• 한국식물병리학회:학술대회논문집
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• 한국식물병리학회 1999년도 임시총회 및 추계 학술발표회
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• pp.53.2-53
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• 1999

• Journal of Microbiology and Biotechnology
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• 제26권11호
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• pp.1993-2005
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• 2016

Bacillus pumilus is one of the most characterized microorganisms that are used for high-level production of select industrial enzymes. A novel B. pumilus SCU11 strain possessing high alkaline protease activity was obtained in our previous work. The culture supernatant of this strain showed efficient dehairing capability with minimal collagen damage, indicating promising potential applications in the leather industry. In this study, the strain's extracellular proteome was identified by LC-MS/MS-based shotgun proteomic analysis, and their related secretory pathways were characterized by BLAST searches. A total of 513 proteins, including 100 actual secreted and 413 intracellular proteins, were detected in the extracellular proteome. The functions of these secreted proteins were elucidated and four complete secretory systems (Sec, Tat, Com, and ABC transporter) were proposed for B. pumilus. These data provide B. pumilus a comprehensive extracellular proteome profile, which is a valuable theoretical and applicative basis for future genetic modifications and development of industrial enzymes.

• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.568-571
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• 1999

Aspergillus oryzae U1521, which was a genetically engineered strain, produced 1,000,600 U per g . glucose of extracellular alkaline protease within 72 h in a submerged fermentation. However, the alkaline protease was not detected during the first 24 h. Northern blot analysis indicated that the enzyme synthesis was repressed at the transcriptional level during the lag period. Both catabolite repression and pH of the growth medium significantly affected the enzyme production. Use of this enzyme as a silver recovery agent from used X-ray film was confirmed by experiments in the shake-flask scale.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.334.1-334.1
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• 2002

Streptomyces scabiei subsp. chosunensis M0137. nonadecanoic acid producer. showed the highest protease activity when grown in OSY medium (oatmeal 1.5%, soybean meal 2%, dried yeast 1 %) supplemented with. glycerol (1 %) and CaCO3 (0.1 %). Two forms of protease(SS-1 and SS-2) were fractionated and purified through Ultrogel AcA 54 gel filtration and DEAE-sepharose CL-6B column chromatography. Both proteases were practically stable in the pH range of 6-10. The optimal pH for the activities of both protease 88-1 and 8S-2 were 7.5 and 8.0. respectively. (omitted)

• 한국미생물·생명공학회지
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• 제28권6호
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• pp.329-333
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• 2000

Alkaline bacterium producing a high pro-tease activity at low temperature was isolated by using enrichment culture from soil samples and identified as Bacil-lus sp. WRD-1 Cell growth was maximal at 10 hours and the optimal initial pH and culture time of culture condition for enzyme production was pH 7 and 10 hours, respectively. Temperature range of high enzyme activity were $10~40^{\circ}C$. The optimal pH and temperature for the enzyme activity were pH9 and $30^{\circ}C$.

• KSBB Journal
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• 제22권4호
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• pp.185-190
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• 2007

Productivity of secreted recombinant protein depends largely on its stability in the extracellular environment with protease. Most hGM-CSF produced by transgenic tobacco cell cultures and secreted to the medium was confirmed to be rapidly degraded by protease in medium. To increase the productivity, therefore, various protein stabilizers such as gelling agents such as carrageenan and alginate, polymers, polyols, and amino acids have been tested. The stability of hGM-CSF in spent medium without cells was improved by the presence of gelling agents. However, the reason for the enhanced production by the addition of gelling agents may be due to the increased expression level and permeability rather than stability. The addition of DMSO inhibited the cell growth, but improved specific yield. The others were not effective for stability as well as hGM-CSF production.

• 대한의생명과학회지
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• 제6권3호
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• pp.209-221
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• 2000

