• 한국식품과학회지
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• 제22권4호
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• pp.393-397
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• 1990

장려품종 콩 19종에 함유된 몇 가지 영양저해 인자의 함량과 리폭시게나아제의 역가를 조사하였다. 복부팽만인자인 라피노오스와 스타키오스의 함량은 각각 $0.74{\sim}1.58%\;3.34{\sim}5.30%$ 범위이었으며 힐, 백운, 장백 등의 품종에서 이 두 종류 당의 합량이 높았다. 콩에 함유된 트립신 저해제는 $21.2{\sim}37.0mg$ TI/g, 피트산 형태로 존재하는 인은 건물 100g 당 $337{\sim}605mg$ 정도로 품종에 따라 큰 차이를 보였다. 한편, 콩에 함유된 리폭시게나아제의 역가는 $163.6{\sim}403.5unit$로 팔달, 장백, S-133등의 품종이 높은 값을 나타내었다.

• 한국약용작물학회지
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• 제19권2호
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• pp.77-82
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• 2011

The antioxidative activity of various enzymatic extracts from Sarcodon aspratus (S. aspratus) was evaluated by measuring 1,1-diphenyl-2-picrylhydrazyl (DPPH), and alkyl radical scavenging activity using an electron spin resonance (ESR) spectrometer. For this study, the S. aspratus were enzymatically hydrolyzed by seven carbohydrases (Viscozyme, Celluclast, Dextrozyme, AMG, Promozyme, Maltogenase, and Termamyl) and eight proteases (${\alpha}$-chymotrypsin, Alcalase, Flavourzyme, Neutrase, papain, pepsin, Protamax, and trypsin). The DPPH radical scavenging activities of Viscozyme and pepsin extracts were the highest, and the half maximal inhibitory concentration ($IC_{50}$) values were 0.896 and 0.734mg/mL, respectively. The Celluclast and trypsin extracts showed the highest scavenging activities on alkyl radical, and their $IC_{50}$ values were 0.278 and 0.575mg/mL, respectively. The Celluclast extracts was decreased cell apoptosis in PC-12 cells against $H_2O_2$-induced oxidative damage. The findings of the present study suggest that enzymatic extracts of S. aspratus exhibit antioxidative activity against oxidative stress on PC-12 cells.

• BMB Reports
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• 제33권2호
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• pp.112-119
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• 2000

Three kinds of serine protease inhibitors, members of the Bowman-Birk trypsin inhibitor, were purified from Dolichos lablab seeds and named Dolichos protease inhibitor 1, 2 and 3 (DI-1, DI-2 and DI-3), respectively. Each inhibitor showed a single band with gel mobility at around 15.9, 12.1 and 14.6 kDa on 20% SDS-PAGE under reducing conditions. To characterize inhibitory specificity, the inhibition constant (Ki) for these inhibitors was measured against several known serine proteases. All three Dolichos protease inhibitors (DI-1, DI-2 and DI-3) inhibited the activity of trypsin and plasmin, but had no effect on thrombin and kallikrein (either for human plasma kallikrein or for porcine pancreas kallikrein). DI-1 inhibited chymotrypsin most effectively (Ki = $3.6{\times}10^{-9}\;M$), while DI-2 displayed inhibitory activity for porcine pancreatic elastase (Ki = $6.2{\times}10^{-8}\;M$). Pre-treatment of the 33 mg/kg of DI-mixture (active fractions from $C_{18}$ open column chromatography that included DI-1, DI-2 and DI-3) inhibited the induction of pseudomonal elastase-induced septic hypotension and prevented an increase in bradykinin generation in pseudomonal elastase-treated guinea pig plasma. Also, the increase of kallikrein activity, by injection of pseudomonal elastase, was inhibited by the pretreatment of the DI-mixture in a guinea pig. Since the DI-mixture had no inhibitory effect on kallikrein activity when Z-Phe-Arg-MCA was used as a substrate in vitro, its inhibitory activity in the pseudomonal elastase-induced septic hypotension model might not be due to a direct inhibition of plasma kallikrein in the activation cascade of the Hageman factor and prekallikrein system. These results suggest that the Dolichos DI-mixture might be used as an inhibitor in pathogenic bacterial protease-induced septic shock.

