• Title/Summary/Keyword: Alkalophilic

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Isolation and Characterization of Cyclodextrin Glycosyl Transferase Producing Alkalophilic Bacillus sp. (Cyclodextrin glycosyltransferase를 생산하는 호알칼리성 Bacillus속 미생물)

  • 유주현;정용준;이정수
    • Microbiology and Biotechnology Letters
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    • v.17 no.2
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    • pp.148-153
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    • 1989
  • A strain of alkalophilic Bacillus sp. YC-335 has been isolated from soil. The strain was capable of producing large amount of cyclodextrin glycosyl transferase (CGTase) in the culture broth. The preferable medium composition has been determined to be as follows : 1.5% soluble starch, 5% corn steep liquor, 0.1% $K_2$HPO$_4$, 0.02%mgSO$_4$.7$H_2O$, 1% CaCO$_3$and 1% Na$_2$CO$_3$(pH 10.3). The highest enzyme production was observed after 48 hours of cultivation at 31$^{\circ}C$. The optimum pH and temperature for the activity of crude enzyme were 6.0 and 5$0^{\circ}C$, respectively. The enzyme was stable between pH 5 and 9, and upto 5$0^{\circ}C$. The enzyme converted starch into $\alpha$-, $\beta$- and ${\gamma}$-CD in the relative amounts of 1:10:1.5, respectively.

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Purification and Characterization of Thermotolerable Alkaline Protease by Alkalophilic Bacillus sp. No. 8-16 (알칼리성 Bacillus sp. No.8-16의 내열ㆍ알칼리성 단백질 분해효소의 정제와 특성)

  • Bae, Moo;Park, Pil-Yon
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.545-551
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    • 1989
  • Thermostable alkaline protease of alkalophilic Bacillus sp. No. 8-16 has been purified, and the properties of the enzyme investigated. The characteristic point of the organism used is especially good growth in alkaline and thermal condition. The alkaline protease of the strain No. 8-16 was purified from crude enzyme by acetone precipitation, CM-cellulose ion exchange chromatography, Sephadex G-100 and Sephadex G-75 gel filtration. Through the series of chromatograpies, the enzyme was purified to homogeneity with specific activity of 37 fold higher than that of the crude broth. Characteristics of the purified enzyme were as follow; $K_m$ value for the enzyme was 1.3 mg/ml, the alkaline protease showed a maximal activity at 7$0^{\circ}C$ and from the pH 6.0 through 12.0, and stable for 1 hr. at 6$0^{\circ}C$. The moleclar weight of the enzyme was estimated to be 33,000 by Sephadex G-100 gel filtration. The activity of the alkaline protease was inhibited by iodoacetic acid and Ag$^+$, Hg$^+$, PMSF (phenylmethylsulfonyl fluoride), and activated by $Ca^{2+}$ and Mn$^{2+}$.

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Production of Intracellular Invertase from Alkalophilic and Thermophilic Bacillus sp. TA-11 in the Recombinant E. coli (재조합 대장균에서 호알칼리성,고온성 Bacillus sp. TA-11의 세포내 Invertase의 생산)

  • Yi, Sung-Hun;Lee, Dae-Hyung;No, Jae-Duck;Lee, Jae-Won;Lee, Jong-Soo
    • Microbiology and Biotechnology Letters
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    • v.34 no.4
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    • pp.318-322
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    • 2006
  • The intracellular invertase gene of alkalophilic and thermophilic Bacillus sp. TA-11 which was isolated from compost was cloned and expressed in E. coil HB101 using pUC19 as a vector. The invertase of the recombinant E. coli (pYC 17) was maximally produced when it was incubated at 37$^{\circ}C$ for 9 h in a SY medium containing 0.25% sucrose, 0.5% yeast extract, 0.1% each of $K_2HPO_4$ and $KH_2PO_4$, with an initial pH of 8.0. The final enzyme activity under the above condition was 47.7 U per ml of cell-free extract.

