• Journal of Microbiology and Biotechnology
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• 제3권4호
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• pp.244-251
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• 1993

A novel system has been developed to produce ${\delta}-aminolevulinic$ acid (ALA) using the intact cells of late logarithmic phase of Rhodocyclus gelatinosus KUP-74. The system was shown optimum yield of extracellular ALA under a condition of anaerobic light irradiation (4 Klux) at $30^{\circ}C$ with no variation in cell mass. The rate of extracellular ALA formation was stimulated by low doses of either $C_4\;or\;C_5$ ALA biosynthetic precursors, where 5 mM ($C_4) and 3 mM ($C_5) of each precursors were appeared to generate the maximum rates of 3.3 and 4.0 nmoles of ALA per 0.35 mg cells per hr, respectively. Half-life of the system was 10 hr in a sense of an ability of portage transport of L-glutamate, and sequential dose of this compound was resulted in promising recovery of the ALA.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.161-161
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• 1998

To achieve the aim of this investigation, the extracellular protease was isolated from bacteria BAC-4, a strain was cultivated in the medium for the production of penicillin acilase in a period of 32 hours. The enzyme was first purified by aceton precipitation method, followed by ion exchange chromatography on DEAE-sephacel column. The highest specific activity of the aceton fraction was found to be 2.19 unit per mg, with degree of purification of 13 times. Further purification of the enzyme on DEAE -sephacel had a specific activity of 58.6 unit per mg and degree of purification of 344 times compared to its crude extract. The optimum pH of the enzyme was 8.4, and the potimum temparature was 37$^{\circ}C$. The K$\_$M/ and $V_{max}$ calculated at experiment conditions were found to be 0.66%(W/V) and 3.61 unit per mL respectively.

• Journal of Microbiology and Biotechnology
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• 제3권3호
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• pp.174-180
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• 1993

A strain SMF14 producing an extracellular $\beta$-lactamase was isolated from soil and identified to be a strain of Streptomyces aureofaciens. $\beta$-Lactamase was purified from the cell free culture broth through batchwise hydroxyapatite adsorption, anion exchange chromatography on DEAE Sephadex A-50, gel filtration on Sephadex G-75, and adsorption chromatography on hydroxyapatite. The molecular mass was estimated to be about 43 kDa by SDS-PAGE. The $\beta$-lactamase had substrate specificity to penicillins and it was inhibited by clavulanic acid, being classified to the group 2a of penicillinase.. The optimal reaction pH and temperature were pH 6.0~7.5 and $50^{\circ}C$. The $K_m, and V_{max}$ values of $\beta$-lactamase for penicillin G were calculated to be 1.72 mM and 5.4$\times$$10^5 \mu M \cdot min^{-1}$, respectively.

• 대한음성언어의학회:학술대회논문집
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• 대한음성언어학회 2016년도 제44차 춘계학술대회
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• pp.41-41
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• 2016

• 약학회지
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• 제43권4호
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• pp.458-463
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• 1999

This study was undertaken to test for antiviral activity of the aqueous extracts prepared from 4 medicinal plants of Korea (Terminalia chebula, Sanguisorba officinalia, Rubus coreanus, Rheum palmatum). Aqueous extracts were assayed for the inhibition of hepatitis B virus (HBV) replication by measurement of HBV DNA and surface antigen (HBsAg) levels in the extracellular medium of HepG2 2.2.15 cells. All extracts decreased the levels of extracellular HBV virion DNA at concentrations ranging from 64 to $128{\;}\mu\textrm{g}/ml$ and inhibited the production of HBsAg dose-dependently. Among the 4 tested plants, Terminalia chebula exhibits the most prominent anti-HBV activities. Our findings suggest that these 4 medicinal plants may have potential to develop as specific anti-HBV drugs in the future.