• Applied Biological Chemistry
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• 제35권4호
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• pp.232-236
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• 1992

생대두를 이용한 발효장류의 개발을 위한 기초조사로서 대두의 처리조건을 찾고저 처리방법 [생콩(A), 수침탈각(B), 건열처리(C) 및 습열처리(D)]을 달리한 대두의 일반성분과 효소활성변화를 검토하였다. 그 결과 일반성분 중 수분함량은 A, D구가 B, C구 보다 $2.0{\sim}3.0%$, 조지방 함량은 A, B구가 C, D구 보다 약 2%, 조단백질 함량은 C, D구가 A, B구 보다 $1.16{\sim}1.74%$, 조섬유 함량은 A구가 B, C, D구 보다 $0.11{\sim}1.41%$ 많았으나, 회분은 B, C, D구가 A구 보다 0.49% 많았다. ${\alpha}-amylase$, ${\beta}-amylase$, protease, lipase 활성도는 생콩(A) 및 수침탈각(B) 구가 전열처리(C) 및 습열처리(D)구 보다 약 $2{\sim}5$배 이상 높았다. Trypsin inhibitor 활성도 크기는 $B(56.7{\sim}119.2%)>A(42.9{\sim}95.6%)>D(32.9{\sim}39.6%)>C(20.8{\sim}38.3%)$구 순이었다.

• 한국식품과학회지
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• 제28권1호
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• pp.34-39
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• 1996

메밀(날 메밀, 볶은 메밀, 찐 메밀)에서 추출, 분리한 crude hemicellulose, alcohol-insoluble hemicellulose, residue 및 고분자 가용성 다당류(분자량 10 KDa 이상)와 저분자 가용성 다당류(분자량 10 KDa 이하)가 in vitro에서 췌장의 소화효소 활성에 미치는 영향을 조사하였다. 다당류-효소액을 $37^{\circ}C$에서 5분간 반응시킨 후 다당류를 제거하고 여액의 소화효소 활성을 측정함으로써 다당류가 소화효소의 활성에 미치는 영향을 조사하였다. 메밀의 crude hemicellulose, alcohol-in-soluble hemicellulose, residue는 ${\alpha}-amylase$ 활성을 저하시켰으며, 고분자 수용성 다당류와 저분자 수용성 다당류는 ${\alpha}-amylase$ 활성을 저해하지 않았다. 또한, 저분자 수용성다당류를 제외한 모든 다당류는 lipase의 활성을 감소시켰다. Crude hemicellulose, alcohol-insoluble hemicellulose, residue와 고분자 수용성 다당류는 trypsin과 chymotrypsin 활성을 현저히 저하시켰으나 저분자 수용성 다당류는 이들의 활성을 약간 감소시키는 것으로 나타났다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권4호
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• pp.277-281
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• 2006

Spherical micro silica sol-gel immobilized enzyme beads were prepared in an emulsion system using cyclohexanone and Triton-X 114. The beads were used for the in situ immobilization of transaminase, trypsin, and lipase. Immobilization during the sol to gel phase transition was investigated to determine the effect of the emulsifying solvents, surfactants, and mixing process on the formation of spherical micro sol-gel enzyme beads and their catalytic activity. The different combinations of sol-gel precursors affected both activity and the stability of the enzymes, which suggests that each enzyme has a unique preference for the silica gel matrix dependent upon the characteristics of the precursors. The resulting enzyme-entrapped micronsized beads were characterized and utilized for several enzyme reaction cycles. These results indicated improved stability compared to the conventional crushed form silica sol-gel immobilized enzyme systems.

• BMB Reports
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• 제28권2호
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• pp.138-142
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• 1995

An anticoagulant/fibrinolytic protease was purified to homogeneity from the earthworm Lumbricus rubellus. The protein was a single chain glycoprotein of 32 kDa that exhibited strong proteolytic activity on human thrombin and fibrin clots. Proteolytic degradation of these plasma proteins by the purified enzyme occurred at a neutral pH range. Among several human plasma proteins tested as possible substrates for the protease reaction, the 32 kDa enzyme specifically hydrolyzed both thrombin and fibrin polymers without affecting other proteins, such as serum albumin, immunoglobulin, and hemoglobin. Treatment of the purified enzyme at neutral pH with either phenylmethylsulfonylfluoride or soybean trypsin inhibitor resulted in a loss of catalytic activity. The enzyme hydrolyzed the chromogenic substrate H-D-Phe-L-Pipecolyl-L-Arg-p-nitroanilide with a $K_m$ value of 1.1 ${\mu}M$ at a neutral pH. These results suggest that the anticoagulant/fibrinolytic enzyme from Lumbricus rubellus is a member of the serine protease family having a trypsin-like active site, and one of the potential clevage sites for the enzyme is the carbonyl side of arginine residues in polypeptide chains.

