• Microbiology and Biotechnology Letters
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• v.31 no.3
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• pp.284-291
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• 2003

Bacillus amyloliquefaciens K-42, which produces strongly a fibrinolytic enzyme, Was isolated from Ganjang, a traditional Korean soy sauce. The fibrinolytic enzyme was purified to homogeneity by ammonium sulfate fractionation, ion-exchange chromatography on DEAE-Sephadex A-50, gel chromatography on Sephadex G-100, and gel chromatography on Sephadex G-75 of the culture filtrate of Bacillus amyloliquefaciens K42. The purified enzyme showed the specific activity of 59.4 units per milligram, which was increased by 17.1 fold over the culture broth. And the molecular weight of purified fibrinolytic enzyme was confirmed to be about 45,000 Dalton by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The enzyme activity was relatively stable at pH 4.0-10.0 and the optimum pH was 8.0. The activity of the purified enzyme was increased by $Mg^{2+}$ , Cu$^{2+}$ but the enzyme was totally inhibited by $Ba^{2+}$ $Hg^{2+}$ In addition, the enzyme activity was potently inhibited by EDTA, EGTA and CDTA. It was concluded that the purified enzyme was a metalloprotease. And Km value was 2.03 mg/ml to fibrin.

• Korean Journal of Microbiology
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• v.47 no.4
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• pp.302-307
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• 2011

Native chitin deacetylase of Aspergillus nidulans was purified to apparent homogeneity by a combination of phenyl-Sepharose and Q-Sepharose column chromatography. In order to analyze the amino acid residues involved in the enzyme activity, the enzyme was chemically modified with chemical agent, which selectively reacted with the specific amino acid residue on the protein. When the enzyme was chemically modified with diethylpyrocarbonate, which specifically reacted with histidine residues on the protein, the activity was eliminated. The chitin deacetylase, chemically modified with 100 ${\mu}M$ modifier at the residue of arginine or tyrosine, has shown to have decreased activities. It was shown that the modification at aspartic acid or glutamic acid did not affect the enzyme activity to a greater extent, which would not implicate that acid amino residues were directly involved in catalytic reaction and would affect on the global structures of the proteins. This results demonstrated that histidine and tyrosine residues of enzyme would participate in an important function of the chitin deacetylase activity.

• Korean Journal of Microbiology
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• v.23 no.1
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• pp.13-19
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• 1985

Acc I endonuclease has been isolated from 300g (wet weight) cells of Acinetobacter calcoaceticus. The cells were broken by using French press at 20, 000p.s.i. After ammonium sulfate fractionation, the enzyme was further purified by heparin agarose, DEAE-sephades, Affi.-gel Blue, phosphocellulose, and hydroxylapatite column chromatography. The purified Acc I endonudlease has a single polypeptide species and its subunit molecular weight was 45,000 ${\pm}$ 1,000 daltons as judged by 10% SDS-polyacrylamide gel electrophoresis. The isolated enzyme was essentially free of contaminating nucleases as judged by homochromatography by using a $^{32}P-labeled$ oligonucleotide. The enzyme showed maximum activity at pH values between 8.0 and 11.0 and in the presence of $MgCl_2$. Acc I endonuclease was maximally active in the absence of NaCl and was completely inhibited at 200 mM NaCl.

• Korean Journal of Microbiology
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• v.27 no.3
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• pp.245-249
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• 1989

Extracellular proteases of Bacillus licheniformis were partially purified using ammonium sulfate fractionation and Sephadex G-75 gel filtration chromatography. The partial purification permited the weparation of two different protease activities, type I and type II. Protease type I is an enzyme with rather high protealytic activity toward dasein and was highly susceptible to organofluoride and EDTA inhibitions. It showed maximal proteolytic activity at pH 7.5 and was rapidly denatured at $71^{\circ}C$. Protease type II is a protease with relatively lower proteolytic activity than the type I. It was also inhibited by 10mM of EDTA and 1mM of PMSF by 30 min incubation. The enzyme showed maximal activity at pH 8.0 and was denatured relatively slowly at $71^{\circ}C$.

• Korean Journal of Microbiology
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• v.21 no.3
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• pp.143-148
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• 1983

The occurrence of PZ-peptidase in Streptococcus sanguis and other oral bacteria was investigated utilizing washed whole cells as the enzyme source and PZ-pentapeptide as its substrate. Under the culture conditions employed in the present study. Streptococcus sanguis strains, fresh isolates as well as laboratory strains, produced a broad range of the enzyme activity (0.5-7.9 unit/mg protein). The strains of both Streptococcus mutans and Lactobacilli showed low levels of activity (0-0.5 unit/mg protein for S. mutans). As compared with the enzyme activities of other bacteria, a moderate range of activity was produced by the strains of Strptococcus mitis nad Strptoccus salivarius. Actinomyces strains, like those of S. sanguis, produced a varying amount of activity (0-9.8 unit/ mg protein). A possible involvement of the oral bacterial PZ-peptidase in the metabolism of human saliva proteins is discussed.

• The Microorganisms and Industry
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• v.20 no.1
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• pp.18-22
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• 1994

효소는 반응을 효율적으로 촉진시킬 뿐만 아니라 기질 및 입체특이성을 갖고 있어 광학활성 화합물을 합성하는데 매우 유용한 촉매이다. 효소 반응은 일반적으로 수용액에서 진해오디지만, 유기 용매를 포함하는 용액에서도 가능하다. 대부분의 유기 화학 반응이 유기 용매에서 진행된다는 점을 생각할 때, 유기 용매에서 효소 반응을 이용한 유기합성이 갈수록 중요해지고 있다. 본 글에서는 유기 용매에서 사용가능한 대표적인 효소인 지질 가수분해 효소(lipase)를 이용한 유기합성에 대해서 간략히 기술하고자 한다.

