• Title/Summary/Keyword: Microbial Biotechnology

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Direct Monitoring of Membrane Fatty Acid Changes and Effects on the Isoleucine/Valine Pathways in an ndgR Deletion Mutant of Streptomyces coelicolor

  • Tae-Rim Choi;Suk Jin Oh;Jeong Hyeon Hwang;Hyun Jin Kim;Nara Shin;Jeonghee Yun;Sang-Ho Lee;Shashi Kant Bhatia;Yung-Hun Yang
    • Journal of Microbiology and Biotechnology
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    • v.33 no.6
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    • pp.724-735
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    • 2023
  • NdgR, a global regulator in soil-dwelling and antibiotic-producing Streptomyces, is known to regulate branched-chain amino acid metabolism by binding to the upstream region of synthetic genes. However, its numerous and complex roles are not yet fully understood. To more fully reveal the function of NdgR, phospholipid fatty acid (PLFA) analysis with gas chromatography-mass spectrometry (GC-MS) was used to assess the effects of an ndgR deletion mutant of Streptomyces coelicolor. The deletion of ndgR was found to decrease the levels of isoleucine- and leucine-related fatty acids but increase those of valine-related fatty acids. Furthermore, the defects in leucine and isoleucine metabolism caused by the deletion impaired the growth of Streptomyces at low temperatures. Supplementation of leucine and isoleucine, however, could complement this defect under cold shock condition. NdgR was thus shown to be involved in the control of branched-chain amino acids and consequently affected the membrane fatty acid composition in Streptomyces. While isoleucine and valine could be synthesized by the same enzymes (IlvB/N, IlvC, IlvD, and IlvE), ndgR deletion did not affect them in the same way. This suggests that NdgR is involved in the upper isoleucine and valine pathways, or that its control over them differs in some respect.

Removal Behavior of Biological Nitrogen and Phosphorus and Prediction of Microbial Community Composition with Its Function, in an Anaerobic-Anoxic System form Weak Sewage

  • LEE, JIN WOO;EUI SO CHOI;KYUNG IK GIL;HAN WOONG LEE;SANG HYON LEE;SOO YOOUN LEE;YONG KEUN PARK
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.994-1001
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    • 2001
  • An easier way of understanding the BNR system was proposed from the study on substrate, nutrient removal tendency, microbial community and its metabolic function by applying the municipal settled sewage. During the anaerobic period, the phosphorus release rate per VFACOD we varied depending on the phosphorus content in the sludge. When the phosphorus content in the sludge was $6\%$ VSS, according to influent VFACOD, the phosphorus release rate and PHA production were $0.35 gPO_4P/gVFACOD$ and 1.0 gPHA/gVFACOD, respectively. The $NO_3N$ requirement for the phosphorus uptake as an electron acceptor was about $0.5 gNO_3N/gPO_4P_{uptake}$ based on the proposed equation with PHA, biomass, production, and the concentration of phosphorus release/uptake. Bacterial-community analysis of the sludge, as determined by FISH and 16SrDNA characterization FISH, revealed that the beta-subclass proteobacteria were the most abundant group ($27.9\%$ of the proteobacteria-specific probe EUB338), and it was likely that representative of the beta-subclass played key roles in activated sludge. The next dominant group found was the gamma-protebacteria ($15.4\%$ of probe EUB338). 16S rDNA clone library analysis showed that the members of${\beta}$- and ${\gamma}$-proteobacteria were also the most abundant groups, and $21.5\%$ (PN2 and PN4) and $15.4\%$ (PN1 and PN5) of total clones were the genera of denitrifying bacteria and PAO, respectively. Prediction of the microbial community composition was made with phosphorus content (Pv, $\%$ P/VSS) in wasted sludge and profiles of COD, PHA, $PO_4P,\;and\;NO_3N$ in an anaerobic-anoxic SBR unit. Generally, the predicted microbial composition based upon metabolic function, i.e., as measured by stoichiometry, is fairly similar to that measure by the unculturable dependent method. In this study, a proposal was made on he microbial community composition that was more easily approached to analyze the reactor behavior.

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Selection and Directed Evolution of New Microbial Biocatalysts and Their Application to Organic Synthesis

  • Asano, Yasuhisa
    • Journal of Applied Biological Chemistry
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    • v.43 no.4
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    • pp.207-210
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    • 2000
  • As a typical example of the screening for a microbial biocatalyst from nature, isolation of nitrilesynthesizing microorganisms, characterization of a new enzyme aldoxime dehydratase, and its function in the aldoxime-nitrile pathway are introduced. Catalytic properties of some of our enzymes were improved through a direct evolutionary approach.

