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Yarrowia lipolytica TH65가 생산하는 Alkaline Proteinase의 정제 및 특성
유춘발,김창화,진영호,진익렬 한국미생물생명공학회 ( 구 한국산업미생물학회 ) 1996 한국미생물·생명공학회지 Vol.24 No.3
최적조건에서 생산한 효소를 40~65% ammonium sulfate 분획, DEAE-cellulose chromatography, Sephadex G-100과 Sephadex G-75 gel filtration을 이용하여 수율 14%, 15배 정제하였다. 정제효소의 분자량은 SDS-PAGE로 측정한 결과 31,500 정도였고, 최적온도와 최적 pH는 각각 40℃와 8.5~9.0이었다. 대부분의 2가 금속이온은 효소활성을 저해하였으며, 특히 Hg^2+, Zn^2+, Co^2+ 등은 강하게 저해하였다. PMSF는 효소활성을 완전히 저해시켰으며, EDTA, EGTA 및 phenanthroline은 부분적으로 저해시켰으나 Ca^2+을 첨가하면 모두 효소활성이 복원되는 것으로 나타나 metalloprotease는 아닌 것으로 생각되었다. 이상의 결과로 보아 본 효모 Yarrowia lipolytica TH65가 생산하는 효소는 alkaline serine proteinase로 생각된다. An alkaline proteinase produced by Yarrowia lipolytica TH65 was purified by 40~65% ammonium sulfate fractionation, DEAE-cellulose chromatography, and gel filtration with Sephadex G-100 and Sephadex G-75. The purified enzyme was shown as a single ban on SDS-PAGE, and its molecular weight 31,500. Optimum temperature and pH were 40℃ and 8.5~9.0, respectively, and the enzyme was stable below 40℃ and in the pH range of 6~8. The enzyme was strongly inhibited by divalent ions, completely by PMSF, and partially by EDTA, EGTA, and phenanthroline. But the inhibitory effect in the presence of EDTA, EGTA and phenanthroline could be reversed by addition of Ca^2+. Thus, these results indicated that the purified enzyme was an alkaline serine proteinase (E.C. 3.4.21.14).
Alkaline protease 분비세균의 분리동정 및 생성조건
陳翼烈,李貞淑 慶北大學校 1990 論文集 Vol.49 No.-
An alkalophilic soil isolate from the suburbs of Taegu city was identified as a faculatively anaerobic rod Aeromonas sp. JS-66, which has secreted an alkaline protease in the alkaline medium containing Na_2CO_3. The optimum culture condition for production of the alkaline protease by this strain JS-66 was as follows; 1.0% sucrose, 0.5% yeast extract, 0.5% polypeptone, 0.1% K_2HPO_4, 0.02% MgSO_4·7H_2O, 0.7% Na_2CO_3, pH 9, 30℃ and 20 hr.
호알카리성 Aeromonas 속 균주가 생성하는 Alkaline Protease의 정제 및 특성
陳翼烈,李貞淑 慶北大學校 1990 論文集 Vol.49 No.-
The alkaline protease from the best-cultured broth of Aeromonas sp. JS-66 was purified about 71 folds with the yield of 1.7% by ammonium sulfate precipitation, dialysis, DEAE-cellulose ion exchange chromatography and repeated gel filtration on Sephadex G-150. Optimal temperature and pH for activity of the purified enzyme were 50℃ and pH9.5. This enzyme was stable to 60℃ and within pH7 to 11. This enzyme was inactivated by Cu^2+, Zn^2+, Fe^2+ and EDTA, but activated by Mn^2+. The molecular weight of the enzyme was estimated about 24,000 by the SDS PAGE analysis.
프탈레이트 에스터를 분해하는 Klebsiella sp. KBO222 의 Protocatechuate dioxygenase 유전자의 클로닝
진익렬,김상헌,김병오,서정훈,박영덕 경북대학교 유전공학연구소 1992 遺傳工學硏究所報 Vol.7 No.1
The recombinant DNA molecules of the insertion of the BamH1-digested fragment of Klebsiella chromosomal DNA into the E. coli plasmid vector molecule, pJL3-TB5, could be transformed into E. coli MC1061. The transformants appeared onto the selective medium with protocatechuate as a sole carbon source. Each colony was inoculated into this selective liquid medium, followed by a shaking overnight cultivation. The supernatant of one of the sonicated cultured cells did show the same activity of this protocatechuate dioxygenase with comparison to that of the original Klebsiella KBO222 cells. Further studies are being focused on the isolation of this gene from the transformant.
