• Food Science and Biotechnology
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• 제18권5호
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• pp.1173-1179
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• 2009

Trypsin inhibitors and lectins are defense proteins produced by many organisms. From Chinese 'Large Black Soybeans', a 60 kDa lectin and a 20 Da trypsin inhibitor (TI) were isolated using chromatography on Q-Sepharose, Mono Q, and Superdex 75. The TI inhibited trypsin and chymotrypsin with an $IC_{50}$ of 5.7 and $5{\mu}M$, respectively. Trypsin inhibitory activity of the TI was stable from pH 3 to 13 and from 0 to $65^{\circ}C$. Hemagglutinating activity of the lectin was stable from pH 2 to 13 and from 0 to $65^{\circ}C$. The TI was inhibited by dithiothreitol, signifying the importance of disulfide bond. The TI and the lectin inhibited HIV-1 reverse transcriptase ($IC_{50}$=44 and $26{\mu}M$), and proliferation of breast cancer cells ($IC_{50}$=42 and $13.5{\mu}M$) and hepatoma cells ($IC_{50}$=96 and $175{\mu}M$). The hemagglutinating activity of the lectin was inhibited most potently by L-arabinose. Neither the lectin nor the TI displayed antifungal activity.

• 한국작물학회지
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• 제26권1호
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• pp.51-55
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• 1981

한국에서 재배되고 있는 대두에서의 trypsin inhibitor variant의 분포를 조사하기 위하여 51 품종의 종실 단백질을 disc electrophoresis에 의하여 분리했다. 9품종이 Rf 0.79 band를, 그리고 42품종이 Rf 0.83 band를 갖고 있었으며, Rf 0.79의 분리비율은 17.6% 이었다. 금강대입(Rf 0.79 band)과 의두(Rf 0.83band)의 F_1은 Rf 0.79 band와 Rf 0.83 band를 모두 갖고 있었으며, F_2에서의 Rf 0.79:Rf 079/Rf 0.83:Rf 083의 분리는 22:53:21로서 이들 soybean trypsin inhibitor variant가 1쌍의 codominant alleles에 의하여 지배됨을 나타냈다.

• 약학회지
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• 제22권2호
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• pp.72-82
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• 1978

This study was undertaken establish the relationship between trypsin inhibitor in raw soybean and antinutritional effect of raw legumes. 1) Among legumes produced in Korea, Glycine max contains a relatively high amount of protein(higher than 40%) compared with kindey bean, sword bean and mung bean and, furthermore, soybean which contains a high amount of protein possesses high trypsin inhibitory activity. 2) Disc electrophoretic pattern exhibited pattern exhibited that the crude protein preparation from Glycine max produced about 9-12 protein bands, and the pattern of electrophoretic mobility was very similar to each other. However, only a few protein bands were observed from the crude protein preparation of yard long bean, sword bean, adzuki bean, mung bean and rice adzuki. From the eluate of the sliced gel, it was confirmed that among those bands, only the fastest moving band contains trypsin inhibitory activity. 3) In chicks fed the normal diet the body weight was increased steady from one week and reached to 40% increase for three weeks but in chick fed raw bean diet, there was no body weight gain until two weeks feeding and only 10-20% of body weight gain was observed at the end of three week feeding. On the other hand, in chicks fed raw bean diet the weight of pancreatic tissue per 100g body weight was increased to about two-fold for two or three weeks but there was no change in liver weight. 4) In the case of amylase secretion from the pancreatic fragment, very strong stimulation on amylase secretion from pancreatic tissue of chicks fed a normal diet was produced by one unit of cholecystokin-pancreozymin. However, no stimulation was observed from pancreatic fragment of chick fed raw bean diet.

• Journal of Pharmaceutical Investigation
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• 제30권3호
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• pp.159-166
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• 2000

Kunitz-type soybean trypsin inhibitor (SBTI) and chondroitin sulfate (A, and C type) were conjugated using sodium periodate method. And the physicochemical, pharmacokinetic properties and immunogenecity of the conjugates (Chon-A-SBTI or Chon-C-SBTI) were characterized. We expected the conjugation using chondroitin sulfate to reduce the immunogenecity and to improve the pharmacological effect. As the results, the mean molecular weight of the conjugate highly increased. After I.V. injection of the radiolabeled conjugates or native SBTI into mice, it was found that native SBTI showed rapid elimination from plasma, whereas Chon-A-SBTI and Chon-C-SBTI were slowly eliminated. Organ distribution of the two agents at 30 min after I.V. injection was different : Chon-A-SBTI or Chon-C-SBTI accumulated to a large extent in the liver (13% in Chon-A-SBTI and 16% in Chon-C-SBTI), whereas native SBTI was taken up more rapidly by the kidney (107% dose/g of tissue) and excreated into the urine (26%). In addition we evaluated the therapeutic value of the conjugates by using the sublethal septic shock model caused by pseudomonal elastase and tested the immunogenecity by passive cutaneous anaphylaxis shock (PCA). The conjugates were more effective than native SBTI against pseudomonal elastase induced septic shock in guinea pig. In case of the conjugates, the pharmacological and therapeutic effect lasted over 3 hours long. In immunogenecity test, both of the conjugates showed the reduction of their immunogenecity, especially Chon-A-SBTI looked most effective.

