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Optimal secretion of alkali-tolerant xylanase in Bacillus subtilis by signal peptide screening

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Abstract

Xylanases are industrially important enzymes for xylan digestion. We experimentally screened over 114 Sec and 24 Tat pathway signal peptides, with two different promoters, for optimal production of an alkaline active xylanase (XynBYG) from Bacillus pumilus BYG in a Bacillus subtilis host. Though both promoters yielded highly consistent secretion levels (0.97 Pearson correlation coefficient), the Sec pathway was found to be more efficient than the Tat pathway for XynBYG secretion. Furthermore, the optimal signal peptide (phoB) for XynBYG secretion was found to be different from the optimal peptides for cutinase and esterase reported in previous studies. A partial least squares regression analysis further identified several statistically important variables: helical properties, amino acid composition bias, and the discrimination score in Signal P. These variables explain the observed 23 % variance in the secretion yield of XynBYG by the different signal peptides. The results also suggest that the helical propensity of a signal peptide plays a significant role in the beta-rich xylanase, but not in the helix-rich cutinase, suggesting a coupling of the conformations between the signal peptide and its cargo protein for optimal secretion.

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Acknowledgments

This study was financially supported by the National Science Foundation of China (31372343 and 3137228). This work was also supported by funding from an innovation project of science and technology plan project of Shaanxi Province, China (2014KTCL02-21), and a Ministry of Agriculture (No. 2013-S16) to X.Z.; National Health and Medical Research Council (1059775 and 1083450) of Australia and Australian Research Council’s Linkage Infrastructure, Equipment and Facilities funding scheme (project number LE150100161) to Y.Z., National Natural Science Foundation of China (61271378) to Y.Y. We also gratefully acknowledge the support of the Griffith University eResearch Services Team and the use of the High Performance Computing Cluster “Gowonda” to complete this research. This research/project has also been undertaken with the aid of the research cloud resources provided by the Queensland Cyber Infrastructure Foundation (QCIF).

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Authors and Affiliations

  1. College of Animal Sciences and Technology, Northwest A&F University, 712100, Yangling, People’s Republic of China

    Weiwei Zhang, Mingming Yang & Xin Zhao

  2. Institute for Glycomics and School of Information and Communication Technology, Griffith University, Parklands Dr., Southport, QLD, 4222, Australia

    Yuedong Yang, Jian Zhan & Yaoqi Zhou

  3. Department of Animal Science, McGill University, Quebec, Canada

    Xin Zhao

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  1. Weiwei Zhang
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Correspondence to Yaoqi Zhou or Xin Zhao.

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Weiwei Zhang and Mingming Yang are co-first authors.

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Zhang, W., Yang, M., Yang, Y. et al. Optimal secretion of alkali-tolerant xylanase in Bacillus subtilis by signal peptide screening. Appl Microbiol Biotechnol 100, 8745–8756 (2016). https://doi.org/10.1007/s00253-016-7615-4

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