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Site-saturation mutagenesis of mutant l-asparaginase II signal peptide hydrophobic region for improved excretion of cyclodextrin glucanotransferase

  • Genetics and Molecular Biology of Industrial Organisms - Original Paper
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

The excretion of cyclodextrin glucanotransferase (CGTase) into the culture medium offers significant advantages over cytoplasmic expression. However, the limitation of Escherichia coli is its inability to excrete high amount of CGTase outside the cells. In this study, modification of the hydrophobic region of the N1R3 signal peptide using site-saturation mutagenesis improved the excretion of CGTase. Signal peptide mutants designated M9F, V10L and A15Y enhanced the excretion of CGTase three-fold and demonstrated two-fold higher secretion rate than the wild type. However, high secretion rate of these mutants was non-productive for recombinant protein production because it caused up to a seven-fold increase in cell death compared to the wild type. Our results indicated that the excretion of CGTase is highly dependent on hydrophobicity, secondary conformation and the type and position of amino acids at the region boundary and core segment of the h-region.

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Acknowledgements

This research was supported by the Universiti Teknologi Malaysia (UTM) under the project “Enhance Extracellular Production of Recombinant Cyclodextrin Glucanotransferase (CGTase) by Random Mutagenesis of Signal Peptide and Optimization of Medium Additives” (Grant No.: Q.J130000.7135.00H76).

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  1. Department of Bioprocess and Polymer Engineering, Faculty of Chemical and Energy Engineering, Universiti Teknologi Malaysia, 81310, Skudai, Johor, Malaysia

    Abbas Ismail & Rosli Md. Illias

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Correspondence to Rosli Md. Illias.

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Ismail, A., Illias, R.M. Site-saturation mutagenesis of mutant l-asparaginase II signal peptide hydrophobic region for improved excretion of cyclodextrin glucanotransferase. J Ind Microbiol Biotechnol 44, 1627–1641 (2017). https://doi.org/10.1007/s10295-017-1980-6

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