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Metabolic Fingerprinting of Food Wastewater Treatment System (식품폐수 처리 단계별 미생물 대사지문)

  • Yoo, Ki-Hwan;Lee, Sang-Hyeon;Lee, Dong-Geun
    • Journal of Environmental Health Sciences
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    • v.34 no.4
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    • pp.327-332
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    • 2008
  • To determine structure and activities of microbial communities in a food wastewater treatment system, biofilm of RABC (rotating activated Bacillus contactor) and samples of aeration tanks were analyzed. Heterotrophic bacterial concentrations were similar between biofilm and stage 1 aeration tank and decreased 2-log at stage 3 aeration tank as dissolved oxygen decreased, however portions of Bacillus groups were increased at stage 3 aeration tank. It was revealed by quantitative and qualitative analysis of metabolic fingerprinting patterns of Biolog GN2 plate that RABC represented much higher activities and a different microbial community structure compared to aeration tanks. Metabolic fingerprinting showed the carbon sources that isolated Bacillus groups could or could not use, were used similarly meaning that not only Bacillus groups but also other microbial groups would contribute to the treatment of wastewater.

Microbial Community of Healthy Thai Vegetarians and Non-Vegetarians, Their Core Gut Microbiota, and Pathogen Risk

  • Ruengsomwong, Supatjaree;La-ongkham, Orawan;Jiang, Jiahui;Wannissorn, Bhusita;Nakayama, Jiro;Nitisinprasert, Sunee
    • Journal of Microbiology and Biotechnology
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    • v.26 no.10
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    • pp.1723-1735
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    • 2016
  • Pyrosequencing analysis of intestinal microflora from healthy Thai vegetarians and non-vegetarians exhibited 893 OTUs covering 189 species. The strong species indicators of vegetarians and non-vegetarians were Prevotella copri and Bacteroides vulgatus as well as bacteria close to Escherichia hermanii with % relative abundance of 16.9 and 4.5-4.7, respectively. Core gut microbiota of the vegetarian and non-vegetarian groups consisted of 11 and 20 different bacterial species, respectively, belonging to Actinobacteria, Firmicutes, and Proteobacteria commonly found in both groups. Two species, Faecalibacterium prausnitzii and Gemmiger formicilis, had a prevalence of 100% in both groups. Three species, Clostridium nexile, Eubacterium eligens, and P. copri, showed up in most vegetarians, whereas more diversity of Collinsella aerofaciens, Ruminococcus torques, various species of Bacteroides, Parabacteroides, Escherichia, and different species of Clostridium and Eubacterium were found in most non-vegetarians. Considering the correlation of personal characters, consumption behavior, and microbial groups, the age of non-vegetarians showed a strong positive correlation coefficient of 0.54 (p = 0.001) to Bacteroides uniformis but exhibited a moderate one to Alistipes finegoldii and B. vulgatus. Only a positive moderate correlation of body mass index and Parabacteroides distasonis appeared. Based on the significant abundance of potential pathogens, the microbiota of the non-vegetarian group showed an abundance of potential pathogen varieties of Bilophila wadsworthia, Escherichia coli, and E. hermannii, whereas that of the vegetarian group served for only Klebsiella pneumoniae. These results implied that the microbiota of vegetarians with high abundance of P. copri and low potential pathogen variety would be a way to maintain good health in Thais.

Isolation, Purification, and Partial Characterization of an AMP Deaminase from Saccharomyces cerevisiae D

  • Kim, Myung-Hee;Lee, Jung-Kee;Kim, Hyung-Kwoun;Oh, Tae-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.9 no.4
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    • pp.429-435
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    • 1999
  • An adenosine 5'-monophosphate deaminase (AMP aminohydrolase, EC 3.5.4.6) was purified to homogeneity from the cell-free extract of Saccharomyces cerevisiae DKCTC7248. The molecular mass of subunit was estimated to be 80 kDa on SDS-PAGE, and that of the holoenzyme was shown to be 240 kDa by gel filtration. The isoelectric point of the enzyme (AMP deaminase D) was determined to be 6.2. The AMP deaminase D was specific towards AMP with an apparent $K_m$ value of 4.1 mM and a Hill coefficient, $n_H$, of 2.2. Both ATP and ADP were positive allosteric effectors of the AMP deaminase D: The apparent $K_{m}$ was decreased to 1.6 mM and 3.3 mM in the presence of 0.1 mM ATP and ADP, respectively, lowering $n_{H}$ to 1.0. Univalent cations like $K^+, Na^+ and Li^ +$ activated the enzyme but some divalent cations such as $Cu^{ 2+} and Cd^{2+}$ showed strong inhibitory effects. This enzyme displayed optimum activity at $30^{\circ}C$ and pH 7.0. In addition, it was stable up to $45^{\circ}C$ and over a wide pH range(pH 5.5-9.0). Amino acid sequences of its N-terminal region were analyzed to be ADYKMQMFADDA.

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