• Journal of Nutrition and Health
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• 제27권7호
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• pp.699-708
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• 1994

This study aimed to verify the nutritional and curative effects of protein hydrolysate in rats with cysteamine-induced duodenal uncer. Duodenal ulcer rat model was established by intraperitoneal injections of cysteamine. Sprague-Dawley, female rats weighing approximately 200g were intraperitoneally injected twice cysteamine(13mg/100g BW) at intervals of 3h per day. This procedure was repeated 3$\times$at intervals of 3d. Animals fed on 10% casein diet for infection periods. After last injection, 4 kinds of diets(10% casein, 20% casein, 10% casein hydrolysate, 20% casein hydrolysate) were given. Gastric montility, trypsin activity in gastrointestinal content, retention rate of nitrogen, plasma total protein, albumin, amino-N, urinary urea nitrogen, creatinine and hydroxyproline were analyzed for nutritional effects of dietary nitrogen levels(10%, 20%) and sources(casein, casein hydrolysate). In duodenal ulcer rat model, there was no differences between 20% casein diet and 20% casein hydrolysate in the view of severeness of ulcer, gastric emptying rate, serum total protein, serum albumin, plasma $\alpha$-amino-N, UUN, creatinine excretion, GFR, nitrogen retention. On the other hand, rats on 10% casein hydrolysate diet group had more curative effect of the ulcer, higher plasma albumin concentration and nitrogen retention than 10% casein diet group. The casein hydrolysate diet group was lower trypsin activity in small intestinal content than the casein diet group, at both nitrogen levels(10%, 20%). The results suggest that protein hydrolysate be applied in diet therapy for the patients with gastrointestinal ulcer.

• International Journal of Industrial Entomology and Biomaterials
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• 제25권2호
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• pp.187-193
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• 2012

Serine proteases are major insect enzymes involved in the digestion of dietary proteins and in the process of blood meal digestion. In this study, cDNA was constructed using the whole body of Laccotrephes japonensis. The flanking sequences of the 5- and 3- end of this gene were characterized by RACE-PCR. Sequence analysis showed that this gene contained a 963-bp ORF encoding 320 amino acids. The deduced amino acid sequence showed 62% identity with the Creontiades dilutus serine protease, 58% with the Lygus lineolaris trypsin precursor, and 54% with the Triatoma infestans salivary trypsin. To assess the expression of the L. japonensis serine protease (JGsp), the JGsp gene was cloned into a baculovirus transfer vector, pBac-1, and expressed in Sf9 cells (Spodoptera frugiperda). SDS-PAGE and western blot analysis have shown that the JGsp recombinant protein was a monomer with a molecular weight of about 32 kDa. Recombinant JGsp has shown activity in the protease enzyme assay using gelatin as a substrate.

• Asian-Australasian Journal of Animal Sciences
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• 제17권6호
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• pp.863-868
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• 2004

Lectin activities and chemical characteristics of Escherichia coli, Lactobacillus spp. and Bifidobacterium spp. originating from the porcine cecal mucosal layer were studied based on hemagglutination assay (HA) and hemagglutination inhibition assay (HIA). Although all the bacterial strains were able to agglutinate erythrocytes of porcine or rabbit origin, much higher HA titers were consistently observed for Lactobacillus spp. than for E. coli or for Bifidobacterium spp. A remarkable reduction in HA titers occurred by the treatment of E. coli and Lactobacillus spp. with protease or trypsin and of Bifidobacterium spp. with protease, trypsin or periodate. There were no significant effects on the HA titers of the three groups of bacteria after the treatment with lipase. Hemagglutination of E. coli was strongly inhibited by D (+)-mannose and D (+)-galactose; Lactobacillus spp. by $\alpha$-L-rhamnose and methyl-$\beta$-galactopyranoside; Bifidobacterium spp. by D (+)-alactose, $\alpha$-L-rhamnose, $\alpha$-L-fucose, L (+)-arabinose, D (+)-mannose, D (-)-fructose at a relatively low concentration (1.43 to 3.75 mg/ml). These results, combined with the enhanced HA activities of the three bacterial strains by modification of rabbit erythrocytes with neuraminidase and abolished HA activity of E. coli after treatment with $\beta$-galactosidase, indicate that it might be the glycoproteinous substances surrounding the surface of the bacterial cells that are responsible for the adhesions of these microorganisms by recognizing the specific receptors on the red blood cell.