• Title, Summary, Keyword: Bacillus subtilis K-20

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Characteristics of Flavor and Functionality of Bacillus subtilis K-20 Chunggukjang (Bacillus subtilis K-20에 의한 청국장의 향미성분 및 기능성식품에 관한 연구)

  • Kim, Young-Sook;Jung, Hyuck-Jun;Park, Young-Sook;Yu, Tae-Shick
    • Korean Journal of Food Science and Technology
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    • v.35 no.3
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    • pp.475-478
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    • 2003
  • Bacillus subtilis K-20 chunggukjang is widely used in making soy sauces and bean pastes which are Korean traditional fermented foods. Bacillus subtilis K-20 chunggukjang was cultured, and fermented at $40^{\circ}C$ and 90% humidity for 96 hr after homogenizing with garlic, garlic and onion, and garlic, onion, and ginger. As a result, a product with pizza flavor and taste was obtained from Bacillus subtilis K-20. This product could be used as a functional food to promote immunity.

Antimicrobial Activities of Viscous Substance from Chongkukjang Fermented with different bacillus spry. (청국장 발효 세균의 종류에 따른 청국장 정절물의 항 미생물 활성에 관한 연구)

  • 윤호경;최희선;허성호;홍정화
    • Journal of Food Hygiene and Safety
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    • v.16 no.3
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    • pp.188-193
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    • 2001
  • To evaluate antimicrobial activities of chongkukjang slime fermented by different strains, growth characteristics were compared using various standard microorganisms with addition of chongkykjang slime. Chonghkjang slime was prepared by fermenting cooked soybean after inoculating with Bacillus circulans K-1, Baciilus spp N-1 and Bacillus subtilis CH-1, respectively. Significant antimicrobial activity was observed by chongkukjang slime on gram positive bacteria (Staphylococcus aureus, Bacillus cereus, Bacillus subtilis, Micrococcus luteus), gram negative bacteria(Escherichia coli O157:H7, Salmonella Typhimurium, Pseudomonas fluorescens), and yeast (Pichia membranaefaciens, Saccharomyces cerevisiae, Candida albicans). In case of B. cereus growth inhibition of 80% was achieved by the addition of chongkukjang slime; on the contrary, to Escherichia coli O157:H7 only 20% inhibition was observed. Slime from Bacillus subtilis CH-1, in particular, inhibition of 40% toward bacteria and yeast, whereas slime from Bacillus circulans K-1, Bacillus spp N-1 showed only 20% inhibition.

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Cloning of Fibrinolytic Enzyme Gene from Bacillus subtilis Isolated from Cheonggukjang and Its Expression in Protease-deficient Bacillus subtilis Strains

  • Jeong, Seon-Ju;Kwon, Gun-Hee;Chun, Ji-Yeon;Kim, Jong-Sang;Park, Cheon-Seok;Kwon, Dae-Young;Kim, Jeong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.1018-1023
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    • 2007
  • Bacillus subtilis CH3-5 was isolated from cheonggukjang prepared according to traditional methods. CH3-5 secreted at least four different fibrinolytic proteases (63, 47, 29, and 20 kDa) into the culture medium. A fibrinolytic enzyme gene, aprE2, encoding a 29kDa enzyme was cloned from the genomic DNA of CH3-5, and the DNA sequence determined. aprE2 was overexpressed in heterologous B. subtilis strains deficient in extracellular proteases using a E. coli-Bacillus shuttle vector. A 29 kDa AprE2 band was observed and AprE2 seemed to exhibit higher activities towards fibrin rather than casein.

Isolation and Identification of Bacillus sp. with High Protease and Amylase Activity from Sunchang Traditional Kochujang

  • Jung, Sung-Tae;Kim, Min-Hwa;Shin, Dong-Hwa;Kim, Yong-Suk
    • Food Science and Biotechnology
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    • v.17 no.3
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    • pp.519-526
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    • 2008
  • To improve the quality of traditional kochujang, strains with high protease and amylase activity were isolated and identified from Sunchang traditional kochujang. Twenty-three strains strongly producing protease and 16 strains strongly producing $\alpha$- and $\beta$-amylase were isolated by using 1% isolated soy protein agar medium and 2% starch agar medium, respectively. Protease activities of the IA7, I5, and IA2 strain were 22.5, 21.2, and 20.6 unit/mL, respectively, and were higher than those of the other strains. Stains with high $\alpha$-amylase activity included K9 (967.8 unit/mL), K14 (828.3 unit/mL), K13 (662.5 unit/mL), K8 (601.5 unit/mL), and K11 (405.9 unit/mL). The $\beta$-amylase activity of the K11 strain was the highest, 34.3 unit/mL, among the isolated strains. Based on morphological, physiological properties, and API 50CHB-kit test for assimilation of 49 carbohydrates, 8 strains selected according to protease, $\alpha$-amylase, and $\beta$-amylase activities were tentatively identified as Bacillus megaterium (IA2), Bacillus subtilis (IA7, 15), Bacillus amyloliquefaciens (K8, K9, K11, and K13), and Bacillus stearothermophillus (K14). The IA7, 15, and K11 strains were finally identified as B. subtilis (99% ID) based on 16S rDNA sequencing.

