• Title/Summary/Keyword: D-serine utilization

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Metagenome Resource for D-Serine Utilization in a DsdA-Disrupted Escherichia coli

  • Lim, Mi-Young;Lee, Hyo-Jeong;Kim, Pil
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.374-378
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    • 2011
  • To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

Characterization of Microbial Communities in a Groundwater Contaminated with Landfill Leachate using a Carbon Substrate Utilization Assay (탄소원 이용도 평가를 활용한 매립지 침출수로 오염된 지하수의 미생물 군집 특성 해석)

  • Koo, So-Yeon;Kim, Ji-Young;Kim, Jai-Soo;Go, Kyung-Seok;Lee, Sang-Don;Cho, Kyung-Suk;Go, Dong-Chan
    • Journal of Soil and Groundwater Environment
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    • v.12 no.2
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    • pp.20-26
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    • 2007
  • The microbial community properties of groundwater samples contaminated with landfill leachates were examined using Ecoplate including 31 sole carbon sources. The samples were KSG1-12 (leachate), KSG1-16 (treated leachate), KSG1-07 (contaminated groundwater), KSG1-08 (contaminated groundwater), and KSG1-13 (uncontaminated groundwater). Among the carbon sources used as substrates, 2-hydroxy benzoic acid, D,L-$\alpha$-glycerol phosphate, and D-malic acid were not utilized in any sample, while D-xylose, D-galacturonic acid, L-aspargine, tween 80, and L-serine were utilized in all 5 samples. The rest of substrates showed very different patterns among the samples. Average well color development (AWCD) analysis demonstrated that the potential activity on 31 substrates was in the order of KSG1-16 > KSG1-12 > KSG1-07 > KSG-08 > KSG1-13, which generally agrees with the degree of pollution, except KSG1-16. Principal component analysis (PCA) on similarity between samples showed two groups (KSG1-12, -07 and -08 vs KSG1-16 and -13), coinciding with contaminated and uncontaminated groups. Shannon index showed that the microbial diversities were similar among the samples.

Analysis and Quantification of Ammonia-Oxidizing Bacteria Community with amoA Gene in Sewage Treatment Plants

  • Hong, Sun Hwa;Jeong, Hyun Duck;Jung, Bongjin;Lee, Eun Young
    • Journal of Microbiology and Biotechnology
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    • v.22 no.9
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    • pp.1193-1201
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    • 2012
  • The analysis and quantification of ammonia-oxidizing bacteria (AOB) is crucial, as they initiate the biological removal of ammonia-nitrogen from sewage. Previous methods for analyzing the microbial community structure, which involve the plating of samples or culture media over agar plates, have been inadequate because many microorganisms found in a sewage plant are unculturable. In this study, to exclusively detect AOB, the analysis was carried out via denaturing gradient gel electrophoresis using a primer specific to the amoA gene, which is one of the functional genes known as ammonia monooxygenase. An AOB consortium (S1 sample) that could oxidize an unprecedented 100% of ammonia in 24 h was obtained from sewage sludge. In addition, real-time PCR was used to quantify the AOB. Results of the microbial community analysis in terms of carbon utilization ability of samples showed that the aeration tank water sample (S2), influent water sample (S3), and effluent water sample (S4) used all the 31 substrates considered, whereas the AOB consortium (S1) used only Tween 80, D-galacturonic acid, itaconic acid, D-malic acid, and $_L$-serine after 192 h. The largest concentration of AOB was detected in S1 ($7.6{\times}10^6copies/{\mu}l$), followed by S2 ($3.2{\times}10^6copies/{\mu}l$), S4 ($2.8{\times}10^6copies/{\mu}l$), and S3 ($2.4{\times}10^6copies/{\mu}l$).

