• Parasites, Hosts and Diseases
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• v.43 no.3 s.135
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• pp.101-110
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• 2005

In this study, the trypsin gene (bgtryp-1) from the German cockroach, Blattella germanica, was cloned via the immunoscreening of patients with allergies to cockroaches. Nucleotide sequence analysis predicted an 863 bp open reading frame which encodes for 257 amino acids. The deduced amino acid sequence exhibited $42-57\%$ homology with the serine protease from dust mites, and consisted of a conserved catalytic domain (GOSGGPLV). bgtryp-1 was determined by both Northern and Southern analysis to be a 0.9 kb, single-copy gene. SDS-PAGE and Western blotting analyses of the recombinant protein (Bgtryp-1) over-expressed in Escherichia coli revealed that the molecular mass of the expressed protein was 35 kDa, and the expressed protein was capable of reacting with the sera of cock-roach allergy patients. We also discussed the possibility that trypsin excreted by the digestive system of the German cockroach not only functions as an allergen, but also may perform a vital role in the activation of PAR-2.

• Biomolecules & Therapeutics
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• v.4 no.4
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• pp.385-390
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• 1996

General pharmacological properties of urinary trypsin inhibitor (UTI) following intravenous administration of 1,000,000 units/kg were examined in terms of effects on central nervous system, cardiovascular system, respiratory system, gastrointestinal system in mice, rats and rabbits. Administration of UTI (1,000,000 units/kg, iv) had no effect on central nervous system; no influences on pentobarbital sleeping time, spontaneous activity, normal body temperature, chemoshock produced by pentylenetetrazole solution, writhing syndromes induced by 0.6% acetic acid solution, and motor coordination of mice. The administration of UTI (1,000,000) units/kg, iv) in rats had no effect on systolic blood pressure and pulse rate. UTI (500,000 units/kg, iv) given to anesthetized rabbits showed no effect on respiratory rate. However, it showed significant elevation of respiratory rate at the concentration of 1,000,000 units/kg. Gastric secretion of rat and intestinal motility of mice were not influenced by the dose of 1,000,000 units/kg. In terms of autonomic nervous system, the material did not show direct effect and inhibitory or augmentative action of histamine- or acetylcholine-induced contraction at the concentration of 2,000 units/ml in the isolated ileum of guinea pig.

• The Korean Journal of Zoology
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• v.38 no.1
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• pp.125-135
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• 1995

붉은지렁이 (Lumbricus rubellus)의 체내에 존재하는 prophenoloxidase-phenoloxidase(prPO$\longrightarrow$PO)의 활성계는 몇 종류의 다른 경로에 의해 활성화 됨을 발견하였다 Propo는 exogenous trypsin $\beta$ 1.3-glucan, Ca2' 이온. lipopolysaccharide (LPS) 및 열처리 등에 의하여 활성도가 증가 되었고 Ca2' 이온이 나머지 4가지 종류의 처리와 함께 병행되었을 때 그 효과가 더욱 증가하였다 Propo의 활성도는 LPS나 Ca2' 이온의 농도가 각각 1 5H 10-s g Lps/r리, 15 mM(Ca2')의 농도에서 propo의 최대활성치를 나타냈으나 그 이상의 농도에서는 propo의 활성이 오히려 감소하였다. LPS. $\beta$ 1,3-glucan 및 Ca2' 이온 등은 trypsin 억제인자인 soybean trypsin inhibitor(571)가 함께 존재할 경우 전혀 propo를 활성화 시킨지 못하는 것으로 미루어 $\beta$ 1,3-glucan 및 Ca2' 이온 등은 체내의 trypsin 유사 효소의 활성을 증가시켜 궁극적으로는 proPO$\longrightarrow$PO의 활성화 반응에 간접적으로 작용한다고 생각되었다. 한편. 571의 존재하에서도 50"C의 열처리는 propo의 활성화에 아주 효과적인 물리적 요인으로 작용하였다. 따라서 열처리는 Ca2'이나 LPS. f 1,3-glucan파는 달리 직접적으로 proPO$\longrightarrow$PO의 활성화 반응에 작용하는 것으로 생각되어 붉은 지렁이의 체내에서 proPO가 활성화되는 괴정(propo-activating system)에는 최소한 2가지 이상의 경로가 있다고 생각된다.생각된다.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.26 no.1
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• pp.51-55
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• 1981

Seed proteins of 51 varieties of soybean were separated by disc electrophoresis. Nine varieties had Rf 0.79 electrophoretic band, and 42 varieties had Rf 0.83 band. The frequency for Rf 0.79 was 0.176 and the frequency for Rf 0.83 was 0.824. The F_1 seeds of crossed between Kumgangdaepp (Rf 0.79) and Uidu (Rf 0.83) possessed both bands. Analysis of 96 F_2 seeds showed a ratio of 22 : 53 : 21 (Rf 0.79 : Rf 0.79/Rf 0.83 : Rf 0.83), suggesting single gene control with two co dominant alleles.

