The invention discloses recombinant 
bacteria used for producing beta-
carotene and a construction method utilizing a 
Crispr-Cas9 technology. A Ku70 
gene is subjected to knockout from 
yarrowia lipolytica, and then phytoene synthase / 
lycopene cyclase (carRA), phytoene desaturase (carB), 
geranyl pyrophosphate synthetase (GGS1) and a 3-hydroxy-3-methyl 
glutaryl coenzyme A reductase (tHMG) 
gene take snfas a target point and are integrated in a 
yarrowia lipolytica 
gene set after knockout of the ku70 gene, wherein the phytoene synthase / 
lycopene cyclase (carRA) and phytoene desaturase (carB) are from 
blakeslea trispora, and the 
geranyl pyrophosphate synthetase (GGS1) and the 3-hydroxy-3-methyl 
glutaryl coenzyme A reductase (tHMG) gene are from the 
yarrowia lipolytica; the 
Crispr-Cas9 technology isutilized for regulating the copy number of the carRA, the carRA, the GGS1 and the tHMG, and the recombinant 
bacteria capable of producing the high-yield beta-
carotene is constructed. After the recombinant 
bacteria is fermented, cultured, extracted and separated, the content of the beta-
carotene can reach the 
dry cell weight of 35 mg / g, and the 
bacterial strain stability is high.