• Journal of Microbiology and Biotechnology
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• 제10권2호
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• pp.181-186
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• 2000

MAPI (microbial alkaline protease inhibitor) was isolated from cultrue broth of Streptomyces chromofuscus SMF28. The Ki values of MAPI for the representative serine proteases such as chymotrypsin and proteinase K were 0.28 and $0.63{\;}\mu\textrm{M}$, respectively, and for the cysteine proteases cathepsin B and papain were 0.66 and $0.28{\;}\mu\textrm{M}$, respectively. These data indicate that MAPI is not a potent selective inhibitor of serine or cysteine proteases. Progress curves for the inhibition of three proteases by MAPI exhibithe characteristic patterns; MAPI exhibited slow-binding inhibition of cathepsin B. It was rapidly associated with chymotrypsin before the addition of substrate and then reactivation of MAPI-inhibited enzyme was investigated in the presence of substrate. On the other hand, MAPI-proteinase K interaction was typical for those classical inhibitors. When MAPI was incubated with trypsin, there was an extensive reduction in the ingibitory activities of MAPI corresponding to 66.5% inactivation of MAPI, indicating that trypsin-like protease may play a role in the decrease of the inhibitory activity during cultivation.

• Biomolecules & Therapeutics
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• 제4권4호
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• pp.385-390
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• 1996

General pharmacological properties of urinary trypsin inhibitor (UTI) following intravenous administration of 1,000,000 units/kg were examined in terms of effects on central nervous system, cardiovascular system, respiratory system, gastrointestinal system in mice, rats and rabbits. Administration of UTI (1,000,000 units/kg, iv) had no effect on central nervous system; no influences on pentobarbital sleeping time, spontaneous activity, normal body temperature, chemoshock produced by pentylenetetrazole solution, writhing syndromes induced by 0.6% acetic acid solution, and motor coordination of mice. The administration of UTI (1,000,000) units/kg, iv) in rats had no effect on systolic blood pressure and pulse rate. UTI (500,000 units/kg, iv) given to anesthetized rabbits showed no effect on respiratory rate. However, it showed significant elevation of respiratory rate at the concentration of 1,000,000 units/kg. Gastric secretion of rat and intestinal motility of mice were not influenced by the dose of 1,000,000 units/kg. In terms of autonomic nervous system, the material did not show direct effect and inhibitory or augmentative action of histamine- or acetylcholine-induced contraction at the concentration of 2,000 units/ml in the isolated ileum of guinea pig.

• BMB Reports
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• 제32권1호
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• pp.86-91
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• 1999

The sprT gene encoding Streptomyces griseus trypsin (SGT) was introduced into Streptomyces lividans TK24 and Streptomyces lividans 1326 to study which strain would be better to overexpress the extracellular proteinase. Various media with different compositions were also used to maximize the productivity of SGT in heterologous hosts. The SGT productivity was best when the transformants of S. lividans TK24 and 1326 were cultivated in R2YE medium, and their relative trypsin activity of the culture broth measured with an artificial chromogenic substrate, N-${\alpha}$-benzoyl-DL-arginine-${\rho}$-nitroanilide, were 382 units/ml and 221 units/ml, respectively. They produced high levels of SGT in GYE medium but relatively lower than those in R2YE medium, and negligible amount of SGT was produced in Ferm, RASF, LIVID, and NDSK media. Considering non-SGT associated activity in Pronase powder, it was estimated that the transformant of S. lividans TK24 can produce SGT in R2YE 3.5 times more than the amount by S. griseus 10137 from which the sprT gene had been originated. The growth of S. lividans reached the maximum level of cell mass at 5 d of culture, but SGT production started in the stationary phase of cell growth and kept increasing until the ninth day of culture in R2YE medium, but in GYE media the productivity reached at the maximum level at 7 d of cultivation.

