• 미생물학회지
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• 제29권6호
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• pp.345-352
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• 1991

A serine proteinase of molecular weight 60 kd was purified from culture supernatant of P. aeruginosa using DEAE-Trisacryl M ion-exchange and AcA 54 gel filtration column chromatography, and the properties of serine proteinase were characterized. By means of SDS-polyacrylamide gel electrophoresis, the molecular weight of the enzyme was 55 kd. The optimal pH for the activity of purified enzyme was 7.5. The activity of the purified enzyme was completely inhibited by Di-isopropylfluorophosphate(DFP) and N-.alpha.-p-tosyl-L-lysine choloromethyl detone(TLCK) but not by other proteinase inhibitors such as E-64, pepstatin A, 1, 10-phenanthroline. The purified enzyme was capable of degrading type I and type IV collagen. Antisera obtained from hymans infected with Pseudomonas aeruginosa reacted to the purified serine proteinase in immunoblots. These results indicate that the purified enzyme is trypsin-like serine proteinase and this enzyme of P. aeruginosa may play an important role in tissue damage as a spreading factor and may be useful for serodiagnosis of Pseudomonas infections.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.381-385
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• 1996

Three distinct plasmids, with approximate molecular weights of 1, 4.5, and 33 megadaltons, were found in Lactococcus lactis subsp. lactis (L. lactis) ML8. Slow acid-producing mutants of L. lactis ML8, isolated by plasmid curing with acriflavine treatment, lacked the 33-megadalton plasmids. The plasmid-cured mutant showed lactose-negative (Lac) characteristics and the alteration of extracellular proteinase pattern. The possible involvement of extracellular proteinase with the 33-megadalton plasmid is highlighted in this research.

• 미생물학회지
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• 제28권1호
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• pp.65-70
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• 1990

방선균의 세포분화와 관련된 연구의 일환으로 Streptomyces fradiae NRRL2702가 생성하는 단백질분해효소 저해물질을 분리정제하여 그 특성을 분석하였다. 즉 S. fradiaesm s일반 환경 조건하에서 단백질분해효소를 생성하나 특정 조건하에서는 그 단백질의 활성을 저해하는 저해제를 생성함을 알았다. 이 저해제의 분자량은 16,800이며 serine proteinasem이 일부만을 저해하는 특징이 있었다. Pronase E의 활성의 저해양상은 competitive inhibition 이었고 열에 대하여 매우 안정함을 알았다.

• 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.

• 한국미생물·생명공학회지
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• 제24권4호
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• pp.452-458
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• 1996

A yeast strain TH65 producing a high level of proteinase under alkaline condition was isolated, and identified as Yarrowia lipolytica by morphological, physiological, and biochemical characteristics. In proteinase productivity, glycerol and glucose among tested carbon sources were very effective, and optimum concentration of glucose was 0.5%. Skim milk was found to be most effective nitrogen source in productivity, and its optimum concentration was 0.6%. But, cysteine, cystine and tryptophane decreased the proteinase productivity. Yeast extract was relatively effective at the range of 0.1-0.5%. The yeast showed maximum production of proteinase at 18$\circ$C, pH 9-11, and cultivation time of 36 hours.

• 한국식품과학회지
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• 제21권1호
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• pp.86-91
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• 1989

Streptococci의 배양중의 염의 농도나 배지내 질소원의 조성 등이 쓴 맛 펩타이드의 형성에 깊이 관여한다고 알려진 세포내 및 세포외 프로테이나제(ICP, ECP)의 역가에 미치는 영향을 조사하기 위해 Streptococcus lactis $ML_8$ 및 Streptococcus cremoris $ML_4$ 두 균주를 NaCl 농도 $0{\sim}4%$, 배지 질소원중 카제인 함량을 $0{\sim}100%$로 달리한 조건에서 배양하고 그 때의 생육도와 균체내 및 균체외 프로테이나제의 역가를 측정하고 또한 같은 균체수에서의 프로테이나제 역가를 비교하기 위해 $10^{10}$ 세포당 효소의 역가도 계산하였다. 배지중 NaCl농도가 증가함에 따라 균체의 생육과 ECP의 역가는 감소했으나 ICP의 경우 $10^{10}$ 세포당 역가는 오히려 증가하였다. 이는 배지중 NaCl이 세포벽과 세포막에 관련된 ECP의 역가에만 직접 영향을 미치고 ICP의 역가에는 큰 영향을 미치지 않음을 시사하는 것으로 생각되었다. 배지의 질소원중 미분해 단백질인 카제인의 함량을 높게 하였을 때 균체의 생육은 낮았으나 효소 생산계 촉진에 의해 ECP의 역가는 두 균주 모두 높아졌다.

