathione contents in selenium-enriched Candida utilis by acid stress.

    Science.gov (United States)

    Zhang, Gao-Chuan; Wang, Da-Hui; Wang, Dong-Hua; Wei, Gong-Yuan

    2017-03-01

    Batch culture of Candida utilis CCTCC M 209298 for the preparation of selenium (Se)-enriched yeast was carried out under different pH conditions, and maximal intracellular organic Se and glutathione (GSH) contents were obtained in a moderate acid stress environment (pH 3.5). In order to elucidate the physiological mechanism of improved performance of Se-enriched yeast by acid stress, assays of the key enzymes involved in GSH biosynthesis and determinations of energy supply and regeneration were performed. The results indicated that moderate acid stress increased the activity of γ-glutamylcysteine synthetase and the ratios of NADH/NAD(+) and ATP/ADP, although no significant changes in intracellular pH were observed. In addition, the molecular mechanism of moderate acid stress favoring the improvement of Se-yeast performance was revealed by comparing whole transcriptomes of yeast cells cultured at pH 3.5 and 5.5. Comparative analysis of RNA-Seq data indicated that 882 genes were significantly up-regulated by moderate acid stress. Functional annotation of the up-regulated genes based on gene ontology and the Kyoto Encyclopedia of Genes and Genome (KEGG) pathway showed that these genes are involved in ATP synthesis and sulfur metabolism, including the biosynthesis of methionine, cysteine, and GSH in yeast cells. Increased intracellular ATP supply and more amounts of sulfur-containing substances in turn contributed to Na2SeO3 assimilation and biotransformation, which ultimately improved the performance of the Se-enriched C. utilis.

  14. Lactobacillus herbarum sp. nov., a species related to Lactobacillus plantarum.

    Science.gov (United States)

    Mao, Yuejian; Chen, Meng; Horvath, Philippe

    2015-12-01

    Strain TCF032-E4 was isolated from a traditional Chinese fermented radish. It shares >99% 16S rRNA sequence identity with L. plantarum, L. pentosus and L. paraplantarum. This strain can ferment ribose, galactose, glucose, fructose, mannose, mannitol, N-acetylglucosamine, amygdalin, arbutin, salicin, cellobiose, maltose, lactose, melibiose, trehalose and gentiobiose. It cannot ferment sucrose, which can be used by L. pentosus, L. paraplantarum, L. fabifermentans, L. xiangfangensis and L. mudanjiangensis, as well as most of the L. plantarum strains (88.7%). TCF032-E4 cannot grow at temperature above 32 °C. This strain shares 78.2-83.6% pheS (phenylalanyl-tRNA synthetase alpha subunit) and 89.5-94.9% rpoA (RNA polymerase alpha subunit) sequence identity with L. plantarum, L. pentosus, L. paraplantarum, L. fabifermentans, L. xiangfangensis and L. mudanjiangensis. These results indicate that TCF032-E4 represents a distinct species. This hypothesis was further confirmed by whole-genome sequencing and comparison with available genomes of related species. The draft genome size of TCF032-E4 is approximately 2.9 Mb, with a DNA G+C content of 43.5 mol%. The average nucleotide identity (ANI) between TCF032-E4 and related species ranges from 79.0 to 81.1%, the highest ANI value being observed with L. plantarum subsp. plantarum ATCC 14917T. A novel species, Lactobacillus herbarum sp. nov., is proposed with TCF032-E4T ( = CCTCC AB2015090T = DSM 100358T) as the type strain.

  15. Pedobacter humi sp. nov., isolated from a playground soil.

    Science.gov (United States)

    Trinh, Huan; Yi, Tae-Hoo

    2016-06-01

    A Gram-stain-negative, aerobic, non-motile, rod-shaped and yellow-pigmented bacterium, designated strain THG S15-2T, was isolated from playground soil in Sindorim-dong, Guro-gu, Seoul, South Korea. According to 16S rRNA gene sequence comparisons, strain THG S15-2T was found to be related most closely to Pedobacter ginsengisoli Gsoil 104T (97.5 % similarity), Pedobacter panaciterrae Gsoil 042T (97.4 %), Pedobacter seoulensis THG-G12T (97.1 %) and Pedobacter caeni LMG 22862T (97.1 %). The level of DNA-DNA relatedness between strain THG S15-2T and its phylogenetically closest neighbours was below 30.0 %. The only isoprenoid quinone detected in strain THG S15-2T was menaquinone-7. The DNA G+C content was 45.9 mol%. The major polar lipid was phosphatidylethanolamine. The major component in the polyamine pattern was sym-homospermidine. The major fatty acids were identified as summed feature 3 (C16:1ω7c and/or C16:1ω6c), iso-C15:0 and C16:0. These data supported the affiliation of strain THG S15-2T to the genus Pedobacter. Strain THG S15-2T was distinguished from related Pedobacter species by physiological and biochemical tests. Therefore, strain THG S15-2T represents a novel species, for which the name Pedobacter humi sp. nov. is proposed. The type strain is THG S15-2T (= KCTC 42735T = CCTCC AB 2015293T).

  16. Bacillus thermotolerans sp. nov., a thermophilic bacterium capable of reducing humus.

    Science.gov (United States)

    Yang, Guiqin; Zhou, Xuemei; Zhou, Shungui; Yang, Dehui; Wang, Yueqiang; Wang, Dingmei

    2013-10-01

    A novel thermotolerant bacterium, designated SgZ-8(T), was isolated from a compost sample. Cells were non-motile, endospore-forming, Gram-staining positive, oxidase-negative and catalase-positive. The isolate was able to grow at 20-65 °C (optimum 50 °C) and pH 6.0-9.0 (optimum 6.5-7.0), and tolerate up to 9.0 % NaCl (w/v) under aerobic conditions. Anaerobic growth occurred with anthraquinone-2,6-disulphonate (AQDS), fumarate and NO3(-) as electron acceptors. Phylogenetic analysis based on the16S rRNA and gyrB genes grouped strain SgZ-8(T) into the genus Bacillus, with the highest similarity to Bacillus badius JCM 12228(T) (96.2 % for 16S rRNA gene sequence and 83.5 % for gyrB gene sequence) among all recognized species in the genus Bacillus. The G+C content of the genomic DNA was 49.3 mol%. The major isoprenoid quinone was menaquinone 7 (MK-7) and the polar lipids consisted of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified phospholipid. The major cellular fatty acid was iso-C16 : 0. On the basis of its phenotypic and phylogenetic properties, chemotaxonomic analysis and the results of physiological and biochemical tests, strain SgZ-8(T) ( = CCTCC AB 2012108(T) = KACC 16706(T)) was designated the type strain of a novel species of the genus Bacillus, for which the name Bacillus thermotolerans sp. nov. is proposed.

