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High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant

BACKGROUND: Several members of the bacterial Halomonadacea family are natural producers of polyhydroxyalkanoates (PHA), which are promising materials for use as biodegradable bioplastics. Type-strain species of Cobetia are designated PHA positive, and recent studies have demonstrated relatively high...

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Autores principales: Christensen, Mikkel, Jablonski, Piotr, Altermark, Bjørn, Irgum, Knut, Hansen, Hilde
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8686332/
https://www.ncbi.nlm.nih.gov/pubmed/34930259
http://dx.doi.org/10.1186/s12934-021-01713-0
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author Christensen, Mikkel
Jablonski, Piotr
Altermark, Bjørn
Irgum, Knut
Hansen, Hilde
author_facet Christensen, Mikkel
Jablonski, Piotr
Altermark, Bjørn
Irgum, Knut
Hansen, Hilde
author_sort Christensen, Mikkel
collection PubMed
description BACKGROUND: Several members of the bacterial Halomonadacea family are natural producers of polyhydroxyalkanoates (PHA), which are promising materials for use as biodegradable bioplastics. Type-strain species of Cobetia are designated PHA positive, and recent studies have demonstrated relatively high PHA production for a few strains within this genus. Industrially relevant PHA producers may therefore be present among uncharacterized or less explored members. In this study, we characterized PHA production in two marine Cobetia strains. We further analyzed their genomes to elucidate pha genes and metabolic pathways which may facilitate future optimization of PHA production in these strains. RESULTS: Cobetia sp. MC34 and Cobetia marina DSM 4741(T) were mesophilic, halotolerant, and produced PHA from four pure substrates. Sodium acetate with- and without co-supplementation of sodium valerate resulted in high PHA production titers, with production of up to 2.5 g poly(3-hydroxybutyrate) (PHB)/L and 2.1 g poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)/L in Cobetia sp. MC34, while C. marina DSM 4741(T) produced 2.4 g PHB/L and 3.7 g PHBV/L. Cobetia marina DSM 4741(T) also showed production of 2.5 g PHB/L from glycerol. The genome of Cobetia sp. MC34 was sequenced and phylogenetic analyses revealed closest relationship to Cobetia amphilecti. PHA biosynthesis genes were located at separate loci similar to the arrangement in other Halomonadacea. Further genome analyses revealed some differences in acetate- and propanoate metabolism genes between the two strains. Interestingly, only a single PHA polymerase gene (phaC(2)) was found in Cobetia sp. MC34, in contrast to two copies (phaC(1) and phaC(2)) in C. marina DSM 4741(T). In silico analyses based on phaC genes show that the PhaC(2) variant is conserved in Cobetia and contains an extended C-terminus with a high isoelectric point and putative DNA-binding domains. CONCLUSIONS: Cobetia sp. MC34 and C. marina DSM 4741(T) are natural producers of PHB and PHBV from industrially relevant pure substrates including acetate. However, further scale up, optimization of growth conditions, or use of metabolic engineering is required to obtain industrially relevant PHA production titers. The putative role of the Cobetia PhaC(2) variant in DNA-binding and the potential implications remains to be addressed by in vitro- or in vivo methods. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01713-0.
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spelling pubmed-86863322021-12-20 High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant Christensen, Mikkel Jablonski, Piotr Altermark, Bjørn Irgum, Knut Hansen, Hilde Microb Cell Fact Research BACKGROUND: Several members of the bacterial Halomonadacea family are natural producers of polyhydroxyalkanoates (PHA), which are promising materials for use as biodegradable bioplastics. Type-strain species of Cobetia are designated PHA positive, and recent studies have demonstrated relatively high PHA production for a few strains within this genus. Industrially relevant PHA producers may therefore be present among uncharacterized or less explored members. In this study, we characterized PHA production in two marine Cobetia strains. We further analyzed their genomes to elucidate pha genes and metabolic pathways which may facilitate future optimization of PHA production in these strains. RESULTS: Cobetia sp. MC34 and Cobetia marina DSM 4741(T) were mesophilic, halotolerant, and produced PHA from four pure substrates. Sodium acetate with- and without co-supplementation of sodium valerate resulted in high PHA production titers, with production of up to 2.5 g poly(3-hydroxybutyrate) (PHB)/L and 2.1 g poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)/L in Cobetia sp. MC34, while C. marina DSM 4741(T) produced 2.4 g PHB/L and 3.7 g PHBV/L. Cobetia marina DSM 4741(T) also showed production of 2.5 g PHB/L from glycerol. The genome of Cobetia sp. MC34 was sequenced and phylogenetic analyses revealed closest relationship to Cobetia amphilecti. PHA biosynthesis genes were located at separate loci similar to the arrangement in other Halomonadacea. Further genome analyses revealed some differences in acetate- and propanoate metabolism genes between the two strains. Interestingly, only a single PHA polymerase gene (phaC(2)) was found in Cobetia sp. MC34, in contrast to two copies (phaC(1) and phaC(2)) in C. marina DSM 4741(T). In silico analyses based on phaC genes show that the PhaC(2) variant is conserved in Cobetia and contains an extended C-terminus with a high isoelectric point and putative DNA-binding domains. CONCLUSIONS: Cobetia sp. MC34 and C. marina DSM 4741(T) are natural producers of PHB and PHBV from industrially relevant pure substrates including acetate. However, further scale up, optimization of growth conditions, or use of metabolic engineering is required to obtain industrially relevant PHA production titers. The putative role of the Cobetia PhaC(2) variant in DNA-binding and the potential implications remains to be addressed by in vitro- or in vivo methods. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12934-021-01713-0. BioMed Central 2021-12-20 /pmc/articles/PMC8686332/ /pubmed/34930259 http://dx.doi.org/10.1186/s12934-021-01713-0 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Christensen, Mikkel
Jablonski, Piotr
Altermark, Bjørn
Irgum, Knut
Hansen, Hilde
High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant
title High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant
title_full High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant
title_fullStr High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant
title_full_unstemmed High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant
title_short High natural PHA production from acetate in Cobetia sp. MC34 and Cobetia marina DSM 4741(T) and in silico analyses of the genus specific PhaC(2) polymerase variant
title_sort high natural pha production from acetate in cobetia sp. mc34 and cobetia marina dsm 4741(t) and in silico analyses of the genus specific phac(2) polymerase variant
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8686332/
https://www.ncbi.nlm.nih.gov/pubmed/34930259
http://dx.doi.org/10.1186/s12934-021-01713-0
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