본 논문은 환자들의 병소에서 채취된 가검물에 존재하는 미생물들 가운데 pretense을 분비하는 균주들을 분리하고, 그 가운데 가장 효소생산력이 우수한 균주를 선정하여 가정용 Protease 세제생산에 활용 가능성을 알아보고자 그들이 생산하는 protease의 기초적 생산조건 및 부분적 효소학적 특성 등에 관한 실험에서 얻은 결과들을 보고하고자 작성되었다. Serratia sp. 2000-1로 동정된 본 실험 균주가 생산하는 protease의 기초적 생산조건과 부분적 효소학적 특성을 조사한 결과 효소생산을 위한 최적 배지에 탄소원과 질소원 그리고 금속염의 종류와 농도는 각각 Glucose 1.5%, C.S.P 2.0%, CaCl$_2$ 0.1%였다. 그리고 최적 배양온도는 3$0^{\circ}C$, 초발 pH는 8.0, 배양시간은 72시간 이었다. Protease의 정제는 ammonium sulfate precipitation, DEAE-cellulose, Sephadex G-200 column을 통하여 분리 정제하였으며 이때 최종 효소수율은 14.4%, 효소 비활성도는 약 29배 증가한 것으로 나타났다. 그리고 효소 작용의 최적온도와 pH는 35$^{\circ}C$와 pH 8.0으로 나타났고, 효소 작용의 열과 pH에 대한 안정성은 4$0^{\circ}C$와 pH 6~10까지는 효소의 활성이 비교적 안정한 것으로 나타났다. 금속이온들 중 $Mg^{2+}$, $Ba^{2+}$, $Ca^{2+}$, Mn$^{2+}$은 효소활성을 촉진하나 Hg$^{2+}$, Ag$^{2+}$, Cu$^{2+}$는 효소활성을 저해하는 것으로 나타났다. 그리고 효소활성 저해제들 중에는 SDS가 가장 강하게 효소활성을 저해하는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제32권1호
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• pp.99-109
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• 2022

This study is the first report on production and characterization of the enzyme from an Ornithinibacillus species. A 4.2-fold increase in the extracellular protease (called L9^T) production from Ornithinibacillus caprae L9^T was achieved through the one-factor-at-a-time approach and response surface methodological optimization. L9^T protease exhibited a unique protein band with a mass of 25.9 kDa upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. This novel protease was active over a range of pH (4-13), temperatures (30-80℃) and salt concentrations (0-220 g/l), with the maximal activity observed at pH 7, 70℃ and 20 g/l NaCl. Proteolytic activity was upgraded in the presence of Ag⁺, Ca²⁺ and Sr²⁺, but was totally suppressed by 5 mM phenylmethylsulfonyl fluoride, which suggests that this enzyme belongs to the serine protease family. L9^T protease was resistant to certain common organic solvents and surfactants; particularly, 5 mM Tween 20 and Tween 80 improved the activity by 63 and 15%, respectively. More importantly, L9^T protease was found to be effective in dehairing of goatskins, cowhides and rabbit-skins without damaging the collagen fibers. These properties confirm the feasibility of L9^T protease in industrial applications, especially in leather processing.

• 한국미생물·생명공학회지
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• 제34권4호
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• pp.323-328
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• 2006

식충식물에서 채취된 시료로부터 protease치 생산균으로 분리된 SH-8은 그람 양성간균으로 16S rRNA외 부분 염기서열에 근거하여 Bacillus속 균주로 확인되었다. Protease 생산을 위한 배지를 제조하기 위해 질소원, 탄소원, 인, 금속이온의 성분을 변화시키면서 균의 성장과 효소 생산성을 비교하였다. 포도당을 탄소원으로 사용하였을 때는 균의 성장은 정상적으로 일어나지만, pretense생산이 완전히 억제되는 것으로 나타났으며, 가용성 전분을 탄소원으로 사용하였을 때 효소 생산성이 가장 높았다. 질소원으로는 yeast extact가 효소 생산에 가장 적합하였으며, 농도가 높으면 효소 생산성이 저하되었다. 한편 2가 금속이온중 $Zn^{2+}$, $Cu^{2+}$, $Co^{2+}$, $Mn^{2+}$을 배지에 첨가하였을 때는 균의 성장이 심하게 저해되었으며, 효소 생산도 되지 않았다. $CaCl_2$를 첨가한 배지에서는 효소 생산성이 증가되었으며, 효소 반응에 $CaCl_2$를 첨가하였을 때도 효소 활성이 증가하였다. 이로보아 $CaCl_2$는 Bacillus sp. SH-8의 protease 생산성과 활성을 모두 증가시키는 것으로 판단된다. 가용성 전분(2%), yeast extract(0.3%), $CaCl_2$(0.3%), $CaCl_2$(0.01%)와 $KH_2PO_4$(0.01%)를 포함하는 것으로 구성된 최적화 배지에서 최대효소 생산성은 435 U/ml로 나타났으며 26시간이 되었을 때 배양액 내 효소활성은 급격히 감소하였다.

• 생명과학회지
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• 제8권3호
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• pp.333-338
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• 1998

Productivity of alkaline proteinase from Yarrowia lipolytica 504D was investigated. For the production fo the enzyme, hemoglobin was the best nitogen source, however, casein and skim milk were also good. All carbon sources inhibited strongly the producitivity of the enzyme. Yeast extract increased the productivity of the enzyme to 220%, but almost mineral salts except monovalant ions decreased it. Based on these results, optimal medium was composed of 1.2% casein, 0.2% glucose, 0.16% yeast extract, and 0.1% ammonium sulfate. the best condition for the production of the enzyme was observed at pH 9 and $20^{\circ}C$ for 42 hours.