• 한국축산식품학회지
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• 제25권2호
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• pp.238-243
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• 2005

산양유 $\beta-casein$의 효소에 의한 가수분해 특성과 가수분해물의 ACE 저해 효과를 측정하고자 산양유의 $\beta-casein$을 양이온 교환 컬럼인 Mono S HR 5/5를 이용하여 분리하였으며 분리된 $\beta-casein$을 동물성 분해효소인 trypsin으로 처리하여 가수분해 특성을 확인하였고 가수분해물의 ACE 저해활성을 측정하였다. Mono S HR 5/5 양이온 교환 컬럼을 이용한 산양유 산 케이신으로 부터 순수한 $\beta-casein$의 분리는 SDS-PAGE를 이용하여 확인한 결과 순수한 $\beta-casein$의 분리가 이루어졌음을 확인할 수 있었다. $\beta-casein$을 $37^{\circ}C4$에서 trypsin으로 처리하여 전기영동으로 확인한 결과 가수분해 직후부터 $\beta-casein$ 위치의 band가 희미해지기 시작하고 저분자량의 band가 나타나기 시작하였으나 120분이 지난 후에는 모든 band가 가수분해되어 사라졌고 산양유에서 분리된 $\beta-casein$을 trypsin으로 처리하여 120분 경과 후 그 가수분해물을 이용하여 ACE 저해효과를 측정한 결과 가수분해하지 않은 $\beta-casein$은 $1.80\pm1.21\%$의 ACE 저해활성을 보였으나 trypsin으로 가수분해하여 ACE 저해 활성을 측정하였을 때 $25.36\pm0.79\%$의 저해 활성을 나타내었으며, trypsin에 의한 $\beta-casein$가수분해물의 $IC_{50}$을 측정한 결과 $308.7\pm2.77({\mu}g/mL)$로 나타났다.

• 한국식품과학회지
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• 제26권4호
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• pp.464-470
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• 1994

환삼덩굴의 생리활성물질을 검색하기 위하여 물과 메탄올로 추출하여 항균성과 trypsin 저해활성 및 항산화 효과를 조사하였으며, 메탄올 추출물은 다시 계통 분획하여 분획별 활성을 조사하였다. 세균 6종과 효모 2종을 대상으로 항균성을 조사한 결과, 물 추출물 보다는 메탄올 추출물의 항균성이 우수하였으며, Escherichia coli를 제외한 모든 시험균주에 항균효과를 나타내었다. 분획별 항균효과를 비교한 결과, 부탄올 분획이 모든 시험균주에 항균효과를 나타내었으며, 균종별로는 Staphylococcus aureus와 Saccharomyces cerevisiae에 대한 항균효과가 우수하였다. 부탄올 분획의 미생물에 대한 최소저해농도(MIC)는 $0.1{\sim}0.4%$ 범위였다. 환삼덩굴 추출물의 trypsin 저해활성을 조사한 결과, 물 추출물은 저해활성을 나타내지 않았으나, 메탄올 추출물은 저해활성을 나타내었다. 분획별 저해활성은 클로로포름 분획에서 가장 높았으며 클로로포름 분획의 50% 저해농도($IC_{50}$)는 1mg/ml였으며, EI(enzyme-inhibitor) 복합체 형성시간은 20분에 90% 이상이었다. 환삼덩굴의 메탄올 추출물은 돈지와 대두유를 기질로한 항산화성 시험에서 과산화물의 생성을 억제하였다. 메탄올 추출물의 분획별 활성은 클로로포름 분획에서 가장 높게 나타났으며, 클로로포름 분획의 농도를 0.2% 첨가하였을 때, 유지의 유도기간은 돈지는 15일, 대두유는 9일로서 산화안정성이 연장되었다.

• Journal of Ginseng Research
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• 제9권1호
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• pp.72-85
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• 1985

Attempts were made to see if feeding of ginseng ethanol extract could affect proper- ties of rat brain lactate dehydrogenase such as specific activity, heat stability, Km for substrate, inactivation by 3-bromopyruvate and trypsin, and immune response. The following results were obtained. Specific activity of LDH was observed to reach maximum in 5 month after birth and then decrease steadily. However, that of LDH from rat fed with ginseng ethanol extract was found in rat fed with ginseng ethanol extract. 3-bromopyruvate was shown to inactivate LDH-5 from old rat fed. Inactivation of LDH-5 by trypsin was remarkable in old rat fed. Km value for pyruvate in old rat fed was remarkably decreased. Cumulative results suggest that ginseng ethanol extract could affect conformational change of LDH responsible for altered properties through unknown mechanism.