프탈레이트 에스터 분해세균의 분리 및 분해효소의 최적 생성조건
진익렬,김병오,박완,김란숙 경북대학교 유전공학연구소 1995 遺傳工學硏究所報 Vol.10 No.1
A strain degrading phthalate ester was isolated from a sludge of Taegu area and identified as a strain of Klebsiella. The optimum culture conditions for the protocatechuate dioxygenase production were oleo investigated. This strain produced the enzyme in question under the shaking cultivation at 30℃ for the 48 hrs in the medium containing 0.1% protocatechuate as the sole carbon source, 0.1% ammonium sulfate and 0.1% yeast extract as the nitrogen source and mineral salt mixture of magnesium sulfate, sodium chloride, calcium chloride, ferric chloride, manganese sulfate, zinc sulfate and cupric sulfate. This enzyme was intracellularly localized and probably linked to cell membrane, and induced by protocatechuate.
유전자조작, 균주분리 상업용 김치 생산과정에서 대장균유사세균의 발생과 억제
이수진 ( Su Jin Lee ),한태원 ( Tae Won Han ),김마리 ( Ma Rie Kim ),설경조 ( Keyung Jo Seul ),박유미 ( Yu Mi Park ),진익렬 ( Ing Nyol Jin ),김사열 ( Sa Youl Ghim ) 한국미생물생명공학회 2009 한국미생물·생명공학회지 Vol.37 No.2
As consumption of kimchi has increased, factories have begun to produce this traditional Korean fermented vegetable dish on a large scale. Following the rise in manufacturing, the hygienic conditions under which commercial kimchi is being made have become an issue. We isolated 17 coliform bacteria from commercial kimchi that had not been fully fermented. These bacteria were partially identified as one of seven different species from three genera by 16S rDNA sequence analysis as follows: Enterobacter intermedius, Ent. cloacae, Ent. amnigenus, Klebsiella terrigena, K. ornithinolytica, K. oxytoca, and Hafnia alvei. Lactobacillus paraplantarum KNUC25 has been isolated from over-fermented Chinese cabbage kimchi and its antimicrobial activity reported in the literature. In our study, the KNUC25 strain showed antibacterial activity against isolated coliform bacteria and some pathogenic coliform bacteria through spot-on-the-lawn tests and viable cell tests. Through development and use of a cell-free supernatant of L. paraplantarum KNUC25, we effectively controlled coliform bacteria in commercial kimchi.
연료용 알콜의 고온생산을 위한 고온성 효모 Saccharomyces cerevisiae F38-1의 분리
김재완,진익렬,서정훈 한국산업미생물학회 1995 한국미생물·생명공학회지 Vol.23 No.5
대구근교의 고온환경에서 고온성, 내당성, 내알콜성, 응집성이 있고 고온에서의 알콜발효능이 우수한 효모균주를 분리, 동정한 결과 Saccharomyces cerevisiae로 판명되어 F38-1로 명명했다. 이 균주는 50%(W/V)의 포도당농도에서는 생육이 가능하고, 40℃에서 알콜농도 6∼10%(V/V)에 이틀간 노출될 때 균의 성장이 저해를 받았으나, 43℃의 고온에서도 생육하며 알콜생성하는 thermotolerant 효모이다. 그리고 20%포도당과 0.2% yeast extract를 포함하는 발효배지를 사용하여 40℃의 고온에서 발효했을 때 9.8%(V/V)의 알콜을 생산하는 우수한 발효율을 나타냈다. A new thermotolerant yeast strain was siolated, and its characteristics have been studied. The strain was identified and named Saccharomyces cerevisiae F38-1. This strain could grow not only at high temperature, but also in high concentrations of sugar and ethanol. S. cerevisiae F38-1 could grow in a medium containing 50% glucose. The isolate produced ethanol at 43℃, but didn't grow at 40℃ in the presence of 8% ethanol. Fermentation studies showed that the isolate ferments 20% glucose to 9.8% (V/V) ethanol at 40℃ in the presence of 0.2% yeast extract.