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Role of two amino acid residues’ insertion on thermal stability of thermophilic α-amylase AMY121 from a deep sea bacterium Bacillus sp. SCSIO 15121

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Abstract

α-Amylases from Bacillus licheniformis (BLA) and Bacillus amyloliquefaciens (BAA) are both important industrial enzymes with high similarity in structure but significant differences in thermostability. The mechanisms underlying this discrepancy are still poorly understood. Here, we investigated the role of two amino acids’ insertion on the thermostability of these two group amylases. A newly obtained thermophilic amylase AMY121 was found much closer to BLA in both primary structure and enzymological properties. Two amino acids’ insertion widespread among BAA group α-amylases was identified as one of the key factors leading to the thermostability differences, since thermostability of insertion mutants (AMY121-EG and AMY121-AA) from AMY121 significantly decreased, while that of deletion mutant from BAA increased. Moreover, we proposed that conformational disturbance caused by insertion mutation might weaken the calcium-binding affinity and consequently decrease the enzyme thermostability.

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Acknowledgments

This research was supported by Administration of Ocean and Fisheries of Guangdong Province (GD2012-D01-002), National High Technology Research and Development Program of China (863 Program, 2012AA092104), Public Science and Technology Research Funds Projects of Ocean (Grant: 201305018), the National Natural Science Foundation of China (Grant NO. 41406193), “Strategic Priority Research Program” of the Chinese Academy of Sciences, Grant NO. XDA10030400, the Knowledge Innovation Program of the Chinese Academy of Sciences (SQ201301) and the Key Research Program of the Chinese Academy of Sciences, Grant NO.KSCX2-EW-B-13.

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Authors and Affiliations

  1. CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, Guangdong Key Laboratory of Marine Materia Medica, RNAM Center for Marine Microbiology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People’s Republic of China

    Lizhen Li, Jian Yang, Jie Li, Lijuan Long, Yunzhu Xiao, Xinpeng Tian, Fazuo Wang & Si Zhang

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

    Lizhen Li & Yunzhu Xiao

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  1. Lizhen Li
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  2. Jian Yang
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  3. Jie Li
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  8. Si Zhang
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Correspondence to Si Zhang.

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Lizhen Li and Jian Yang contributed equally to this work.

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Li, L., Yang, J., Li, J. et al. Role of two amino acid residues’ insertion on thermal stability of thermophilic α-amylase AMY121 from a deep sea bacterium Bacillus sp. SCSIO 15121. Bioprocess Biosyst Eng 38, 871–879 (2015). https://doi.org/10.1007/s00449-014-1330-2

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