• Title, Summary, Keyword: Bacillus subtilis

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Cloning and Expression of A Liquefying $\alpha$-Amylase Gene from Bacillus amyloliquefaciens in Bacillus subtilis (Bacillus amyloliquefaciens 액화형 $\alpha$-amylase 유전자의 클로닝 및 Bacillus subtilis에서의 발현)

  • 김사열;송방호;이인구;서정환;홍순덕
    • Microbiology and Biotechnology Letters
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    • v.14 no.6
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    • pp.479-485
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    • 1986
  • A 5200 basepair DNA fragment containing the Bacillus amyloliquefaciens amyE gene, encoding liquefying $\alpha$-amylase (1,4-$\alpha$-1)-glucan glucanohydrolase, EC 3.2.1.1), has been inserted into BamHI site of the pUB110 and the hybrid plasmid was designated as pSKS3. The pSKS3 was transformed into the Bacillus subtilis KM2l3 as a host which is a saccharifying $\alpha$-amylase deficient mutant of Bacillus subtilis NA64, and the plasmid in the transformed cell was expressed $\alpha$-amylase production and kanamycin resistance. The $\alpha$-amylase production of the transformed cell was reduced to one fifth of that of the donor strain. The Bacillus subtilis KM2l3 tarring pSKS3 indicated that the amyE gene product is a polypeptide which has the same electrophoretic mobility with that of the Bacillus amyloliquefaciens, but different from the saccharifying $\alpha$-amylase of Bacillus subtilis NA64. It means that the amyE gene of pSKS3 originales from the Bacillus amyloliquefaciens.

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High-Level Expression and Secretion of Bacillus pumilus Lipase B26 in Bacillus subtilis Chungkookjang

  • Lee, Mi-Hwa;Song, Jae-Jun;Choi, Yoon-Ho;Hong, Seung-Pyo;Rha, Eu-Gene;Kim, Hyung-Kwoun;Lee, Seung-Goo;Poo, Har-Young;Lee, Sang-Chul;Seu, Young-Bae;Sung, Moon-Hee
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.892-896
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    • 2003
  • High-level expression of the lipase B26 gene from Bacillus pumilus was achieved using Bacillus subtilis Chungkookjang isolated from the Korean traditional fermented bean paste, Chungkookjang. For the secretory production of recombinant lipase B26 in a Bacillus host system, pLipB26 was constructed by ligating the lipase B26 gene into the recently designed Escherichia coli-Bacillus shuttle vector, pLipSM, and that was then transformed into B. subtilis Chungkookjang. Among the various vector, medium, and host combinations, B. subtilis Chungkookjang harboring the pLipB26 exhibited the highest lipase activity in PY medium, and B. subtilis Chungkookjang secreted two times more enzymes than B. subtilis DB 104 under the same condition. When B. subtilis Chungkookjang harboring the pLipB26 was cultured in a 5-1 jar-fermentor containing 21 of a PY medium, the maximum lipase activity (140 U/ml) and production yield (0.68 g/l) were obtained during the late exponential phase from a cell-free culture broth. Although B. subtilis Chungkookjang also secreted extracellular proteases at the late exponential phase, these results suggested the potential of B. subtilis Chungkookjang as a host for the secretory production of foreign proteins.

Expression of Bacillus macerans Cyclodextrin Glucanotransferase in Bacillus subtilis

  • Kim, Chang-Sup;Han, Nam-Soo;Kweon, Dae-Hyuk;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.230-233
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    • 1999
  • A plasmid vector was constructed for the expression and secretion of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in Bacillus subtilis. The vector, pUBACGT, was composed of the ribosome-binding sequence, signal sequence, and cgt gene from B. macerans under the control of amyR2, the promoter of amyE gene coding for $\alpha$-amylase from B. subtilis var. natto. Bacillus subtilis LKS88, a mutant strain lacking genes for an amylase and two proteases, was used as a host for the transformation of the plasmid vector. The transformants were selected on kanamycin-containing Luria-Bertani plates. The starch hydrolyzing activity was observed on the starch-containing plates by the iodine method and cyclodextrin-forming activity was detected in the culture medium. A SDS-PAGE analysis showed that most of the expressed CGTase in the recombinant B. subtilis was secreted into the medium at a high expression level.

