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Isolation and Characterization of Bacillus sp. BS-214 Producing Pectinase
Jeon,Beong Sam,Cha,Jae Young,Lee,Young Choon 한국생명과학회 2000 한국생명과학회 학술발표회 Vol.27 No.-
A bacterial strain BS-214 producing extracellular pectinase was isolated from soil. The isolated bacterium was identified as a strain of Bacillus sp. based on the morphological, biochemical, and physiological characteristics. Cell growth and pectinase activity of Bacillus sp. BS-214 were reached to maxium in the culture condition of pH 8.5 at 40℃. Production of pectinase by the strain was the highest when polygalacturonic acid was added to culture medium as a carbon source, and its optimal concentration was 1%. Also, yeast extract was used as the best nitrogen source for the production of pectinase by the concentration of 0.25%. Decomposition of a constituent of Edzeworthia papyrifera by the strain was observed by scanning electron microscope.
Jeon, Beong-Sam,Park, Jeong-Won,Shin, Gab-Gyun,Kim, Beom-Kyu,Kim, Hee-Kyu,Song, Jae-Young,Cho, Young-Su,Cha, Jae-Young Korean Society of Life Science 2004 생명과학회지 Vol.14 No.3
초호열성 고세균 Thermococcus litoralis 유래의 4-$\alpha$-glucanotransferase는 클로닝 되어 염기배열이 밝혀졌으며, 대장균에서 발현되었다 발현된 이 효소는 기능적인 면에서는 D-enzyme과 유사하지만 아미노산 배열에서는 큰 차이점을 나타내었다. 이 효소는 cycloamylose를 생산하는 새로운 기능적인 특성을 가지고 있어 당대사 관련 효소 단백질에 관한 연구의 중요성 때문에 산업적으로 많은 각광을 받고 있다. 본 연구는 초호열성 고세균 T. litoris 유래 4-$\alpha$-glucanotransferase 유전자를 부위 특이적 변이 방법으로 재조합하여 lac와 T7프로모터를 이용해서 대장균 발현 벡터 시스템에서 대량발현 시켰다. 대장균에서 대량 발현된 재조합 효소 단백질은 열처리, 501빌Butyl-Toyopearl, Mono Q 크로마티그래피 방법에 의하여 간단히 정제되었다. 정제된 재조합 효소 단백질은 본래의 효소 단백질과 같은 기능을 가지고 있는 것으로 확인되었다. The gene encoding a extremely thermostable 4-$\alpha$-glucanotransferase from a hyperthermophilic archaeon, Thermococcus litoralis, was cloned, sequenced and expressed in Escherichia coli. The amino acid sequence of the enzyme was distantly related to other functionally-related ones, such as D-enzymes. The enzyme is of industrial interest because of a novel activity of producing cycloamylose and is also important for fundamental studies of protein, sugar-metabolizing enzymes. In this paper, the overexpression of 4-$\alpha$-glucanotransferase in E. coli was carried out expression vector system with lac and T7 promoters. The enzyme was successfully overexpressed, and purified by the heat treatment of a cell-free extract, successive Butyl-Toyopearl and Mono Q chromatographies. The purified recombinant enzyme showed the same specific activity and the same mobility in SDS-PAGE as natural enzyme.
Purification and Some Properties of an Extracellular Pectinase from Bacillus sp. BS-214
Jeon, Beong-Sam,Song, Jae-Young,Lee, Gang-Deog,Kim, Beom-Kyu,Cha, Jae-Young,Lee, Young-Choon Korean Society of Life Science 2000 Journal of Life Science Vol.10 No.1
Pectinase was isolated from culture medium of Bacillus sp. BS-214 and purified 105-fold with 3.4% yield by ammonium sulfate precipitation, gel filteration using Sephadex G-75 and DEAE-cellulose followed by gel filteration through Sephadex G-100. The molecular weight of the purified enzyme was estimated to be about 43 kDa on SDS-PAGE and by gel filtration, indicating that the enzyme is a monomer. the optium pH and temperature of the enzyme were 9.0 and 55$^{\circ}C$, respectively. the enzyme was stable at 60$^{\circ}C$ for 30min and in a pH range from 7.5 to 10.5 for 12 h ant 4$^{\circ}C$. The enzyme activity was highly enhanced by Ca2+, and also K+, Li+ and Na+showed a positive effect, while stongly inhibited by Zn2+ and Hg2+.