• 한국작물학회지
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• 제49권5호
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• pp.419-422
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• 2004

Soybean is a major source of protein meal in the world. Kunitz trypsin inhibitor (KTI) protein is responsible for the inferior nutritional quality of unheated or incompletely heated soybean meal. The objective of this research was to identify RAPD markers linked to KTI protein allele using bulked segregant analysis. Cultivar Jinpumkong2 (TiTi) was crossed with C242 (titi, absence of KTI protein) and F. seeds were planted. The $F_1$. plants were grown in the greenhouse to produce $F_2$ seeds. Each $F_2$ seed from $F_1$. plants was analysed electrophoretically to determine the presence of the KTI protein band. The present and absent bulks contained twenty individuals each, which were selected on the basis of the KTI protein electrophoresis, respectively. Total 94 $F_2$ individuals were constructed and 1,000 Operon random primers were used to identify RAPD primers linked to the Ti locus. The presence of KTI protein is dominant to the lack of a KTI protein and Kunitz trypsin inhibit protein band is controlled by a single locus. Four RAPD primers (OPAC12, OPAR15, OPO12, and OPC08) were linked to the Ti locus. RAPD primer OPO12 was linked to Ti locus, controlling kunitz trypsin inhibitor protein at a distance of 16.0 cM. This results may assist in study of developing fine map including Ti locus in soybean.

• Preventive Nutrition and Food Science
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• 제10권1호
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• pp.6-10
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• 2005

The trypsin inhibitor activity (TIA) of various soybean cultivars was evaluated by measuring the inhibition of trypsin activity using N-benzoyl-DL-arginine-p-nitro-anilide (BAPNA) as the substrate. The TIA values of eleven white shelled soybean cultivars including a glyphosate-tolerant soybean (16.58 to 17.90㎎/g) were not significantly different among cultivars. Black shelled soybeans had higher TIA values, ranging from 40.09 to 52.11㎎/g, compared to white shelled soybeans (p<0.05). When the TIA of commercially processed soybean foods were determined, no TIA was detected in soysauce, tofu and soybean paste. During conventional moist heating, the IT/sub 50/ (Time required to reach 50% inhibition of TIA) values were decreased as heating temperature and cooking pressure increased. The IT/sub 50/ values of moist heating were estimated to be 91.68, 37.71 and 19.50 min at 60, 80 and 100℃, respectively. The IT/sub 50/ value of microwave cooking was 4.75 min at medium heat, while that of the pressure cooking at 120℃ was only 2.62min. Moreover, there was a negative relationship between temperature and IT/sub 50/ values (R=0.92, p<0.01). The TIA of soybean sprouts was completely inactivated after heating at 100℃ for 5 min, although fresh soybean sprouts showed one fifth of the TIA value of white shelled soybeans. Based on our results, pressure cooking is the most effective cooking method to reduce TIA in soybeans.

• Applied Biological Chemistry
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• 제33권2호
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• pp.109-115
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• 1990

대두 STI의 열처리에 의한 불활성화에 L-cysteine 및 sodium sulfite의 첨가 효과를 조사하고, 대두 품종별 STI활성을 측정하였으며 활성도와 단백질 및 cysteine함량 그러고 소화율과의 관계를 비교하였다. 열처리에 의한 STI의 불활성화에 L-cysteine과 sodium sulfite의 첨가는 불활성화 효과를 크게 증가시켰으며 대두 품종간 불활성화 정도는 차이를 보이지 않았으며 L-cysteine과 sodium sulfite의 첨가는 불활성화된 STI의 재활생화를 크게 억제하였다. 품종별 STI의 활성도는 대두분말 g당 $64.7{\sim}86.47\;TIU$의 범위에 있었으며, 장백>힐>장엽, 광교>단엽>백운>단경>팔달, 새알, 덕유>황금 이었으며, 단백질 함량과 STI의 활성도와는 상관관계(r=-0.192)가 없는 것으로 나타났다. 품종별 cysteine 함량은 힐, 장백, 단경, 단엽, 황금, 백운, 장엽, 새알, 덕유, 광교, 활달의 순서이었으며, cysteine의 함량은 $73.5{\sim}40.0\;{\mu}mole/g$ 대두분 이었다. 또한 cysteine함량과 STI 활성도 사이에 정의 상관(r=0.6568)을 나타냈다. 품종별 소화율은 광교, 백운, 팔달, 단경, 새알, 힐, 장엽, 덕유, 황금, 장백, 단엽의 순서이었으며 $81.9{\sim}76.7%$ 정도이었다. 또한 소화율과 STI활성도 사이에는 부의 상관(r=-0.7695)을 나타내었다.