• KSBB Journal
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• v.21 no.1 s.96
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• pp.20-27
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• 2006

The fibrinolytic activities of soluble proteins extracted from seeds of Coix lacryma-jobi L., Carthamus tinctorius L. and Malva venicillata L. were studied. Fibrinolytic activity of extract from C. lacryma-jobi L. showed 1.3 times higher than plasmin used as positive control. The fibrinolytic enzyme was confirmed and extracted directly from seed of C. lacryma-jobi L. by a fibrin zymography. The protein was composed of a single polypeptide and its apparent molecular weight was found to be 7.8 kDa, as judged by SDS-polyacrylamide gel electrophoresis. The effect of temperature for the proteolytic enzyme activity were stabilized above $50^{\circ}C$ and then dramatically decreased. Also, the enzyme activity was clearly inhibited by APMSF, PMSF and TPCK, suggesting that it is a member of the chymotrypsin-like serine pretense. In addition, effects of gamma-irradiated on seed of each plants were revealed that 8 Gy and 64 Gy were higher than others. This result shown that gamma-irradiation of seeds were capable to increase the fibrinolytic activity. All these results suggest the pretense is a fibrinolytic enzyme belong to a family of chymotrypsin-like serine pretense.

• Proceedings of the Korea Water Resources Association Conference
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• 2009.05a
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• pp.2132-2137
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• 2009

갈대(Phragmites communis Trin.)란 염분이 있는 곳에서 자라는 염생식물로서 우리나라 전역에 분포하고 있다. 지난 50년 동안 우리나라의 여러 습지에 걸쳐서 갈대는 우점종으로 자라왔고, 육지와 수중서식지에서 갈대의 확산범위는 증가하고 있다. 갈대의 확산은 다른 습지 식물의 서식지를 파괴하고, 갈대가 번식하면 동 식물들의 번식 자체가 어려울 뿐 아니라 갈대숲에 포식자가 늘어나 살아가기 어려운 환경으로 변하기때문에 갈대를 체계적으로 관리할 수 있는 방안이 마련되어야 한다. 본 연구는 우점종이 다른 두 습지에서 갈대군집의 성장률을 관찰하고, 토양의 화학적 분석과 식물의 생리적 분석을 통해 갈대군집 성장에 미치는 영양염류의 영향을 규명하였다. 연구 대상지는 한강하구에 위치한 장항습지와 성동습지로서 동일하게 갈대가 분포하며, 장항습지에는 줄 군락이 성동습지에는 새섬매자기 군락이 우점하고 있다. 분석 항목은 이화학적 항목을 비롯하여 용존유기탄소(DOC, dissolved organic carbon), 체외미생물효소활성도(Extracellular enzyme activities), 암모니아성 질소($NH_4^+$), 질산성 질소($NO_3^-$)을 분석하였다. 실험결과, 두 습지 갈대의 성장은 7월부터 9월에 증가하였고 성동습지의 토양성분이 점토질로 형성되어 높은 수분함량과 유기물함량을 유지하고 있기 때문에 갈대의 밀도가 높고 성장률이 활발한 것으로 나타났다. 또한 미생물활성과 환경인자간 양의 상관관계를 보아 환경인자들이 미생물 활성을 자극하고 미생물들은 식물의 성장을 촉진하여 영향을 주며, 반면 식물 뿌리는 enzyme을 생성하는 미생물에게 C 삼출물을 공급해 enzyme 활성에 영향을 미칠 것으로 사료된다.

• Microbiology and Biotechnology Letters
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• v.34 no.2
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• pp.121-128
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• 2006

The purified xylanase from Bacillus alcalophilus AX2000 was modified with various chemical modifiers to determine amino acid residues in the active site of the enzyme. Treatment of the enzyme with group-specific reagents such as carbodiimide or N-bromosuccinimide resulted in complete loss of enzyme activity. These results suggested that these reagents reacted with glutamic acid or aspartic acid and tryptophan residues located at or near the active site. In each case, inactivation was performed by pseudo first-order kinetics. Inhibition of enzyme activity by carbodiimide and N-bromosuccinimide showed non-competitive and competitive inhibition type, respectively. Addition of xylan to the enzyme solution containing N-bromosuccinimide prevented the inactivation, indicating the presence of tryptophan at the substrate binding site. Analysis of kinetics for inactivation showed that the loss of enzyme activity was due to modification of two glutamic acid or aspartic acid residues and single tryptophan residue.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.5
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• pp.839-843
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• 2001

The effect of gamma-irradiation on the hydrolytic enzyme activities of some Koran soybean-based fermented foods was studied. Doenjang (soybean paste), kanjang (soy sauce), kochujang (red pepper paste), chungkukjang and meju were prepared and irradiated at 0, 5, 10 and 20 kGy. Then activities of protease, amylase, lipase and fibrinolytic enzyme were determined. Hydrolytic enzyme activities of meju, chungkukjang and doenjang were relatively higher than those of kanjang and kochujang. Amylase, protease and lipase activities were not affected by 10 kGy and were slightly (about 10%) inactivated by 20 kGy of gamma irradiation, with no statistical significance. Fibrinolytic enzyme was stable in all treatments.