Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

Abstract

Abstract Astaxanthin is a value-added ketocarotenoid with great potential in nutraceutical and pharmaceutical industries. Genetic engineering of heterologous hosts for astaxanthin production has attracted great attention. In this study, we assessed some key factors, including codon usage of the expressed genes, types of promoters, bacterial strains, and culture media, for engineered Escherichia coli to produce astaxanthin. The effect of codon usage was shown to be related to the types of promoters. E. coli DH5α was superior to other strains for astaxanthin production. Different culture media greatly affected the contents and yields of astaxanthin in engineered E. coli. When the expression cassette containing GadE promoter and its driving genes, HpCHY and CrBKT, was inserted into the plasmid pACCAR16ΔcrtX and expressed in E. coli DH5α, the engineered strain was able to produce 4.30 ± 0.28 mg/g dry cell weight (DCW) or 24.16 ± 2.03 mg/L of astaxanthin, which was a sevenfold or 40-fold increase over the initial production of 0.62 ± 0.03 mg/g DCW or 0.61 ± 0.05 mg/L. Graphical Abstract

Keywords

• Astaxanthin
• Escherichia coli
• Bacterial strains
• Codon usage
• Medium