• Title/Summary/Keyword: enrichment culture

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Screening of Exiguobacterium acetylicum from Soil Samples Showing Enantioselective and Alkalotolerant Esterase Activity

  • Hwang Bum-Yeol;Kim Ji-Hyun;Kim Juhan;Kim Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.367-371
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    • 2005
  • About 3,000 bacterial colonies with esterase activities were isolated from soil samples by enrichment culture and halo-size on Luria broth-tributyrin (LT) plates. The colonies were assayed for esterase activity in microtiter plates using enantiomerically pure (R)- and (S)-2-phenylbutyric acid resorufin ester (2PB-O-res) as substrates. Two enantioselective strains (JH2 and JH13) were selected by the ratio of initial rate of hydrolysis of enantiomerically pure (R)- and (S)-2-PB-O-res. When cell pellets were used, both strains showed high apparent enantioselectivity ($E_{app}>100$) for (R)-2PB-O-res and were identified as Exiguobacterium acetylicum. The JH13 strain showed high esterase activity on p-nitrophenyl acetate (pNPA), but showed low lipase activity on p-nitrophenyl palmitate (pNPP). The esterase was located in the soluble fraction of the cell extract. The crude intracellular enzyme preparation was stable at a pH range from 6.0 to 11.0.

Improvement of Aspergillus niger 55, a Raw Corn Meal Saccharifying Enzyme Hyperproducer, through Mutation and Selective Screening Techniques (옥수수 生 전분 당화 효소 高 생산성 변이주 개발)

  • Oh, Sung-Hoon;O, Pyong-Su
    • Microbiology and Biotechnology Letters
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    • v.19 no.2
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    • pp.140-146
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    • 1991
  • Mutation experiments were performed to select the mutant of Aspergillus niger 55, which had lost almost all the ability to produce transglucosidases but retained that of high productivity of raw meal saccharifying enzyme, by means of successive induction with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG), ultraviolet(UV) light, and ${\gamma}$-rays. Also, we used the mutant enrichment techniques, such as liquid culture-filtration procedure and differential heat sensitivity of conidia, in order to increase the possibility of obtaining a mutant. The glucoamylase productivity of mutant PFST-38 was 11 times higher than that of the parent strain. The mutant PFST-38 was morphologically identical to the parent strain, except for the size of conidia, the tendency to form conidia and the lenght of conidiophore. Asp. niger mutant PFST-38 apeared to be useful for the submerged production of the raw corn meal saccharifying enzyme.

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Isolation and Characteris tics of Polyvinyl Alcohol Degrading Bacteria (폴리비닐 알콜 분해균주의 분리 및 특성)

  • 정선용;조윤래;김정목;조무환
    • Microbiology and Biotechnology Letters
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    • v.20 no.1
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    • pp.96-101
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    • 1992
  • Two strains of polyvinyl alcohol (PVA) utilizing bacteria were isolated from the waste water and soil. These strains, G5Y and PW, were able to utilize PVA symbiotically as a carbon source, but could not utilize PVA separately. In the mixed culture of these strains, 0.5 percent of PVA was almost completely degraded in 3 days. Effect of degree of PVA polymerization on the its utilization was examined, and there was no remarkable difference among three kind of PVA (PVA 500, 1500, a d 2000). These bacteria were able to utilize PV,4 in the desizing waste water of factory as well as enrichment PVA medium. These strains, C5Y and PW, were identified as Pseudomonas cepucia and Pseudomonus pseudomallei, respectively, based on morpholofical and biological characteristics.

