• Title, Summary, Keyword: Trypsin Inhibitor

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A Simple Method for Detection of Trypsin Inhibitors in Soybean (Glycine max) (대두 Trypsin Inhibitor의 간이검정법)

  • Jo, Ku-Hyung;Lee, Chun-Yung;Hong, Jong-Uek;Kim, In-Soo
    • Korean Journal of Food Science and Technology
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    • v.18 no.5
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    • pp.339-344
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    • 1986
  • The specific reaction of trypsin inhibitors with trypsin to form stable complexes was successfully applied for detection of trypsin inhibitors in soybean. Soybean extract was treated with $Ca^{++}$ to remove globulin fraction, followed by digestion with trypsin and fractionated by chromatography on Sephadex G-50. The void volume fraction contained the trypsin-trypsin inhibitor complexes as well as trypsin. The trypsin inhibitors were then detected by their molecular weight differences on SDS-polyacrylamide gel electrophoresis, in which the complexes dissociate into trypsin and its inhibitors. With the method proposed, trypsin inhibitors were indentified by their ability forming the stable complexes with trypsin and their anti-tryptic moiety. The formation of the complexes with trypsin was further confirmed by two dimensional electrophoresis and DEAE-Sephadex A-25 chromatography. Employing the proposed method, it was found that soybean (Glycine max cv. Hill) contained 7 trypsin inhibitors.

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Trypsin Inhibitor from Streptomyces sp. (Part 2) Biological Activities or the Inhibitor (Streptomyces 속 균주가 생성하는 Trypsin Inhibitor (제2보) 저해물질의 생물학적 작용상)

  • Yi, Dong-Heui;Seu, Jung-Hwn
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.283-288
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    • 1982
  • Trypsin inhibitor produced by Streptomyces sp. was investigated its reactive characteristics against trypsin. The mode of inhibition against trypsin was mixed type of non-competitive and competitive with casein, and enzyme-inhibitor complex was formed rapidly. The inhibitory activity was increased by the addition of isoleucine and depressed by silver, mercuric or cupric ion. And when egg albumin or hemoglobin was used as substrate for trypsin, the inhibition ratio was changed. The inhibitor inhibited coagulation of blood of bovine.

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Purification and Characterization of Trypsin Inhibitor from Alismatis Rhizoma (택사(Alismatis Rhizoma) trypsin inhibitor의 정제와 특성)

  • 박종옥;이인섭
    • Journal of Life Science
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    • v.12 no.2
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    • pp.151-157
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    • 2002
  • A trypsin inhibitor was isolated and purified from Azismatis Rhizoma which has been used as a galenic for diuretic and antiphlogistic. Purification was carried out by 0-80% saturated ammonium sulfate salting out, DEAE- cellulose ion exchange chromatogrphy, Sephadex G-150 gel filtration. The molecular weight of Alismatis Rhizoma trypsin inhibitor(ARTI) was estimated to be about 23,000 Da by gel filtration and SDS-PAGE, it must be monomer. ARTI was stable at 0~6$0^{\circ}C$, but at higher temperature its activity was decreased about 35%. When benzoyl-dl-arginine p-nitroanilide was used as a substrate of trypsin, half-maximal inhibition of ARTI was observed at 0.071 $\mu$M. ARTI inhibited the hydrolysis of trypsin non-competitively and Km value was 0.81 $\mu$M.

Production and Purification of Trypsin Inhibitor from Streptomyces S-217 (Streptomyces S-217에 의한 Trypsin 저해물질의 생산 및 정제)

  • 류병호;이주화;신동분;김동석
    • Microbiology and Biotechnology Letters
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    • v.20 no.5
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    • pp.534-542
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    • 1992
  • Trypsin inhibtor produced by Streptomyces sp. S-217 was purified by solvent extraction and various column chromatographies. and physico-chemical properties of the inhibitor were investigated. Inhibitor complex was formed for incubation of 10 min. Streptomyces 5-217 showed the highest production of trypsin inhibitor when it was cultivated at $37^{\circ}C$ for 66 hr in the medium containing 2% mannitol & 0.9% peptone, pH 7.0. Trypsin inhibitor was purified by column chromatography and high performance liquid chromatography. Trypsin inhibitor indicated the maxium wavelength at 215 nm and solubilities in water, methanol and dimethyl sulfoxide were 95, 70 and 75%, respectively. The concentration of 50% inhibition ($IC^{50}$) was 15 $\mu$g/ml. The inhibitor was stable on heating at $100^{\circ}C$ for 60 min in pH 5~9 and was more stable in alkaline region than acidic region.

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Purification and Characterization of Trypsin Inhibitor from Alismatis Rhizoma and its Binding Protein, 10-Formyltetrahydrofolate Dehydrogenase (택사(Alismatis Rhizoma)로부터 트립신 저해제의 정제와 특성 규명 및 이와 결합하는 단백질, 10-Formyltetrahydrofolate Dehydrogenase에 관한 연구)

  • Kim, Ji-Man;Park, Jong-Ok;Shin, Young-Hee
    • YAKHAK HOEJI
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    • v.52 no.1
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    • pp.79-84
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    • 2008
  • Alismatis Rhizoma has been used as diuretics and antiphlogistics in the Chinese oriental medicine. A trypsin inhibitor was isolated from Alismatis Rhizoma using DEAE ion exchange column, trypsin affinity column, and FPLC chromatography, and its activity and characteristics were studied. The purifed Alismatis Rhizoma trypsin inhibitor (ARTI) was estimated to be about 22 kDa. The sequence determination on N-terminal amino acid residues and 84 amino acid residues has been completed, yet no homology has been found with trypsin inhibitors reported at NCBI. ARTI did not show inhibitory activities on chymotrypsin and elastase, however it exhibited a significant inhibitory activity on bovine trypsin, and formed a complex with rat liver 10-formyltetrahydrofolate dehydrogenase.

