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Engineering the carotenoid biosynthetic pathway in Rhodothermus marinus for lycopene production

Kristjansdottir, Thordis ; Ron, Emanuel Y.C. LU ; Molins-Delgado, Daniel LU ; Fridjonsson, Olafur H. ; Turner, Charlotta LU ; Bjornsdottir, Snaedis H. ; Gudmundsson, Steinn ; van Niel, Ed W.J. LU ; Karlsson, Eva Nordberg LU and Hreggvidsson, Gudmundur O. (2020) In Metabolic Engineering Communications 11.
Abstract

Rhodothermus marinus has the potential to be well suited for biorefineries, as an aerobic thermophile that produces thermostable enzymes and is able to utilize polysaccharides from different 2nd and 3rd generation biomass. The bacterium produces valuable chemicals such as carotenoids. However, the native carotenoids are not established for industrial production and R. marinus needs to be genetically modified to produce higher value carotenoids. Here we genetically modified the carotenoid biosynthetic gene cluster resulting in three different mutants, most importantly the lycopene producing mutant TK-3 (ΔtrpBΔpurAΔcruFcrtB::trpBcrtBT.thermophilus). The genetic modifications and subsequent structural analysis of carotenoids... (More)

Rhodothermus marinus has the potential to be well suited for biorefineries, as an aerobic thermophile that produces thermostable enzymes and is able to utilize polysaccharides from different 2nd and 3rd generation biomass. The bacterium produces valuable chemicals such as carotenoids. However, the native carotenoids are not established for industrial production and R. marinus needs to be genetically modified to produce higher value carotenoids. Here we genetically modified the carotenoid biosynthetic gene cluster resulting in three different mutants, most importantly the lycopene producing mutant TK-3 (ΔtrpBΔpurAΔcruFcrtB::trpBcrtBT.thermophilus). The genetic modifications and subsequent structural analysis of carotenoids helped clarify the carotenoid biosynthetic pathway in R. marinus. The nucleotide sequences encoding the enzymes phytoene synthase (CrtB) and the previously unidentified 1′,2′-hydratase (CruF) were found fused together and encoded by a single gene in R. marinus. Deleting only the cruF part of the gene did not result in an active CrtB enzyme. However, by deleting the entire gene and inserting the crtB gene from Thermus thermophilus, a mutant strain was obtained, producing lycopene as the sole carotenoid. The lycopene produced by TK-3 was quantified as 0.49 ​g/kg CDW (cell dry weight).

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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Carotenoids, Genetic engineering, Lycopene, Rhodothermus marinus
in
Metabolic Engineering Communications
volume
11
article number
e00140
publisher
Elsevier
external identifiers
  • scopus:85089023950
  • pmid:32793416
ISSN
2214-0301
DOI
10.1016/j.mec.2020.e00140
language
English
LU publication?
yes
id
a2878706-435a-4358-a2f9-fbc4d2f279a2
date added to LUP
2020-08-11 11:26:48
date last changed
2021-04-06 01:11:45
@article{a2878706-435a-4358-a2f9-fbc4d2f279a2,
  abstract     = {<p>Rhodothermus marinus has the potential to be well suited for biorefineries, as an aerobic thermophile that produces thermostable enzymes and is able to utilize polysaccharides from different 2nd and 3rd generation biomass. The bacterium produces valuable chemicals such as carotenoids. However, the native carotenoids are not established for industrial production and R. marinus needs to be genetically modified to produce higher value carotenoids. Here we genetically modified the carotenoid biosynthetic gene cluster resulting in three different mutants, most importantly the lycopene producing mutant TK-3 (ΔtrpBΔpurAΔcruFcrtB::trpBcrtB<sub>T.thermophilus</sub>). The genetic modifications and subsequent structural analysis of carotenoids helped clarify the carotenoid biosynthetic pathway in R. marinus. The nucleotide sequences encoding the enzymes phytoene synthase (CrtB) and the previously unidentified 1′,2′-hydratase (CruF) were found fused together and encoded by a single gene in R. marinus. Deleting only the cruF part of the gene did not result in an active CrtB enzyme. However, by deleting the entire gene and inserting the crtB gene from Thermus thermophilus, a mutant strain was obtained, producing lycopene as the sole carotenoid. The lycopene produced by TK-3 was quantified as 0.49 ​g/kg CDW (cell dry weight).</p>},
  author       = {Kristjansdottir, Thordis and Ron, Emanuel Y.C. and Molins-Delgado, Daniel and Fridjonsson, Olafur H. and Turner, Charlotta and Bjornsdottir, Snaedis H. and Gudmundsson, Steinn and van Niel, Ed W.J. and Karlsson, Eva Nordberg and Hreggvidsson, Gudmundur O.},
  issn         = {2214-0301},
  language     = {eng},
  publisher    = {Elsevier},
  series       = {Metabolic Engineering Communications},
  title        = {Engineering the carotenoid biosynthetic pathway in Rhodothermus marinus for lycopene production},
  url          = {http://dx.doi.org/10.1016/j.mec.2020.e00140},
  doi          = {10.1016/j.mec.2020.e00140},
  volume       = {11},
  year         = {2020},
}