Codon optimization, promoter and expression system selection that achieved high-level production of Yarrowia lipolytica lipase in Pichia pastoris

Enzyme and Microbial Technology
Wen-Jing ZhouLi-Hong Miao

Abstract

Lipase (EC 3.1.1.3) stands amongst the most important and promising biocatalysts for industrial applications. In this study, in order to realize a high-level expression of the Yarrowia lipolytica lipase gene in Pichia pastoris, we optimized the codon of LIP2 by de novo gene design and synthesis, which significantly improved the lipase expression when compared to the native lip2 gene. We also comparatively analyzed the effects of the promoter types (PAOX1 and PFLD1) and the Pichia expression systems, including the newly developed PichiaPink system, on lipase production and obtained the optimal recombinants. Bench-top scale fermentation studies indicated that the recombinant carrying the codon-optimized lipase gene syn-lip under the control of promoter PAOX1 has a significantly higher lipase production capacity in the fermenter than other types of recombinants. After undergoing methanol inducible expression for 96h, the wet cell weight of Pichia, the lipase activity and the protein content in the fermentation broth reached their highest values of 262g/L, 38,500U/mL and 2.82g/L, respectively. This study has not only greatly facilitated the bioapplication of lipase in industrial fields but the strategies utilized, such as de novo g...Continue Reading

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Citations

Apr 28, 2017·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Juan C VilladaWendel Batista da Silveira
Apr 8, 2021·Journal, Genetic Engineering & Biotechnology·Selfela Restu AdinaEsti Puspitasari
May 25, 2021·Journal of Microbiology and Biotechnology·Han Byeol Jeong, Hyung Kwoun Kim
Sep 14, 2021·Frontiers in Bioengineering and Biotechnology·Ting MiaoWei Jiang

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