• Title/Summary/Keyword: Artificial protease

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Isolation and Characterization of a Protease-Producing Bacterium, Bacillus amyloliquefaciens P27 from Meju as a Probiotic Starter for Fermented Meat Products

  • Lee, Mi-Sun;Lee, Na-Kyoung;Chang, Kyung-Hoon;Choi, Shin-Yang;Song, Chi-Kwang;Paik, Hyun-Dong
    • Food Science of Animal Resources
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    • v.30 no.5
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    • pp.804-810
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    • 2010
  • This study was performed to select protease-producing Bacillus sp. as a potential probiotic starter for fermented meat products. In order to isolate protease-producing bacterium from meju, measured the diameter of the clear zone on agar plate (TSA, 1% (w/v) skim milk) and analyzed for intracellular protease activity, then 10 Bacillus-like strains were isolated. Three Bacillus-like strains (P19, P27, and P33) among 10 strains were able to tolerate in acidic condition (TSB, pH 2.5, 2 h incubation). These 3 strains were showed antimicrobial activity against food-borne pathogenic bacteria. These vegetative cells of 3 strains were showed a survival rate of 0.04% to 0.08% under the artificial gastric acidic condition (TSB, pH 2.5 with 1% (w/v) pepsin), but spore-forming cells were 56.29% to 84.77%. Vegetative cells of 3 strains were the least bile-resistant, while spore-forming cells of 3 strains showed higher survival rate more than 76% under artificial bile condition (TSB, 0.1% (w/v) oxgall bile). In these strains, P27 strain was finally selected as a good probiotic strain. P27 strain was tentatively identified as Bacillus amyloliquefaciens by API CHB kit and 16S rDNA sequence analysis. The results of this study suggest that B. amyloliquefaciens P27 can be used as a potential probiotic starter for fermented meat product.

The Optimal Culture Condition for the Collagenolytic Protease Production from Vibrio vulnificus CYK279H (Vibrio vulnificus CYK 279H에서 Collagenolytic Protease 생산을 위한 최적배양조건)

  • 강성일;김영문;장영부;임동중;공재열
    • KSBB Journal
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    • v.19 no.4
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    • pp.295-300
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    • 2004
  • A marine bacterium for producing an collagenolytic protease was isolated from the southern sea of Korea and identified as Vibrio vulnificus and named as Vibrio vulnificus CYK279H. This strain producing an collagenolytic protease was showed high activity toward collagen and gelatin as substrate. The optimum initial pH, NaCl, and temperature for cell growth and protease production was 7.5, 2.0% and 25$^{\circ}C$, respectively. Optimization for collagenolytic protease production was composed of 0.3% D-galactose, 0.6% yeast extract, 4.0% gelatin, 0.2% (NH$_4$)$_2$SO$_4$, and 0.2 mM ferric citrate in artificial sea water. The maximum protease production was required gelatin and yeast extract. The collagenolytic protease production by Vibrio vulnificus CYK279H reached a maximum of 73 unit/l after the cultivation for 18 h under the optimized medium.

The Effects of Protease and Lipase on the Detergency of Fabrics (프로테아제와 리파제가 직물의 세척에 미치는 영향)

  • Lee, Jeong-Sook;Chung, So-Wha
    • Fashion & Textile Research Journal
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    • v.2 no.4
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    • pp.339-345
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    • 2000
  • The effects of protease and/or lipase on the removal of protein soil and oily soil were investigated in this study. Cotton, rayon, nylon, and PET fabrics were soiled by padding of fresh bovine blood and spotting of mixed artificial sebum evenly. The soiled fabrics were aged at $130^{\circ}C$ for 30 minutes. The fabrics were washed by using Terg-O-Tometer at various conditions. Protease and/or lipase were added in the alcohol ethoxylate (AE) detergent solution. The removal efficiency was evaluated by analysis of protein and/or oil on the fabrics before and after washing, respectively. The detergency of protein and/or oil on the fabrics was discussed with enzyme concentration, washing time, washing temperature, pH of washing solution and fiber characteristics. The hydrolysis of protease improved effectively the removal of oil as well as protein by increasing removal of protein-oil mixed soil at the same time. The effect of lipase added detergent solution was slightly shown on the removal of oil and/or protein. The removal of mixed soils from cotton fabrics was very low because of large amount of residual soils caused by the physical characteristics of cotton fiber.

