• 한국발생생물학회지:발생과생식
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• 제7권2호
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• pp.113-118
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• 2003

포유동물의 성숙한 난포의 난포액 속에는 여러 종류의 단백질 분해효소가 있으며 이들은 난포의 형성과 퇴화 및 난자의 성숙과 배란 등의 다양한 변화에 중요한 역할을 하는 것으로 여겨진다. 난포액 속의 단백질 가수분해효소 중에는 serine proteinase가 비교적 잘 알려져 있으나 다른 효소 특히, caseinolytic enzyme에 대해서는 거의 알려져 있지 않다. 본 연구에서는 사람의 난포액을 재료로 하여 caseinolytic enzyme의 존재 여부 및 동 효소의 특성을 알아보고자 하였다. 사람의 난포액과 혈청 그리고 사람의 제대혈을 $\alpha$-casein을 기질로 하는 zymography 방법으로 분석 한 결과 분자량 80 kDa의 매우 강한 caseinolytic activity를 지니는 효소 단백질과 분자량 78 kDa의 비교적 약한 caseinolytic activity를 갖는 단백질 등 두 개의 caseinolytic enzyme이 관찰되었다. 이 caseinase들의 특성을 알아보기 위해 phenylmethylsulfonyl fluoride(PMSF), soybean trypsin inhibitor(SBf), 1,10-phenanthroline, E-64 그리고 ethylenediamine tetraacetic acid(EDTA)를 zymouaphy의 substrate buffer에 처리한 결과 EDTA와 SBTI에 의해서 80 kDa와 78 kDa caseinase의 활성이 억제되었다 이로 미루어 80 kDa와 78 kDa caseinase는 trypsin-like enzyme인 것으로 추측된다. 한편 사람 난포액의 zymouaphy 수행시에 5 mM의 EDTA가 첨가된 substrate buffer에 CaC $l_2$, MgC $l_2$, MnC $l_2$ ZnC $l_2$를 각각 0에서 10 mM의 농도별로 처리한 결과 모두 5 mM 농도에서 가장 높은 caseinase 활성을 보였다. 금속 이온의 첨가없이 EDTA만 처리한 대조군의 경우 caseinase의 활성은 나타나지 않았다. 이로 미루어 80 kDa 및 78 kDa caseinase는 효소 활성을 위해 이가 금속 양이온을 필요로 하는 것으로 여겨진다.

• 한국수산과학회지
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• 제46권4호
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• pp.351-358
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• 2013

A protease inhibitor from fish eggs was fractionated using chromatographic methods. The fractionation efficiency was evaluated in terms of specific inhibitory activity (SIA, U/mg), purity (fold), total inhibitory activity (TIA, U), and recovery (%). The protease inhibitor (PI) from egg extracts of skipjack tuna (ST Katsuwonus pelamis), yellowfin tuna (YT Thunnus albacares) and Alaska pollock (AP Theragra chalcogramma) was fractionated using Sephadex G-50 gel filtration and DEAE-Sepharose CL-6B anion exchange chromatography based on protein size exclusion and net charge, respectively. Fractions exhibiting strong inhibitory activity were contained in the 30-50 kDa fraction on gel filtration and in the range of 0.4-0.7 M NaCl gradient fraction on anion exchange chromatography. The respective TIA and percent recovery of the fraction obtained with gel filtration toward trypsin and $N{\alpha}$-benzoyl-L-arginine-p-nitroanilide (BAPNA) were 2,758.7 U and 29.6% for ST, 1,005.5 U and 25.6% for YT, and 1,267.5 U and 26.0% for AP. Gel filtration chromatography was more effective at fractionating PI than using ion exchange chromatography. These results suggest that fish eggs act as serine protease inhibitors and might be useful for protease inhibition in foodstuffs.

• 한국식품과학회지
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• 제22권6호
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• pp.668-674
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• 1990