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Fermentation and Quality Characteristics of Cheonggukjang Fermented with Bacillus subtilis BC-P1 (Bacillus subtilis BC-P1 균주를 이용하여 제조한 청국장의 발효 및 품질 특성분석)

  • Park, Sung-Yong;Bang, Mi-Ae;Oh, Boung-Jun;Park, Jeong-Hoon;Song, Won-Seob;Choi, Kyung-Min;Choung, Eui-Su;Boo, Hee-Ock;Cho, Seung-Sik
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.262-269
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    • 2013
  • The object of this study was to improve the quality of Cheonggukjang with new starter, Bacillus subtilis BC-P1. Twenty strains were isolated from the commercial cheonggukjang and 1 Bacillus strain (BC-P1) with protease activity was selected. The 16S rRNA gene sequence revealed that the BC-P1 was closely related to B. subtilis with 99% homology. The quality characteristics of chunggukjang fermented with B. subtilis BC-P1, Bacillus nato (PC) and commercial chunggukjang (NC) were investigated. The characteristics of fermentation were determined by protease, lipase, xylanase, chitinase, and fibrinolytic activities, reducing sugar, nutrient composition and amino acid contents of cheonggukjang sample. Cheonggukjang fermented with B. subtilis BC-P1 showed the strongest fibrinolytic, xylanase, and chitinase activities. Reducung sugar contents of Cheonggukjang samples were $30.16{\pm}2.11$ mg/g (NC), $28.56{\pm}1.52$ mg/g (PC), $32.39{\pm}1.87$ mg/g (BC-P1). And their total amino acid contents were 338.99 mg% (NC), 445.19 mg% (PC), 741.35 mg% (BC-P1). These results suggested that B. subtilis BC-P1 was suitable to be used as a starter to enhance the quality and effects of cheonggukjang.

Effect of Meju Shapes and Strains on the Chemical Composition of Soybean Paste (Bacillus속과 Aspergillus oryzae로 만든 메주가 개량식 된장의 성분에 미치는 영향)

  • Seo, Jeong-Sook;Man, Eun-Mi;Lee, Taik-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.15 no.4
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    • pp.1-9
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    • 1986
  • The mashes of soybean paste were preparea using the conventional meju fermented naturally by wild microoganisms or the new types of meju fermented by pure cultures of Aspergillus oryzae, Bacillus natto and B. subtilis to elucidate changes during the aging period. The results obtained are as follows ; The soybean paste made with conventional meiu and Asp. oryzae meju showed higher content of amino nitrogen than those of B, natto and B. subtilis meju. Soybean paste made with conventional meju contained a little more content of total and reducing sugars than other soybean pastes. ph during aging period was higher than 5.0 for the Asp. oryzae paste while less than 4.5 for B. subtilis paste. Aspartic acid. threonine, serine, glutamic acid, glycine, alanine, cystine, valine, methionne, leucine and histidine for Asp. oryzae paste ; tyrosine, arginine and proline for conventional meju paste; and isoleucine and phenylalanine for B. subtilis paste were found to be peak amount 90 days after the preparation. The content of total free amino acid was high in the order of Asp. oryzae paste, conventional paste, B. natto paste and B. subtilis paste.

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Optimization of \beta-mammanase Production from Bacillus subtilis JS-1. (\beta-Mannanase를 생산하는 Bacillus subtilis JS-1의 분리 및 효소 생산성)

  • 임지수;정진우;이종수;강대경;강하근
    • Microbiology and Biotechnology Letters
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    • v.31 no.1
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    • pp.57-62
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    • 2003
  • A bacteria strain producing extracellular $\beta$-mannanase was isolated from soil and was identified as Bacillus subtilis by 16S rRNA sequence comparison and biochemical determinations. The optimum pH and temperature for the $\beta$-mannanase activity were 5.0 and 5.5$^{\circ}C$, respectively. The zymogram technique revealed a single protein band exhibiting $\beta$-mannanase activity from the culture supernatant. The molecular mass of the enzyme was estimated at approximately 130 kDa. The addition of 0.5% lactose or 0.5% locust bean gum to the LB medium caused to Increase significantly the $\beta$-mannanase productivity from Bacillus subtilis JS-1. The cells grown on LB medium supplemented with lactose produced maximal enzyme activity at the stationary phase. In contrast to this, the $\beta$-mannanase was induced at the logarithmic phase from the cells grown on LB medium supplemented with locust bean gum. The discrepancy in induction times suggests that $\beta$-mannanase was induced by different induction mechanisms depending on the carbon sources in Bacillus subtilis JS-1 .

