• Natural Product Sciences
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• 제11권4호
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• pp.207-212
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• 2005

Tetragonia tetragonoides (Aizoaceae) has been known as an anti-cancer agent. The activation of proteinase-activated receptor-2 (PAR-2) by trypsin appears to play a role in inflammation. In the present study, we examined the inhibitory effects of Tetragonia tetragonoides water extract (TTWE) on the production of tumor necrosis $factor-{\alpha}\;(TNF-{\alpha})$ and tryptase in trypsin-stimulated human leukemic mast cells (HMC-1) expressing PAR-2. HMC-1 cells were stimulated with trypsin in the presence or absence of TTWE (10, 100, and $1000\;{\mu}g/ml$). The level of $TNF-{\alpha}$ secretion from HMC-1 cells was measured by enzyme-linked immunosorbent assay (ELISA). $TNF-{\alpha}$ and tryptase mRNA expression were examined by reverse transcription-PCR. Also, extracellular signal-regulated kinese (ERK) activation was assessed by Western blot analysis. Trypsin activity was measured using the substrate Bz-DL-Arg-p-nitroanilide (BAPNA). It was observed that $TNF-{\alpha}$ secretion, tryptase mRNA and $TNF-{\alpha}$ mRNA expression in trypsin-stimulated HMC-1 cells were inhibited by pretreatment of TTWE ($1000\;{\mu}g/ml$). Furthermore, the pretreatment of TTWE ($1000\;{\mu}g/ml$) resulted in the reduction of ERK phosphorylation and trypsin activity. These results suggest hat TTWE might have the inhibitory effects on the PAR-2-dependent inflammation processes and it is likely to function as PAR-2 antagonist.

• Journal of Microbiology
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• 제38권3호
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• pp.160-168
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• 2000

A-factor I a microbial hormone that can positively control cell differentiation leading to spore formation and secondary metabolite formation in Streptomyces griseus. to identify a protease that is deeply involved in the morphological and physiological differentiation of Streptomyces, the proteases produced by Streptomyces griseus IFO 13350 and its A-factor deficient mutant strain, Streptomyces griseus HH1, as well as Streptomyces griseus HH1 transformed with the afsA gene were sturdied. In general Streptomyces griseus showed a higher degree of cell growth and protease activity in proportion to its ability to produce a higher amount of A-factor. In particular, the specific activity of the trypsin of Streptomyces griseus IFO 13350 was greatly enhanced more than twice compared with that of Streptomyces griseus HH1 in the later stage of growth. The specific activity of the metalloprotease of Streptomyces griseus HH1 was greatly enhanced more than twice compared with that of Streptomyces griseus IFO 13350, and this observation was reversed in the presence of thiostreptione, However, Streptomyces griseus HH1 transformed with the afsA gene showed a significantly decreased level of trypsin and metalloprotease activity compared with that of the HH1 strain. There was no significant difference between Streptomyces griseus IFO 13350 and HH1 strain in their chymotrypsin and thiol protease activity, yet the level of leu-amionpeptidase activity was 2 times higher in Streptomyces griseus HH1 than in strain IFO 13350 . Streptomyces griseus HH1 harboring afsA showed a similar level of enzyme activity , however, all the three protease activities sharply increased and the thiol protease activity was critically increased at the end of the fermentation. When a serine protease inhibitor, pefabloc SC, and metalloprotease inhibitor, EDTA, were applied to strain IFO 13350 to examine the in vivo effects of the protease inhibitors on the morpholofical differentiation, the formation of aerial meycelium and spores was delayed by two or three days.

• Journal of Nutrition and Health
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• 제4권4호
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• pp.25-28
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• 1971

The effects of additives for food and drug upon the tryptic hydrolysis of casein an a Synthetic substrate, $N^{\alpha}-Benzoyl-L-arginine$ ethylester (BAEE) in vitro has been studied. The results of this study were summarized as follows 1) It was found that the action of inhibition became stronger in the following order: Methyl parabene>Rose Bengal> Phloxine> Sod. DHA> Erythrosine by the colorimetric method using BAEE. These results also showed that other additives had no effect on the activity of trypsin. 2) All samples tested showed respectively same tendency using casein in this method. But the activity by Erythrosine and Sod. DHA was slightly increased in this experiment.