• Parasites, Hosts and Diseases
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• 제44권4호
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• pp.321-330
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• 2006

The pathogenic mechanism of granulomatous amebic encephalitis (GAE) and amebic keratitis (AK) by Acanthamoeba has yet to be clarified. Pretense has been recognized to play an important role in the pathogenesis of GAE and AK. In the present study, we have compared specific activity and cytopathic effects (CPE) of purified 33 kDa serine proteinases from Acanthamoeba strains with different degree of virulence (A. healyi OC-3A, A. lugdunensis KA/E2, and A. castelianii Neff). Trophozoites of the 3 strains revealed different degrees of CPE on human corneal epithelial (HCE) cells. The effect was remarkably reduced by adding phenylmethylsulfonylfluoride (PMSF), a serine proteinase inhibitor. This result indicated that PMSF-susceptible proteinase is the main component causing cytopathy to HCE cells by Acanthamoeba. The purified 33 kDa serine proteinase showed strong activity toward HCE cells and extracellular matrix proteins. The purified proteinase from OC-3A, the most virulent strain, demonstrated the highest enzyme activity compared to KA/E2, an ocular isolate, and Neff, a soil isolate. Polyclonal antibodies against the purified 33 kDa serine proteinase inhibit almost completely the proteolytic activity of culture supernatant of Acanthamoeba. In line with these results, the 33 kDa serine proteinase is suggested to play an important role in pathogenesis and to be the main component of virulence factor of Acanthamoeba.

• Journal of Microbiology and Biotechnology
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• 제25권10호
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• pp.1734-1741
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• 2015

Few research had investigated the secretion of phospholipase and aspartyl proteinase from Candida spp. causing infection in females with type 2 diabetes mellitus. This research aimed to investigate the prevalence of vulvovaginal candidiasis (VVC) in diabetic versus non-diabetic women and compare the ability of identified Candida isolates to secrete phospholipases and aspartyl proteinases with characterization of their genetic profile. The study included 80 females with type 2 diabetes mellitus and 100 non-diabetic females within the child-bearing period. Candida strains were isolated and identified by conventional microbiological methods and by API Candida. The isolates were screened for their extracellular phospholipase and proteinase activities by culturing them on egg yolk and bovine serum albumin media, respectively. Detection of aspartyl proteinase genes (SAP1 to SAP8) and phospholipase genes (PLB1, PLB2) were performed by multiplex polymerase chain reaction. Our results indicated that vaginal candidiasis was significantly higher among the diabetic group versus nondiabetic group (50% versus 20%, respectively) (p = 0.004). C. albicans was the most prevalent species followed by C. glabrata in both groups. No significant association between diabetes mellitus and phospholipase activities was detected (p = 0.262), whereas high significant proteinase activities exhibited by Candida isolated from diabetic females were found (82.5%) (p = 0.000). Non-significant associations between any of the tested proteinase or phospholipase genes and diabetes mellitus were detected (p > 0.05). In conclusion, it is noticed that the incidence of C. glabrata causing VVC is increased. The higher prevalence of vaginal candidiasis among diabetics could be related to the increased aspartyl proteinase production in this group of patients.

• 한국동물학회:학술대회논문집
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• 한국동물학회 1996년도 한국생물과학협회 국제학술대회
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• pp.204.1-204
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• 1996

No Abstract, See Full Text

• Journal of Microbiology
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• 제35권2호
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• pp.109-116
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• 1997

An extracellular proteinase of Candida albicans was purified by a combination of 0~75% ammonium sulfate precipitation, DEAE Sepharose Fast Flow ion exchange chromatography, and Sephacryl S-200 HR molecular sieve chromatography. Its mlecular weight was approximately 41 kDa on SDS-PAGE and isoelectric point was 4.4. The enzyme was inhibited by pepstain A. Optimum enzyme activity ranged from pH 2.0 to 3.5 with its maximum at pH 2.5 and a temperature of 45$^{\circ}C$. The addition of divalent cations, $Ca^{2+}$, Zn$^{2+}$ and $Mg^{2+}$, resulted in no significant inhibition of enzymatic activity. However, some inhibitory effects were observed by Fe$^{2+}$, Ag$^{2+}$ and Cu$^{2+}$. With BSA as substrate, an apparent $K_m$ was determined to be 7$\times$10$^{-7}$ M and $K_i$, using pepstatin A as an inhibitor, was 8.05$\times$10$^{-8}$ M. N-terminal amino acid sequence was QAVPVTLXNEQ. Degradation of BSA and fibronectin was shown but not collagen, hemoglobin, immunoglobulin G, or lysozyme. The enzyme preferred peptides with Glu and Leu at the P$_1$ position, but the enzyme activity was highly reduced when the P$_2$ position was phe or pro. This enzyme showed antigenicity against sera of patients with candidiasis.