Genetic manipulation of Rhodobacter sphaeroides for improved production of coenzyme Q10

Abstract:

[Objective] The objective of this study was to improve the production of coenzyme Q10 of Rhodobacter sphaeroides by replacing crtB (encoding phytoene synthase) with ubiC and ubiA (encoding chorismate pyruvate-lyase and 4-hydroxybenzoate: polyprenyldiphosphate 3-polyprenyltransferase, respectively) from Escherochia coli DH5α. [Methods] The plasmid for gene replacement was constructed with a suicide pSUP202 as vector, and its insertions, including the up and down streams of crtB in R. sphaerodies 2.4.1, spectinomycin resistance gene, ubiC and ubiA from E. coli DH5α were obtained by PCR. RT-PCR was used to detect the transcription of genes. HPLC was used to quantify the production of coenzyme Q10. [Results] The gene replacement mutant of R. sphaerodies was constructed, in which the crtB was replaced by ubiC and ubiA. The transcription of heterologous genes was confirmed by RT-PCR. The productivity of gene engineered strain was 1.6 fold of wide type. [Conclusion] The strain of R. sphaerodies with improved production of coenzyme Q10 was obtained successfully, and the ubiC and ubiA from E. coli could transcript with its native promoter in R. sphaerodies.