• Bulletin of the Korean Chemical Society
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• 제34권10호
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• pp.2973-2977
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• 2013

In this study, we developed a liquid crystal (LC)-based optical sensor for monitoring enzymatic activity through orientational changes in liquid crystals (LCs) coupled to the properties of a poly-${\small{L}}$-lysine (PLL)-based polymeric membrane. We prepared a PLL-based polymeric membrane at the planar interface between the thermotropic liquid crystal and aqueous phases. The PLL-based polymeric membrane was obtained by contacting the PLL solution with water immiscible LCs, 4-cyano-4'-pentyl-biphenyl (5CB) doped with adipoyl chloride. We then investigated the membrane properties by examining the permeability of the membrane to phospholipids, 1,2-didodecanoyl-rac-glycero-3-phosphocholine (DLPC). The permeability of the membrane to transport phospholipids was monitored through the orientational transition of 5CB in contact with the dispersions of DLPC. Since trypsin can enzymatically catalyze the hydrolysis of PLL, we incubated an aqueous trypsin solution with the membrane for 2 h at room temperature to cause an increase in the permeability of the polymeric membrane to DLPC. As a result, a bright to dark optical shift of LCs was observed, which implied that an enzymatic reaction between trypsin and PLL-based membrane occurred. Two control experiments using chymotrypsin and bovine serum albumin (BSA) revealed no sign of improved permeability based on the orientational transition of LCs.

• 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.

• BMB Reports
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• 제32권6호
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• pp.573-578
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• 1999

Treatment of Hafnia alvei aspartase with limited tryptic digestion resulted in a marked increase in enzymatic activity. The activation required a few minutes to attain maximum level and, thereafter, the activity gradually decreased to complete inactivation. The degree of cleavage associated with the activation was extremely small as judged by SDS-PAGE. Upon activation, the optimum pH and temperature were essentially unchanged. When trypsin-activated enzyme was denatured in 4 M guanidine-HCI followed by removal of the denaturant by dilution, the restoration of activity was similar (40%) to that of the native enzyme, indicating a degree of stability. The $pK_a$ obtained on the acidic side and the $pK_b$ obtained on the basic side of trypsin-activated aspartase were 6.6 and 8.6, respectively, the same as those of the native aspartase, indicating that aspartase may exist in a stable conformation after limited tryptic digestion. These results indicate that the activation of H. alvei may be mediated by a conformational change away from the active site of individual subunits.

• 한국임상수의학회지
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• 제18권1호
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• pp.7-13
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• 2001

Depending on the degree of exocrine pancreatic insufficiency, there are inconsistent values in the serum trypsin-like immunoreactivity (TLI), hematology, and serology. To determine the correlation between those values and complete pancreatic insufficiency, 10 pancreatectomized dogs as the treated group and 10 sham operation dogs as control group were used. The entire treated group showed significant decrease in serum TLI level from 2.63$\pm$0.20 ng/ml (day 0) to 1.81$\pm$0.43 ng/ml at day 1 after the pancreatectomy (p<0.05) and the decline maintained till the end of the experiment. Blood glucose level gradually increased, but albumin level showed significant decrease (p<0.05) at day 1 and maintained the decline. Three clinical signs were observed such as depression, vomiting, and diarrhea. The entire treated group showed diarrhea throughout the experiment, but depression and vomiting were gradually diminished. The activity of serum alkaline phosphatase in the treated group increased till day 2 then decreased, on the other hand, control group showed continuous increase till day 4, and then decreased. Only experimental group showed the increased alanine aminotransferase activity at day 1. Serum lipase activity in the treated group jumped up at day 1 and then dropped down, which was even lower than the control. Regarding serum amlyase activity gradually decreased. According to these results, the assay of trypsin-like immunoreactivity showed the consistent result, so it suggests that TLI is an useful tool to determine the pancreatic exocrine function and possible diagnosis of exocrine pancreatic insufficiency compared to the simple assay of plasma amylase and lipase.