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Overproduction of Streptomyces griseus Protease A and B Induces Morphological Changes in Streptomyces lividans

  • Chi, Won-Jae;Kim, Jung-Mee;Choi, Si-Sun;Kang, Dae-Kyung;Hong, Soon-Kwang
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1077-1086
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    • 2001
  • The sprA and sprB gene encoding chymotrypsin-like proteases Streptomyces griseus protease A (SGPA) and Streptomyces griseus protease B (SGPB) and the sprT gene that encodes Streptomyces griseus trypsin (SGT) were cloned from Streptomyces griseus ATCC10137 and overexpressed in Streptomyces lividans TK24 as a heterologous host. The chymotrypsin activity of tole culture broth measured with the artificial chromogenic substrate , N-succinyl-ala-ala-pro-phe-p-nitroanilide, was 10, 14 and 14 units/mg in the transformants haboring the sprA, sprB and sprD genes, respectively. The growth of S. lividans reached the maximum cell mass after 4 days of culture, yet SGPA and SGPD production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The trypsin activity of the culture broth measured with the artificial chromogenic substrate , N-${\alpha}$-benzoyl-DL- arginine-p-nitroanilide , was 16 units/mg and SGT production started in the stationary phase of cell growth and kept increasing for up to 10 days of culture in an R2YE medium. The introduction of the sprA gene into S, lividans TK24 triggered the biosynthesis of pigmented antibiotics, actinorhodin and undecylprodigiosin, and induced significant morphological changes in the colonies in Benedict, R2YE, and R1R2 media. In addition, the introduction of the sprT gene also induced morphological changes in the colony shape without affecting the antibiotic production, thereby implying that certain proteases would appear to play very important and specific roles in secondary-metabolites formation and morphological differentiation in Streptomyces.

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Characterization and optimum production condition of extracellular protease from Pseudoalteromonas donghaensis HJ51 (Pseudoalteromonas donghaensis HJ51의 체외 단백질 분해효소 특성과 생산 조건)

  • Oh, Ji-Sung;Choi, Yoon-Soo;Roh, Dong-Hyun
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.75-80
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    • 2015
  • Pseudoalteromonas donghaensis HJ51, isolated from the East Sea, has been reported as a novel strain to produce extracellular protease. Crude supernatant was used to determine optimal activity and optimal production conditions for the enzyme. It was found that the optimal temperature and pH of the protease were $40^{\circ}C$ and pH 7.5-10.5, respectively. The enzyme activity was kept to 88% at the pH 11. In metal requirement analysis, the enzyme exhibited the highest activity when 10 mM $Fe^{3+}$ was supplied. While supplementation of additional carbon sources used in study showed no positive effect on cell growth and enzyme activity, the addition of beef extract, tryptone, or casamino acids instead of peptone of PY-ASW containing 1% glucose increased enzyme production to 21, 7, 4%, respectively. Taken together these properties, the enzyme produced from P. donghaensis HJ51 can be applied to the industries that require protease activity under alkaline pH and low temperature.

Artificial Metalloproteases with Broad Substrate Selectivity Constructed on Polystyrene

  • Ko, Eun-Hwa;Suh, Jung-Hun
    • Bulletin of the Korean Chemical Society
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    • v.25 no.12
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    • pp.1917-1923
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    • 2004
  • Although the proteolytic activity of the Cu(II) complex of cyclen (Cyc) is greatly enhanced upon attachment to a cross-linked polystyrene (PS), the Cu(II)Cyc-containing PS derivatives reported previously hydrolyzed only a very limited number of proteins. The PS-based artificial metalloproteases can overcome thermal, mechanical, and chemical instabilities of natural proteases, but the narrow substrate selectivity of the artificial metalloproteases limits their industrial application. In the present study, artificial metalloproteases exhibiting broad substrate selectivity were synthesized by attaching Cu(II)Cyc to a PS derivative using linkers with various structures in an attempt to facilitate the interaction of various protein substrates with the PS surface. The new artificial metalloproteases hydrolyzed all of the four protein substrates (albumin, myoglobin, ${\gamma}$-globulin, and lysozyme) examined, manifesting $k_{cat}/K_m$ values of 28-1500 $h_{-1}M_{-1}$ at 50 $^{\circ}C$. The improvement in substrate selectivity is attributed to steric and/or polar interaction between the bound protein and the PS surface as well as the hydrophobicity of the microenvironment of the catalytic centers.

Studies on the Efficiency of the Artificial Diet for the Silkworm, Bombyx mori L. (누에인공사료의 사료효율증진에 관한 연구)

  • 김주읍
    • Journal of Sericultural and Entomological Science
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    • v.28 no.2
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    • pp.38-47
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    • 1986
  • Silkworm(Bombix mori) were reared with modified artificial diets which were mixed with, as additives, leaf powders of Erigeron canadensis L., Cassia tora L., Cyperus anuricus Var.Laxus and Vigna Sinensis NEDL. The effects of additives on silkworm characteristisc of tested plants were summarized as follows ; 1. About 2-5% addition on dry weight base of leaf powders of E. canadensis, C. tora, C. anuricus or V.sinensis to the basic artificial diet promoted feeding response and digestion and resulted in good practical silkworm characteristics. The addition of V.sinensis and C.anuricus showed especially good effects. 2. The syneristic effect between different plant species was not recognized based on the feeding response and digestion of silkworm reared with various combinations of 2-4 different plant additives. 3. Electrophoretic zymograms of estrase, protease and phosphatase on haemolymph, intestine and silkgland were significantly different among treatments. In general, 1 or 2 more electrophoretic bands were detected when feeding response and digestion were promoted. 4. Contents of starch, crude fat, crude protein and inorganic base were apparently higher in the tested plants than in mulberry leaves. However, no volatile ingredent which is directly realted with feeding response was identified.

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