Enhanced Production of Astaxanthin by Metabolically Engineered Non-mevalonate Pathway in Escherichia coli

Abstract

Astaxanthin is one of the major carotenoids used in pigment has a great economical value in pharmaceutical markets, feeding, nutraceutical and food industries. This study was to increase the production of astaxanthin by co-expression with transformed Escherichia coli using six genes involved in the non-mevalonate pathway. Involved in the non-mevalonate biosynthetic pathway of the strain Kocuria gwangalliensis were cloned dxs, ispC, ispD, ispE, ispF, ispG, ispH and idi genes in order to increase astaxanthin production from the transformed E. coli. And co-expression with the genes to compared the amount of astaxanthin production. This engineered E. coli, containing both the non-mevalonate pathway gene and the astaxanthin biosynthesis gene cluster, produced astaxanthin at 1,100 µg/g DCW (dry cell weight), resulting in approximately three times the production of astaxanthin.

Biotechnological advances for improving natural pigment production: a state-of-the-art review

“… E. coli Gene screening (four CrtZ and twelve CrtW genes) and gene combination Shake flask 1.99 mg/g DCW Scaife et al ( 2012 ) E. coli Genomic integration ( crtE , crtB , crtI , crtY and crtZ genes from P. ananatis as well as crtW148 gene from N. punctiforme PCC 73102) and promoter engineering Shake flask 1.4 mg/g DCW Lemuth et al ( 2011 ) E. coli Optimization of gene expression via ribosome-binding site combinatorics Shake flask 5.8 mg/g DCW Zelcbuch et al ( 2013 ) E. coli Gene mining of Crt E and Crt Z genes from Sphingomonas sp. ATCC 55669 Shake flask 6.6 mg/g DCW Ma et al ( 2016 ) E. coli Combinatorial expression of different β -carotene ketolase and ketolases Shake flask 7.4 ± 0.3 mg/g DCW Lu et al ( 2017 ) E. coli Pathway engineering- metabolic engineering of DXP pathway by introduction of genes from Kocuria gwangalliensis , as well as introduction of astaxanthin downstream biosynthetic pathway from Paracoccus haeundaensis Shake flask 1100 μg/g DCW Jeong et al ( 2018 ) E. coli Optimization of gene codon, promoters, strain species and culture media Shake flask 4.30 ± 0.23 mg/g DCW 24.16 ± 2.03 mg/L Li and Huang ( ...…”

Spatiotemporal Regulation of Astaxanthin Synthesis in S. cerevisiae

“…Microbial biosynthesis has recently emerged as a promising alternative production route to alga extraction and chemical synthesis. Heterologous production of astaxanthin has been achieved in Escherichia coli, − Saccharomyces cerevisiae, − Yarrowia lipolytica, − Kluyceromyces marxianus, , and Corynebacterium glutamicum, , with enhancing the supply of β-carotene, balancing the hydroxylation and ketolation activities of β-carotene by screening, engineering, and expression adjustment of β-carotene hydroxylase and ketolase, uncovering new engineering targets by random mutagenesis as common strategies. Although extensive efforts have been made, the astaxanthin yields in engineered chassis systems remain relatively low compared to the natural producer Haematococcus pluvialis, which could accumulate astaxanthin up to 4% of dry weight .…”