• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.651-654
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• 2000

A fermentation study of the recombinant Escherichia coli system has been tired to overproduce a recombinant Paragonimus westermani cysteine proteinase, rPwCP1. Using the modified LB media, main cultures and chemical induction with IPTG were carried out to examine the possibility of secretory protein overproduction at high cell density culture. As a result, the target protein of rPwCP1, purified by metal affinity chromatography and gel filtration, has been shown at 50.8 kDa on SDS-PAGE, and its final concentration turns out to be 350mg/L.

• Journal of Food Hygiene and Safety
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• v.34 no.6
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• pp.557-561
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• 2019

This study investigated the efficacy of DNA extraction methods for real-time PCR detection of foodborne pathogenic bacteria in livestock manure composts. Livestock manure composts were inoculated with Escherichia coli O157:H7, Salmonella Typhimurium, Listeria monocytogenes, Bacillus cereus and incubated in enrichment broth. For DNA extraction, enriched samples were treated following boiling method, by chloroform, C₁₈ powder, and proteinase K. As a result, 4 species of bacteria were detected by real-time PCR when subjected to boiling for 30 min and treated with proteinase K. These results suggest that detection of foodborne pathogens by real-time PCR from livestock manure composts could be applicable using effective DNA extraction methodology such as the boiling method or proteinase K.

• Tuberculosis and Respiratory Diseases
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• v.40 no.1
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• pp.43-51
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• 1993

Background: The polymerase chain reaction (PCR) is a very sensitive method for the detecting of mycobacterial DNA. There are many reports revealing the efficacy of PCR for the diagnosis of M. tuberculosis, but there are many different methods for DNA extraction from Mycobacterium tuberculosis. Bead beater method is a very useful method for DNA extraction from clinical spectimens, but its procedures are relatively complicated and time-consuming. So we studied other methods for the DNA extraction from Mycobacterium tuberculosis $H_{37}Rv$ and some clinical specimens (5 smear positive sputa and 5 smear negative CSF). Method: We extracted the mycobacterial DNA with 6 different methods from H37Rv strain and clinical specimens. The methods included SDS-microwave oven method, NaOH lysis method, Triton X-100-Proteinase K method, Lysis buffer method, SDS-proteinase K method and bead beater method. The target DNA was 123bp of IS6110 and was detected by examination of ethidium bromide-stained agarose gels. Results: Among 6 methods, SDS-proteinase K method, bead beater method, lysis buffer method and triton X-100-proteinase K method were excellent, but SDS-proteinase K method was the best method in the aspect of simplicity and cost-effectiveness. Conclusion: We suggest that SDS-porteinase K method is a simple and convinient method and might be the best method for the extraction of mycobacterial DNA.

• Journal of Plant Biotechnology
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• v.6 no.3
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• pp.145-150
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• 2004

Cabbage plants were transformed with the potato proteinase inhibitor II (PINII) gene, bar gene, and hpt gene using Agrobacterium. The expression of the PINII gene was driven by its own promoter which was wound-inducible. Ten transgenic plants were obtained from medium containing hygromycin as a selection antibiotic. The integration and expression of PINII and bar genes were confirmed by Southern and Northern hybridization. Growth and development of diamondback moths (Plutella xylostella) and tobacco cutworm (Spodoptera litura) larvae were examined on $T_1$ plants. The weight of the larvae and pupae of these two insects grown on transgenic plants was not different compared to those grown on wild type plants. However, the pupation and emergence rate of diamondback moths and tobacco cutworms fed on some transgenic plants was lower than on wild type plants. These results suggest that the PINII transgene under the control of a wound-induced promoter may be used for control of insects in transgenic cabbage through reduction of insect progeny number.

• The Korean Journal of Zoology
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• v.29 no.3
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• pp.171-180
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• 1986

The effects of proteases, neuraminidase and EDTA on adhesion of amoebae on the substratum, ultrastructure and biochemical composition of the cell surface were studied by concanavalin A (con A) cytochemistry and SDS PAGE. By con A cytochemistry the glycocalyx of the plasmalemma was easily subdivided into outer filamentous (F) layer and the inner amorphous (A) layer. On treatment with neuraminidase, amoebae attached to the substratum and spreaded better than untreated cells exposing the more con A binding sites in A- and F-layer. When the cells were treated with trypsin or proteinase K, cells stayed unattached for 12 and 48 hr, respectively. Con A binding sites of A layer and all of those glycoproteins were removed by proteinase K. On the other hand, trypsin damaged all of the con A binding sites in both A- and F-layer without significant change in PAS-stained profile of the plasmalemma. Some of the mucopolysaccharides of the cell surface were released by these enzymes and EDTA. When the cells were incubated with monovalent con A they did not attch on the substratum and cytolysed. From these results adhesion of amoebae on the substratum appears to be mediated by the interaction of the glycoproteins and mucopolysaccharides of the A layer.

