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Characterization of a thermostable alkaline phytase from Bacillus licheniformis ZJ-6 in Pichia pastoris

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Abstract

A novel neutral phytase gene (phyC) from Bacillus licheniformis was cloned and expressed in Pichia pastoris under the control of AOX1 promoter. The gene is 1,146 bp in size and encodes a polypeptide of 381 amino acids. The recombinant PhyCm (rePhyCm), driven by the Saccharomyces cerevisiae α-mating factor, was secreted into culture medium. After 0.5% methanol induction for 96 h, the activity of rePhyCm in culture supernatant reached 0.23 U/ml. The optimum temperature and pH of purified rePhyCm were 60°C and 7.5, respectively. The rePhyCm was stable in a wide pH range of 5.0–9.0, especially for alkaline pH. The residual activities of rePhyCm retained over 80% after being incubated at pH 5.0–9.0, 37°C for 1 h in the presence of 1 mM CaCl2. Interestingly, supplemental Ca2+ upgraded both the thermostability and pH stability of rePhyCm. Substrate specificity of rePhyCm, effects of metal ions and chemicals on phytase activity were also investigated in current study.

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Acknowledgments

This work was supported by the National High-Tech Research and Development Plan (2007AA100601), and was also supported by the Science and Technology Department of Zhejiang Province of China (2006C12036) and the Shandong Science and Technology Development Project of People’s Republic of China (2009GG10009001). The authors gratefully acknowledge Miss Shirley for her useful comments on this work and critical reading of the manuscript.

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Authors and Affiliations

  1. Microbiology Division, Institute of Feed Science, College of Animal Science, Zhejiang University, 310029, Hangzhou, China

    Qian Wang, Shi-Jun Fu & Jian-Yi Sun

  2. College of Life Science, Zhejiang University, 310058, Hangzhou, China

    Xiao-Yan Weng

  3. Key Laboratory of Preventive Veterinary Medicine and Animal Biotechnology, Shandong Binzhou Animal Science and Veterinary Medicine Academy, 256600, Binzhou, China

    Shi-Jun Fu

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  1. Qian Wang
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Correspondence to Xiao-Yan Weng.

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Wang, Q., Fu, SJ., Sun, JY. et al. Characterization of a thermostable alkaline phytase from Bacillus licheniformis ZJ-6 in Pichia pastoris . World J Microbiol Biotechnol 27, 1247–1253 (2011). https://doi.org/10.1007/s11274-010-0574-5

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  • DOI: https://doi.org/10.1007/s11274-010-0574-5

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