Comparative analysis of volatile and non-volatile flavor compounds in rice paste made by α-amylase according to cultivars (α-amylase를 이용하여 제조한 쌀 페이스트의 품종에 따른 휘발성비휘발성 향미성분 비교분석)

  • Son, Eun Young;Kim, Hye Won;Kim, Sun Ah;Lee, Sang Mi;Paek, Se Hee;Kim, Sun Hee;Seo, Yong Ki;Park, Hye-Young;Oh, Sea-Kwan;Kim, Young-Suk
    • Journal of Applied Biological Chemistry
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    • v.60 no.3
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    • pp.283-291
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    • 2017
  • Rice that the half of population in the world eats as a staple food is mostly produced and consumed in Asia. However, its consumption is nowadays decreasing mainly due to diet diversity. Accordingly, some attempts are in demand to enhance the utilization of rice. In this study, profiling of volatile and non-volatile flavor components in rice pastes obtained by ${\alpha}$-amylase was performed and compared according to nine different rice cultivars domestically cultivated in Korea using gas chromatography-mass spectrometry combined by solid phase microextraction and gas chromatography-time of flight-mass spectrometry after a derivatization, respectively. In total, 46 volatile compounds identified included 6 alcohols, 6 aldehydes, 4 esters, 4 furan derivatives, 4 ketones, 1 acid, 1 sulfur-containing compound, 7 hydrocarbons, 5 aromatics and 8 terpenes. The non-volatile flavor components found were composed of 12 amino acids, 6 sugars and 4 sugar alcohols. In principal component analysis, rice paste samples could be discriminated according to cultivars on the score plots of volatile and non-volatile flavor compounds. In particular, some volatile compounds such as pentanal and 4,7-dimethylundecane could contribute to distinguish Senong 17 white and Senong 17 brown, whereas ethanol, 6-methylhep-5-en-2-one, and tridecane could be highly related to the discrimination of Iipum from other cultivars. Among non-volatile compounds, some amino acids such as glycine, serine and ${\gamma}$-aminobutyric acid and some sugars such as sucrose and fructose were mainly responsible for the discrimination of Danmi from the other cultivars. On the other hand, galactose, arabitol and mannose were more closely related to Senong 17 white than Senong 17 brown.

Effects of Dietary Levels of Corn Distiller's Dried Grains with Solubles and Phytase on Performance and Nutrient Utilization of Broilers (옥수수 주정박의 수준과 Phytase의 첨가 유무가 육계의 생산성과 영양소 이용률에 미치는 영향)

  • Hong, E.C.;Na, S.H.;Yu, D.C.;Kim, H.K.;Park, M.N.;Jung, K.C.;Choo, H.J.;Park, H.D.;Chung, W.T.;HwangBo, J.
    • Korean Journal of Poultry Science
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    • v.36 no.3
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    • pp.247-255
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    • 2009
  • This work was conducted to investigate the effects of the addition level of Corn distiller's dried grains with solubles (CDDGS) and phytase in broiler diets on the broiler performance and the ileal amino acid digestibility. Three eighty four Ross broiler (one-day-old) were assigned randomly to 8 treatments groups (12 birds/pen, 4 pens/treatment) as $4{\times}2$ complex factors of CDDGS 4 treatments (0, 5, 10 and 15%) and phytase 2 treatments (0, 500 FTU/kg), and fed experimental diets for 6 weeks (starter, 0~2 wk; grower, 2~4 wk; finisher, 4~6 wk). After the feeding trial was finished, 12 broilers (6 weeks) with similar weights were selected from all treatments to investigate the digestibility of nutrients and amino acids for the first week. The experimental diets contained similar ME and CP of all treatments. Body weight gains and feed intakes were high at phytase treatments (P>0.05), but there was no difference in feed intake of the chicks. The amino acid digestibilities of diets were improved for iso-leucine, lysine, threonine, valine, aspartic acid, proline, serine and tyrosine (P<0.05). Finally, this work showed that growth performance and amino acid digestibility were improved by the addition of CDDGS and phytase in broiler diets.