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곤충 병원성 곰팡이 Beauveria bassiana로부터 Protease의 정제와 특성
고휘진,김현규,김범기,강선철,권석태 ( Hwi Jin Ko,Hyun Kyu Kim,Beom Gi Kim,Sun Chul Kang,Suk Tae Kwon ) 한국응용생명화학회 1997 Applied Biological Chemistry (Appl Biol Chem) Vol.40 No.5
Extracellular protease (bassiasin I), from the culture filtrate of entomopathogenic fungus Beauveria bassiana ATCC7159, was successively purified by precipitation with ammonium sulfate followed by DEAE-Sephadex A-50, CM-cellulose and Hydroxyapatite column chromatography. A typical procedure provided 41-fold purification with 13.6% yield. The molecular weight of the purified protease (bassiasin I) was found to be approximately 32,000 by SDS-PAGE. Isoelectric-focusing analysis of the enzyme showed a pI of 9. 5. NH₂-terminal sequence of the protease showed homology with those of the fungal proteases. The enzyme has an optimal pH for activity at 10.5 and is stable over pH 5.0-11.0. The maximum activity of the enzyme was at 60-65℃, and approximately 20% activity remained at 60℃ after 120 min. The protease was inhibited by phenylmethylsulfonyl fluoride (PMSF) and diisopropyl fluorophosphate (DIFP).
곤충 병원성 곰팡이 Beauveria bassiana 로부터 Protease 의 최적 생산
고휘진(Hwi Kyung Kim),김현규(Hyun Kyu Kim),강선철(Sun Chul Kang),권석태(Suk Tae Kwon) 한국응용생명화학회 1996 Applied Biological Chemistry (Appl Biol Chem) Vol.39 No.6
We investigated the optimal condition for the production of extracellular protease(a cuticle-degrading protease) from entomopathogenic fungus Beauveria bassiana(ATCC7159) in liquid medium by adding of gelatin, bovine serum albumin(BSA), casein and polypeptone. The optimal induction medium for production of extracellular proteases is composed of 0.5% polypeptone, trace elements and 50 mM potassium phosphate(pH 6.0). In this condition, the production of extracellular proteases increased rapidly after the 24hrs, peaking at the third day and there was little inductive effect in culture broth more than pH 7.0. The proteases were inhibited by phenyl methyl sulfonyl fluoride(PMSF). High activity of protease was showed both range of pH 8.5 and 11.5 and also detected by three different portions of slice gel derived from nondenaturing isoelectricfocusing gel. At least three different extracellular proteases are produced in optimal production medium when polypeptone is used as the sole carbon and nitrogen source.
조상원,고휘진,박태현 한국생물공학회 2021 Biotechnology and Bioprocess Engineering Vol.26 No.1
Several studies investigated the development of a non-invasive and early diagnostic method for cancer. Recently, the volatile organic compounds (VOCs) exhaled from cancer patients have been shown to act as a potential biomarker for cancer. However, the gas profile of breath is highly variable and not conducive to identification of biomarkers accurately. Accordingly, we identified a cancerspecific VOC as a biomarker using an in vitro cancer system in order to exclude the variables from patient condition. VOCs were collected from the headspace gas of lung cancer cell line (SK-MES) and normal lung cell line (MRC-5), followed by analysis using solid-phase microextraction (SPME)-GC/MS. As a result, 2-ethyl-1-hexanol was detected from headspace gas of SK-MES and decided as a potential biomarker for lung cancer. By screening human olfactory receptor libraries to detect a lung cancer biomarker, human olfactory receptor 4D11P (hOR4D11P) was identified as a receptor specific for 2-ethyl-1-hexanol and expressed heterologously in HEK293 cells. Additionally, we generated nanovesicles containing hOR4D11P from HEK293 cells and confirmed the expression and functionality of hOR4D11P in nanovesicles.