• Applied Biological Chemistry
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• 제20권2호
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• pp.182-187
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• 1977

본 실험은 대두(大豆)의 발아(發芽)에 따른 trypsin inhibitor activity(TIA)와 protein pattern의 변화에 관하여 조사함과 아울러 그 상관관계를 살퍼보기위한 목적으로 시도되었다. 자엽의 TIA는 발아(發芽)과정중(中) 조금 감소하였고, 배축에서는 급격히 감소해서 발아(發芽) 제 7일에는 제2일의 50%이하로 줄었다. 그리고 발아(發芽) 제 2,3,4일의 defatted dry sample에서 배축은 자엽보다 더 높은 TUI를 보여줬으며 배축의 수분함량이 자엽에 비해 높으므로 fresh sample당(當) TIA는 자엽보다 더 낮았다. 그리고 protein pattern은 자엽과 배축에 있어서 분자량이 큰 15S등의 polymer (band 1), 11S (band 2) 그리고 7S (band 3)가 감소했고 Rm치(値) 자엽의 band 5를 비롯하여 $0.5{\sim}1.0$사이에 높은 mobility를 가진 물질들이 생성(生成)되었다. 자엽의 band 4와 배축의 band 6은 거의 변하지 않았다. 그리고 발아에 따라 TIA의 감소와 저 장단백질의 감소는 있었으나 그 감소량이 서로 일치하지는 않았다.

• 한국식품영양과학회지
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• 제7권2호
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• pp.7-12
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• 1978

본(本) 실험(實驗)은 한국산(韓國産) 대두(大豆) 5 품종(品種)(수원62호, 수원81호, 수원 82호, 수원 83호, 경남 3 호)의 수용(水溶), 단백질(蛋白質)을 gel filtration하여 trypsin inhibitor를 fractionation하여 대두품종별(大豆品種別) trypsin inhibitor의 pattern을 조사(調査)하고 또 이의 내열성(耐熱性)을 살펴보기 위한 목적(目的)으로 시도하여 다음과 같은 결과(結果)를 얻었다. 대두(大豆)의 수용성(水溶性) 단백질(蛋白質)을 추출(抽出)하여 sephadex G-75로 column chromatography하여 분획한 결과(結果) 3개(個)의 peak를 얻었으며, TIA는 두번째 peak 부근에 걸쳐 있었고 trypsin inhibitor는 모두 4개의 fraction(FI, FII, FIII, FIV)으로 나뉘어졌으며, trypsin inhibitor pattern은 수원81호, 수원82호, 수원83호는 FIII의 TIA가 높았으며, 경남 3호 및 수원62호는 FIV에서 높게 나타났다. gel filtration에서 분획(分劃)한 4개의 fraction에 있어서 단백질(蛋白質) mg당(當) TIA는 모두 FIV가 가장 높게 나타났으며 이 fraction의 내열성(耐熱性)은 $100^{\circ}C$에서 20분간(分間) 가열처리(加熱處理)를 기준으로 했을 때 대체로 수원81호가 가장 내열성(耐熱性)이 약했으며 경남 3 호, 수원38호, 수원82호, 수원62호의 순서로 내열성(耐熱性)이 강하게 나타났고 fraction별(別)로는 수원82호를 제외하고는 모두 FIV가 가장 내열성(耐熱性)이 강하게 나타났으며 대체로 FI의 내열성(耐熱性)이 약하였으며 경남 3 호의 FI은 $100^{\circ}C$ 20분간(分間) 처리(處理)에서 완전(完全)히 소실되었다.

• 한국식품과학회지
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• 제12권1호
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• pp.24-33
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• 1980

한국산 두류 중 대두를 비롯한 팥, 녹두, 강낭콩에 대하여 trypsin 저해(沮害) 활성(活性)과 적혈구(赤血球) 응집(凝集) 활성(活性)을 측정하고 이들을 가열 처리에 의한 파괴 효과를 실험한 결과는 다음과 같다. 1. 각 두류의 건물(乾物) 중량 당 저해(沮害) 활성도(活性度) (% [TU] inhibited/mg)는 대두가 가장 높아 79.9를 나타낸 반면 팥, 강낭콩은 각각 46.4, 43.2로 대두의 반정도이었으며 녹두는 17.7로 제일 적었다. 이를 단백질 중량당으로 환산하면 팥이 제일 높아 222를 보였고 그 다음이 대두 194, 강낭콩 170의 순이었으며 녹두는 제일 낮아 75를 보였다. 2. 열에 의한 trypsin 저해(沮害) 활성도(活性度)의 파괴율은 팥, 녹두가 각각 $85{\sim}87,\;87{\sim}94\;%$로 높았고 강낭콩, 대두는 각각 $76{\sim}79,\;67{\sim}72\;%$로 낮았다 그러나 boiling과 autoclaving 간의 가열 방법에 따른 차이는 발견할 수 없었다. 3. 각 두류의 토끼 혈액에 대한 적혈구(赤血球) 응집(凝集) 활성(活性)(hemagglutinating unit/g)은 팥에서는 나타나지 않았고 강남콩이 제일 많아 건물 1g당 48,300을 보인데 비해 대두는 18,000, 녹두는 136이었다. 이를 단백질 중량당으로 환산하면 마찬가지 경향을 보여 강남콩 190,600, 대두 43,700, 녹두 581로 나타났다. 이들 두류의 적혈구(赤血球) 응집(凝集) 활성(活性)은 가열 처리에 의하여 모두 파괴되었다.