  17. The effect silage aditives supplementation on dynamic fermentation process, quality and aerobic stability of corn silage

    Directory of Open Access Journals (Sweden)

    Václav Pyrochta

    2008-01-01

    Full Text Available In the experiment, the effect of additives supplementation on the fermentation quality of corn silage was examined, compared with the untreated control (K. The aditive „A“ contained bacterial component of (Propionibactrium acidipropionici – MA126/4U 3*1010 and Lactobacillus plantarum – MA18/5U. The effective substances of bacterial inoculants „B“, selected were bacterial strains of (Lactobacillus casei ssp. rhamnosus LC – 705 DSM 7061 4*1011, Propionibacterium freudenreichii spp. shermanii JS DSM 6067 2-4*1011. There were used as effective substances of bacterial inoculants „C“ lactic bacteria and enzyme (Lactobacillus plantarum CCM 3769 1.67*1010, Lactococcus lactis CCM 4754 1.67*1010, Enterococcus faecium CCM 6226 1.67*1010, Pediococcus pentosaceus CCM 3770 1,67*1010, cellulase, hemicellulase, sodium benzoate. They were applied in the dose of prescript by producer. At conservations with all aditivum were statistically significant (P < 0.01 increase of lactic acid formation from 55.31±9.72 g/kg DM of control silage to 59.60±10.84 g/kg DM aditivum „A“, 59.36±10.04 g/ kg DM aditivum „B“ rather to 60.74±9.90 g/kg DM aditivum „C“. Aditives „A“ and „B“ were statistically significant (P < 0.01 increase propoinic acid and total fermentation acid content in silages occured. The fermentation characteristics in the microbial aditivum silages by us were more favourable. The date of fermentation was statistically significant (P < 0.01 increase the contents of acetic acid from 45.49±2.83 g/kg DM of 4st day to 63.07±4.25 g/kg DM of 32ndday rather to 67.70±2.94 g/kg DM of 64st day. There were statistically significant (P < 0.01 increase contents of acetic acid and total acid content. The date of fermentation was statistically significant (P < 0.01 degressive of pH.

  18. Influence of ensiling, exogenous protease addition, and bacterial inoculation on fermentation profile, nitrogen fractions, and ruminal in vitro starch digestibility in rehydrated and high-moisture corn.

    Science.gov (United States)

    Ferraretto, L F; Fredin, S M; Shaver, R D

    2015-10-01

    or inoculated with 1 of 3 microbial inoculants and with or without exogenous protease addition. The inoculant treatments contained (1) Lactobacillus buchneri 40788 and Pediococcus pentosaceus, (2) L. buchneri 40788, and (3) a mixture of P. pentosaceus and Propionibacterium freudenreichii. Protease, but not inoculation, increased ivSD by 7.5 percentage units (44.4 vs. 51.9%). Protease addition increased ivSD in rehydrated corn and HMC. Microbial inoculation improved fermentation profiles but did not affect ivSD.

  19. Effect of applying inoculants with heterolactic or homolactic and heterolactic bacteria on the fermentation and quality of corn silage.

    Science.gov (United States)

    Arriola, K G; Kim, S C; Adesogan, A T

    2011-03-01

    This study examined the effect of applying different bacterial inoculants on the fermentation and quality of corn silage. Corn plants were harvested at 35% DM, chopped, and ensiled in 20-L mini silos after application of (1) deionized water (CON) or inoculants containing (2) 1 × 10(5) cfu/g of Pediococcus pentosaceus 12455 and Propionibacteria freudenreichii (B2); (3) 4 × 10(5) cfu/g of Lactobacillus buchneri 40788 (BUC); or (4) 1 × 10(5) cfu/g of Pediococcus pentosaceus 12455 and 4 × 10(5) cfu/g of L. buchneri 40788 (B500). Four replicates of each treatment were weighed into polyethylene bags within 20-L mini silos. Silos were stored for 575 d at ambient temperature (25°C) in a covered barn. After silos were opened, aerobic stability, chemical composition, and yeast and mold counts were determined. The DNA in treated and untreated silages was extracted using lysozyme/sodium dodecyl sulfate lysis and phenol/chloroform and used as a template for a conventional PCR with primers designed on the 16S rRNA gene to detect the presence of L. buchneri in all silage samples. Acetic acid concentration was greater in B2 silages versus others (6.46 vs. 4.23% DM). Silages treated with BUC and B500 had lower pH and propionic acid concentration and greater lactic acid concentration than others. The B500 silage had the greatest lactic:acetic acid ratio (1.54 vs. 0.41), and only treatment with BUC reduced DM losses (5.0 vs. 14.3%). Yeast and mold counts were less than the threshold (10(5)) typically associated with silage spoilage and did not differ among treatments. Consequently, all silages were very stable (>250 h). Aerobic stability was not improved by any inoculant but was lower in B500 silages versus others (276 vs. 386 h). The conventional PCR confirmed the presence of similar populations of L. buchneri in all silages. This may have contributed to the prolonged aerobic stability of all silages. Copyright © 2011 American Dairy Science Association. Published by Elsevier

  20. Bacillus oleivorans sp. nov., a diesel oil-degrading and solvent-tolerant bacterium.

    Science.gov (United States)

    Azmatunnisa, M; Rahul, K; Subhash, Y; Sasikala, Ch; Ramana, Ch V

    2015-04-01

    Two Gram-stain-positive, diesel oil-degrading, solvent-tolerant, aerobic, endospore-forming, rod-shaped bacteria were isolated from a contaminated laboratory plate. Based on 16S rRNA gene sequence analysis, strains JC228(T) and JC279 were identified as belonging to the genus Bacillus within the family Bacillaceae of the phylum Firmicutes and were found to be most closely related to Bacillus carboniphilus JCM 9731(T) (98.1% 16S rRNA gene sequence similarity) and shared Bacillus . The DNA-DNA hybridization value between the two strains was 88±2%. Strain JC228(T) showed 23.4±1% reassociation (based on DNA-DNA hybridization) with B. carboniphilus LMG 18001(T). The DNA G+C content of strains JC228(T) and JC279 was 39 and 38.4 mol%, respectively. Both strains were positive for catalase and oxidase activities, and negative for hydrolysis of starch and Tween 80. Strains JC228(T) and JC279 grew chemoorganoheterotrophically with optimum growth at pH 7 (range pH 7-9.5) and 35 °C (range 25-40 °C). Diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified phospholipid (PL2) were the major polar lipids. Major cellular fatty acids were iso-C(15 : 0), anteiso-C(15 : 0), iso-C(17 : 0) and C(16 : 0). Whole-cell hydrolysates contained l-alanine, d-alanine, d-glutamic acid and meso-diaminopimelic acid. Both strains utilized diesel oil as sole carbon and energy source. The results of physiological, biochemical, chemotaxonomic and molecular analyses allowed clear differentiation of strains JC228(T) and JC279 from their closest phylogenetic neighbours. Therefore strains JC228(T) and JC279 represent a novel species of the genus Bacillus , for which the name Bacillus oleivorans sp. nov. is proposed. The type strain is JC228(T) ( = LMG 28084(T) = CCTCC AB 2013353(T)).

