• Title/Summary/Keyword: Substrate culture

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Laboratory Scale Preparation of S-Adenosyl-L-Methionine from Yeast (효모로부터 S-Adenosyl-L-Methionine의 실험실 규모 생산)

  • 이종남;류양욱;최명언
    • Microbiology and Biotechnology Letters
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    • v.19 no.6
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    • pp.588-591
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    • 1991
  • S-adenosyl-L-methionine (SAM) is essential substrate for biological methylation reactions. The present work describes a reoptimized procedure of SAM preparation in laboratory scale by the method of yeast fermentation. The fermentation medium enriched with methionine and the culture conditions were reoptimized. The isolation steps consisted of 5 steps including extractions, precipitations, and chromatography. This improved procedure over original method provides relatively high yield of biologically active product within a 4 day-period.

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Effect of Concentration of Nutrient Solution on Early Yield and Fruit Qualify of Tomato in Substrate Culture (고형배지경에서 배양액농도가 토마토 초기수량 및 품질에 미치는 영향)

  • 노미영;이용범;박권우;권영삼
    • Proceedings of the Korean Society for Bio-Environment Control Conference
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    • 1994.05a
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    • pp.121-122
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    • 1994
  • 배양액농도는 작물의 양분과 수분흡수에 영향을 주고 생육, 수량 및 품질에 미치는 영향이 큰 근권제어요인이다. 순수수경재배에서는 작물종류별 배양액농도에 대한 연구가 많이 되어 있으나 배지경에서 배지종류별 작물에 따른 적정배양액 농도에 대한 연구는 드물다. 특히 적정배양액 농도는 작물종류, 품종, 생육단계에 따라 다르고, 생산물의 품질에도 지대한 영향을 미치는 것으로 밝혀지고 있다. 따라서 토마토에서 배지종류별(펄라이트, 버미큘라이트, 피트모스) 육모기와 정식후 배양액농도에 따른 토마토 수량 및 품질에 미치는 영향을 밝히고져 실험을 수행하였다. (중략)

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Isolation and Characterization of Enterobacter sp. Producing Galacto-oligosaccharides

  • YANG, JI-WON;HYUN-JAE SHIN;SANG-PIL YEOM;BYUNG-DAE YUN;MIN-HONG KIM
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.343-348
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    • 1994
  • Enterobacter sp. producing -$\beta$-galactosidase with high transgalactosylation activity was isolated from dairy wastewater. The isolate had common biochemical features to E. aerogenes and E. cloacae. Enzyme production increased as the cell mass increased with optimum enzyme activity of 0.21 Unit/mg-protein (o-nitro-phenyl-$\beta$ -D-galactoside (ONPG) as substrate) until 8 hr of culture. Whole cells permeabilized by toluene were used to produce galacto-oligosaccharide. Optimum toluene concentration, temperature and pH for -$\beta$-galactosidase activity of permeabilized whole cells were 10% (v/v), $50^{\circ}C$ and 6.0, respectively. A maximum of 38% (w/w) of galacto-oligosaccharide was obtained with lactose concentration of 20% (w/w) at $40^\{\circ}C$ and pH 6.0.

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Optimization of the Functional Expression of Coprinus cinereus Peroxidase in Pichia pastoris by Varying the Host and Promoter

  • Kim, Su-Jin;Lee, Jeong-Ah;Kim, Yong-Hwan;Song, Bong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.966-971
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    • 2009
  • Peroxidase from Coprinus cinereus (CiP) has attracted attention for its high specific activity and broad substrate spectrum compared with other peroxidases. In this study, the functional expression of this peroxidase was successfully achieved in the methylotrophic yeast Pichia pastoris. The expression level of CiP was increased by varying the microbial hosts and the expression promoters. Since a signal sequence, such as the alpha mating factor of Saccharomyces cerevisiae, was placed preceding the cDNA of the CiP coding gene, expressed recombinant CiP (rCiP) was secreted into the culture broth. The Mut Pichia pastoris host showed a 3-fold higher peroxidase activity, as well as 2-fold higher growth rate, compared with the $Mut^s $ Pichia pastoris host. Furthermore, the AOX1 promoter facilitated a 5-fold higher expression of rCiP than did the GAP promoter.

Copolyester of 3-Hydroxybutyrate and 3-Hydroxyvalerate Produced by Methylobacterium sp. GL-10 (Methylobacterium sp. GL-10이 생산하는 3-Hydroxybutyrate와 3-Hydroxyvalerate의 Copolyester)

  • 이호재;박진서;이용현
    • Microbiology and Biotechnology Letters
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    • v.19 no.1
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    • pp.94-99
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    • 1991
  • - The further study for the identification of the previously reported pink-pigmented facultative methylotrophic bacterium (PPFM) GL-10 was carried out. The PPFM GL-10 was Gram nagative, rod, and motile by a single polarly inserted flagellum. The colonies were smooth, pink, circular, along with convex with entire margin. The isolate could utilize C1 compounds and a variety of multicarbon substrates as sole carbon and energy source. The isolate was obligately aerobic, and exhibited both catalase and oxidase activities. The deoxyribonucleic acid base composition was 65-67 mol% guanine plus cytosine. The isolate was mostly identical with Methylobacterium extorquens and named Methylobacterium sp. strain GL-10. Methylobacterium GL-10 accumulated a copolyester of 3-hydroxybutyrate and 3-hydroxyvalerate (poly-3HB/3 HV) when grown in nitrogen-free culture media containing sodium propionate as substrate at the second polyester accumulation stage. The composition of copolyester, as determined from $^1h$ NMR spectra, was 23 mol% of 3-hydroxyvalerate (3HV).