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점토로부터 철불순물의 생물학적 제거에 미치는 탄소원의 영향

  • Lee, Eun-Young;Cho, Kyung-Suk;Ryu, Hee-Wook;Bae, Moo
    • Microbiology and Biotechnology Letters
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    • v.25 no.6
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    • pp.552-559
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    • 1997
  • Fe (III) impurities in clay could be microbially removed by inhabitant dissimilatory Fe (III) reducing microorganisms. Insoluble Fe (III) in clay particles was leached out as soluble reductive form, Fe (II). The microorganisms removed from 10 to 45% of the initial Fe (III) when each sugar was supplemented to be in ranges of 1 - 5 % (w/w; sugar/clay). The microorganisms reduced 2.1 - 12.8 mol of Fe (III) per 100 mol of carbon in sugars metabolized when sugars such as glucose, maltose, and sucrose were used as sole carbon source. Bacillus sp. IRB-W and Pseudomonas sp. IRB-Y were isolated from the enrichment culture of the clay. The isolates were considered to participate in metabolizing organic compounds to fermentative intermediates with relatively little Fe (III) reduction at initial Fe (III) reduction process. By the microbial treatment, the whiteness of the clay was increased form 63.20 to 79.64, whereas the redness was obviously decreased form 13.47 to 3.55. This treatment did not cause any unfavorable modifications in mineralogical compositions of the clay.

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Alcohol Dehydrogenase Active on Furfuryl Alcohol from Pseudomonas sp. (Part 1) Purification and Properties of Alcohol Dehydrogenase (Pseudomonas 속균이 생산하는 Alcohol Dehydrogenase에 관한 연구 (제1보) Alcohol Dehydrogenase 정제와 일반적성질)

  • ;Hirosake Okadar
    • Microbiology and Biotechnology Letters
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    • v.8 no.1
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    • pp.27-32
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    • 1980
  • We isolated a strain of Pseudomonas sp. from soil to utilize furfuryl alcohol as a carton source by enrichment culture. Alcohol dehydrogenase from this bacteria was purified 700-fold by Sephadex G-200 and affinity column chromatography to be homogeneous by electrophoresis and analytical centrifugation. This enzyme had a molecular weight of 120,000 and was composed of four subunits consisting of 266 amino acid residues. The optimal pH of the enzyme was pH 8.5 to 9, and the optimal temperature was, 45$^{\circ}C$. This enzyme was stable at 55$^{\circ}C$, but lost 80% of its activity in 10min at 6$0^{\circ}C$.

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Screening and Characterization of Thermotolerant Alcohol-producing Yeast

  • Sohn, Ho-Yong
    • Journal of Microbiology and Biotechnology
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    • v.4 no.3
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    • pp.215-221
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    • 1994
  • Two strains of yeast (RA-74-2 and RA-912) showing superior fermenting ability at a high temperature were isolated from soils and wastewaters by an enrichment culture method. Based on the morphological and physiological charateristics, the two strains were identified as Saccharomyces cerevisiae and Kluyveromyces marxianus, respectively. RA-74-2 was able to grow upto $43^{\circ}C$ and sustain similar fermenting ability in the temperatures range from 30 to $40^{\circ}C$. In addition, the sugar- and ethanol-tolerance of RA-74-2 were 30% (w/v) glucose and 10% (v/v) ethanol, which appeared to be higher than those of nine other industrial yeast strains currently being used in the alcohol factories. The thermotolerant ethanol fermenting yeast RA-912 showed identical growth in the temperatures range from 35 to $45^{\circ}C$ and was resistant to various heavy metals. The quality and quantity of byproducts of the isolated yeast strains in fermentation broth after fermentation at $40^{\circ}C$ and $45^{\circ}C$ were similiar with those obtained at $30^{\circ}C$. These results show that RA-74-2 can be adopted for the ethanol fermentation process where the expenses for cooling system is significant, and suggest that RA-912 may be applied in either SSF(simultaneous saccharification and fermentation) or Flash-fermentation process and RA-912 may be used as a gene donor for the development of thermotolerant ethanol-fermenting yeasts.

