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High-level secretive expression of a novel achieved Talaromyces cellulolyticus endo-polygalacturonase in Pichia pastoris by improving gene dosage for hydrolysis of natural pectin

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Abstract

Pectin is a type of complex hydrophilic polysaccharide widely distributed in plant resources. Thermal stable pectinase has its advantage in bioapplication in the fields of food processing, brewing, and papermaking, etc. In this study, we enzymatically characterized a putative endo-polygalacturonase TcPG from a Talaromyces cellulolyticus, realized its high-level expression in Pichia pastoris by in vitro constructing of a series of multi-copy expression cassettes and real time quantitative PCR screening. The secretive expression level of TcPG was nonlinear correlated to the gene dosage. Recombinants with five-copy TcPG gene in the host genome showed the highest expression. After cultivation in a bioreactor for about 96 h, the enzyme activity reached 7124.8 U/mL culture. TcPG has its optimal temperature of 70 °C. Under the optimized parameters, the pectin could be efficiently hydrolyzed into oligosaccharides.

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Acknowledgements

We thank Dr. JK YANG for her help with the conduct of the experiments. This work was financially supported by the project (No. DY135-B2-06) from China Ocean Mineral Resources R & D Association.

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  1. College of Biology and Pharmaceutical Engineering, Wuhan Polytechnic University, Wuhan, 430023, China

    Xiao-Bo Peng, Guang-Jun Chen, Zhen-Gang Han & Jiang-Ke Yang

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  1. Xiao-Bo Peng
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  2. Guang-Jun Chen
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  3. Zhen-Gang Han
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Correspondence to Jiang-Ke Yang.

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Peng, XB., Chen, GJ., Han, ZG. et al. High-level secretive expression of a novel achieved Talaromyces cellulolyticus endo-polygalacturonase in Pichia pastoris by improving gene dosage for hydrolysis of natural pectin. World J Microbiol Biotechnol 35, 84 (2019). https://doi.org/10.1007/s11274-019-2657-2

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