Production of Astaxanthin Using CBFD1/HFBD1 from Adonis aestivalis and the Isopentenol Utilization Pathway in Escherichia coli

Bioengineering · 2 authors, 1 centre

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TL;DR

This study engineered *E. coli* strains with different isoprenoid pathways to produce the antioxidant astaxanthin, finding that the isopentenol utilization pathway (IUP) dramatically outperformed the native and overexpressed MEP pathways in minimal media, yielding a 13-fold higher total carotenoid titre.

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In complex LB media, the trcMEP strain was most productive, reaching a carotenoid titre of 6.05 ± 0.95 mg/L. In minimal M9 media, the IUP strain was vastly superior, producing 11.3 ± 0.55 mg/L of total carotenoids, a 13-fold increase over the wild-type strain. The work demonstrates a potential method for microbial astaxanthin bioproduction using inexpensive minimal media.

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other

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biomedical research

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ORIGINAL PAPER

PMC10525928

Production of Astaxanthin Using CBFD1/HFBD1 from Adonis aestivalis and the Isopentenol Utilization Pathway in Escherichia coli

Bioengineering

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ABSTRACT

Abstract Astaxanthin is a powerful antioxidant and is used extensively as an animal feed additive and nutraceutical product. Here, we report the use of the β-carotene hydroxylase (CBFD1) and the β-carotene ketolase (HBFD1) from Adonis aestivalis , a flowering plant, to produce astaxanthin in E. coli equipped with the P. agglomerans β-carotene pathway and an over-expressed 4-methylerythritol-phosphate (MEP) pathway or the isopentenol utilization pathway (IUP). Introduction of the over-expressed MEP pathway and the IUP resulted in a 3.2-fold higher carotenoid content in LB media at 36 h post-induction compared to the strain containing only the endogenous MEP. However, in M9 minimal media, the IUP pathway dramatically outperformed the over-expressed MEP pathway with an 11-fold increase in total carotenoids produced. The final construct split the large operon into two smaller operons, both with a T7 promoter. This resulted in slightly lower productivity (70.0 ± 8.1 µg/g·h vs. 53.5 ± 3.8 µg/g·h) compared to the original constructs but resulted in the highest proportion of astaxanthin in the extracted carotenoids (73.5 ± 0.2%). Keywords: carotenoids, hydrophobic products, Adonis aestivalis , Escherichia coli , isopentenol utilization pathway (IUP)

CITATION EXPORT

Jared H Roth, Valerie C A Ward. Production of Astaxanthin Using CBFD1/HFBD1 from Adonis aestivalis and the Isopentenol Utilization Pathway in Escherichia coli. Bioengineering. (2023). https://doi.org/10.3390/bioengineering10091033

AUTHORS

2 · 1 CENTRES

Jared H Roth, Valerie C A Ward

1Department of Chemical Engineering, University of Waterloo, Waterloo, ON N2L 3G1, Canada