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Optimization of a hybrid bacterial/Arabidopsis thaliana fatty acid synthase system II in Saccharomyces cerevisiae

Pozdniakova, Tatiana A. ; Cruz, João P. ; Silva, Paulo César ; Azevedo, Flávio ; Parpot, Pier ; Domingues, Maria Rosario ; Carlquist, Magnus LU and Johansson, Björn LU (2023) In Metabolic Engineering Communications 17.
Abstract

Fatty acids are produced by eukaryotes like baker's yeast Saccharomyces cerevisiae mainly using a large multifunctional type I fatty acid synthase (FASI) where seven catalytic steps and a carrier domain are shared between one or two protein subunits. While this system may offer efficiency in catalysis, only a narrow range of fatty acids are produced. Prokaryotes, chloroplasts and mitochondria rely instead on a FAS type II (FASII) where each catalytic step is carried out by a monofunctional enzyme encoded by a separate gene. FASII is more flexible and capable of producing a wider range of fatty acid structures, such as the direct production of unsaturated fatty acids. An efficient FASII in the preferred industrial organism S. cerevisiae... (More)

Fatty acids are produced by eukaryotes like baker's yeast Saccharomyces cerevisiae mainly using a large multifunctional type I fatty acid synthase (FASI) where seven catalytic steps and a carrier domain are shared between one or two protein subunits. While this system may offer efficiency in catalysis, only a narrow range of fatty acids are produced. Prokaryotes, chloroplasts and mitochondria rely instead on a FAS type II (FASII) where each catalytic step is carried out by a monofunctional enzyme encoded by a separate gene. FASII is more flexible and capable of producing a wider range of fatty acid structures, such as the direct production of unsaturated fatty acids. An efficient FASII in the preferred industrial organism S. cerevisiae could provide a platform for developing sustainable production of specialized fatty acids. We functionally replaced either yeast FASI genes (FAS1 or FAS2) with a FASII consisting of nine genes from Escherichia coli (acpP, acpS and fab -A, -B, -D, -F, -G, -H, -Z) as well as three from Arabidopsis thaliana (MOD1, FATA1 and FATB). The genes were expressed from an autonomously replicating multicopy vector assembled using the Yeast Pathway Kit for in-vivo assembly in yeast. Two rounds of adaptation led to a strain with a maximum growth rate (μmax) of 0.19 h−1 without exogenous fatty acids, twice the growth rate previously reported for a comparable strain. Additional copies of the MOD1 or fabH genes resulted in cultures with higher final cell densities and three times higher lipid content compared to the control.

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author
; ; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
E. coli, FASI, FASII, Fatty acid synthase, Metabolic engineering, Saccharomyces cerevisiae
in
Metabolic Engineering Communications
volume
17
article number
e00224
pages
10 pages
publisher
Elsevier
external identifiers
  • scopus:85162945328
  • pmid:37415783
ISSN
2214-0301
DOI
10.1016/j.mec.2023.e00224
language
English
LU publication?
yes
id
ab64bf10-151b-41d6-b8c6-0937a7513411
date added to LUP
2023-08-20 11:49:01
date last changed
2024-06-15 06:12:35
@article{ab64bf10-151b-41d6-b8c6-0937a7513411,
  abstract     = {{<p>Fatty acids are produced by eukaryotes like baker's yeast Saccharomyces cerevisiae mainly using a large multifunctional type I fatty acid synthase (FASI) where seven catalytic steps and a carrier domain are shared between one or two protein subunits. While this system may offer efficiency in catalysis, only a narrow range of fatty acids are produced. Prokaryotes, chloroplasts and mitochondria rely instead on a FAS type II (FASII) where each catalytic step is carried out by a monofunctional enzyme encoded by a separate gene. FASII is more flexible and capable of producing a wider range of fatty acid structures, such as the direct production of unsaturated fatty acids. An efficient FASII in the preferred industrial organism S. cerevisiae could provide a platform for developing sustainable production of specialized fatty acids. We functionally replaced either yeast FASI genes (FAS1 or FAS2) with a FASII consisting of nine genes from Escherichia coli (acpP, acpS and fab -A, -B, -D, -F, -G, -H, -Z) as well as three from Arabidopsis thaliana (MOD1, FATA1 and FATB). The genes were expressed from an autonomously replicating multicopy vector assembled using the Yeast Pathway Kit for in-vivo assembly in yeast. Two rounds of adaptation led to a strain with a maximum growth rate (μmax) of 0.19 h<sup>−1</sup> without exogenous fatty acids, twice the growth rate previously reported for a comparable strain. Additional copies of the MOD1 or fabH genes resulted in cultures with higher final cell densities and three times higher lipid content compared to the control.</p>}},
  author       = {{Pozdniakova, Tatiana A. and Cruz, João P. and Silva, Paulo César and Azevedo, Flávio and Parpot, Pier and Domingues, Maria Rosario and Carlquist, Magnus and Johansson, Björn}},
  issn         = {{2214-0301}},
  keywords     = {{E. coli; FASI; FASII; Fatty acid synthase; Metabolic engineering; Saccharomyces cerevisiae}},
  language     = {{eng}},
  publisher    = {{Elsevier}},
  series       = {{Metabolic Engineering Communications}},
  title        = {{Optimization of a hybrid bacterial/<i>Arabidopsis thaliana</i> fatty acid synthase system II in<i> Saccharomyces cerevisiae</i>}},
  url          = {{http://dx.doi.org/10.1016/j.mec.2023.e00224}},
  doi          = {{10.1016/j.mec.2023.e00224}},
  volume       = {{17}},
  year         = {{2023}},
}