Abstract
A novel gene (Ba-ega) of Bacillus sp. AC-1, encoding an endoglucanase (Ba-EGA), was cloned and expressed in Escherichia coli. Ba-ega, containing a 1,980-bp open reading frame (ORF), encoded a protein of 659 amino acids and had a molecular mass of 74.87 kDa. Ba-EGA was a modular enzyme composed of a family-9 glycosyl hydrolase catalytic module (CM9) and a family-3 carbohydrate-binding module (CBM3). To investigate the functions of the CBM3 and CM9, a number of truncated derivatives of Ba-EGA were constructed, and all were active. The catalytic module (rCM9) alone was less stable at high temperature than the recombinant Ba-EGA (rBa-EGA). The temperature stability for the complex of rCM9 and rCBM3 was still lower than rBa-EGA, but higher than rCM9 alone. These observations indicated the existence of a non-covalent interaction between CM9 and CBM3 that might strengthen the stability of CM9. However, this interaction is not strong enough to mimic the protective effect of the CBM in the wild-type enzyme.
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Acknowledgment
This work was supported by the grants from the National Natural Science Foundation of China (no. 30370336), the Major State Basic Research Development Program of China (nos. 2003CB716006 and 2004CB719702), and the Creation Foundation from Shanghai Institutes for Life Sciences. Shuang Zhang and Qiu-yu Yin contributed equally to this work.
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Zhang, S., Yin, Qy., Li, Yh. et al. Molecular and biochemical characterization of Ba-EGA, a cellulase secreted by Bacillus sp. AC-1 from Ampullaria crosseans . Appl Microbiol Biotechnol 75, 1327–1334 (2007). https://doi.org/10.1007/s00253-007-0961-5
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DOI: https://doi.org/10.1007/s00253-007-0961-5