대두, 수수, 대두-수수 혼합곡물 템페를 인도네시아의 전통적인 종균(LARU ; mixed cultures of Rhizopus oligosporus)를 사용하여 제조하였다. 발효가 끝난 후 대두 템페와 대두-수수 혼합템페는 단백질과 섬유질의 양이 발효시키지 않은 대조군 시료보다 증가하였고, 지방 함량은 수수템페와 대두-수수 혼합템페의 경우 약간 증가하였다. 발효가 진행함에 따라 3가지 형태의 템페 모두가 pH, 수용성 고형분, 수용성 질소 함량이 증가한 반면 총고형분 함량에는 큰 변화가 없었다. 트립신억제제 활성(TIA)과 피트산 함량은 발효 32시간 후 감소되었다. 이는 Rhizopus oligosporus 균이 대두를 발효시키는 과정에서 트립신억제제와 피트산을 분해하는 분해능을 가지고 있음을 제시하는 결과이다. 티아민과 나이아신 함량이 3가지 형태의 템페 모두에서 증가하였다. 32시간 발효 후 대두, 수수, 대두-수수 혼합템페 모두에서 총아미노산 함량은 감소하였고 아미노산 중 리신, 발린, 티로신, 알라닌 등이 증가하였고 세린과 글루탐산 이 감소하였다.

• 한국식품과학회지
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• 제30권5호
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• pp.1012-1017
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• 1998

진품콩에 함유된 lipoxygenase, urease, trypsin inhibitor, 탄닌, 피트산을 60, 80, $100^{\circ}C$로 열처리 하는 중의 함량변화를 조사하였다. lipoxygenase-1은 $60^{\circ}C$로 가열할 경우 100 분까지 활성이 있었으나, $80^{\circ}C$, $100^{\circ}C$에서는 20분과 10분 후에 활성이 없어졌다. lipoxygenase-2와 lipoxygenase-3의 활성은 없었다. urease활성은 $60^{\circ}C$로 100분을 가열하여도 초기와 비슷하였으나, $80^{\circ}C$에서는 20분, $100^{\circ}C$에서는 10분만에 완전히 불활성화 되었다. 트립신 저해제는 $60^{\circ}C$에서는 비교적 서서히 파괴되어 100분 후에는 58.6%가 활성이 없어졌으며, $80^{\circ}C$와 $100^{\circ}C$의 경우는 가열 초기에 급격히 파괴되지만, $20{\sim}30$분 후부터는 파괴 속도가 느려지면서 $80^{\circ}C$에서 100 분간 가열한 후에는 78.1%, $100^{\circ}C$에서 50분간 가열한 후에는 91.9%가 활성이 없어졌다. 열처리한 진품 콩에 있는 탄닌 함량은 열처리 중 다소 증가되었다. 피트산 양은 $60^{\circ}C$로 열처리 할 때 증가되었고, $80^{\circ}C$에서는 거의 변화가 없었고, $100^{\circ}C$로 50분 가열하였을 때는 감소하였다.

• 한국수산과학회지
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• 제48권2호
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• pp.178-186
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• 2015

Protease inhibitors (PI) of trypsin and papain as target proteases from the roe of bastard halibut Paralichthys olivaceus were fractionated out using ammonium sulfate precipitation (A), DEAE 650M anion exchange chromatography (D), and Sephacryl S-300 gel filtration (S). The recovery percentages of the fractions with the strongest inhibitory activity for each fractionation method were 13% for the A4 fraction, 21.2% for the D3 fraction, and 21.3% for the S2 fraction, with specific inhibitory activities of the fractions toward trypsin and casein of 168, 139, and 218 U/mg, respectively, while no inhibition of papain was observed. The $IC_{50}$ for the trypsin-specific substrate $N{\alpha}$-benzoyl-$\small{L}$-arginine-p-nitroanilide (BAPNA) was 0.65, 1.55, 2.26, and 2.85 mg/mL for the A4, S2, A3, and D3 fractions, respectively. These results suggest that chromatographic fractionation methods (D and S) based on the molecular mass and charge of the protein were more effective at fractionating PI than was ammonium sulfate precipitation based on protein solubility, and that the bastard halibut roe extract acts as a serine protease inhibitor. Therefore, the PI fraction from fish roe might be useful for inhibiting proteases in foodstuffs, and could constitute an alternative food-grade inhibitor for the surimi industry.

• 한국수산과학회지
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• 제46권2호
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• pp.119-128
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• 2013

A protease inhibitor was fractionated from fish eggs using methods based on protein solubility. Fractionation efficiency was evaluated with regard to percent recovery and total inhibitory activity (U). The fractionation of protease inhibitor (PI) from egg extracts of skipjack tuna (ST, Katsuwonus pelamis), yellowfin tuna (YT, Thunnus albacores), and Alaska pollock (AP, Theragra chalcogramma) was performed by precipitation with cold acetone or ammonium sulfate (AS). Fractions exhibiting the strongest inhibitory activity contained 20-40% (v/v) cold acetone or 40-60% saturated AS fractions. AS fractionation was more effective in isolating PI than was precipitation with acetone. The total inhibitory activity and percent recovery of fraction obtained with AS 40-60% toward trypsin and $N{\alpha}$-benzoyl-L-arginine-p-nitroanilide (BAPNA) were 4,976 U and 24.2% for ST, 3,331 U and 38.1% for YT, and 4,750 U and 43.8% for AP, respectively. In comparisons against six commercial proteases, 40-80% AS fractions, made by combining the 40-60% and 60-80% AS fractions from fish egg extract, exhibited the strongest inhibition of trypsin when using a casein substrate. These results suggest that fish eggs act as serine protease inhibitors and may be useful for protease inhibition in foodstuffs.

