• Proceedings of the Korean Society of Plant Pathology Conference
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• 1999.10a
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• pp.53.2-53
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• 1999

• Journal of Microbiology and Biotechnology
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• v.26 no.11
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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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• v.9 no.5
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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.

• Proceedings of the PSK Conference
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• 2002.10a
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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)

• Microbiology and Biotechnology Letters
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• v.28 no.6
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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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• v.22 no.4
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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.

• Biomedical Science Letters
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• v.6 no.3
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• pp.209-221
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• 2000

The purpose of this study was to investigate the practical availability of pretense production that can be used at home after isolating Serratia sp.2000-1 which produced extracellular pretense from clinical specimen. Basic production conditions and partial enzymatic characteristics of pretense produced by Serratia sp. 2000-1 was as follows: The kind and concentration of carbohydrate, nitrogen and metal salts for optimal enzyme production condition were each identified as the concentration of 1.5% glucose, 2.0% CSP, and 0.1% CaCl$_2$, and the optimal temperature, time and initial pH for culture were each 3$0^{\circ}C$, 72 hours, and pH 8.0. The final enzymatic yeild that was purified by 3 steps with ammonium sulfate precipitation (45~80%), DEAE-cellulose column chromatography, and Sephadex G-200 gel chromatography was 14.4%, and enzyme inactivity rate increased approximately 291314s. The optimal temperature and pH for purified pretense activity were 35$^{\circ}C$ and pH 7.0~8.0, and purified pretense activity was relatively stable by 4$0^{\circ}C$ at pH 6~10 for 30 min, however heating at 6$0^{\circ}C$ for 30 min, it liminated detectable pretense activity. The pretense activity was activated by $Mg^{2+}$, $Ba^{2+}$, $Ca^{2+}$, Mn$^{2+}$, but inactiviaed by Hg$^{2+}$, Ag$^{2+}$, Cu$^{2+}$, and the pretense activity was inhibited strongly by SDS among enzyme activity inhibitors. Further study is required to evaluate the practical availability of pretense production that can be used at home by isolating Serratia sp. from more clinical specimen and examining pretense more in details.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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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.

• Microbiology and Biotechnology Letters
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• v.34 no.4
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• pp.323-328
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• 2006

A bacterium producing the extracellular protease was isolated from insect-eating plant and has been identified as a member of the genus Bacillus based on partial 165 rRNA sequences. In order to develop the medium composition, effects of ingredients including nitrogen sources, carbon source, metal ions and phosphate were examined for protease production of the isolate, SH-8. Soluble starch increased the protease productivity, while glucose repressed it. Yeast extract was effective nitrogen source for enzyme production, but the pretense production of Bacillus sp. SH-8 was reduced by large amount of yeast extract. The calcium was found to induce pretense activity as well as protease productivity. However, cell growth and enzyme production was completely inhibited by divalent ions such as $Zn^{2+}$, $Cu^{2+}$, $Co^{2+}$ and $Mn^{2+}$. The maximum protease productivity was reached 435 unit/ml in the optimized medium consisting of soluble starch (2%), yeast extract (0.3%), $CaCl_2$ (0.3%), $K_2HPO_4$ (0.01%) and $KH_2PO_4$ (0.01%). The pretense activity of culture filtrate was dramatically decreased after incubation for 26 h.

• Journal of Life Science
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• v.8 no.3
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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.