• 한국의류학회지
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• 제37권7호
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• pp.852-863
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• 2013

We investigate the immobilization of trypsin on chitosan nonwoven using glutaraldehyde (GA). The conditions for trypsin on chitosan nonwoven and GA cross-linking were optimized depending on different conditions. The order of GA cross-linking was determined by the activity of immobilized trypsin. The characteristics of chitosan nonwoven were examined by Fourier-transform infrared (FT-IR) and surface morphology analyses (SEM). Results showed that the optimal treatment conditions for trypsin on chitosan nonwoven were as follows: pH 8.5; temperature $37^{\circ}C$; trypsin concentration 15% (o.w.f); and treatment time 60 min. Those for GA cross-linking were: pH 10.0; GA concentration 3% (v/v); and treatment time 120 min. FT-IR analysis showed that GA was cross-linked on chitosan nonwoven. The SEM analysis also showed that trypsin was immobilized on chitosan nonwoven.

• Journal of Yeungnam Medical Science
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• 제4권1호
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• pp.17-23
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• 1987

백서심장조직을 trypsin과 collagenase 두가지의 효소를 사용하여 분리하면서 각 효소의 분리효과와 두가지 효소의 복합적 세포분리효과를 조사하였으며 이와 동시에 dimethyl sulfoxide가 세포분리과정 및 단기간 배양중에 세포에 미치는 효과를 조사한 결과를 다음과 같이 요약할 수 있다. 1. 백서심장조직에서 세포를 분리할 때 $4^{\circ}C$ trypsin 18시간 전처치후 $37^{\circ}C$ collagenase 처치한 군에서 세포생존율 세포수확량 및 심근세포 생존율이 가장 높았다. 즉 단일 효소 처리보다는 효소 복합연속 처리가 더 효율적이었다. 2. $37^{\circ}C$ trypsin만으로 세포분리를 하였을 때 세포생존율과 수확량이 가장 낮았다. 3. 백서심장세포의 분리과정과 초기 배양 과정에 세포회복이나 보호에 대한 DMSO의 효과는 인정되지 않았으며 오히려 세포파괴 효과가 높음을 알 수 있었다.

• 한국양식학회지
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• 제21권3호
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• pp.184-189
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• 2008

흰다리새우 Litopenaeus vannamei 적정 사육기법을 개발하고자 사육수온(20, 23과 $26^{\circ}C$)에 따른 간췌장, 전장과 중장에서의 trypsin 활성 및 먹이섭취량을 조사하였다. $26^{\circ}C$에서의 흰다리새우 간췌장내의 효소활성은 $23^{\circ}C$ 구간보다 2배, $20^{\circ}C$ 구간보다 3배의 활성을 보였다. 전장에서의 효소활성은 사육수온에 관계없이 300 nmol/mg/min 과 340 nmol/mg/min로 유사한 최대 활성을 보였으나 먹이섭취 후 최대 활성이 나타나는 시간은 수온이 높을수록 짧아지는 것으로 나타났다. 중장에서의 효소활성은 먹이섭취량과 관계없이 $26^{\circ}C$ 구간에서 가장 높게 기록되어 수온이 높을수록 중장에서의 효소활성 또한 높아지는 것으로 나타났다. 체중에 대한 전장과 중장의 무게 비율의 변화, 즉 섭취된 먹이량에 의한 비율변화를 조사한 결과 전장에서의 최대 먹이섭취는 $23^{\circ}C$ 구간으로 체중의 0.6%를 섭취하였으며, $26^{\circ}C$는 0.5% 그리고 $20^{\circ}C$에서는 0.4%의 먹이를 섭취한 것으로 나타났다. 본 연구에서 최대먹이섭취는 $23^{\circ}C$에서 일어났지만 수온이 높은 구간인 $26^{\circ}C$에서 효소의 활성은 가장 높았고 배설률 또한 높아서 먹이의 잔류시간이 가장 짧은 것으로 나타났다. $20^{\circ}C$ 구간에서는 가장 적은 양의 먹이를 섭취하였고 효소활성 또한 가장 낮았으나 소화관내 먹이 잔류율이 실험기간 동안 가장 높은 것으로 나타났다. 각 수온에서 먹이섭취량에 따른 소화효소의 활성변화와 배설률에 따른 먹이 잔류시간을 고려하여 먹이공급량과 시간을 조절한다면 최적의 성장을 유도할 수 있는 방법으로 제시될 수 있다는 가능성을 확인하였다.