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Appl Microbiol Biotechnol. 2019 Dec;103(23-24):9393-9399. doi: 10.1007/s00253-019-10182-w. Epub 2019 Oct 31.

Pathway engineering for efficient biosynthesis of violaxanthin in Escherichia coli.

Author information

1
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa, 921-8836, Japan. mtake@ishikawa-pu.ac.jp.
2
Institute of Health Sciences, Ezaki Glico Co., Ltd., Osaka, 555-8502, Japan.
3
Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa, 921-8836, Japan.
4
Division of Food Function and Chemistry, Research Institute for Production Development, Kyoto, 606-0805, Japan.
5
Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology (AIST), Tsukuba, 305-8566, Japan.
6
Department of Applied Chemistry and Biotechnology, Chiba University, Chiba, 263-8522, Japan.

Abstract

Carotenoids are naturally synthesized in some species of bacteria, archaea, and fungi (including yeasts) as well as all photosynthetic organisms. Escherichia coli has been the most popular bacterial host for the heterologous production of a variety of carotenoids, including even xanthophylls unique to photosynthetic eukaryotes such as lutein, antheraxanthin, and violaxanthin. However, conversion efficiency of these epoxy-xanthophylls (antheraxanthin and violaxanthin) from zeaxanthin remained substantially low. We here examined several factors affecting their productivity in E. coli. Two sorts of plasmids were introduced into the bacterial host, i.e., a plasmid to produce zeaxanthin due to the presence of the Pantoea ananatis crtE, crtB, crtI, crtY, and crtZ genes in addition to the Haematococcus pluvialis IDI gene, and one containing each of zeaxanthin epoxidase (ZEP) genes originated from nine photosynthetic eukaryotes. It was consequently found that paprika (Capsicum annuum) ZEP (CaZEP) showed the highest conversion activity. Next, using the CaZEP gene, we performed optimization experiments in relation to E. coli strains as the production hosts, expression vectors, and ribosome-binding site (RBS) sequences. As a result, the highest productivity of violaxanthin (231 μg/g dry weight) was observed, when the pUC18 vector was used with CaZEP preceded by a RBS sequence of score 5000 in strain JM101(DE3).

KEYWORDS:

Escherichia coli; Pathway engineering; Violaxanthin; Zeaxanthin epoxidase

PMID:
31673744
DOI:
10.1007/s00253-019-10182-w

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