Diversion of the Long-Chain Acyl-ACP Pool in Synechocystis to Fatty Alcohols through CRISPRi Repression of the Essential Phosphate Acyltransferase PlsX
Abstract
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, 10.3mgg-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.
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Algal strains such as Chlamydomonas reinhardtii and Synechocystis sp. have been modified to improve resistance to environmental stressors and boost growth rates by manipulating pathways involved in light harvesting.
“Algal strains such as Chlamydomonas reinhardtii and Synechocystis sp. have been modified to improve resistance to environmental stressors and boost growth rates by manipulating pathways involved in light harvesting [119-123].”
From A Review of Algae-Based Carbon Capture, Utilization, and Storage (Algae-Based CCUS) (2024) · cites this paper as [122]
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- Authors
- Kaczmarzyk, D., Cengic, I., Yao, L., & Hudson, E. P.
- Affiliations
- Science for Life Laboratory (SE)
- Corresponding author
- Hudson, Elton P.
- Venue
- Metabolic Engineering
- Publisher
- Elsevier BV
- Published
- 2018-01
- Volume
- 45
- Pages
- 59-66
- ISSN
- 1096-7176 (ISSN-L)
- OpenAlex
- W2773548304
- Cited by
- 120
- References
- 45
- Citations by year
- 2026: 6 · 2025: 10 · 2024: 10 · 2023: 8 · 2022: 15 · 2021: 15 · 2020: 25 · 2019: 15 · 2018: 16
- FWCI
- 6.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Metabolic Engineering (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Stiftelsen för Strategisk Forskning (RBP14-0013)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biochemistry › Lipid metabolism and biosynthesis
- Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
- Keywords
- Synechocystis sp. PCC 6803, CRISPR interference, octadecanol, phosphatidic acid synthesis, membrane homeostasis, reactive oxygen species, heterologous expression
- MeSH terms
- Acyltransferases, Bacterial Proteins, Fatty Alcohols, Plant Proteins, Oryza, Synechocystis, Marinobacter, CRISPR-Cas Systems
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