  1. Nocardioides albidus sp. nov., an actinobacterium isolated from garden soil.

    Science.gov (United States)

    Singh, Hina; Du, Juan; Trinh, Huan; Won, KyungHwa; Yang, Jung-Eun; Yin, ChangShik; Kook, MooChang; Yi, Tae-Hoo

    2016-01-01

    A novel bacterial strain, designated THG-S11.7T, was isolated from garden soil in Incheon, South Korea. Cells of the strain were Gram-stain-positive, aerobic, non-motile cocci, and were catalase- and oxidase-positive. Colonies of the strain were white. Strain THG-S11.7T grew optimally at 28 °C, at pH 7.0 and in the presence of 2.0 % NaCl. 16S rRNA gene sequence analysis indicated that the strain was a member of the genus Nocardioides. Strain THG-S11.7T showed a 16S rRNA gene sequence similarity of 98.2 % to Nocardioides kongjuensis KCTC 19054T, 98.0 % to Nocardioides caeni KCTC 19600T, 97.9 % to Nocardioides daeguensis KCTC 19799T, 97.8 % to Nocardioides nitrophenolicus KCTC 047BPT, 97.6 % to Nocardioides aromaticivorans KACC 20613T, 97.5 % to Nocardioides simplex KACC 20620T and 97.0 % to Nocardioides ginsengisoli KCTC 19135T. DNA-DNA relatedness values between strain THG-S11.7T and the closest phylogenetic neighbours were below 45.0 % and the DNA G+C content of strain THG-S11.7T was 72.2 mol%. Strain THG-S11.7T was characterized chemotaxonomically as having ll-diaminopimelic acid in the cell-wall peptidoglycan and menaquinone MK-8(H4) as the predominant isoprenoid quinone. The major phospholipid was determined to be diphosphatidylglycerol. The major cellular fatty acids of strain THG-S11.7T were iso-C15 : 0, C16 : 0 and iso-C16 : 0. Based on the phenotypic, genotypic and phylogenetic analyses, it is proposed that the isolate represents a novel species of the genus Nocardioides, for which the name Nocardioides albidus sp. nov. is proposed. The type strain is THG-S11.7T ( = KCTC 39607T = CCTCC AB 2015297T).

  2. Purification and characterization of a novel chlorpyrifos hydrolase from Cladosporium cladosporioides Hu-01.

    Science.gov (United States)

    Gao, Yan; Chen, Shaohua; Hu, Meiying; Hu, Qiongbo; Luo, Jianjun; Li, Yanan

    2012-01-01

    Chlorpyrifos is of great environmental concern due to its widespread use in the past several decades and its potential toxic effects on human health. Thus, the degradation study of chlorpyrifos has become increasing important in recent years. A fungus capable of using chlorpyrifos as the sole carbon source was isolated from organophosphate-contaminated soil and characterized as Cladosporium cladosporioides Hu-01 (collection number: CCTCC M 20711). A novel chlorpyrifos hydrolase from cell extract was purified 35.6-fold to apparent homogeneity with 38.5% overall recovery by ammoniumsulfate precipitation, gel filtration chromatography and anion-exchange chromatography. It is a monomeric structure with a molecular mass of 38.3 kDa. The pI value was estimated to be 5.2. The optimal pH and temperature of the purified enzyme were 6.5 and 40°C, respectively. No cofactors were required for the chlorpyrifos-hydrolysis activity. The enzyme was strongly inhibited by Hg²⁺, Fe³⁺, DTT, β-mercaptoethanol and SDS, whereas slight inhibitory effects (5-10% inhibition) were observed in the presence of Mn²⁺, Zn²⁺, Cu²⁺, Mg²⁺, and EDTA. The purified enzyme hydrolyzed various organophosphorus insecticides with P-O and P-S bond. Chlorpyrifos was the preferred substrate. The Km and Vmax values of the enzyme for chlorpyrifos were 6.7974 μM and 2.6473 μmol·min⁻¹, respectively. Both NH2-terminal sequencing and matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometer (MALDI-TOF-MS) identified an amino acid sequence MEPDGELSALTQGANS, which shared no similarity with any reported organophosphate-hydrolyzing enzymes. These results suggested that the purified enzyme was a novel hydrolase and might conceivably be developed to fulfill the practical requirements to enable its use in situ for detoxification of chlorpyrifos. Finally, this is the first described chlorpyrifos hydrolase from fungus.

  3. Roseomonas chloroacetimidivorans sp. nov., a chloroacetamide herbicide-degrading bacterium isolated from activated sludge.

    Science.gov (United States)

    Chu, Cui-Wei; Chen, Qing; Wang, Cheng-Hong; Wang, Hong-Mei; Sun, Zhong-Guan; He, Qin; He, Jian; Gu, Jin-Gang

    2016-05-01

    A Gram-negative, aerobic, short rod-shaped, pink-pigmented, non-motile bacterium, designated BUT-13(T), was isolated from activated sludge of an herbicide-manufacturing wastewater treatment facility in Jiangsu province, China. Growth was observed at 0-5.5 % NaCl, pH 6.0-9.0 and 12-37 °C. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain BUT-13(T) is a member of the genus Roseomonas, and shows high sequence similarities to R. pecuniae N75(T) (98.0 %) and R. rosea 173-96(T) (97.5 %), and lower (Roseomonas species. Chemotaxonomic analysis revealed that strain BUT-13(T) possesses Q-10 as the predominant ubiquinone; summed feature 8 (C18:1 w7c and/or C18:1 w6c; 38.8 %), C18:0 (16.6 %), C16:0 (15.2 %), summed feature 3 (C16:1 ω6c and/or C16:1 ω7; 7.9 %) and C18:1 w9c (4.7 %) as the major fatty acids. The polar lipids were found to consist of two aminolipids, a glycolipid, a phospholipid, a phosphoglycolipid, phosphatidylcholine, phosphatidylethanolamine and diphosphatidylglycerol. Strain BUT-13(T) showed low DNA-DNA relatedness with R. pecuniae N75(T) (45.2 %) and R. rosea 173-96(T) (51.2 %). The DNA G+C content was determined to be 67.6 mol%. Based on the phylogenetic analysis, DNA-DNA hybridization and chemotaxonomic analysis, as well as biochemical characteristics, strain BUT-13(T) can be clearly distinguished from all currently recognised Roseomonas species and should be classified as a novel species of the genus Roseomonas, for which the name Roseomonas chloroacetimidivorans sp. nov. is proposed. The type strain is BUT-13(T) (CCTCC AB 2015299(T) = JCM 31050(T)).