• KSBB Journal
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• v.23 no.4
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• pp.350-354
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• 2008

Magnetically separable immobilized trypsin was developed and their biocatalytic activity was evaluated for the different immobilization media. The activity, recyclability, pH effect, and stability of immobilized enzymes were evaluated for the different supporting media. The biocatalytic activity of immobilized trypsin was highest with magnetically separable polyaniline (PAMP), and Vm and Km of PAMP were 0.169 mM/min and 0.263 mM respectively. With increasedpH, the biocatalytic activity increased for all supporting materials used. Immobilized enzymes were recycled and recycle activities were over 90% of their original activity after ten times reuse. The operational stabilities of enzymes were greatly improved with enzyme immobilization.

• 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.

• 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 Microbiology and Biotechnology
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• v.28 no.9
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• pp.1482-1492
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• 2018

Fusarium oxysporum trypsin (FOT) is a fungal serine protease similar to mammal trypsin. The FOT could be successfully expressed in Pichiapastoris by engineering the natural propeptide APQEIPN. In this study, we constructed two recombinant enzymes with engineered amino acid sequences added to the N-terminus of FOT and expressed in P. pastoris. The N-terminal residues had various effects on the structural and functional properties of trypsin. The FOT, and the recombinants TE (with peptide YVEF) and TS (with peptide YV) displayed the same optimum temperature ($40^{\circ}C$) and pH (8.0). However, the combinants TE and TS showed significantly increased thermal stability at $40^{\circ}C$ and $50^{\circ}C$. Moreover, the combinants TE and TS also showed enhanced tolerance of alkaline pH conditions. Compared with those of wild-type FOT, the intramolecular hydrogen bonds and the cation ${\pi}$-interactions of the recombinants TE and TS were significantly increased. The recombinants TE and TS also had significantly increased catalytic efficiencies (referring to the specificity constant, $k_{cat}/K_m$), 1.75-fold and 1.23-fold than wild-type FOT. In silico modeling analysis uncovered that the introduction of the peptides YVEF and YV resulted in shorter distances between the substrate binding pocket (D174, G198, and G208) and catalytic triad (His42, Asp102, and Ser180), which would improve the electron transfer rate and catalytic efficiency. In addition, N-terminal residues modification described here may be a useful approach for improving the catalytic efficiencies and characteristics of other target enzymes.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.35 no.2
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• pp.171-175
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• 1990

One thousand and seven hundreds and six soybean land reces and 103 wild soybeans of Korea, and 167 cultivars and 71 wild soybeans of exotic were analyzed using polyacrylamide gel electrophoresis to study the distribution of trypsin inhibitor phenotypes. The four ti/ti lines were observed only in Korean native land races. The Ti$\^$*/a allele of cultivar was found to be the highest in Chinese soybean (0,9888) than in Korean soybean(0.8347) and Japanese soybean (0.5954). The heterozygosity of typsin inhibitor in Korean lines occured in relatively high ratio, The percentages of heterozygosity of the Korean land race and wild soybean are 3.6% (N=61) and 9.7%(N=10), respectively.

• KSBB Journal
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• v.22 no.4
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• pp.260-264
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• 2007

Peptides are frequently studied as candidates for new drug development. Recently, synthesized peptide library is screened for a certain functionality on a microarray biochip format. In this study, in order to replace the conventional cellulose membrane with glass for a microarray chip substrate for peptide library screening, we modified the glass surface from amines to thiols and covalently immobilized the peptides. Using trypsin-FITC (fluorescein isothiocyanate) conjugate that could specifically bind to a trypsin binding domain consisting of a 7-amino acid peptide, we checked the degree of surface modification. Because of the relatively lower hydrophilicity and reduced surface roughness, the conjugation reaction to the glass required a longer reaction time and a higher temperature. It took approximately 12 hr for the reaction to be completed. From the fluorescence signal intensity, we could differentiate between the target and the control peptides. This difference was confirmed by a separate experiment using QCM. Furthermore, a smaller volume and higher concentration of a spot showed a higher fluorescence intensity. These data would provide the basic conditions for the development of microarray peptide biochips.