Optimization Strategy of Specific Knockout of Key POX3 Gene in Γ-Decalactone Metabolism of Yarrowia Lipolytica

College of Animal Science and Technology, Sichuan Agricultural University, Chengdu 625014, China

^* Corresponding author: zhangyudie@stu.sicau.edu.cn

Abstract

As an important spice raw material, γ-decalactone is widely used in the food industry. Six peroxisome acyl-CoA oxidases (POX) are rate-limiting enzymes for γ-decalactone metabolism in Yarrowia lipolytica. However, there are many unsolved problems in the function of each member of this family, which limits the strain optimization and the industrial production capacity efficiency of γ-decalactone. In this study, based on the conservation analysis of the ORF and Flanking sequences of POX1-POX6, the gene knockout verification primers with high specificity were designed based on the Cre / LoxP system. Based on the CRISPR / Cas9 system, two target sites with high specificity were screened to achieve specific knockout of the POX3 gene which is in Yarrowia lipolytica, aiming to explore the function of POX3 and subsequently refine the production process of γ-decalactone. The results of this study provide a new genetic engineering design idea for the microbial production capacity of γ-decalactone, deeply reveal the function of the POX gene family, help optimize the production efficiency of γ-decalactone, and lay a theoretical foundation for industrial application.