  4. Roseomonas rhizosphaerae sp. nov., a triazophos-degrading bacterium isolated from soil.

    Science.gov (United States)

    Chen, Qing; Sun, Li-Na; Zhang, Xiao-xia; He, Jian; Kwon, Soon-Wo; Zhang, Jun; Li, Shun-peng; Gu, Jin-gang

    2014-04-01

    A novel aerobic, non-spore-forming, non-motile, catalase- and oxidase-positive, Gram-stain-negative, coccoid to short-rod-shaped bacterial strain, designated YW11(T), was isolated from soil under long-term application of triazophos. The strain was able to hydrolyse triazophos. Strain YW11(T) grew at 15-40 °C (optimum at 28 °C), at pH 5.0-8.0 (optimum at pH 7.5) and with 0-5.0 % (w/v) NaCl (optimum at 0.5 %). The major respiratory quinone was ubiquinone 10 (Q-10) and the major cellular fatty acids were C18 : 1ω7c, C16 : 0, C18 : 1 2-OH and C18 : 0. The genomic DNA G+C content of strain YW11(T) was 69.6±0.5 mol%. The major polar lipids were phosphatidylglycerol, phosphatidylethanolamine, diphosphatidylglycerol, phosphatidylcholine, an unknown glycolipid and two unknown aminolipids. Phylogenetic analysis based on 16S rRNA gene sequence comparison revealed that strain YW11(T) was a member of the genus Roseomonas, and showed the highest sequence similarity to Roseomonas cervicalis KACC 11686(T) (97.9 %) and Roseomonas aestuarii KACC 19645(T) (97.8 %) and then to Roseomonas ludipueritiae KACC 13843(T) (96.9 %). Strain YW11(T) showed low DNA-DNA relatedness with R. cervicalis KACC 11686(T) (32.3±2.9 %), R. aestuarii KACC 16549(T) (28.2±2.6 %) and R. ludipueritiae KACC 13843(T) (30.2±2.6 %). Based on the results of phylogenetic analysis and DNA-DNA hybridization, the whole-cell fatty acid composition as well as biochemical characteristics, strain YW11(T) was clearly distinguished from all recognized species of the genus Roseomonas and should be assigned to a novel species of the genus Roseomonas, for which the name Roseomonas rhizosphaerae sp. nov. is proposed. The type strain is YW11(T) ( = KACC 17225(T) = CCTCC AB2013041(T)).

  5. Heterologous expression, biochemical characterization, and overproduction of alkaline α-amylase from Bacillus alcalophilus in Bacillus subtilis

    Directory of Open Access Journals (Sweden)

    Li Jianghua

    2011-10-01

    Full Text Available Abstract Background Alkaline α-amylases have potential applications for hydrolyzing starch under high pH conditions in the starch and textile industries and as ingredients in detergents for automatic dishwashers and laundries. While the alkaline α-amylase gains increased industrial interest, the yield of alkaline α-amylases from wild-type microbes is low, and the combination of genetic engineering and process optimization is necessary to achieve the overproduction of alkaline α-amylase. Results The alkaline α-amylase gene from Bacillus alcalophilus JN21 (CCTCC NO. M 2011229 was cloned and expressed in Bacillus subtilis strain WB600 with vector pMA5. The recombinant alkaline α-amylase was stable at pH from 7.0 to 11.0 and temperature below 40°C. The optimum pH and temperature of alkaline α-amylase was 9.0 and 50°C, respectively. Using soluble starch as the substrate, the Km and Vmax of alkaline α-amylase were 9.64 g/L and 0.80 g/(L·min, respectively. The effects of medium compositions (starch, peptone, and soybean meal and temperature on the recombinant production of alkaline α-amylase in B. subtilis were investigated. Under the optimal conditions (starch concentration 0.6% (w/v, peptone concentration 1.45% (w/v, soybean meal concentration 1.3% (w/v, and temperature 37°C, the highest yield of alkaline α-amylase reached 415 U/mL. The yield of alkaline α-amylase in a 3-L fermentor reached 441 U/mL, which was 79 times that of native alkaline α-amylase from B. alcalophilus JN21. Conclusions This is the first report concerning the heterologous expression of alkaline α-amylase in B. subtilis, and the obtained results make it feasible to achieve the industrial production of alkaline α-amylase with the recombinant B. subtilis.

  6. Rhizobium populi sp. nov., an endophytic bacterium isolated from Populus euphratica.

    Science.gov (United States)

    Rozahon, Manziram; Ismayil, Nurimangul; Hamood, Buayshem; Erkin, Raziya; Abdurahman, Mehfuzem; Mamtimin, Hormathan; Abdukerim, Muhtar; Lal, Rup; Rahman, Erkin

    2014-09-01

    . The type strain is K-38(T) ( = CCTCC AB 2013068(T) = NRRL B-59990(T) = JCM 19159(T)).