• Korean Journal of Food Science and Technology
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• v.7 no.4
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• pp.229-237
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• 1975

An attempt was made to utilize the enzyme produced by Asp. oryzae as meat tenderizer. The production, purification, and various properties of proteinase produced by Asp. oryzae were investigated. Results obtained are as follow; 1. A strain which had the highest proteolytic activity was selected among 9 Aspergillus species. 2. Culture medium consisted of wheat bran 10g, 2% glucose, 0.03% urea and 0.1% $MgSO_4$ (pH 6.5). Mold was incubated at $30^{\circ}C$ for 3 days. 3. Enzyme extract from culture medium were fractionated with ammonium sulfate and purified by Sephadex G-75 column chromatography. 4. When pH of reaction mixture was controlled, maximal activity of proteinase by Asp. oryzae was obtained at pH 3, pH 6.6, $8.4{\sim}8.5$ and pH 10.0 to 10.5. Those results were interpreted to show that enzyme consists of acid proteinase, neutral proteinase and alkaline proteinase. Enzyme was stable at pH 6 to 10. 5. Opt. temperature for proteinase activity was $50^{\circ}C$, but enzyme was stable up to $40^{\circ}C$. 6. The proteinase was inhibited by $Ag^+$. It was also inhibited by EDTA. 7. When myofibrillar proteins were treated by proteinase from Asp. oryzae, ATPase activities of myofibrillar proteins changed remarkably. Accordingly, it was concluded that proteinase produced by Asp. oryzae were able to be used as meat tenderizer.

• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1444-1448
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• 2009

Potentilla chinensis Ser. (Rosaceae) has long been used for a remedy of diarrhea and inflammation in Korea. In this study, the anti-inflammatory effects of the Potentillae chinensis Herba water extract (PCX) was investigated in proteinase-activated receptor-2 (PAR2)-mediated rat paw edema. Paw edema was induced by injection of trypsin or trans-cinnamoyl-LIGRLO-$NH_2$ (tc-$NH_2$) into the hind paw of rats. PCX (10, 50, 100 and 200 mg/kg) was orally administered 1 h before the induction of inflammation. At doses of 50, 100 and 200 mg/kg, PCX showed significant inhibition on both change in paw volume and vascular permeability. PCX (100 mg/kg) significantly inhibited PAR2 agonists-induced myeloperoxidase (MPO) activity in paw tissue. These results indicate that PCX has an anti-inflammatory action in PAR2-mediated paw edema.

• Korean Journal of Medicinal Crop Science
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• v.12 no.1
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• pp.47-52
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• 2004

The root of Patrinia villosa Jussieu (Valerianaceae) has long been used for treatment of infectious diseases in Korea. In this study, the anti-inflammatory effect of the Patrinia villosa root water extract (PVWX) was investigated in proteinase-activated receptor-2 (PAR2)-mediated rat paw edema. Paw edema was induced by injection of trypsin or $trans-cinnamoyl-LIGRLO-NH_2\;(tc-NH_2)$ into hindpaw of rats. PVWX. (10, 50, 100 and 200 mg/kg) was orally administered 1 h before the induction of inflammation. At doses of 50, 100 and 200 mg/kg, PVWX. showed significant inhibition on both change in paw volume and vascular permeability. PVWX. (100 mg/kg) significant1y inhibited PAR2 agonists-induced myeloperoxidase (MPO) activity in paw tissue. These results indicate that PVWX has an anti-inflammatory action in PAR2-mediated paw edema.

• Korean Journal of Food Science and Technology
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• v.22 no.5
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• pp.569-574
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• 1990

The influence of NaCl and pH on degradation of chicken breast muscle myofibrillar proteins by porcine leukocyte lysosomal proteinases was investigated. The degradation reactions were carried out at $38^{\circ}C$ for 24hours under different conditions. The degradation of myofibrillar proteins by leukocyte lysosomal enzymes at various pH values was limited to partial hydrolysis. Reactions at higher pH values resulted in lower molecular weight degradation products while reactions at lower pH resulted in higher molecular weight degradation products. When NaCl was added into the reaction mixture, enzyme activities of degradation were increased at all pH values studied, as evidenced by NPN-analysis and SDS-PAGE. More severe degradation was observed with higher salt concentration. The concentration of 0.5M NaCl in the reaction mixture gave more degradation of myosin heavy chain by enzyme than that of 0.1M NaCl.

• Korean Journal of Microbiology
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• v.36 no.1
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• pp.14-19
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• 2000

Algal lytic enzyme, an extracellular enzyme, was purified from the culture filtrate of Penicillium oxalicum(HCLF-34) by ultrafiltration, gel filtration chromatography, and anion exchange chromatography. The enzyme has a molecular mass of approximately 22 kDa, an it is a monomer by renaturation SDS-PAGE. The amino acid sequences of the enzyme was revealed to be NH2-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, had about 84% identity with the mature light chain of aspergillopepsin II precursor and 81% identity with the mature protein of the acid proteinase EapC precursor.