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Diversion of the long-chain acyl-ACP pool in Synechocystis to fatty alcohols through CRISPRi repression of the essential phosphate acyltransferase PlsX
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0001-8317-1654
KTH, Centres, Science for Life Laboratory, SciLifeLab.
KTH, School of Biotechnology (BIO), Proteomics and Nanobiotechnology. KTH, Centres, Science for Life Laboratory, SciLifeLab.ORCID iD: 0000-0003-1899-7649
2018 (English)In: Metabolic engineering, ISSN 1096-7176, E-ISSN 1096-7184, Vol. 45, p. 59-66Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
Academic Press, 2018. Vol. 45, p. 59-66
Keywords [en]
Acyl-ACP, Acyltransferase, CRISPRi, Cyanobacteria, Fatty alcohols, Membranes
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:kth:diva-220198DOI: 10.1016/j.ymben.2017.11.014ISI: 000424292100007Scopus ID: 2-s2.0-85036651462OAI: oai:DiVA.org:kth-220198DiVA, id: diva2:1167349
Funder
Swedish Foundation for Strategic Research , RBP14-0013Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Note

QC 20171218

Available from: 2017-12-18 Created: 2017-12-18 Last updated: 2018-03-13Bibliographically approved

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Kaczmarzyk, DanutaCengic, IvanaYao, LunHudson, Elton P.

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