  7. Rhizobium tarimense sp. nov., isolated from soil in the ancient Khiyik River.

    Science.gov (United States)

    Turdahon, Maripat; Osman, Ghenijan; Hamdun, Maryam; Yusuf, Khayir; Abdurehim, Zumret; Abaydulla, Gulsumay; Abdukerim, Muhtar; Fang, Chengxiang; Rahman, Erkin

    2013-07-01

    A Gram-negative, non-motile, pale-yellow, rod-shaped bacterial strain, PL-41(T), was isolated from Populus euphratica forest soil at the ancient Khiyik River valley in Xinjiang Uyghur Autonomous Region, People's Republic of China. Strain PL-41(T) grew optimally at 30 °C and pH 7.0-8.0. The major quinone was Q-10. The predominant cellular fatty acids of strain PL-41(T) were summed feature 8 (comprising C18 : 1ω7c and C18 : 1ω6c), C16 : 0 and C19 : 0 cyclo ω8c. Polar lipids of strain PL-41(T) include two unidentified aminophospholipids (APL1, 2), two unidentified phospholipids (PL1, 2), phosphatidylcholine and three unidentified lipids (L1-3). Strain PL-41(T) showed 16S rRNA gene sequence similarity of 97.0-97.5 % to the type strains of recognized species of the genus Rhizobium. Phylogenetic analysis of strain PL-41(T) based on the sequences of housekeeping genes recA and atpD confirmed (similarities are less than 90 %) its position as a distinct species of the genus Rhizobium. The DNA G+C content was 57.8 mol%. DNA-DNA relatedness between strain PL-41(T) and the type strains of Rhizobium huautlense S02(T), Rhizobium alkalisoli CCBAU 01393(T), Rhizobium vignae CCBAU 05176(T) and Rhizobium loessense CCBAU 7190B(T) were 33.4, 22.6, 25.5 and 45.1 %, respectively, indicating that strain PL-41(T) was distinct from them genetically. Strain PL-41(T) also can be differentiated from these four phylogenetically related species of the genus Rhizobium by various phenotypic properties. On the basis of phenotypic properties, phylogenetic distinctiveness and genetic data, strain PL-41(T) is considered to represent a novel species of the genus Rhizobium, for which the name Rhizobium tarimense sp. nov. is proposed. The type strain is PL-41(T) ( = CCTCC AB 2011011(T) = NRRL B-59556(T)).

  8. Sphingomonas gei sp. nov., isolated from roots of Geum aleppicum.

    Science.gov (United States)

    Zhu, Lingfang; Si, Meiru; Li, Changfu; Xin, Kaiyun; Chen, Chaoqiong; Shi, Xu; Huang, Ruijun; Zhao, Liang; Shen, Xihui; Zhang, Lei

    2015-04-01

    A yellow-pigmented bacterium, designated strain ZFGT-11(T), was isolated from roots of Geum aleppicum Jacq. collected from Taibai Mountain in Shaanxi Province, north-west China, and was subjected to a taxonomic study by using a polyphasic approach. Cells of strain ZFGT-11(T) were Gram-stain-negative, strictly aerobic rods that were surrounded by a thick capsule and were motile by means of a single polar flagellum. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain ZFGT-11(T) was a member of the genus Sphingomonas and was closely related to Sphingomonas naasensis KACC 16534(T) (97.6% similarity), Sphingomonas kyeonggiense JCM 18825(T) (96.8%), Sphingomonas asaccharolytica IFO 15499(T) (96.7%) and Sphingomonas leidyi DSM 4733(T) (96.6%). The predominant respiratory quinone was ubiquinone-10 (Q-10) and the major cellular fatty acids were summed feature 8 (comprising C(18 : 1)ω7c and/or C(18 : 1)ω6c), C(17 : 1)ω6c, C(14 : 0) 2-OH, C(16 : 0) and C(15 : 0) 2-OH. The major polyamine of strain ZFGT-11(T) was sym-homospermidine. Phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylmonomethylethanolamine, phosphatidylcholine, sphingoglycolipid, two unidentified aminoglycolipids, two unidentified phospholipids and two unidentified lipids were detected in the polar lipid profile. The DNA G+C content was 66.8 mol%. DNA-DNA relatedness for strain ZFGT-11(T) with respect to its closest phylogenetic relative S. naasensis KACC 16534(T) was 26.2±4.8% (mean±SD). On the basis of data from the present polyphasic taxonomic study, strain ZFGT-11(T) is considered to represent a novel species of the genus Sphingomonas , for which the name Sphingomonas gei sp. nov. is proposed. The type strain is ZFGT-11(T) ( = CCTCC AB 2013306(T) = KCTC 32449(T) = LMG 27608(T)). © 2015 IUMS.

  9. Halomonas songnenensis sp. nov., a moderately halophilic bacterium isolated from saline and alkaline soils.

    Science.gov (United States)

    Jiang, Juquan; Pan, Yuanyuan; Hu, Shaoxin; Zhang, Xiaoxia; Hu, Baozhong; Huang, Haipeng; Hong, Shan; Meng, Jing; Li, Cheng; Wang, Kaibiao

    2014-05-01

    A moderately halophilic bacterium (strain NEAU-ST10-39T) was isolated from saline and alkaline soils in the oilfield of Daqing City, Heilongjiang Province, China. The strain was strictly aerobic, Gram-stain-negative, rod-shaped and motile by peritrichous flagella. Its colonies were yellow. It grew at NaCl concentrations of 0.2-15% (w/v) (optimum 4%, w/v), at temperatures of 4-40 °C (optimum 35 °C) and at pH 5-10 (optimum pH 7). It did not produce acids from sugars or alcohols. Its DNA G+C content was 57.4 mol%. Phylogenetic analyses based on 16S rRNA gene sequences and concatenated 16S rRNA, gyrB and rpoD gene sequences indicated that it belonged to the genus Halomonas in the class Gammaproteobacteria. The most phylogenetically related species were Halomonas axialensis, Halomonas meridiana and Halomonas aquamarina, whose types shared 98.3% (16S rRNA), 82.7% (gyrB) and 83.9-84.5% (rpoD) sequence similarity with strain NEAU-ST10-39T. The results of DNA-DNA hybridization assays showed 20±2%-50±1 % relatedness between strain NEAU-ST10-39T and the most closely related species including Halomonas axialensis DSM 15723T, Halomonas meridiana DSM 5425T, Halomonas aquamarina DSM 30161(T), Halomonas johnsoniae DSM 21197T, Halomonas stevensii DSM 21198T, Halomonas nanhaiensis CCTCC AB 2012911(T), Halomonas hamiltonii DSM 21196T and Halomonas arcis CGMCC 1.6494T. The major fatty acids were C18 : 1ω7c (47.2%), C16:1ω7c and/or C16:1ω6c (18.9%) and C16:0 (16.3%), the only respiratory quinone detected was ubiquinone 9 and polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, two unknown phospholipids and three unknown lipids. The new isolate is proposed to represent a novel species with the name Halomonas songnenensis sp. nov., NEAU-ST10-39T (=CGMCC 1.12152T=DSM 25870T) being the type strain.