Purification and Characterization of Fibrinolytic Enzyme Produced by Bacillus subtilis K7 Isolated from Korean Traditional Soy Sauce (한국재래간장 발효균 Bacillus subtilis K7 유래의 혈전용해 Protease의 정제 및 특성)

  • Kim, Doo-Young;Lee, Eun-Tag;Kim, Sang-Dal
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.176-182
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    • 2003
  • An alkaline fibrinolytic protease-producing bacteria was isolated front Korean traditional soy sauce and identified as Bacillus subtilis K7 from the results of analyses of its morphological and physiological properties, $API^{\circledR}$, and Biolog system. The enzyme was purified by 75% ammonium sulfate fractionation, QAE-Sephadex anion and SP-Sephadex cation exchange column chromatography and Sephadex G-100 gel filtration. The specific activity of the purified enByme was 233.9 unit/mg protein and the yield of enzyme was 3.8%. The homogeneity of the purified enzyme was confirmed by polyacrylamide gel electrophoresis. Molecular mass of the enzyme was estimated about 21,500 Da by SDS-polyacrylamide get electrophoresis and gel chromatography. The optimum temperature and pH for the enzyme activity were $40^{\circ}C$ and 9.0, respectively. The enzyme was stable in a pH range of 5.0 to 12.0, and 60% of its activity was lost on heat treatment at $50^{\circ}C$ for 20 min. The activity of the purified enzyme was inhibited by the presence of $Fe^{2+},\;Ag^{2+},\;Cu6{2+}$, iodoacetate, ethylene diamine tetraacetic acid (EDTA), and trans-1,2-diaminocycloheane-N,N,N',N'-tetraacetic acid (CDTA). The results indicates that the enzyme requires a metal ion for its enzymatic activity.

Variation of fibrinolytic enzyme activity produced Bacillus subtilis by gene cloning (유전자 cloning에 의한 Bacillus subtilis의 fibrinolytic enzyme 활성 변화)

  • 이홍석;유천권;이철수;강상모
    • Microbiology and Biotechnology Letters
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    • v.28 no.1
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    • pp.14-20
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    • 2000
  • The transformation of Bacillus subtilis K-54 and J-10 was carried out with constructed vectors containing structure and enhancer genes of aprN and prtR, to increase their fibrinolytic enzyme activity. Bands for the aprN and prtR genes were identified from B. subtilis J-10 by PCR that was carried out with the constructed primers for the genes. In addition, the gene fragments contained promoter site based on the results of analysing their nucleotide sequence. The two gene fragments, aprN and prtR, obtained by the PCR, were, then, inserted to vector such as T-vector and E.coli/Bacillus shuttle vector. The constructed vector were designated as pAPR2 (aprN), pENC2 (prtR) and pFLA1 (aprN and prtR), respectively. The constructed vector was used for transformation of the strains of B.subtilis J-10 and B. subtilis K-54 and the fribrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and the fibrinolytic activity of the transformed strains was investigated. The introduction of the vector, pAPR2 and pFLA1, resulted in the increase of fibrinolyitic enzyme activity in B. subtilis J-10 by 27.3% and 16%, respectively. However, the introduction of pENC2 to B. subtilis J-10 did not seem to induce increase of the enzyme activity. The strain of B.subtilis K-54 transformed with pENC2 showed an increased fibrinolytic activity by 5 folds compared with that of the original strain of B. subtilis K-54.

