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Characterization of cold adapted and ethanol tolerant β-glucosidase from Bacillus cellulosilyticus and its application for directed hydrolysis of cellobiose to ethanol

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논문

Characterization of cold adapted and ethanol tolerant β-glucosidase from Bacillus cellulosilyticus and its application for directed hydrolysis of cellobiose to ethanol

학술지

International journal of biological macromolecules

저자명

Wu, Jian; Geng, Alei; Xie, Rongrong; Wang, Hongcheng; Sun, Jianzhong

초록

<P><B>Abstract</B></P> <P>A &beta;-glucosidase gene from <I>Bacillus cellulosilyticus</I> was expressed in <I>Escherichia coli</I> BL21 and characterized. The recombinant enzyme (BcBgl1A) showed the optimal activity at 40&deg;C, and the half-life of BcBgl1A activity at 40&deg;C was 24h. BcBgl1A was sensitive to heat and exhibited obvious cold-adapted activity, retained 27.2% and 46.5% of the optimal activity at 4 and 20&deg;C, respectively. The best activity was observed at pH 7.0 and stable over the range of pH 6.0&ndash;8.0. The activity was significantly enhanced by Fe<SUP>2+</SUP>, Fe<SUP>3+</SUP> and Mn<SUP>2+</SUP>, also showed a high level of NaCl tolerance. BcBgl1A exhibited high activity using both pNPG and cellobiose as substrates, the <I>V</I> <SUB>max</SUB> for pNPG and cellobiose was 66.2 and 151.5U/mg of protein, respectively, and its <I>K</I> <SUB>m</SUB> values were 2.97 and 10.4mM, respectively. Catalysis efficiency (<I>K</I> <SUB>cat</SUB>/<I>K</I> <SUB>m</SUB>) of 45.8s<SUP>&minus;1</SUP> (cellobiose) and 70.3s<SUP>&minus;1</SUP> (pNPG). It displayed high tolerance to glucose and ethanol, the <I>K<SUB>i</SUB> </I> and IC50 value was 200mM and 15% (v/v), respectively. Additionally, the BcBgl1A was found to be highly efficient for conversion of cellobiose (100g/L) to ethanol by simultaneous saccharification and fermentation process, achieved an ethanol content of 47.2g/l corresponding to 87.9% of the theoretical ethanol yield. The results suggest that BcBgl1A has a potential for biotechnological applications in the bioconversion of lignocellulosic materials.</P>

발행연도

2018

발행기관

Elsevier

ISSN

0141-8130

ISSN

1879-0003

109

페이지

pp.872-879

주제어

Bacillus cellulosilyticus; β-glucosidase; Ethanol and salt tolerant; Cold adapted; Cellobiose hydrolysis

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1 2023-12-11

논문; 2018-04-01

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