Mode of Action and Chemical Modification of an Alkaline Xylanase (CX-III) from Alkalophilic Cephalosporium sp. RYM-202 (호알카리성 Cephalosporium sp. RYM-202로부터 분리된 alkaline xylanase (CX-III)의 작용 양상 및 화학적 변환)

  • Kang, Myoung-Kyu;Maeng, Pil-Jae;Rhee, Young-Ha
    • The Korean Journal of Mycology
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    • v.24 no.4 s.79
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    • pp.255-264
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    • 1996
  • The hydrolysis products formed from birchwood xylan by the action of an alkaline xylanase (CX-III) from alkalophilic Cephaloxporium sp. RYM-202 were xylobiose and xylooligosaccharides polymerized with more than 4 sugar molecules. This enzyme was not active on xylobiose but readily attacked xylotriose accumulating xylobiose as a major product. The predominant end-products from xylotetraose by CX-III were xylobiose and xylotriose. These results indicate that the enzyme is typically endo-type xylanase possessing transglycosidase activity. Chemical modification of CX-III with N-bromosuccinimide revealed that two tryptophan residues per molecule of CX-III were essential for its catalytic activity on xylan. On the other hand, iodoacetamide and diethylpyrocarbonate did not influence the activity of the enzyme, suggesting that cysteine and histidine residues are not involved in the active site of this alkaline xylanase.

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Effect of pHs on Morphological and Cultural Characteristics of Alkalophilic Coryneform Bacteria TU-19 (호알칼리성 Coryneform Bacteria TU-19의 형태적, 배양적 특성에 미치는 pH효과)

  • Choi, Myoung-Chul;Yang, Jae-Sub;Hwang, Cher-Won;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.41 no.5
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    • pp.337-341
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    • 1998
  • The morphological and cultural characteristics of alkalophilic Coryneform bacteria TU-19 were investigated at various pHs. This bacterium showed normal growth pattern at $pH\;9.0{\sim}10.0$, but the cell growth was completely inhibited at extreme pH (12.0 or more). Interestingly, at pH 8.0 the morphology of the bacterial cells seems to form convoluted filaments during the exponential growth phase while at pH 10.0, the optimal pH for the growth of this organism, the bacteria grew with variable paired or single forms, and straight rods during growth stages. Growing in alkaline media $(pH\;9.0{\sim}11.0)$, it adjusted the pH of the culture media to around pH 8.5 by itself.

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Purification of Alkaline Restriction Endonuclease from Alkalophilic Bacillus sp. 8-13 (제한효소 생성능을 지닌 알칼리성 Bacillus sp. 8-13 균주로부터 알칼리성 제한효소의 정제)

  • Bae, Moo;Lee, Jee-Eun;Park, Kyoung-Sook;Lee, Kang-Man
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.289-294
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    • 1992
  • Twenty-four bacterial strains among alkalophillic bacteria isolated from soil samples were examined for the presence of type II restriction endonuclease in aerobic culture. One strain was found to contain specific enzyme to cleave lambda DNA. The characteristics of this microorganism is the ability to grow well in alkalophilic and high temperature condition, that is at pH 10.3 and $50^{\circ}C$. This strain was tentatively identified to Bacillus alkaloPhilus subsp. halodurans when morphological, physiological and biochemical characteristics were examined. The enzyme was purified from crude extract by streptomycin sulfate, ammonium sulfate precipitation, which was followed by DEAE-cellulose and phosphocellulose ion exchange column chromatography, and the subunit molecular weight was about, 32,000 daltons by polyacrylamide gel electrophoresis containing 0.1% SDS.

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Isolation and Growth Characteristics of Alkalophilic Bacillus sp. for Removal of Anthraquinone Dye. (Anthraquinone계 염료의 제거를 위한 호알칼리성 Bacillus sp.의 분리와 성장 특성)

  • 김정목
    • Microbiology and Biotechnology Letters
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    • v.29 no.2
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    • pp.67-71
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    • 2001
  • Isolation and Growth Characteristics of AIkalophilic Bacillus sp. for Removal of Anthraquinone Dye. Kim, Jeong-Mog. School of Environmental Information, Taekyeung College, Kyungsan, 712-850, Korea -Alkalophilic strain degrading and decolorizing anthraquinone dye, Remazol brilliant blue R was isolated from natural system and named as Bacillus sp. ARB!. The optimal temperature and pH of Bacillus sp. ARBI were 35°C and 9.0, respectively. The pH of culture media during the fermentation were changed from 10 and 10.5 of initial values to 9.3 and 9.4 after 40 hrs, respectively. Decolorization efficiency in aerobic shaking culture of Bacillus sp. ARBI was markedly higher than that in standing culture. At the optimal culture condition, decolorization efficiency by the Bacillus sp. ARBl was 93% after 32 hrs batch culture. In the case of batch culture using real dye processing wastewater, dye decolorization efficiency of Bacillus sp. ARBl was 78% after 40 hrs.