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Inhibitory Effects of Acetic Acid and Temperature on Growth of Campylobacter jejuni ATCC 33291

  • Kim, Wang-june;Shin, Soon-Young;Hwang, Han-Joon
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.934-939
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    • 2001
  • The growth inhibition of Campylobacter jejuni ATCC 33291 in the presence of $1\%$ acetic acid at 4, 25, and $42^{\circ}C$, followed by $25^{\circ}C$ and $4^{\circ}C$, at pH 5.5 and pH 6.5, and by the addition of $1\%$ acetic acid aat 4, 25, and $42^{\circ}C$ were determined to be 22, 8.5, and 1.4 min, respectively, in an FBP-SBB medium. The D values of C. jejuni were increased by the addition of chicken and did not follow the linear relationship observed in the FBP-SBB media without chicken. When using distilled water instead of FBP-SBB in the model system, the death rate of C. jejuni was dramatically accelerated. The injured or low cell numbers that were impossible to enumerate using the plate count method, were detected by a polymerase chain reaction and enrichment culture procedure. These results suggested that acetic acid is reliable and effective as a disinfectant, however, it is necessary to take additional care at refrigeration temperatures due to the potential of injred cells during poultry processing.

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Production of Single-Cell Protein from Methanol (Part 1) Isolation of Methanol-Utilizing Microorgamism and Composition of Medium (Methanol을 이용한 단세포단백질의 생산에 관한 연구 (제 1 보) Methanol 이용 미생물의 분리 및 배지조성)

  • 유주현;정건섭;변유량
    • Microbiology and Biotechnology Letters
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    • v.7 no.2
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    • pp.65-70
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    • 1979
  • By the successive enrichment culture, methanol-utilizing bacteria of 213 strains were isolated from soil samples collected from various places. Among them one strain showing excellent growth was selected. The organism isolated was obligate methylotroph and identified as Methylomonas methanolica on the basis of its mophological and physiological characteristics of the cell. The medium have been to be collected for the maximum biomass productivity. The microorganism was capable of growing satisfactorily on a medium containing only methanol 0.8% (v/v), ammonium sulfate 0.6%, magnesium sulfate 0.1%, phosphate salts, but did not require growth factor.

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Screening and Purification of a Novel Transaminase Catalyzing the Transamination of Aryl ${\beta}-Amino$ Acid from Mesorhizobium sp. LUK

  • Kim, Ju-Han;Kyung, Do-Hyun;Yun, Hyung-Don;Cho, Byung-Kwan;Kim, Byung-Gee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1832-1836
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    • 2006
  • Mesorhizobium sp. LUK, which utilizes 3-amino-3-phenylpropionic acid as the sole source of nitrogen with high enantioselectivity (E(S)>100), was isolated using enrichment culture. The enzyme involved in the utilization of (S)-3-amino-3-phenylpropionic acid was confirmed to be a transaminase and was purified by 235-folds with a specific activity of 0.72 U/mg. The molecular weight of the purified protein was ca. 47 kDa and the active enzyme was determined as a dimer on gel filtration chromatography. The N-terminal sequence was obtained from the purified protein. Spontaneous decarboxylation of produced ${\beta}-keto$ acids was observed during the chiral resolution of 3-amino-3-phenylpropionic acid.

Isolation of a Pseudomonas sp. Capable of Utilizing 4-Nonylphenol in the Presence of Phenol

  • Chakraborty Joydeep;Dutta Tapan K.
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1740-1746
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    • 2006
  • Enrichment techniques led to the isolation of a Pseudomonas sp. strain P2 from municipal waste-contaminated soil sample, which could utilize different isomers of a commercial mixture of 4-nonylphenol when grown in the presence of phenol. The isolate was identified as Pseudomonas sp., based on the morphological, nutritional, and biochemical characteristics and 16S rDNA sequence analysis. The ${\beta}$-ketoadipate pathway was found to be involved in the degradation of phenol by Pseudomonas sp. strain P2. Gas chromatography-mass spectrometric analysis of the culture media indicated degradation of various major isomers of 4-nonylphenol in the range of 29-50%. However, the selected ion monitoring mode of analysis of biodegraded products of 4-nonylphenol indicated the absence of any aromatic compounds other than those of the isomers of 4-nonylphenol. Moreover, Pseudomonas sp. strain P2 was incapable of utilizing various alkanes individually as sole carbon source, whereas the degradation of 4-nonylphenol was observed only when the test organism was induced with phenol, suggesting that the degradation of 4-nonylphenol was possibly initiated from the phenolic moiety of the molecule, but not from the alkyl side-chain.