  10. Streptomyces luozhongensis sp. nov., a novel actinomycete with antifungal activity and antibacterial activity.

    Science.gov (United States)

    Zhang, Renwen; Han, Xiaoxue; Xia, Zhanfeng; Luo, Xiaoxia; Wan, Chuanxing; Zhang, Lili

    2017-02-01

    A novel actinomycete strain, designated TRM 49605(T), was isolated from a desert soil sample from Lop Nur, Xinjiang, north-west China, and characterised using a polyphasic taxonomic approach. The strain exhibited antifungal activity against the following strains: Saccharomyces cerevisiae, Curvularia lunata, Aspergillus flavus, Aspergillus niger, Fusarium oxysporum, Penicillium citrinum, Candida albicans and Candida tropicalis; Antibacterial activity against Bacillus subtilis, Staphylococcus epidermidis and Micrococcus luteus; and no antibacterial activity against Escherichia coli. Phylogenetic analysis based on 16S rRNA gene sequences affiliated strain TRM 49605(T) to the genus Streptomyces. Strain TRM 49605(T) shows high sequence similarities to Streptomyces roseolilacinus NBRC 12815(T) (98.62 %), Streptomyces flavovariabilis NRRL B-16367(T) (98.45 %) and Streptomyces variegatus NRRL B-16380(T) (98.45 %). Whole cell hydrolysates of strain TRM 49605(T) were found to contain LL-diaminopimelic acid as the diagnostic diamino acid and galactose, glucose, xylose and mannose as the major whole cell sugars. The major fatty acids in strain TRM 49605(T) were identified as iso C16:0, anteiso C15:0, C16:0 and Summed Feature 5 as defined by MIDI. The main menaquinones were identified as MK-9(H4), MK-9(H6), MK-9(H8) and MK-10(H6). The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylinositol and phosphatidylinositol mannoside. The G+C content of the genomic DNA was determined to be 71.2 %. The DNA-DNA relatedness between strain TRM 49605(T) and the phylogenetically related strain S. roseolilacinus NBRC 12815(T) was 60.12 ± 0.06 %, which is lower than the 70 % threshold value for delineation of genomic prokaryotic species. Based on the phenotypic, chemotaxonomic and phylogenetic data, strain TRM 49605(T) (=CCTCC AA2015026(T) = KCTC 39666(T)) should be designated as the type strain of a novel species of the genus

  11. 柠檬酸钠促进S-腺苷蛋氨酸和谷胱甘肽联合高产%Increased Co-production of S-adenosylmethionine and Glutathione by Sodium Citrate Addition

    Institute of Scientific and Technical Information of China (English)

    王玉磊; 朱健; 卫功元; 许宏庆; 汪成富

    2013-01-01

    S-adenosylmethionine (SAM) and glutathione (GSH) are both important small S-contained compounds in cells.The effects of sodium citrate on the fermentative co-production of SAM and GSH with Candida utilis CCTCC M 209298 were investigated in flasks.Sodium citrate was found to be beneficial for the high co-production of SAM and GSH.The response surface analysis was applied in the optimization of sodium citrate concentration and addition time,and a strategy of 10 g/L sodium citrate addition at 6 h was predicted by a statistical model and verified to be the best approach for increased co-production of SAM and GSH.Based on the results derived from the kinetic analysis on the batch fermentation processes,intracellular levels of NADH and ATP could be significantly improved by sodium citrate,and which in turn provided essential energy substance needed for the over-production of SAM and GSH.The results also provide a potential approach for efficient production of analogical useful chemicals biosynthesized with the consumption of energy.%考察了柠檬酸钠对S-腺苷蛋氨酸(SAM)和谷胱甘肽(GSH)联产发酵的影响,发现柠檬酸钠有利于SAM和GSH的联合高产.采用响应面分析法对柠檬酸钠浓度及其添加时间进行优化,模型预测和验证实验结果均表明在联产发酵6h时一次性添加10 g/L柠檬酸钠的效果最佳.通过对SAM和GSH联产发酵过程进行分析,发现柠檬酸钠能够显著提高胞内NADH和ATP的水平,为SAM和GSH的过量合成提供了足够的能量物质,也为类似耗能化合物的生物合成及其发酵高产提供了可行的优化策略.

  12. Brachybacterium horti sp. nov., isolated from garden soil.

    Science.gov (United States)

    Singh, Hina; Du, Juan; Yang, Jung-Eun; Shik Yin, Chang; Kook, MooChang; Yi, Tae-Hoo

    2016-01-01

    A bacterial strain, THG-S15-4T, was isolated from garden soil taken from the Guro-gu district of Seoul, Republic of Korea. Strain THG-S15-4T was Gram-stain-positive, facultatively anaerobic, coccus-shaped and non-motile, forming white colonies. The strain grew optimally at 25-37 °C, at pH 7.0 and in the presence of 0-2.0 % (w/v) NaCl. Phylogenetic analysis, based on 16S rRNA gene sequences, showed that strain THG-S15-4T was affiliated to species of the genus Brachybacterium, and the most closely related species were Brachybacterium rhamnosum KCTC 9917 T (98.5 % sequence similarity) and Brachybacterium squillarum KCTC 19899T (96.9 % sequence similarity). The DNA-DNA relatedness between strain THG-S15-4T and B. rhamnosum KCTC 9917 T was found to be below 20.0 %. The DNA G+C content was determined to be 69.5 mol%. The major isoprenoid quinone detected was MK-7. Strain THG-S15-4T was characterized chemotaxonomically as having meso-diaminopimelic acid in the cell-wall peptidoglycan. The polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol, unidentified glycolipids and an unidentified polar lipid. The major fatty acids were found to be iso-C16 : 0 and anteiso-C15 : 0. The results of physiological and biochemical tests enabled strain THG-S15-4T to be differentiated phenotypically from species of the genus Brachybacterium with validly published names. Therefore, it is suggested that this newly isolated organism represents a novel species, for which the name Brachybacterium horti sp. nov. is proposed. The type strain is THG-S15-4T ( = KCTC 39563T = CCTCC AB 2015116T).

