• Biomolecules & Therapeutics
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• v.3 no.2
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• pp.165-170
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• 1995

The biochemical properties of the fibrinolytic protease of 50,800 Da isolated from the venom of Kgdistrodon blomhoffi brevicaudus were characterized. The enzyme hydrolyzed the carboxyl side of arginine in the synthetic chromogenic peptides, N-Benzoyl-Phe-Val-Arg-pNA and N-p-Tosyl-Gly-Pro-Arg-pNA, and the enzyme activity was inhibited by phenylmethylsulfonylfluoride indicating that the enzyme belongs to the serine protease family. The pretense showed maximum activity at pH 7.5 and inhibited by ZnCl$_2$, CuSO$_4$, but not by soybean trypsin inhibitor, pepstatin A, 2-mercaptoethanol and EDTA. The fm value determined with N-p-Tosyl-Gly-Pro-Arg-pNA was 0.2 mM. The thrombolytic activity of the purified enzyme was evaluated by platelet aggregation test in rabbits. While the platelet count ratio in blood of the rabbits injected with thrombin alone declined from 1.0 to 0.6 within 7 min and maintained around 0.6 for 24 hours thereafter, the ratio rapidly recovered from around 0.6 to 0.8 in 1 hr, to 1.0 in 24 hrs when the rabbits were sequentially treated with thrombin and the purified enzyme. The result showed that the serine protease from A. blomhoffi brevicoudus of 50,800 Da had a thrombolytic activity in vivo and the enzyme might be developed as a therapuetic agent for the treatment of thrombic disease.

• The Korean Journal of Mycology
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• v.36 no.2
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• pp.183-188
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• 2008

A fibrinolytic serine protease was purified from the fruiting bodies of an edible mushroom, Albatrellus confluens. The enzyme had a molecular mass of 30086.41 Da, as measured by MALDI-TOF mass spectrometry. The N-terminal amino acid sequence of the enzyme was Glu-Thr-Val-Thr-Glu-Thr-Thr-Ala -Pro-Trp-Gly-Leu-Ser-Arg-Ile. It displayed optimal activity at $50^{\circ}C$ and within a pH range of $8.0{\sim}10.0$, suggesting that the enzyme is an alkaline protease. The enzyme was stable up to $30^{\circ}C$. The enzyme displayed a strong substrate specificity for the synthetic peptide, N-Suc-Ala-Ala-Pro-Phe pNA. The enzyme activity was completely inhibited by addition of PMSF, indicating that the enzyme is a serine protease. No inhibition was observed following addition of E-64, pepstatin, or EDTA. The activity of the purified enzyme was decreased in the presence $Fe^{2+}$ or $Co^{2+}$, and the enzyme was completely inhibited by addition of $Hg^{2+}$. From these results, we propose that Albatrellus confluens could be used for biofunctional foods development and has potential therapeutic value for the treatment of vascular diseases.

• The Korean Journal of Zoology
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• v.32 no.1
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• pp.40-47
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• 1989

Alteration of intracellular calcium ion Concentration by adding of either calcium ionophore A23187 or EGTA in culture medium at 24 hr after cell plating resulted in remarkable changes in the progression of differentiation of chick embryo myoblast. When separated myoblast proteins using two-dimensional gel electrophoresis, synthesis patterns of several proteins changed upon the addition of either A23187 or EGTA. Treatment of A23187 and calciumactivated neutral protease at 24 hr after initial plating caused an increase in the rate of fusion compared to control culture. However, EGTA inhibited the myoblast fusion to a marked degree. A23187 treated at 24hr also increased the activity of protein kinase C during the fusionprogressed period. It seems that intracellular calcium ion plays an important role in the myoblast differentiation in vitro together with the protein kinase C and calcium-activated neutral protease.

• The Korean Journal of Physiology and Pharmacology
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• v.13 no.1
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• pp.49-54
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• 2009

The mushroom Cordyceps militaris has been used for a long time in eastern Asia as a nutraceutical and in traditional Chinese medicine as a treatment for cancer patients. In the present study, a cytotoxic antifungal protease was purified from the dried fruiting bodies of C. militaris using anion-exchange chromatography on a DEAE-Sepharose column. Electrophoretic analyses indicated that this protein, designated C. militaris protein(CMP), has a molecular mass of 12 kDa and a pI of 5.1. The optimum conditions for protease activity were a temperature of $37^{\circ}C$ and pH of $7.0{\sim}9.0$. The enzyme activity was specifically inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride. Amino acid composition of intact CMP and amino acid sequences of three major peptides from a tryptic digest of CMP were determined. CMP exerted strong antifungal effect against the growth of the fungus Fusarium oxysporum, and exhibited cytotoxicity against human breast and bladder cancer cells. These results indicate that C. militaris represents a source of a novel protein that might be applied in diverse biological and medicinal applications.