• Food Science and Biotechnology
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• 제17권1호
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• pp.161-165
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• 2008

The trypsin inhibitor activities (TIA) of various potato cultivars were evaluated by measuring the inhibition of trypsin inhibitor activity using N-benzoyl-DL-arginine-p-nitroanilide (BAPNA) as substrate. The TIA values of 5 potato cultivars (1.99 to 2.88 mg/g) were significantly different among cultivars (p<0.05). When the TIA values of commercially processed potatoes were determined, no TIA was detected. During cooking, the $IT_{50}$ (time required to reach 50% inhibition of TIA) values were decreased as heating temperature and time increased. The ITso of moist heating was estimated to be 0.34 min at $100^{\circ}C$, whereas for deep-fat frying the $IT_{50}$ was 0.13 min at $180^{\circ}C$ and 5.28 min for oven baking at $100^{\circ}C$. The $IT_{50}$ value of microwave cooking was 0.194 min at medium heat, and which was similar to that of pressure cooking at $120^{\circ}C$ (0.185 min). Moreover, there was a negative relationship between temperature (${\geq}80^{\circ}C$) and $IT_{50}$ values ($R^2=0.99$, p<0.01). The TIA of potato was completely inactivated by moist heating at $100^{\circ}C$ within 5 min, whereas the pressure cooking at $120^{\circ}C$ and deep-fat frying at $180^{\circ}C$ within 60 and 30 sec, respectively. Based on our results, deep-fat frying is the most effective cooking method to reduce TIA in potatoes.

• 한국미생물·생명공학회지
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• 제17권5호
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• pp.529-532
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• 1989

A strain of Streptomyces sp. (YS-221-B) extracellularly produced an inhibitory substance for $\alpha$-D-Glucosidase. The substance was purified 96-fold from culture filtrate by dialysis, heat treatment, adsorption on active carbon, Bio-Gel P-10 and Sephadex G-75 column chromatography with yield of 9.2%. The substance was stable in pH range from 7.0 to 11.0 at 37$^{\circ}C$, and a treatment at 10$0^{\circ}C$ for 20 min diminished only 15% of the original activity. The inhibitor was not inactivated by the treatment of $\alpha$-, $\beta$-amylases, glucoamylases, trypsin and chymotrypsin but inactivated by pyoteases from Streptomyces griseus and Tritirachium album.

• 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.

• The Korean Journal of Pain
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• 제26권4호
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• pp.356-360
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• 2013

Background: Nerve injury sometimes leads to chronic neuropathic pain associated with neuroinflammation in the nervous system. In the case of chronic neuropathic pain, the inflammatory and algesic mediators become predominant and result in pain hypersensitivity following nervous system damage. It is well known that urinary trypsin inhibitor (ulinastatin, UTI) has an anti-inflammatory activity. Recently, the neuroprotective action of UTI on the nervous system after ischemic injury has been reported. Thus, we evaluated the neuroprotective effect of ulinastatin in a rat model of neuropathic pain. Methods: Neuropathic pain was induced with L5 spinal nerve ligation (SNL) in male Sprague-Dawley rats weighing 100-120 g. The rats were divided into 3 groups, with n = 8 in each group. The rats in the control group (group 1) were administered normal saline and those in group 2 were administered UTI (50,000 U/kg) intravenously through the tail vein for 3 days from the day of SNL. Rats in group 3 were administered UTI (50,000 U/kg) intravenously from the $5^{th}$ day after SNL. The paw withdrawal threshold was measured using the von Frey test for 3 days starting from the $5^{th}$ day after SNL. Results: The paw withdrawal thresholds were significantly increased in the rats of group 2 compared to the other groups (P < 0.05). Conclusions: Ulinastatin, which was administered for 3 days after SNL, increased the paw withdrawal threshold and it could have a neuroprotective effect in the rat model of neuropathic pain.