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Appl Microbiol Biotechnol. 2017 Sep;101(18):6879-6889. doi: 10.1007/s00253-017-8420-4. Epub 2017 Jul 29.

Improved fermentative L-cysteine overproduction by enhancing a newly identified thiosulfate assimilation pathway in Escherichia coli.

Author information

1
Innovation Medical Research Institute, University of Tsukuba, Tsukuba, Japan.
2
Research Institute for Bioscience Products and Fine Chemicals, Ajinomoto Co., Kawasaki, Japan.
3
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan.
4
MS Business Unit, Shimadzu Co., Nakagyo, Japan.
5
Innovation Medical Research Institute, University of Tsukuba, Tsukuba, Japan. ohtsu.iwao.fm@u.tsukuba.ac.jp.
6
Graduate School of Biological Sciences, Nara Institute of Science and Technology, Ikoma, Japan. ohtsu.iwao.fm@u.tsukuba.ac.jp.

Abstract

Sulfate (SO42-) is an often-utilized and well-understood inorganic sulfur source in microorganism culture. Recently, another inorganic sulfur source, thiosulfate (S2O32-), was proposed to be more advantageous in microbial growth and biotechnological applications. Although its assimilation pathway is known to depend on O-acetyl-L-serine sulfhydrylase B (CysM in Escherichia coli), its metabolism has not been extensively investigated. Therefore, we aimed to explore another yet-unidentified CysM-independent thiosulfate assimilation pathway in E. coli. ΔcysM cells could accumulate essential L-cysteine from thiosulfate as the sole sulfur source and could grow, albeit slowly, demonstrating that a CysM-independent thiosulfate assimilation pathway is present in E. coli. This pathway is expected to consist of the initial part of the thiosulfate to sulfite (SO32-) conversion, and the latter part might be shared with the final part of the known sulfate assimilation pathway [sulfite → sulfide (S2-) → L-cysteine]. This is because thiosulfate-grown ΔcysM cells could accumulate a level of sulfite and sulfide equivalent to that of wild-type cells. The catalysis of thiosulfate to sulfite is at least partly mediated by thiosulfate sulfurtransferase (GlpE), because its overexpression could enhance cellular thiosulfate sulfurtransferase activity in vitro and complement the slow-growth phenotype of thiosulfate-grown ΔcysM cells in vivo. GlpE is therefore concluded to function in the novel CysM-independent thiosulfate assimilation pathway by catalyzing thiosulfate to sulfite. We applied this insight to L-cysteine overproduction in E. coli and succeeded in enhancing it by GlpE overexpression in media containing glucose or glycerol as the main carbon source, by up to ~1.7-fold (1207 mg/l) or ~1.5-fold (1529 mg/l), respectively.

KEYWORDS:

Escherichia coli; L-Cysteine; L-Cysteine production; Sulfur assimilation; Thiosulfate sulfurtransferase

PMID:
28756590
DOI:
10.1007/s00253-017-8420-4
[Indexed for MEDLINE]

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