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Cloning, Expression, Sequence Analysis, and Partial Characterization of Two Alkaline β-1, 4-endoglucanases of Phaeosphaeria sp. LH21 from Deep-Sea Mud

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Abstract

Here we cloned and expressed two alkaline β-1, 4-endoglucanases of Phaeosphaeria sp. LH21 from deep-sea mud. The two enzymes shared 71 and 63 % of identities with their known β-1, 4-endoglucanases, respectively. According to the primary and spatial structures, the potential active sites of one of the two enzymes could be Asp122 and Asp11, while the other enzyme could be Asp16. The enzymatic properties of their recombinant enzymes from Pichia pastoris GS115 showed that they were optimally active at pH 8 and 60–65 °C, exhibited >90 % residual relative activities at pH 3–10, and obtained relative activities >75 % at pH 5–10.

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Acknowledgments

This research was supported by the National Basic Research Program of China (973, Program NO. 2012CB721103), National High Technology Research and Development Program of China (863, Program NO. 2012AA022206), National Natural Science Foundation of China (31360217), and Fundamental Research Funds for the Central Universities.

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Authors and Affiliations

  1. State key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China

    Xi-Hua Zhao, Dong-Zhi Wei & Wei Wang

  2. College of Life Science, Jiangxi Normal University, Nanchang, 330022, China

    Xi-Hua Zhao

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  1. Xi-Hua Zhao
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  2. Dong-Zhi Wei
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  3. Wei Wang
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Correspondence to Dong-Zhi Wei or Wei Wang.

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Zhao, XH., Wei, DZ. & Wang, W. Cloning, Expression, Sequence Analysis, and Partial Characterization of Two Alkaline β-1, 4-endoglucanases of Phaeosphaeria sp. LH21 from Deep-Sea Mud. Appl Biochem Biotechnol 173, 1295–1302 (2014). https://doi.org/10.1007/s12010-014-0924-0

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  • DOI: https://doi.org/10.1007/s12010-014-0924-0

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