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Mutation on N-terminus of polyhydroxybutyrate synthase of Ralstonia eutropha enhanced PHB accumulation

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Abstract

Polyhydroxyalkanoate (PHA) synthase is the central enzyme involved in the biosynthesis of PHA, a family of bacterial biodegradable polyesters. Due to its high variability, the N-terminal fragment of this enzyme was previously considered as unnecessary for a functionally active enzyme. In this study, polyhydroxybutyrate synthase from Ralstonia eutropha (PhbCRe) with a deletion on N-terminal 88 amino acid residues showed a significant reduced activity, as reflected by only 1.5% PHB accumulation compared with the wild type which produced 58.4% PHB of the cell dry weight. Whilst several site-specific mutagenesis results revealed the amphiphilic α-helix assembled by the amino acid region, D70–E88 played an important role in both maintaining the PHB synthase activity and regulating molecular weight and polydispersity of accumulated PHB homopolymer.

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Acknowledgements

This research was supported by the National Nature Science Foundation for Distinguished Young Scholar (grant no. 30225001). The State 863 Funds (no. 2002A213051) also contributed financially to this study. We are also in debt to Professor S. Taguchi of Hokkaido University/Japan and Professor B. Rehm of Massey University/New Zealand for their critical comments and suggestions to improve this paper. Their expert views have contributed to the quality of this paper. The plasmid pBHR68 and the host strain R.eutropha PHB−4 were kindly provided by Dr. B. Rehm and Professor A. Steinbüchel of the Westfälische Wilhelm-Univerität Münster, Münster, Germany. We also thank Jing-Yu Chen and Andrew R. Chang for their assistance in the manuscript preparation.

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  1. MOE Laboratory of Protein Science, Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, 100084, China

    Zhong Zheng, Ming Li, Xiao-Jing Xue, Hong-Lei Tian, Zhi Li & Guo-Qiang Chen

  2. Life Sciences Division, Graduate School at Shenzhen, Tsinghua University, Guangdong, 518055, China

    Ming Li

  3. Multidisciplinary Research Center, Shantou University, Guangdong, 515063, China

    Guo-Qiang Chen

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  1. Zhong Zheng
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Correspondence to Guo-Qiang Chen.

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Zhong Zheng and Ming Li contributed equally to this study.

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Zheng, Z., Li, M., Xue, XJ. et al. Mutation on N-terminus of polyhydroxybutyrate synthase of Ralstonia eutropha enhanced PHB accumulation. Appl Microbiol Biotechnol 72, 896–905 (2006). https://doi.org/10.1007/s00253-006-0371-0

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