• Natural Product Sciences
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• v.11 no.4
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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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• v.38 no.3
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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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• v.4 no.4
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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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• v.34 no.10
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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.

• YAKHAK HOEJI
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• v.22 no.2
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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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• v.32 no.6
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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.

• Journal of Veterinary Clinics
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• v.18 no.1
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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.

• Journal of the Korean Society of Clothing and Textiles
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• v.37 no.7
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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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• v.4 no.1
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• pp.17-23
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• 1987

New born rat heart cells were dissociated using trypsin and/or collegenase to elucidate the dissociation efficiency of these two enzymes. And the effect of dimethyl sulfoxide during and immediately after cell dissociation was also investigated to clarify the so-called protective activity of dimethyl sulfoxide on cell performance. The results can be summarized as follows. 1. Cold trypsin 18 hours pretreatment followed by warm collagenase treatment resulted best cell viability and cell yield. 2. Single, warm trypsin treatment gave the poorest result. 3. Dimethyl sulfoxide did not seem to play any protective role during or immediately after rat heart cell dissociation. It had very damaging effect on rat heart cells.

• Journal of Aquaculture
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• v.21 no.3
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• pp.184-189
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• 2008

Tryptic enzyme activities in the digestive glands (hepatopancreas) and digestive tracts of Pacific white shrimps Litopenaeus vannamei were assayed at three water temperature regimes. At $26^{\circ}C$, trypsin activity in the hepatopancreas was 200% higher than at $23^{\circ}C$ and 300% higher than at $20^{\circ}C$. The highest foregut trypsin activity levels showed no significant difference in the temperature regimes, but the time between peaks in foreguts and midguts shortened at higher temperature. In the midgut, the level of enzyme activity was highest at $26^{\circ}C$ regardless of the amount of ingested feed. The ratio of foregut and/or midgut dry weight to the body wet weight indicated feed movement through the digestive track directly and gave accurate account about the feeding mechanism. Maximum feed ingestion in the foregut was equivalent to 0.6% of the body weight (wet weight) at $23^{\circ}C$, 0.4% of the body weight at $20^{\circ}C$, and 0.5% of the body weight at $26^{\circ}C$. In view of the temperatures chosen for this study, although maximum ingestion was observed at $23^{\circ}C$, the shrimps showed highest enzyme activity, but lowest feed retention time at $26^{\circ}C$ and lowest enzyme activity, but highest retention time at $20^{\circ}C$.