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Purification of Three Extracellular Proteases from Alkalophilic Coryneform Bacteria TU-19 (호알칼리성 Coryeform bacteria TU-19가 생산하는 세종류의 균체외 단백질분해호소의 정제)

  • Choi, Myoung-Chul;Yang, Jae-Sub;Kang, Sun-Chul
    • Applied Biological Chemistry
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    • v.38 no.6
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    • pp.534-540
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    • 1995
  • Alkalophilic coryneform bacteria TU-19 isolated from soil extracellularly produced at least three proteases (Protease I, II, and III). Investigating the cultural conditions related to the enzyme production of this bacterial cell, the optimum pH and temperature were 10.0 and $30^{\circ}C$, respectively. In order to purify these enzymes from the 2 day culture broth ammonium sulfate fractionation, gel filtration and QAE-Sephadex column chromatography were performed step by step. And then these three proteases were purified to near homogeneity by judging from SDS-PAGE pattern, and had the molecular weights of 120, 80, and 45 kilodaltons, respectively. The optimum pH and temperature for the enzyme activity of Protease I and II were 10.5 and $45^{\circ}C$, respectively, and Protease II were 11.0 and $50^{\circ}C$. And the enzymes were completely inhibited by PMSF suggesting serine protease, but not affected by pCMB. 1,10-phenanthroline, IAA, and EDTA.

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Identification of Alkalophilic Bacillus sp. S-1013 Producing Non-Cariogenicity Sugar Fuc($1{\to}4$)gaINAc($2{\to}6$)NeuAc and Optimization of Culture Condition for Its Production (비우식성 당 Fuc($1{\to}4$)gaINAc($2{\to}6$)NeuAc를 생산하는 호알칼리성 Bacillus sp. S-1013의 동정 및 생산조건의 최적화)

  • Ryu Il-Hwan;Kim Sun-Sook;Lee Kap-Sang;Lee Eun-Sook
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.235-243
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    • 2006
  • The study was performed to identification of producing microbe Non-Cariogenicity Sugar (NCS; Fuc($1{\to}4$) gaINAc($2{\to}6$)NeuAc) with anti-caries activity, and to optimization of production condition. A typical strain which produced the NCS was identified alkalophilic Bacillus sp. S-1013 through the results of morphological, biochemical and chemotaxonomic characteristics and 16S rDNA sequencing. The optimal medium composition for the maximal production of the NCS from alkalophilic Bacillus sp. S-1013 was as follow: soluble starch 30 g, dextrin 15 g, yeast extract 5 g, peptone 10 g, $K_{2}HPO_4$ 2 g in a liter of distilled water. Optimal temperature and pH were 25 and 11.0, respectively. The highest production of NCS was shown 60 hrs cultivation using the optimal medium, and then NCS productivity and dry cell weight of culture broth increased 4.24 and 2.67 time than initial medium, respectively.

Production and Characteristics of Lytic Enzyme against Streptococcus mutans Cell Wall from Alkalophilic Bacillus sp. 4830 (호알카리성 Bacillus sp. 4830이 생산하는 Streptococcus mutans 세포벽 분해효소의 분리와 특성)

  • Kim, Yun-Keun;Bai, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.35 no.6
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    • pp.1143-1149
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    • 2003
  • To elucidate a method of preventing dental caries, strains producing lytic enzymes were isolated and their characteristics were investigated. Among 5,00 alkalophilic strains isolated from soil, 22 strains showed lytic activity against Streptococcus mutans. Strain No. 4830, with the highest lytic activity, was selected for further study. Strain 4830 showed 94% sequence homology with the 16S rDNA sequence of Bacillus alcalophilus, but it was concluded to be different from Bacillus alcalophilus because of its biochemical characteristics. The strain was named Bacillus sp. 4830. The lytic enzyme from Bacillus sp. 4830 was purified by ethanol precipitation and CM-agarose column chromatography. The molecular weight of the lytic enzyme was determined to be 28 kDa by SDS-PAGE. The lytic enzyme was stable between pH 5.0 and pH 11 and up to $40^{\circ}C$. The optimal pH and temperature for the lytic activity was 9.0 and $50^{\circ}C$, respectively.