Optimum Condition for Pigment Production and Antioxidative Activity of the Products by Bacillus subtilis DC-2 with Response Surface Methodology (반응표면 분석에 의한 Bacillus subtilis DC-2의 색소생성 및 그 생성물의 항산화성에 대한 최적조건)

  • 최웅규;지원대;정현채;최동환;정영건
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.4
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    • pp.620-624
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    • 1997
  • The conditions for color intensity and electron donating ability to $\alpha$,$\alpha$-diphenyl-$\beta$- picryl-hydrazyl (DPPH) of Bacillus subtilis DC-2 were investigated. Temperature, pH and cultivation time were chosen as three factors, and the optimal conditions of color intensity and DPPH was determined with response surface methodology. Color intensity was affected by cultivation temperature(p<0.1). DPPH was influenced by cultivation temperature(p<0.05) and pH(p<0.1). But cultivation time was affected neither color in- tensity nor DPPH. Optimal conditions of color intensity with Bacillus subtilis DC-2 were appeared at cultivation temperature of 39.$25^{\circ}C$, pH 8.83 and cultivation time of 84.41hrs. Optimal conditions of DPPH with Bacillus subtilis DC-2 were revealed at cultivation temperature of 39.19$^{\circ}C$, pH 8.84 and cultivation time of 82.21hrs.

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Antioxidant Activity of Soybean Yogurt added Tomato Extract by Bacillus subtilis and Lactobacillus plantarum (Bacillus subtilis와 Lactobacillus plantarum에 의한 토마토 첨가 두유 요구르트의 항산화 활성)

  • Yang, Ming;Kwak, Jung Soon;Jang, Seri;Jia, Yuan;Park, Inshik
    • The Korean Journal of Food And Nutrition
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    • v.26 no.2
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    • pp.280-286
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    • 2013
  • This study was performed to investigate the effect of types of microorganisms on the antioxidant activity of soybean yogurt by a single or mixed culture of Bacillus subtilis and Lactobacillus plantarum isolated from chunggukjang and kimchi. The fermented soybean milk by Bacillus subtilis exhibited the highest values of total polyphenol, DPPH radical scavenging activity, reducing power and nitrite scavenging activity compared to those of Lactobacillus plantarum or mixed culture of Bacillus subtilis/Lactobacillus plantarum. As the amount of tomato extract was added to the soybean milk, various antioxidant parameters, such as total polyphenol, DPPH radical scavenging activity, reducing power and nitrite scavenging activity, were linearly increased.

Comparison of Characteristics of Koji Manufactured with Bacillus subtilis B-4 and Aspergillus oryzae F-5 (Bacillus subtilis B-4와 Aspergillus oryzae F-5로 제조한 코오지의 특성 비교)

  • Kwon, Dong-Jin
    • Korean Journal of Food Science and Technology
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    • v.34 no.5
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    • pp.873-878
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    • 2002
  • In order to industrialize traditional Meju-tasting Koji, the characteristics of Koji manufactured with bacteria and fungi found in traditional Meju were investigated. Bacillus subtilis B-4 and Aspergillus oryzae F-5 showing high enzyme activities including those of amylase and protease were used. L-value of Koji manufactured with B. subtilis B-4 had darker color and higher enzyme production than A. oryzae F-5 made one. B. subtilis B-4 made Koji showed higher enzyme production and sensony evaluation score than A. oryzae F-5 Koji. A. oryzae F-5 Koji showed superior color to B. subtilis B-4 Koji. Activity in color, capacity of enzyme production, viable cell count, and sensory evaluation of water activity controlled Koji was superior to the uncontrolled one.

Bacillus subtilis as a Tool for Screening Soil Metagenomic Libraries for Antimicrobial Activities

  • Biver, Sophie;Steels, Sebastien;Portetelle, Daniel;Vandenbol, Micheline
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.850-855
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    • 2013
  • Finding new antimicrobial activities by functional metagenomics has been shown to depend on the heterologous host used to express the foreign DNA. Therefore, efforts are devoted to developing new tools for constructing metagenomic libraries in shuttle vectors replicatable in phylogenetically distinct hosts. Here we evaluated the use of the Escherichia coli-Bacillus subtilis shuttle vector pHT01 to construct a forest-soil metagenomic library. This library was screened in both hosts for antimicrobial activities against four opportunistic bacteria: Proteus vulgaris, Bacillus cereus, Staphylococcus epidermidis, and Micrococcus luteus. A new antibacterial activity against B. cereus was found upon screening in B. subtilis. The new antimicrobial agent, sensitive to proteinase K, was not active when the corresponding DNA fragment was expressed in E. coli. Our results validate the use of pHT01 as a shuttle vector and B. subtilis as a host to isolate new activities by functional metagenomics.