Diversion of the long-chain acyl-ACP pool in Synechocystis to fatty alcohols through CRISPRi repression of the essential phosphate acyltransferase PlsX

Kaczmarzyk, Danuta (author): KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab

Cengic, Ivana (author): KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab

Hudson, Elton P. (author): KTH,Proteomik och nanobioteknologi,Science for Life Laboratory, SciLifeLab

(creator_code:org_t)

Academic Press, 2018
2018
English.
In: Metabolic engineering. - : Academic Press. - 1096-7176 .- 1096-7184. ; 45, s. 59-66

Related links:: https://urn.kb.se/re...; show more...; https://doi.org/10.1...; show less...

Abstract Subject headings

Fatty alcohol production in Synechocystis sp. PCC 6803 was achieved through heterologous expression of the fatty acyl-CoA/ACP reductase Maqu2220 from the bacteria Marinobacter aquaeolei VT8 and the fatty acyl-ACP reductase DPW from the rice Oryza sativa. These platform strains became models for testing multiplex CRISPR-interference (CRISPRi) metabolic engineering strategies to both improve fatty alcohol production and to study membrane homeostasis. CRISPRi allowed partial repression of up to six genes simultaneously, each encoding enzymes of acyl-ACP-consuming pathways. We identified the essential phosphate acyltransferase enzyme PlsX (slr1510) as a key node in C18 fatty acyl-ACP consumption, repression of slr1510 increased octadecanol productivity threefold over the base strain and gave the highest specific titers reported for this host, 10.3 mg g−1 DCW. PlsX catalyzes the first committed step of phosphatidic acid synthesis, and has not been characterized in Synechocystis previously. We found that accumulation of fatty alcohols impaired growth, altered the membrane composition, and caused a build-up of reactive oxygen species.

By the author/editor: Kaczmarzyk, Danu ...; Cengic, Ivana; Yao, Lun; Hudson, Elton P.

About the subject

NATURAL SCIENCES: NATURAL SCIENCES; and Biological Scien ...; and Biochemistry and ...

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