• The Korean Journal of Zoology
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• v.39 no.1
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• pp.65-74
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• 1996

Treatment of regenerating amphibian limbs with retinoic acid (RA) is known to induce paftern duplication, which is closely related to the extent of dedifferentiation. In the present study, the activities of trypsin- and chymotrypsin-like proteases are examined to delineate a possible role in the process of dedifferentiation in the regenerating limbs of urodeles, the Korean salamander (Hynobius leechii) and the Mexican axolod (Ambystoma mexicanum). Specifically, we were interested to know if there is any correlation between trypsin- and chymotrypsin-like protease activities and the state of dedifferentiation which is augmented by RA treatment. We were also interested in expoloring if there is any species-specific difference in the profile of enzyme activities during limb regeneration. The results showed that the activities of these two enzymes reached a peak level at dedifferentiation stage, and RA treatment caused elevation of their activities, especially in the case of trypsin-like protease. The increase of trypsin-like protease activity after RA treatment was pronounced in the Korean salamander, which might reflect a species-specific responsiveness to RA. The present results imply that trypsin and chymotrypsin or similar proteases may play an active role in the process of dedifferentiation in regenerating limbs, and that trypsin or trypsin-like eryrymes might be involved in the RA-evoked enhancement of dedifferentiation which precedes overt pattern duplication.

• Journal of Plant Biology
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• v.37 no.2
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• pp.175-181
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• 1994

해녀콩(Canavalia lineata) 유식물 자엽에서 N-$\alpha$-benzoyl-DL-arginine p-nitroanilide hydrolase(BApNAase)를 부분정제하여 그 성질을 밝혔다. 부분정제한 BApNAase는 purification fold가 77.5이었고 회수율은 7%이었으며 비활성도는 7.75 unit/mg이었다. BApNAase의 분자량은 200 kD이고 젤라틴분해효소 P3인 것으로 밝혀졌으며 최적 pH는 9.5이었다. BApNAase의 Vmax와 Km은 각각 15.5 unit/mg와 1.6 mM로 최대반응속도가 동물의 트립신보다 7배 가량 낮은 반면에 N-$\alpha$-benzoyl-DL-arginine p-nitroanilide(BApNA)에 대한 기질 친화성은 4배 가량 높았다. 또한, BApNAse는 1 mM의 phenylmethanesulfonyl fluoride(PMSF)에 의해 90%나 크게 억제된 반면 aprotinin에 의해서는 크게 억제되지 않아 트립신과는 다른 serine proteinase로 판단되었으며, 효소활성은 Ca2+과 Mg2+에 의해 다소 증가하나 Mn2+, Hg2+, Zn2+에 의해 크게 억제되었다.

• Proceedings of the Membrane Society of Korea Conference
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• 1991.10a
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• pp.45-46
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• 1991

분리막 공정은 상변화를 일으키지 않기 때문에 장치비가 저렴하며, 열 및 화학적으로 민감한 물질, 특히 생체물질 등을 다루는데 적합하다. 또한 최종 생성물질을 정제하는 방법으로는 이온교환 크로마토그래피, 전기 영동 및 친화성 크로마토그래피(affinity chromatography)가 있다. 이중 친화성 크로마토그래피는 가장 선택적이고, 단일 장치로서 기존의 분리방법에 비해 약 1000배 가까이 정제할 수 있다. 본 실험에서는 트립신 억제제인 m-aminobenzamidine을 포함하고 있는 수용성 고분자를 제조하여 트립신을 선택적으로 분리하고자 하였으며, 본 연구 필요한 분리막은 막제조가 용이하며 비용이 저렴한 cellulose acetate막을 사용하였다. CA 막의 세공크기는 겔화매질의 에탄올 농도로 조절하였다. Casting 용액의 조성은 다음 표에 간략하게 나타내었다.

