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Functional role of β domain in the Thermoanaerobacter tengcongensis glucoamylase

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Abstract

Thermoanaerobacter tengcongensis MB4 glucoamylase (TteGA) contains a catalytic domain (CD), which is structurally similar to eukaryotic GA, and a β domain (BD) with ambiguous function. Firstly, BD is found to be essential to TteGA activity because CD alone could not hydrolyze soluble starch. However, starch hydrolysis activity, similar to that of intact TteGA, was restored to CD in the presence of BD. Secondly, BD is found to be an important helper in the correct folding of CD because CD was mainly expressed in the inclusion bodies on its own in Escherichia coli. By contrast, intact TteGA, BD, and CD combined with BD could be expressed as soluble proteins. Additionally, BD is essential to the thermostability of TteGA because CD displayed lower thermostability compared with the intact TteGA and exhibited enhanced thermostability in the presence of BD in vitro. Truncation of TteGA or mutagenesis of the residues that participate in the interdomain interaction at its BD also led to the reduced thermostability of TteGA.

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Acknowledgments

This work was supported by the National High Technology Research and Development Program of China (863 Program, 2012AA022203), and the National Basic Research Program (973 Program, 2011CBA00800). Q.W. is supported by the Bairenjihhua Program of the Chinese Academy of Sciences. The authors are grateful to Prof. Keqian Yang for comments and discussions of this paper, and Dr. Ence Yang for critically reading this paper.

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Authors and Affiliations

  1. Department of Industrial Microbiology and Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Zilong Li, Pingying Wei, Peng He & Ning Jiang

  2. Graduate University, Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Zilong Li & Pingying Wei

  3. The State Key Laboratory of Microbial Metabolism and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200030, People’s Republic of China

    Hairong Cheng

  4. Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, Tianjin, 300308, China

    Qinhong Wang

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  1. Zilong Li
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Correspondence to Ning Jiang.

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Li, Z., Wei, P., Cheng, H. et al. Functional role of β domain in the Thermoanaerobacter tengcongensis glucoamylase. Appl Microbiol Biotechnol 98, 2091–2099 (2014). https://doi.org/10.1007/s00253-013-5051-2

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