Characterization and Pharmacological Effect of Mung Bean Trypsin Inhibitor (녹두(Vigna radiata L.) Trypsin Inhibitor의 정제 및 약물학적 특성)

  • 문성은;신영희
    • Journal of Life Science
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    • v.12 no.5
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    • pp.528-534
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    • 2002
  • A kypsin inhibitor was isolated and purified from Mung bean (Vigna radiata L. wilczek) which has been used as a galenic and traditional food. In addition, we evaluated the pharmacological effect of the mung bean trypsin inhibitor (MBTI) using septic shock induced guinea pig model. Purification was carried out by Sephadex G-50 gel filtration, DEAE-cellulose ion exchange chromatography, and trypsin affinity column. The molecular weight of MBTI was estimated to be about 8,000 Da by 20% SDS-PACE under reducing condition. The chemically determined partial amino acid sequences of the purified MBTI perfectly coincide with those of previously reported MBTI which is BBI type trypsin inhibito. (Bowman-birk inhibitor type). These results suggest that the purified MBTI is authentic. Hypotension shock was prevented by the pretreatment of the MBTI (10 mg/kg of the body weight) on the septic shock guinea pig model caused by psedomonal elastase.

Effects of Trypsin Inhibitors on Oleic acid Induced Acute Pancreatitis in Dogs (개에서 Oleic acid로 유발시킨 급성췌장염에 대한 Trypsin inhibitor의 투여효과)

  • 윤영민;최희인;조명행
    • Biomolecules & Therapeutics
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    • v.5 no.2
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    • pp.158-164
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    • 1997
  • To investigate the effects of trypsin inhibitors, aprotinin and urinary trypsin inhibitor (UTI), on the cute pancreatitis, this study was carried out in dogs of acute pancreatitis induced by oleic acid (0.28 mg/kg). Administration with aprotinin and UTI seemed to have a therapeutic effect on the clinical sign, ultrasonographic finding, histopathologic finding. But in amylase and lipase activity, there were no significant differences among three groups.

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Bowman-Birk type proteinase inhibitors from soybean : Isolation and partial characterization (대두 Bowman-Birk형 proteinase inhibitor들의 분리 및 성질)

  • Choi, Ki-Bong;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.33 no.4
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    • pp.287-292
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    • 1990
  • Eight(I through VII) of Bowman-Birk proteinase inhibitor have been isolated from soybean with DEAE-Sephadex A-50. Inhibitor VII was a typical BBTI, showing high cysteine content(17%/mole) ud low trypsin to chymotrypsin inhibiting activity(TIA/CIA= 1.0) with the independent reactive site to each enzyme. Dissociation constant of trypsin-BBTI and chymotrypsin-BBTl complexes were $9.17{\times}10^{-9}M$ and $5.14{\times}10^{-8}M$, respectively. Inhibitor Vll was extremely heat stable. Six hours heat treatment at $100^{\circ}C$ caused only 50% decrease in it's original inhibiting activity. Except inhibitor III,6 other isoinhibitors differed from a typical BBTI in TIA/CIA, values, ranging from 3 to 29.

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A New Soybean Cultivar "Gaechuck#2": Yellow Soybean Cultivar with Lipoxygenase2,3-free and Kunitz Trypsin Inhibitor-free

  • Chung, Jong Il
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.612-615
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    • 2009
  • Lipoxygenase and Kunitz trypsin inhibitor protein are the main antinutritional factor in mature soybean seed. A new soybean cultivar, "Gaechuck#2" with yellow seed coat, lipoxygenase2,3-free and Kunitz trypsin inhibitor protein-free was developed. It was selected from the population derived from the cross between "Jinpumkong2ho" and C242. Plants of "Gaechuck#2" have determinate growth habit with purple flowers, tawny pubescence, yellow seed coat, yellow hilum, oval leaflet shape and brown pods at maturity. Seed protein and oil content on a dry weight basis were 40.7% and 18.7%, respectively. It has shown a resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and soybean pod borer. Gaechuck#2 matured in 4 October with plant height of 54cm and a 100-seed weight of 24.4g. Average Yield of Gaechuck#2 was 230 - 250 kg/10a in 2005 - 2007.

A New Soybean Cultivar "Gaechuck#1" : Black Soybean Cultivar with Lipoxygenase2,3-free, Kunitz Trypsin Inhibitor-free and Green Cotyledon

  • Chung, Jong Il
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.603-606
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    • 2009
  • Lipoxygenase and Kunitz trypsin inhibitor protein of mature soybean [Glycine max (L.) Merr.] seed are main anti-nutritional factors in soybean seed. A new soybean cultivar, "Gaechuck#1" with the traits of black seed coat, green cotyledon, lipoxygenase2,3 and Kunitz trypsin inhibitor protein free was developed. It was selected from the population derived the cross of "Gyeongsang#1" and C242. Plants of "Gaechuck#1" have a determinate growth habit with purple flowers, brown pubescence, black seed coat, black hilum, oval leaflet shape and brown pods at maturity. Seed protein and oil content on dry weight basis have averaged 39.1% and 16.2%, respectively. It has shown resistant reaction to soybean necrosis, soybean mosaic virus, Cercospora leaf spot and blight, black root rot, pod and stem blight, and soybean pod borer. "Gaechuck#1" matured on 5-10 October with a plant height of 50 cm. The 100-seed weight of "Gaechuck#1" was 23.2g. Yield of "Gaechuck#1" was averaged 2.2 ton/ha from 2005 to 2007.