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Purification and Charactedrization of Cysteine Desulfhydrase from Streptomyces albidoflavus SMF301

  • Ryu, Jae-Gon;Kang, Sung-Gyun;Kim, In-Seop;Rho, Young-Taik;Lee, Sang-Hee;Lee, Kye-Joon
    • Journal of Microbiology
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    • v.35 no.2
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    • pp.97-102
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    • 1997
  • Cysteine desulfhydrase (EC 4.4.1.1.) was purified from the culture supernatant of Streptomyces albidoflavus SMF301 by hydroxyapatite, gel filtration and Resource Q ion-exchange chromatography with a purification fold of six identical subunits. The enzyme was stabilized by dithiothreitol and pyridoxal 5'-phosphate during the purification procedures. The optimum pH and temperature were pH 8.6 and 35$^{\circ}C$, respectively. The N-terminal amino acid sequence was identified as A-P-L-P-T-A-D-V-R-S-D-P-G-Y-R-E-W-L-G-E-A-V. The purified cystein desulfhydrase had a high substrate specificity toward cysteine, and exhibited no cystahionine $\gamma$-lyase activity. The $K_m$ value for cysteine was determined to be 0.37 mM.

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Identification of to Hexapeptides that Render C2 Myoblasts the Resistant Menadione-induced Cell Death

  • Hwang, Sung-Ho;Kim, Min-Jeong;Lim, Jeong-A;Woo, Joo-Hong;Kim, Hye-Sun
    • Animal cells and systems
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    • v.12 no.1
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    • pp.35-39
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    • 2008
  • Menadione induced cell death in cultured C2 myoblasts. By screening synthetic peptide libraries composed of random sequence of hexapeptides, we identified the hexa-peptides pool of(Ala/Ile)-(Ile/Met)-Val-Ile-Asp-(Met/Ser)-$NH_2$ that protected the myoblasts against menadioneinduced cell death. Pre-incubation with the hexapeptide pool reduced the number of cells detached from culture dish substrate and increased the ratio of relative viability against menadione. In addition, the peptides strongly increased the expression of Bcl-2, an anti-apoptotic protein. These results suggest that the hexapeptides might enhance the resistance to cell death against menadione by increasing the expression of Bcl-2.

Production and Characterization of Antimicrobial Sophorolipids

  • Kim, Young-Bum;Kim, Gab-Jung;Kim, Eun-Ki
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.220-223
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    • 2000
  • Sophorolipid, a glycolipid type biosurfactant produced from Candida Bombicola, inhibited the growth of bacteria and fungi. Between two types of sophorolipid, acid and lactone form, the latter had more strong antimicrobial activity. Culturing parameters including oil substrate, initial glucose concentration, aeration, culture mode, affected the ratio of two forms. The fatty acid moety varied depending on the oil having different fatty acid chains. Minimum inhibitory concentration of lactonic sophorolipid toward Propionibacterium acne was below 0.5ppm. This result indicated the potentials of sophorolipid as antimicrobial agent in various area including cosmetics.

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Hydrolysis of the Nitrile group in $\alpha$-Aminophenylacetonitrile by Nitrilase;Development of a New Biotechnology for Stereospecific Production of S-$\alpha$-Phenylglycine

  • Choi, Soo-Young;Goo, Yang-M
    • Archives of Pharmacal Research
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    • v.9 no.1
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    • pp.45-47
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    • 1986
  • Phenylglycine was obtained as the sole metabolite when .alpha.-aminophenylacetonitrile was ted to the culture broth of Aspergillus furmigatus furmigatus. The isolated phenylglycine showed L-configuration with 80% optical purity. Examination of the hydrolysis of the substrate to phenylglycine with cell free extracts, and the supernatant fraction and the particulate fraction both of which were obtained after ultracentrifugation of the cell free extract at 100,000g, indicated that the nitrile group hydrolyzing enzymes, nitrilase existed not only in cytoplasm, but in microsome fractions.

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Properties of the Endonuclease Secreted by Human B Lymphoblastic IM9 Cells

  • Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • v.31 no.1
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    • pp.106-110
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    • 1998
  • We have employed a DNA-native-polyacrylamide gel electrophoresis (DNA-native-PAGE) assay system to characterize the enzyme activity of the endonuclease secreted by human B lymphoblastic IM9 cells. Experimental results clearly demonstrated that the endonuclease activity of IM9 cell culture medium is distinct from that of DNase I in the DNA-native-PAGE assay system. Immunoprecipitation analysis using anti-DNase I antibody showed that the secreted endonuclease is not recognized by the antibody. The secreted endonuclease was estimated using supercoiled plasmid DNA as a substrate. The pH optimum required for the catalytic activity was determined to be in the range of pH 6.6-7.4. No significant difference in the endonuclease secretion was observed by stimulation of the IM9 cells with interferon-${\gamma}$ or interleukin-$1{\beta}$.

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