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Characteristics of Protease Produced by Bacillus subtilis PCA 20-3 isolated from Korean Traditional Meju (전통 메주로부터 분리한 Bacillus subtilis PCA 20-3 유래의 Protease 생산과 특성)

  • Lim, Seong-Il;Kim, Hyun-Kyu;Yoo, Jin-Young
    • Korean Journal of Food Science and Technology
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    • v.32 no.1
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    • pp.154-160
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    • 2000
  • Protease production and its characteristics were investigated with Bacillus subtilis PCA20-3 which was isolated from Korean traditional meju. The optimum culture conditions of Bacillus subtilis PCA20-3 for the production of the protease were as follow: 0.2% soytone, 2% starch, 0.1% $(NH_4)_2SO_4,\;0.2%\;CaCl_2,\;0.01%\;yeast\;extract,\;0.1%\; K_2HPO_4,\;0.1%\;KH_2PO_4,\;pH\;7.0,\;30^{\circ}C$ and 20 hrs. The optimum pH and temperature for enzyme activity of protease producing Bacillus subtilis PCA20-3 were pH 8.0-10.0 and $55^{\circ}C$, respectively. The enzyme was relatively stable at pH $6.0{\sim}11.0$ and at temperature below $50^{\circ}C$. The activity of the enzyme was inhibited by $Fe^(2+)\;and\;Cu^(2+)$. 2 mM phenymethanesulfonyl fluoride inhibited 89.2% of enzyme activity. This indicates that the enzyme is serine protease. The $K_m$ value was $5\;{\times}\;10^{-4}\;M,\;V_{max}\;value\;was\;100\;{\mu}g/min$. This enzyme hydrolyzed casein more rapidly than bovine serum albumin.

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Characteristics of Antifungal Bacterium, Bacillus subtilis YS1 and It′s Mutant Induced by Gamma Radiation (온천수로부터 분리한 항진균세균의 특성 및 감마선$(Co^{60})$ 조사를 이용한 돌연변이체 유기)

  • 이영근;김재성;송인근;정혜영;장화형
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.305-311
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    • 2001
  • Antifungal bacterium, Bacillus subtilis YS1 was isolated from Yusong hot spring showed broad antifungal spectrum against 12 kinds of plant pathogenic fungi and Candida albicans, animal pathogen. From the gamma($Co^{60}$) radiation sensitivity test, $D_10$ value was 2.08 kGy and it survived above 20 kGy of radiation dose. Several mutants were induced by gamma radiation. Among them, YS1-1009 mutant showed resistance against tebuconazole of herbicide, increased activity against Botryoshaeria dothidea and ligninase activity. YS67 mutant was antifungal deficient auxotrophic mutants(trp-pro-or arg-ura-). From this results, it suggested that gamma irradiation could be useful method for mutant induction.

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Two- Dimensional Electrophoresis Analysis of Proteins; Bacillus subtilis LTD and Its Antifungal Activity Deficient Mutant

  • Lee, Young-Keun;Dinh, Le Thi;Jang, Yu-Sin;Chung, Hye-Young;Chang, Hwa-Hyoung
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.487-493
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    • 2004
  • To investigate the antifungal activity related protein in pesticidal bacteria, a bacterial strain LTD was isolated from soil collected at Gimje in Jeonbuk province, Korea, and identified as Bacillus subtilis LTD based on a API50 CHB kit and 168 rDNA sequencing. It has an antifungal activity against 9 plant pathogenic fungi in a paper disc assay. The antifungal activity- deficient mutant, B. subtilis mLTD was induced at a 5 kGy dose of $^{60}Co$ gamma radiation. Using the two-dimensional electrophoresis and the matrix assisted laser desorption ionization time-of-flight mass spectrometry, the comparison analysis of proteins between the wild and mutant were performed. A major intracellular serine proteinase IspA (MW: 32.5 kDa), a NAD (P) H dehydrogenase (MW: 20.0 kDa), and a stage II sporulation protein AA, SpoIIAA (MW: 14.3kDa) were detected only in the B. subtilis LTD. These results suggested that the functions of these proteins found only in the B. subtilis LTD could. be closely related to the antifungal activity against plant pathogenic fungi.

Characterization of Bacteriocin from Bacillus subtilis cx 1 (Bacillus subtilis cx1이 생산하는 박테리오신의 특성)

  • 김수인;장지윤;김인철;장해춘
    • Microbiology and Biotechnology Letters
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    • v.29 no.1
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    • pp.50-55
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    • 2001
  • A new bacteriocin produced by Bacillus subtilis cx1, was partially purified and characterized. The bactericoin from B. subtilis cx1 was stable in the range of pH 2.5-9.5. B. subtilis csx1 retained its antimicrobial activity to long-term exposure at $-20^{\circ}C$ and $-70^{\circ}C$. However, B. subtilis cx1 was inactivated completely within 15 min over $60^{\circ}C$ and lost 50% of its antimicrobial activity within 15 min at $50^{\circ}C$, B. subtilis cx1 was inactivated by protease, trypsin, proteinase K and carboxypeptidase, which indi-cates its protein nature. Direct detection of the antimicrobial activity on Tricine -SDS-PAGE suggested an apparent molecular mass of about 9,500 dalton.

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