Abstract: This disclosure belongs to the field of genetic engineering technology, and in particular relates to a Zymomonas mobilis chassis cell, a construction method therefor and use thereof. The chassis cell uses a recombinant Zymomonas mobilis strain with the Cas12a gene integrated into a genome as a starting strain, and integrates the RecET gene into a genome of the recombinant Zymomonas mobilis strain to obtain a chassis cell with improved efficiency of large-fragment gene editing. The present disclosure provides an efficient editing method for directional editing of large fragments of the Zymomonas mobilis genome, which is suitable for the construction of chassis cells requiring frequent gene editing, and greatly reduces the time of gene editing of chassis cells; moreover, the genome of the chassis cell is greatly simplified, which improves the biosynthesis efficiency of biological components and pathways and demonstrates good industrial application value.

Abstract: A genetically engineered strain of Z. mobilis and uses thereof are provided. The genetically engineered strain named ZMNP is obtained by knockouts of four endogenous plasmids of pZM32, pZM32, pZM36, pZM33, and pZM39 from strain ZM4-Cas12a. ZMNP contains a reduced genome and has a series of excellent traits, such as high transformation efficiency, enhanced tolerance to inhibitors, improved capability to use secondary mother liquor and other excellent performance. ZMNP can be used as a chassis cell to construct diverse cell factories for biochemical production using different feedstocks especially the lignocellulosic biomass.