• Mycobiology
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• 제31권4호
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• pp.209-213
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• 2003

Aspergillus flavus K-03 isolated from poultry forming soil in Korea was studied for its ability to produce extracellular proteases on basal medium containing 2%(w/v) chicken feathers. The fungus was observed to be a potent producer of such enzymes. Keratinolytic enzyme secretion was the best at 15 days of incubation period at pH 9 and temperature $40^{\circ}C$. No relationship existed between the enzyme yield and increase of biomass. Enzyme production was suppressed by exogenous sugars in descending order arabinose>maltose>mannose>fructose. But glucose did not influence the enzyme activity. The keratinolytic enzyme released by the fungus demonstrated the ability to decompose keratin substrates as chicken feather when exogenous glucose was present. The keratinolytic activity was inhibited by $HgCl_2$ and serine-protease inhibitors such as phenymethylsulfonyl fluoride(100%), chymostain(88%), crystalline soybean trypsin inhibtor(80%), antipain(45%) and aprotinin(40%), and was not by cystein-protease and aspartyl-protease inhibitors. The enzyme activity is only partially inhibited by metallo-protease inhibitor. Thus, the enzyme secreted by A. flavus K-03 belongs to the alkaline serine-type protease.

• 한국미생물·생명공학회지
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• 제47권1호
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• pp.96-104
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• 2019

Beauveria bassiana, one of the most common entomopathogenic fungi, has been isolated, pre defined and characterized in-house from soil of tea cultivation area. Experiments have been performed to verify the presence of chitinase as intracellular metabolite and its release as extracellular product rendering the spores with biopesticide activity. Although there are many responsible enzymes for the pest killer action of B. bassiana, binding property of chitinase depending on presence as well as absence of serine supplemented in the media has been studied with respect to the production and kinetics. A programmed investigation conclusively indicates that the isolated spore (hyphae) of B. bassiana has been metabolically enriched with the enzyme chitinase in presence of an externally added amino acid serine with its inhibitory kinetics.

• 한국지하수토양환경학회지:지하수토양환경
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• 제14권6호
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• pp.45-52
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• 2009

나프탈렌 분해균주인 Pseudomonas aeruginosa CZ6을 오염된 토양에서 분리하였으며 분리된 균주는 결정상태의 나프탈렌에 부착하고 그 주변에 extracellular polymeric substance를 분비하는 특성을 가졌다. LB, YM과 MSM 배지를 사용하여 배지의 종류에 따른 biofilm 생성량을 측정한 결과, LB 배지에서 biofilm이 가장 많이 생성되는 것으로 나타났다. 나프탈렌을 기질로 한 배양조건에서 균주는 기질의 농도 영향을 크게 받지 않고 0.10% 기질, 150 rpm 조건에서 최적 배양조건은 $30^{\circ}C$, pH 7로 나타났다. 두 가지 토양에서 배지의 종류에 따른 나프탈렌의 분해특성을 관찰한 결과 초기에는 MSM 배지에서 나프탈렌이 가장 많이 분해가 되었다. 그러나 생물이용성이 제한을 받는 조건에서 LB 배지의 나프탈렌이 가장 빨리 제거가 되었다. 이런 결과는 biofilm의 형성과 extracellular polymeric substance 생성이 토양에 흡착된 잔류 나프탈렌의 생물학적 이용성을 향상시키기 때문인 것으로 사료된다.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.430-434
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• 2004

Three extracellular protease-deficient Bacillus subtilis strains were transformed with the plasmid pCK98 containing the endo-$\beta$-1,4-glucanase (Eng) gene of B. subtilis BSE616. The three transformants, B. subtilis DB104 (pCK98), WB600 (pCK98) and WB700 (pCK98), produced the same high level of enzyme activity and showed similar patterns of cell growth and enzyme production. When B. subtilis DB 104 (pCK98), a two-extracellular protease deficient strain, was cultured for 22 h, almost all the secreted enzyme was found to be in the completely cleaved form by both activity staining and Western blotting studies. B. subtilis WB600 (pCK98), a six-extracellular protease-deficient strain, produced a partially cleaved form in addition to the intact form of the enzyme, although the degree of internal cleavage of the enzyme was greatly reduced. With B. subtilis WB700 (pCK98), a seven-extracellular protease-deficient strain, almost all the enzyme was produced as the intact uncleaved form. This study illustrates that a role of the V pr protease is to degrade foreign proteins produced in B. subtilis and WB700 is a suitable expression system for producing the intact form of the Eng and other foreign proteins that may lose at least part of their efficacy due to internal proteolytic cleavage.

• 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.