  13. Thauera humireducens sp. nov., a humus-reducing bacterium isolated from a microbial fuel cell.

    Science.gov (United States)

    Yang, Gui-Qin; Zhang, Jun; Kwon, Soon-Wo; Zhou, Shun-Gui; Han, Lu-Chao; Chen, Ming; Ma, Chen; Zhuang, Li

    2013-03-01

    A Gram-negative, rod-shaped, non-spore-forming bacterium, designated SgZ-1(T), was isolated from the anode biofilm of a microbial fuel cell. The strain had the ability to grow under anaerobic condition via the oxidation of various organic compounds coupled to the reduction of anthraquione-2,6-disulfonate (AQDS) to anthrahydroquinone-2,6-disulfonate (AHQDS). Growth occurred in TSB in the presence of 0-5.5 % (w/v) NaCl (optimum 0-1 %), at 10-45 °C (optimum 25-37 °C) and at pH 6.0-10.0 (optimum 8.0-8.5). Based on 16S rRNA gene sequence similarity, strain SgZ-1(T) belonged to the genus Thauera. The highest level of 16S rRNA gene sequences similarity (96.7 %) was found to be with Thauera aminoaromatica S2(T) and Thauera selenatis AX(T), and lower values were obtained when compared with other recognized Thauera species. Chemotaxonomic analysis revealed that strain SgZ-1(T) contained Q-8 as the predominant quinone, and putrescine and 2-hydroxyputrescine as the major polyamines. The major cellular fatty acids (>5 %) were C16 : 1ω6c and/or C16 : 1ω7c (44.6 %), C16 : 0 (18.8 %), and C18 : 1ω6c and/or C18 : 1ω7c (12.7 %). Based on its phenotypic and phylogenetic properties, chemotaxonomic analysis and the results of physiological and biochemical tests, strain SgZ-1(T) ( = KACC 16524(T) = CCTCC M 2011497(T)) was designated the type strain of a novel species of the genus Thauera, for which the name Thauera humireducens sp. nov. was proposed.

  14. 添加大豆分离蛋白对羊乳干酪风味物质的影响%Influence of soy protein isolate addition to volatile compounds of goat milk cheese

    Institute of Scientific and Technical Information of China (English)

    祝敏; 杜金华; 吴晓英

    2013-01-01

    In this study,with the addition of different proportion (0,4% and 10% m/m) of fresh goat milk and Soy protein isolate (SPI),high concentration direct-set lactic bacterial including Lactococcus lactis and Leuconostoc,and Propionibacterium freudenreichii ssp.Shermanii were used as the raw material,main culture and secondary culture,respectively,to make goat milk and SPI mixed cheese.Volatile compounds of the cheese were enriched by solid-phase microextraction (SPME) and determined by GC/MS after the cheese maturation.The influence of SPI to volatile compounds of cheese was studied and the appropriate ratio of SPI addition was determined.The result indicated that the major volatile compounds of cheese were acid compounds,following the ketone,alcohol,aldehyde,and ester compounds.SPI addition reduced the contents of acid and ketone compounds of cheese significantly and increased the contents of alcohol compounds.The complexity of the cheese flavor was enhanced when SPI addition was 4%,with a rich cream,butter aroma and less special flavor of goat milk,phenol and pyrrole compounds appeared as well.%以新鲜羊乳为原料,直投式乳酸菌混合菌种为主发酵剂,费氏丙酸杆菌为次级发酵剂,添加不同比例(0、4%、10% m/m)大豆分离蛋白(soy protein isolate,SPI)制作混合型干酪.完全成熟之后利用固相微萃取(SPME)萃取富集干酪挥发性物质、气相色谱-质谱联用(GC/MS)检测干酪风味物质.以研究添加大豆分离蛋白对干酪风味物质的影响,确定适宜添加比例.实验结果表明:干酪中主要风味物质是酸类,其次是酮,醇,醛,酯类化合物;添加大豆分离蛋白使干酪中酸类、酮类化合物种类及含量明显减少;醇类化合物增加.大豆分离蛋白添加比为4%时,干酪有浓郁的奶香及黄油香气,羊乳膻味减少,并检出苯酚、吡咯等化合物,提高了干酪风味物质种类的丰富性.

  15. Inoculants for ensiling low-dry matter corn crop: a midlactation cow perspective.

    Science.gov (United States)

    Nikkhah, A; Ghaempour, A; Khorvash, M; Ghorbani, G R

    2011-10-01

    In many regions, optimum dry matter (DM) content of corn crop pre-ensilage cannot be ensured for management, agronomical and climatic reasons. Under such conditions, corn crops are harvested at low DM, and are easily exposed to unfavourable fermentation pathways and plant spoilage and wastage. Thus, it is a major question for dairy agriculturists whether certain microbial inoculants application to low-DM corn crop pre-ensilage affects silage quality and cow performance. The objective was to determine effects of adding microbial inoculants to low-DM corn crop at ensiling on silage quality, rumen fermentation and milk production of eight Holstein cows fed the treated silages. Whole corn plant was harvested at milk stage of maturity with 204 g DM/kg of fresh crop, cut to a theoretical particle length of 2 cm, filled in 60 t bunker silos, and treated layer by layer with either no inoculant (control), inoculant 'E' (100 000 cfu/g of fresh crop) containing mainly Lactobacillus plantarum, inoculant 'B' (100 000 cfu) containing mainly Pediococcus pentosanus, Lactobacillus plantarum and Propionibacter freudenreichii or a mixture of inoculants 'E' and 'B' (200,000 cfu). Inoculants were mixed with water and sprayed on thin layers of corn chops layer by layer followed by rolling to ensure proper oxygen outage and even microbial distribution throughout the plants. Eight multiparous lactating Holstein cows at 100 ± 20.5 days in milk were used in a replicated 4 × 4 Latin square design with four 20-day periods including 14 days of adaptation and 6 days of sampling. Dietary treatments were mixed rations containing corn silages with or without the inoculants. The basal diet contained 32.9% corn silage, 14.3% alfalfa hay and 52.8% concentrate on a DM basis. Inoculants did not affect silage pH or content of DM, CP, lactate, acetate, ash and total volatile fatty acids (VFA). Applying 'B' to corn crop resulted in higher water soluble carbohydrates (47.7 g/kg vs 29.8 g/kg) and lower

  16. Chryseobacterium chengduensis sp. nov. isolated from the air of captive giant panda enclosures in Chengdu, China*#

    Institute of Scientific and Technical Information of China (English)

    Rong SHE; Zhong-xiang HAO; Lu LUO; Hong LIAO; Zhen-rong CHEN; Guo-quan HAN; San-jie CAO; Rui WU; Qi-gui YAN; Rong HOU; Cai-fang WEN; Li-xin XI; Shan ZHAO