• 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.38 no.5
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• pp.382-386
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• 1995

The potato proteinase inhibitor II (PI-II) protein contains chymotrypsin and trypsin inhibitory site. Among several PI-II genes isolated from genomic library, amino acid sequence deduced from PI-IIT gene has 84% identity with that of the polypeptide chymotrypsin inhibitor (PCI). Therefore a gene fragment having homology with the PCI was cloned into a vector using polymerase chain reaction(PCR) from the potato proteinase inhibitor IIT gene. Two different primers were utilized for cloning; primer A contains NdeI restriction site and 30 nucleotides, which has AUG N-terminal methionine codon, primer B contains BclI restriction site and 28 nucleotides, which has TAG translation stop codon. After PCR, about 160 bp-long DNA fragment was cloned into pRT146, derivative of pUC118, and sequenced. The sequenced NdeI/BclI fragment was moved to pET3a, containing bacteriophage T7 promoter and terminator. The expressed proteins in E. coli BL2l(DE3) were determined on a polyacrylamide gel containing sodium dodecyl sulfate. The expected size of protein deduced from the sequenced gene fragment is about 6,500 dalton whose size was similar to the IPTG-induced protein (6,000 dalton) on a gel. However the expression level was much lower than expected.

• 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.

• Korean Journal of Food Science and Technology
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• v.22 no.7
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• pp.805-811
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• 1990

Hydrolysis of soybean proteins was carried out by immobilized trypsin and/or alpha-chymotrypsin. The partially hydrolyzed products of soybean proteins were evaluated for their molecular weights and molecular charges by using Ferguson's plot. The ratio of average molecular weights to average molecular charges($\bar{M}/log\;\bar{Y}_o$) of modified soybean proteins could be used to predict functional properties such as solubility, water holding capacity, oil holding capacity, and emulsifying ability. The low ratio of modified soybean proteins indicated high solubility. while the high ratio showed high water holding capacity. The appropriate ranges of the ratios were necessary for maximun oil holding capacity and emulsifying ability.

• Food Science of Animal Resources
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• v.29 no.2
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• pp.238-244
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• 2009

Food allergy is a serious nutritional problem in both children and adults. Therefore, food allergenicity reduction methods are greatly needed. The allergenicity is altered by various manufacturing processes, and the digestibility of food proteins can be affected by food processing. This study was conducted to investigate the effect of in-vitro digestibility on the allergenicity of autoclaved market pork sausages using pepsin (30min) and trypsin (5, 30, 60, 90, and 120min). The binding ability of the porcine serum albumin (PSA) from sausages A and B significantly decreased by about 30 and 23%, respectively, after autoclave treatment (121; 5, 10, and 30 min). After the pepsin and trypsin treatments, the binding ability of products A and B at 30 min decreased. These competitive indirect enzyme-linked immunosorbent assay (ci-ELISA) results corresponded well with the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting results. The results demonstrated that the allergenicity of pork sausages considerably decreased after autoclave treatment, and were also maintained or decreased after enzyme treatment. Accordingly, autoclave treatment represents a promising processing technology for the reduction of the allergenicity of diverse food products.

• Journal of the korean veterinary medical association
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• v.47 no.12
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• pp.1086-1090
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• 2011

외인성 췌장부전은 췌장에서 분비되는 소화효소의 부족한 합성 및 분비에 기인한다. 대부분의 개체에서 보이는 임상증상은 체중감소, 다량의 연변, 지방변이다. 혈청 고양이 유사트립신 면역활성 농도검사는 외인성 췌장부전 진단에 있어 매우 유용한 검사법이다. 외인성 췌장부전 환자의 치료는 췌장 소화효소의 공급이다. 대부분의 외인성 췌장부전 환자에서 심각한 혈청 코발라민의 감소와 동반되기 때문에 코발라민의 투여는 반드시 필요하다. 마지막으로 개에 비해 흔하게 확인되진 않지만, 만성적인 연변 및 체중감소를 보이는 환자에서는 진단에 있어 외인성 췌장부전에 대한 고려가 필요하다.

• Journal of the Korean Society of Food Science and Nutrition
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• v.18 no.4
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• pp.410-422
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• 1989

Soybean protein isolates were acylated with succinic anhydride and partially hydrolyzed with trypsin. Chemical modification decreased protein contents of samples and, in amino acid composition, tyrosine was increased comparatively. And lysine was increased remarkably by partial proteolysis. Succinylation and trypsin treatment increased the aqueous solubility and shifted the isoelectric potint that showed high pH-dependence of protein solubility. Protein solubility was influenced by salt concentration such as $NaCl,\;CaCl_2,\;NaNO_3$ and $NaH_2PO_4$. Chemical modification increased the absorption of oil and water, emulsification properties and foam capacity, but decreased foam stability, ultraviolet absorbance and bulk density. Protein-protein Interaction between soybean protein isolates and beef protein increased the emulsifying activity, emulsifying activity index and foaming properties, but it didn't have any influence on emulsion stability.