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Biotechnology and bioprocess engineering v.14 no.3, 2009년, pp.266 - 273  
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Construction and Characterization of Novel Chimeric $\beta$-glucosidases with Cellvibrio gilvus (CG) and Thermotoga maritima (TM) by Overlapping PCR

Kim, Jong-Deog    (Department of Biotechnology, Research Center on Anti-Obesity and Health Care (RCAOHC), Chonnam National University   ); Seo, Hyo-Jin    (Department of Biotechnology, Research Center on Anti-Obesity and Health Care (RCAOHC), Chonnam National University   ); Kiyoshi, Hayashi    (National Food Research Institute  );
  • 초록

    In order to create novel $\beta$ -glucosidase constructs, 8 kinds of chimeric $\beta$ -glucosidases were constructed using overlapping polymerase chain reaction (PCR) based on Celvibrio gilvus (CG) and Thermotoga maritima (TM) genes. Two kinds of novel chimeric $\beta$ -glucosidases (No. 6 and No. 8 type) were selected and their properties characterized. SDS-PAGE analysis showed that both constructs had a molecular mass of 80 kDa. The optimum pH of No. 6 chimeric $\beta$ -glucosidase was found to be 3.0 and 5.0, showing varying maximum activity according to the buffer used. No. 8 chimeric enzyme was found to be optimally active at a pH of 4.5 and the optimum temperature of No.6 and No.8 chimeric $\beta$ -glucosidases was reported to be $60^{\circ}C$ , respectively. The Km values of both novel chimeric enzymes were calculated to be 0.012 mM and 0.0082 mM, respectively and the characteristics of the novel chimeric enzymes were to lie between those of the parental enzymes.


  • 주제어

    Chimeric $\beta$-glucosidase .   overlapping PCR .   Cellvibrio gilvus (CG) .   Thermotoga maritima (TM) .   kinetic parameter.  

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