    2016-01-01

    A Gram-negative, aerobic, non-motile, rod-shaped bacterial strain, designated 25-1T,was isolated from the air inside giant panda enclosures at the Chengdu Research Base of Giant Panda Breeding, China. Strain 25-1T grew optimaly at pH 7.0–8.0, at 28–30 °C and in the presence of NaCl concentrations from 0.0% to 0.5 %. 16S rRNA gene sequence analysis indicated that strain 25-1T belongs to the genusChryseobacterium within the familyFlavo-bacteriaceae and is related most closely toC.carnisG81T(96.4% similarity),C. lathyri RBA2-6T (95.8% similarity), and C. zeae JM1085T (95.8% similarity). Its genomic DNA G+C molar composition was 36.2%. The major celular fatty acids wereiso-C15:0 (44.0%),iso-C17:0 3OH (19.8%) and C16:1ω7c/16:1ω6c (12.7%). The only isoprenoid quinone was menaquinone 6 (MK-6). The major polar lipids were phosphatidylethanolamine, two unidentified amino lipids and two unidentified lipids. The DNA–DNA relatedness between strain 25-1TandC. lathyri RBA2-6T was 38%. Phenotypic, genotypic, and phylogenetic characteristics indicated that strain 25-1T is a novel member of the genusChryseobacterium, for which the nameC. chengduensis sp. nov. is proposed. The type strain is 25-1T(CCTCC AB2015133T=DSM 100396T).%题目:分离自成都大熊猫兽舍空气中的一株金黄杆菌属新种的鉴定目的:鉴定菌株25-1T是否是金黄杆菌属的一个新种。创新点:首次从空气中分离到金黄杆菌属的新种。方法:革兰氏染色镜检;磷钨酸染色然后透射电镜观察菌株25-1T形态结构;全自动生理生化鉴定系统(Phoenix™-100)与传统生理生化反应管相结合;H890气象色谱仪进行脂肪酸组分分析;薄板双相层析进行极性脂组分分析;反相高压液相色谱分析法进行呼吸琨组分分析;熔解温度法检测G+C摩尔含量;16S rRNA序列测定及系统发生分析。结论:根据传统特征分类研究结果(形态特征、培养特性和生理生化特征)

  17. Improvement of fermentor facility to enhance liquid fermentation level of membrane-bound lipase produced by Rhizopus chinensis%发酵罐装置改进提高华根霉膜结合脂肪酶液态发酵水平

    Institute of Scientific and Technical Information of China (English)

    朱增亮; 王栋; 徐岩

    2013-01-01

    研究主要利用7L通风搅拌式发酵罐,基于华根霉膜结合脂肪酶液态发酵的生物学特性和要求对发酵装置进行改进,促进菌体聚集和对橄榄油的利用,以提高其发酵产酶水平.与许多丝状真菌液态发酵类似,华根霉脂肪酶的液态发酵水平与菌丝形态密切相关.在发酵罐中加入套筒改变发酵液流型,促使华根霉菌丝在套筒上聚集生长;添加斜叶搅拌桨增强轴向流,促进橄榄油在发酵液中的分散和利用.最终华根霉膜结合脂肪酶单位菌体酶活达到283.16 U/g,菌体干重量达到14.01 g/L,发酵单位体积总活力达到3967.07 U/L,是未改进前发酵水平的4.2倍.%Membrane-bound lipase with high synthetic activity produced by Rhizopus chinensis (CCTCC 2010021) could catalyze the synthesis of ester in non-aqueous phase and had potential applications in the production of flavor and fragrance compounds, biodiesel, and so on. However, it was difficult to improve the fermentation level of this lipase because of the culture characteristics of this filamentous fungus. Based on the biological characteristics and requirements of Rhizopus chinensis, the improvement of 7 L mechanical stirring fermentor facilities was carried out to promote the aggregation of mycelium and the olive oil was used for increasing the lipase fermentation level. Similar to other filamentous fungus, the lipase liquid fermentation level with Rhizopus chinensis was closely related to the mycelium morphologies. The aggregate growth of mycelia was achieved by changing the fermentation broth rheology with a sleeve; Olive oil in the fermentation broth was more evenly distributed due to the increase of axial flow of broth by adding an oblique leaves impeller, and which improved the induction effect. Ultimately the synthetic activity of membrane-bound lipase produced by Rhizopus chinensis reached 283.16 U/g, dry cell weight reached 14.01 g/L, the total activity per unit volume of

  18. Phylogenetic analyses of the genus Glaciecola: emended description of the genus Glaciecola, transfer of Glaciecola mesophila, G. agarilytica, G. aquimarina, G. arctica, G. chathamensis, G. polaris and G. psychrophila to the genus Paraglaciecola gen. nov. as Paraglaciecola mesophila comb. nov., P. agarilytica comb. nov., P. aquimarina comb. nov., P. arctica comb. nov., P. chathamensis comb. nov., P. polaris comb. nov. and P. psychrophila comb. nov., and description of Paraglaciecola oceanifecundans sp. nov., isolated from the Southern Ocean.

    Science.gov (United States)

    Shivaji, Sisinthy; Reddy, Gundlapally Sathyanarayana

    2014-09-01

    Phylogenetic analyses of the genus Glaciecola were performed using the sequences of the 16S rRNA gene and the GyrB protein to establish its taxonomic status. The results indicated a consistent clustering of the genus Glaciecola into two clades, with significant bootstrap values, with all the phylogenetic methods employed. Clade 1 was represented by seven species, Glaciecola agarilytica, G. aquimarina, G. arctica, G. chathamensis, G. mesophila, G. polaris and G. psychrophila, while clade 2 consisted of only three species, Glaciecola nitratireducens, G. pallidula and G. punicea. Evolutionary distances between species of clades 1 and 2, based on 16S rRNA gene and GyrB protein sequences, ranged from 93.0 to 95.0 % and 69.0 to 73.0 %, respectively. In addition, clades 1 and 2 possessed 18 unique signature nucleotides, at positions 132, 184 : 193, 185 : 192, 230, 616 : 624, 631, 632, 633, 738, 829, 1257, 1265, 1281, 1356 and 1366, in the 16S rRNA gene sequence and can be differentiated by the occurrence of a 15 nt signature motif 5'-CAAATCAGAATGTTG at positions 1354-1368 in members of clade 2. Robust clustering of the genus Glaciecola into two clades based on analysis of 16S rRNA gene and GyrB protein sequences, 16S rRNA gene sequence similarity of ≤95.0 % and the occurrence of signature nucleotides and signature motifs in the 16S rRNA gene suggested that the genus should be split into two genera. The genus Paraglaciecola gen. nov. is therefore created to accommodate the seven species of clade 1, while the name Glaciecola sensu stricto is retained to represent species of clade 2. The species of clade 1 are transferred to the genus Paraglaciecola as Paraglaciecola mesophila comb. nov. (type strain DSM 15026(T) = KMM 241(T)), P. agarilytica comb. nov. (type strain NO2(T) = KCTC 12755(T) = LMG 23762(T)), P. aquimarina comb. nov. (type strain GGW-M5(T) = KCTC 32108(T) = CCUG 62918(T)), P. arctica comb. nov. (type strain BSs20135(T) = CCTCC