• Fashion & Textile Research Journal
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• v.8 no.2
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• pp.239-244
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• 2006

For studies of fibrinolytic enzyme strain K-54 was isolated from the Korean traditional food chungkook-jang. Isolated strains K-54 was identified as Bacillus subtilis. The molecular weight of fibrinolytic enzyme from B. subtilis K-54 was 27 kDa. Optimum temperature for fibrinolytic enzyme of B. subtilis K-54 was $50-70^{\circ}C$ and optimum pH for producing the enzyme of this strain was ranging from 8 to 12. Also, it was found out enzyme activity was completely inhibited by 1mM PMSF. The result indicated this enzyme was thermo-stable alkaline serine protease with strong fibrinolytic activity. The wool and silk were treated with protease of B. subtilis K-54. As a result, the property of dyeing of wool fabrics was increased. By the increasing of treatment time became smoothened. But the change of mechanical properties were not changed.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.7-14
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• 2006

Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium ($0.7%\;K_{2}HPO_{4},\;0.2%\;KH_{2}PO_{4},\;0.1%$ fructose, 1.2% whole chicken feather, $0.01%\;Na_{2}CO_3$, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under $30^{\circ}C$, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with $(NH_4)_{2}SO_4$ and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of $30{\sim}50^{\circ}C$ and pH $6.0{\sim}12.0$.

• Korean Journal of Food Science and Technology
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• v.20 no.4
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• pp.488-492
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• 1988

The physicochemical properties of viscosity, color, acidity and volume of anchovy extract were measured for their changes during extraction with alkali solution and/or proteolytic enzymes. The dried anchovies were ground in 0.3N NaOH solution followed by hydrolysis with neutral or alkaline protease and centrifuged to obtain anchovy extract. The results showed that the volume of supernatant after centrifugation increased from 70% of water only extraction to 89% by combined alkali-enzyme treatment. Titratable acidity of the extract showed a tendency of a little increase while viscosiy decreased with prolonged enzymic hydrolysis. Changes in Hunter value of 'L', 'a', 'b' showed that the extract became darker and less yellowish as protease treatment prolonged.

• Journal of the Korea Fashion and Costume Design Association
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• v.11 no.1
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• pp.43-51
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• 2009

The cotton, wool, cotton/wool blended(80:20) and tencel fabrics were treated with low temperature oxygen or argon plasma, enzymes(cellulase or protease), or oxygen plasma-enzyme and examined for their weight loss and conditions for treatment for the environment friendly finishing. In the plasma treatment argon gas had better effect on the weight loss than oxygen gas did and the weight loss of all the fabrics was increased as increasing discharge power and discharge time. The weight loss of cotton, wool, cotton/wool blended(80:20) fabrics decreased in a large measure after 1 hr but that of tencel didn't decrease after 1 hr. In case of cellulose fibers oxygen gas plasma induced chemical functional groups on the surface of substrate more than argon gas plasma did so the weight loss of wool was larger than that of cotton, tencel fabrics in oxygen plasma-enzyme treatment. The weight loss of cotton and tencel fabrics decreased the initial stage because oxygen plasma pre-treatment caused cross linking as well as etching effect but argon plasma pre-treatment didn't. The plasma pre-treatment cleared the way for enzyme treatment on the whole but oxygen plasma pre-treatment bear in hand the increase of weight loss more or less because of the cross linking on the surface of cellulose fibers. The appropriate conditions for plasma treatment was 10-1Torr, 40W for 30minutes and for cellulase treatment were enzyme concentration of $3g/{\ell}$, pH 5, $60^{\circ}C$ for 1hr and for protease treatment were enzyme concentration of $4g/{\ell}$ pH 8, $60^{\circ}C$ for 1hr.

• Journal of Plant Biology
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• v.38 no.1
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• pp.1-10
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• 1995

Photoinhibition and its recovery of spectrally adapted Anacystis nidulans were studied. Phycocyanin and Chl content and phycocyanin/Chl ratio were increased in cells grown under blue-green light compared with those grown in white light. Photosynthetic activities of white light and blue-green light grown cells were reduced by 50% after 15 min and 10 min of photoinhibitory light treatment (1.2 mmol·m-2s-1), respectively, largely due to the decline of PSII activities. However, their activities were recovered fully after 30 min incubation under weak light. Treatment of rifampicin and chloramphenicol magnified the photoinhibitory effects and suppressed the recovery with disappearance of susceptibility to photoinhibition and delayed the recovery process, indicating no significant differences in phosphorylation, dephosphorylation and protease activity between two cells. Therefore, it is suggested that the increased sensitivity of blue-green adapted cells might be attributed to the decline of protein synthesis, and phosphorylation-dephosphorylation of protein and protease activity might be involved in the recovery process.

• Korean Journal of Food Science and Technology
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• v.32 no.4
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• pp.867-874
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• 2000

Kochujang, which is one of most favorable fermented hot seasonings in Korea, has swelling problem and color changes during distribution. To stop the gas formation, heat treatments were conducted at $50^{\circ}C\;to\;70^{\circ}C$ for 5 to 15 min. at the beginning or after 40 days fermentation. Yeast were not detected at $60^{\circ}C$ for 15min. heat treatment and bacteria were not effected in number that is concerned in fermentation of kochujang. At above heat treatment, there was no effect ${\alpha}-amylase$ but ${\beta}-amylase$ activity was increased in heat treated kochujang. The acid protease activity was higher than nonheat treated kochujang after 60 days fermentation but neutral protease decreased after 80 days fermentation. The L, a and b value by Hunter were gradually decreased during fermentation and ${\Delta}E$ value showed difference in kochujang heated before fermentation. The total hydroxy methyfurfural content was increased, and carotenoid and capsanthin in heated kochujang decreased during fermentation.