• 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 Veterinary Science
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• 제21권1호
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• pp.12.1-12.6
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• 2020

Porcine deltacoronavirus (PDCoV) is an emerging swine enteric coronavirus that causes diarrhea in piglets. However, the biological characteristics of PDCoV are unclear. In this study, the hemagglutination (HA) abilities of two PDCoV strains (CH-01 and HNZK-04) were investigated. Our results showed that PDCoV has the ability to agglutinate rabbit erythrocytes after virion pretreatment with trypsin or neuraminidase. Additionally, the HA assay results showed a significant positive correlation with the infectious viral titer. Our results suggest that assessing the HA activity of PDCoV may be a useful diagnostic method for investigating and surveilling PDCoV infections.

• 한국축산식품학회지
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• 제31권6호
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• pp.886-892
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• 2011

This study investigated the ACE-inhibitory effect of yogurt beverage fortified with hydrolysates as well as the suitability of hydrolysates as a nutraceutical additive to yogurt beverage. Three whey protein hydrolysates hydrolyzed by alcalase, protamex, and trypsin were each added to yogurt beverage at concentrations of 1.25, 2.5, and 5 mg/mL. Yogurt beverage fortified with 2.5 mg/mL of hydrolysates had 61-69% ACE-inhibitory activity, whereas yogurt beverage fortified with 5 mg/mL of hydrolysates showed 74% ACE-inhibitory activity. There were no significant differences in ACE-inhibitory activity between the alcalase or protamex hydrolysates during storage; however, trypsin hydrolysate exhibited significant differences. On the other hand, physicochemical characteristics such as pH (3.47-3.77), titratable acidity (0.81-0.84%), colority, viable cell count, and sensory qualities were not significantly different among the tested yogurt beverage samples during storage. These results showed that yogurt beverage fortified with whey protein hydrolysates maintained antihypertensive activity and underwent no unfavorable changes in physicochemical characteristics regardless of enzyme type.

• 농업생명과학연구
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• 제46권5호
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• pp.111-125
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• 2012

Lipoxygenase(LOX)와 kunitz trypsin inhibitor(KTI) 단백질이 결핍된 non-GM콩(개척#2, 진양콩 및 CJ#1)의 가공적성을 알아보기 위하여 일반콩(태광콩)을 대조로 하여 간장을 제조하여 이화학적 특성 및 항산화 활성을 비교하였다. 원료콩의 일반성분은 시료간에 비슷한 조성을 보였으나, 총 페놀 및 플라보노이드 함량은 태광콩에 비해 LOX와 KTI 단백질 결핍콩에서 유의적으로 높았다. 간장 중 총당 및 환원당 함량은 태광콩 간장이 가장 높았으며, 총 질소 및 아미노태 질소 함량은 진양콩 간장이 가장 높았다. 무기물은 진양콩 및 CJ#1 간장, 아미노산은 3종의 LOX 및 KTI 단백질 결핍콩 간장이 태광콩 간장에 비해 높은 함량이었다. 간장의 갈색도는 진양콩 간장이 가장 높았으며, 황색도는 진양콩 간장이 태광콩 간장과 비슷한 수준이었다. 간장의 종합적인 기호도는 단맛과 구수한 맛이 높았던 진양콩 간장의 점수가 가장 높았다. 간장의 총 페놀 및 플라보노이드 함량은 LOX와 KTI 단백질 결핍콩 간장이 태광콩 간장에 비해 유의적으로 높았다. ABTS 및 DPPH 라디칼 소거활성은 총 페놀 함량에 의존적이었으며, FRAP법에 의한 환원력은 개척#2 및 CJ#1 간장이 태광콩에 비해 유의적으로 높았으며, $Fe^{2+}$ 킬레이트 활성은 태광콩 간장이 여타 간장의 활성에 비해 높았으나, 진양콩 간장과 비슷한 수준이었다. 따라서 LOX와 KTI 단백질 결핍콩으로 제조한 간장은 일반콩 간장에 비해 영양적 성분 및 항산화 활성이 높아 기능성 가공식품 소재로써 활용 가능성이 있는 것으로 판단된다.