Hypoglycemic Effect of Mushroom Fermented Milk in Streptozotocin-Induced Diabetic Rats
Cha, Jae-Young,Jeon, Beong-Sam,Park, Jeong-Won,Shin, Gab-Gyun,Kim, Beom-Kyu,Kim, Hee-Kyu,Cho, Young-Su Korean Society of Life Science 2004 생명과학회지 Vol.14 No.4
버섯 추출물을 첨가하여 유산 발효시킨 버섯발효유와 이때 사용된 버섯 추출물 및 버섯발효유 상등액의 항당뇨 효과를 규명하고자 Sprague-Dawley 수컷에 streptozotocin을 50 mg/kg body weight씩 복강내 주사하여 당뇨를 유발시켜 검토하였다. 버섯발효유, 상등액 및 버섯 추출물을 식이 중에 10% (v/w)씩 동량 첨가 한 식이를 3주간 급여한 후 혈당치, 인슐린 농도 및 경구당부하실험을 실시하였다. 버섯발효유의 이화학적 성분을 분석 한 결과 단백질 2.87%, 지방 0.09%, 탄수화물 6.0%, 식이섬유 0.3%, 락토스 2.01%, 슈크로스 1.23% 및 칼슘과 철 성분을 각각 95.9 및 0.08 mg/l00 g함유하고 있었다. Streptozotocin-유발 대조군 당뇨쥐에 비해 버섯발효유 투여군에서 현저한 혈당강하 효과가 있었으며, 이러한 효과는 인슐린 농도증가에 의한 것으로 나타났다. 또한, 버섯발효유 상등액 및 버섯 추출물에도 혈당강하 효과가 있는 것으로 나타났다. 실험 종료시점에 실시한 경구당부하실험에서도 버섯발효유, 상등액 및 추출물 순으로 현저한 효과를 보였다. 이상의 결과로 볼 때 혈당강하 효과가 있는 버섯과 유산균을 접목한 버섯발효유 제조는 이들 상호간의 시너지 효과에 의해 당뇨성 흰쥐에서 현저한 항당뇨 효과를 발휘하였다. Nutritional concentrations by chemical analyses of mushroom fermented milk were protein 2.87%, fat 0.09%, carbohydrates 6.0%, dietary fiber 0.3%, lactose 2.01%, sucrose 1.23%, calcium 95.9 mg/100 g and iron 0.08 mg/100 g. The present study was undertaken to investigate the hypoglycemic effects of the equal volume of either water (streptozotocin (STZ)rontrol rats), mushrooms water-extract (STZ-extrart fed rats), mushroom fermented milk product (STZ-mushroom yogurt fed rats) or mushroom fermented milk supernatant (STZ-supernatant fed rats) (10%, v/w), in STZ-induced diabetic rats for 3 week period. The mushroom fermented milk given to the STZ-diabetic rats decreased the blood glucose significantly and increased the blood insulin, compared with the STZ-control rats. The supernatant and mushroom water extract also slightly retarded the development of hyperglycemia in the STZ-diabetic rats. Taken together the results, the mushroom yogurt may have a potential for the hypoglycemic effect in the STZ-diabetic rats.
Kim, Beom-Kyu,Jeon, Beong-Sam,Cha, Jae-Young,Park, Jeong-Won,Kim, Sam-Woong,Kim, Ji-Yoon,Park, Yong-Lark,Cho, Young-Su,Song, Jae-Young Korean Society of Life Science 2003 생명과학회지 Vol.13 No.6
한천, 키틴, 셀룰로오스, 자일란, 만난과 같은 복합다당류들에 대한 분해능을 나타내는 효소들을 생산하는 해양미생물을 홍조류인 Porphyra dentata로부터 분리하였다. 분리균 BK1은 그람음성, 호기성 간균으로 DNA의 G+C함량은 51.6 mol%를 나타내었다. 주요 isoprenoid quinine 구성 성분은 ubiquinone-8로 확인되었고, 주요 세포 지방산은 C16:0, C16:1 w6c, C18:1 w7c로 밝혀졌다. 16S rRNA서열의 비교분석 결과는 분리균 BK1이 Pseudomonas 속의 일원인 것으로 확인되었다. 이러한 결과를 바탕으로 분리균 BKl은 Pseudomonas sp. BKl으로 명명하였다. A marine bacterium (strain BKl) that produces extracellular enzymes capable of decomposing complex polysac-charides, such as agar, chitin, carboxymethylcellulose, xylan and mannan, was isolated from the marine red alga Porphyra dentata. Strain BKl was gram-negative, aerobic, catalase- and oxidase-positive, polarly flagellated bacilli that produce gelatinase and urease, but not decarboxylases. The G+C content of the DNA was 51.6 mol%. The major isoprenoid quinone component was identified as an ubiquinone-8, and the major cellular fatty acids were C16:0, C16:1 w6c and C18:1 w7c. Comparative 16S rRNA sequence analysis placed strain BK1 with members of the genus Pseudomonas. On the basis of phenotypic and genotypic data, the strain BK1 was shown to be a member of the subgroup of Pseudomonas, and named as Pseudomonas sp. BK1.
Optimal Fermentation Conditions for Enhanced GlutathioneProduction by Saccharomyces cerevisiae FF-8
Jae-Young Cha1,,Jin-Chul Park,Beong-Sam Jeon,Young-Choon Lee,Young-Su Cho 한국미생물학회 2004 The journal of microbiology Vol.42 No.1
The influence of feedstock amino acids, salt, carbon and nitrogen sources on glutathione production by Saccharomyces cerevisiae FF-8 was investigated. Glucose, yeast extract, KH2PO4, and L-cysteine were found to be suitable feedstock. Highest glutathione production was obtained after cultivation with shaking for 72 h in a medium containing glucose 3.0% (w/v), yeast extract 3.0%, KH2PO4 0.06% and L-cysteine 0.06%. The glutathione concentration achieved using this medium increased 2.27-fold to 204 mg/l compared to YM basal medium.
Optimal Fermentation Conditions for Enhanced Glutathione Production by Saccharomyces cerevisiae FF-8
Cha, Jae-Young,Park, Jin-Chul,Jeon, Beong-Sam,Lee, Young-Choon,Cho, Young-Su The Microbiological Society of Korea 2004 The journal of microbiology Vol.42 No.1
The influence of feedstock amino acids, salt, carbon and nitrogen sources on glutathione production by Saccharomyces cerevisiae FF -8 was investigated. Glucose, yeast extract, KH$_2$PO$_4$, and L-cysteine were found to be suitable feedstock. Highest glutathione production was obtained after cultivation with shaking for 72 h in a medium containing glucose 3.0% (w/v), yeast extract 3.0%, KH$_2$PO$_4$ 0.06% and L-cysteine 0.06%. The glutathione concentration achieved using this medium increased 2.27-fold to 204 mg/l compared to YM basal medium.