Biosynthesis of Poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from CO2 by a Recombinant Cupriavidusnecator

The copolyester of 3-hydroxybutyrate (3HB) and 3-hydoxyhexanoate (3HHx), PHBHHx, is one of the most practical kind of bacterial polyhydroxyalkanoates due to its high flexibility and marine biodegradability. PHBHHx is usually produced from vegetable oils or fatty acids through β-oxidation, whereas bi...

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Autores principales:	Kenji Tanaka, Kazumasa Yoshida, Izumi Orita, Toshiaki Fukui
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	biodegradable plastic PHBHHx CO<sub>2</sub> <i>Cupriavidus necator</i> hydrogen-oxidizing bacterium Technology T Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/9b205fed16704397af1b10a1e215cbb6
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spelling	oai:doaj.org-article:9b205fed16704397af1b10a1e215cbb62021-11-25T16:46:37ZBiosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>10.3390/bioengineering81101792306-5354https://doaj.org/article/9b205fed16704397af1b10a1e215cbb62021-11-01T00:00:00Zhttps://www.mdpi.com/2306-5354/8/11/179https://doaj.org/toc/2306-5354The copolyester of 3-hydroxybutyrate (3HB) and 3-hydoxyhexanoate (3HHx), PHBHHx, is one of the most practical kind of bacterial polyhydroxyalkanoates due to its high flexibility and marine biodegradability. PHBHHx is usually produced from vegetable oils or fatty acids through β-oxidation, whereas biosynthesis from sugars has been achieved by recombinant strains of hydrogen-oxidizing bacterium <i>Cupriavidus necator</i>. This study investigated the biosynthesis of PHBHHx from CO<sub>2</sub> as the sole carbon source by engineered <i>C. necator</i> strains. The recombinant strains capable of synthesizing PHBHHx from fructose were cultivated in a flask using complete mineral medium and a substrate gas mixture (H<sub>2</sub>/O<sub>2</sub>/CO<sub>2</sub> = 8:1:1). The results of GC and <sup>1</sup>H NMR analyses indicated that the recombinants of <i>C. necator</i> synthesized PHBHHx from CO<sub>2</sub> with high cellular content. When 1.0 g/L (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> was used as a nitrogen source, the 3HHx composition of PHBHHx in the strain MF01∆B1/pBBP-ccr<sub>Me</sub>J4a-emd was 47.7 ± 6.2 mol%. Further investigation demonstrated that the PHA composition can be regulated by using (<i>R</i>)-enoyl-CoA hydratase (PhaJ) with different substrate specificity. The composition of 3HHx in PHBHHx was controlled to about 11 mol%, suitable for practical applications, and high cellular content was kept in the strains transformed with pBPP-ccr<sub>Me</sub>J<sub>Ac</sub>-emd harboring short-chain-length-specific PhaJ.Kenji TanakaKazumasa YoshidaIzumi OritaToshiaki FukuiMDPI AGarticlebiodegradable plasticPHBHHxCO<sub>2</sub><i>Cupriavidus necator</i>hydrogen-oxidizing bacteriumTechnologyTBiology (General)QH301-705.5ENBioengineering, Vol 8, Iss 179, p 179 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	biodegradable plastic PHBHHx CO<sub>2</sub> <i>Cupriavidus necator</i> hydrogen-oxidizing bacterium Technology T Biology (General) QH301-705.5
spellingShingle	biodegradable plastic PHBHHx CO<sub>2</sub> <i>Cupriavidus necator</i> hydrogen-oxidizing bacterium Technology T Biology (General) QH301-705.5 Kenji Tanaka Kazumasa Yoshida Izumi Orita Toshiaki Fukui Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>
description	The copolyester of 3-hydroxybutyrate (3HB) and 3-hydoxyhexanoate (3HHx), PHBHHx, is one of the most practical kind of bacterial polyhydroxyalkanoates due to its high flexibility and marine biodegradability. PHBHHx is usually produced from vegetable oils or fatty acids through β-oxidation, whereas biosynthesis from sugars has been achieved by recombinant strains of hydrogen-oxidizing bacterium <i>Cupriavidus necator</i>. This study investigated the biosynthesis of PHBHHx from CO<sub>2</sub> as the sole carbon source by engineered <i>C. necator</i> strains. The recombinant strains capable of synthesizing PHBHHx from fructose were cultivated in a flask using complete mineral medium and a substrate gas mixture (H<sub>2</sub>/O<sub>2</sub>/CO<sub>2</sub> = 8:1:1). The results of GC and <sup>1</sup>H NMR analyses indicated that the recombinants of <i>C. necator</i> synthesized PHBHHx from CO<sub>2</sub> with high cellular content. When 1.0 g/L (NH<sub>4</sub>)<sub>2</sub>SO<sub>4</sub> was used as a nitrogen source, the 3HHx composition of PHBHHx in the strain MF01∆B1/pBBP-ccr<sub>Me</sub>J4a-emd was 47.7 ± 6.2 mol%. Further investigation demonstrated that the PHA composition can be regulated by using (<i>R</i>)-enoyl-CoA hydratase (PhaJ) with different substrate specificity. The composition of 3HHx in PHBHHx was controlled to about 11 mol%, suitable for practical applications, and high cellular content was kept in the strains transformed with pBPP-ccr<sub>Me</sub>J<sub>Ac</sub>-emd harboring short-chain-length-specific PhaJ.
format	article
author	Kenji Tanaka Kazumasa Yoshida Izumi Orita Toshiaki Fukui
author_facet	Kenji Tanaka Kazumasa Yoshida Izumi Orita Toshiaki Fukui
author_sort	Kenji Tanaka
title	Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>
title_short	Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>
title_full	Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>
title_fullStr	Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>
title_full_unstemmed	Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>
title_sort	biosynthesis of poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from co<sub>2</sub> by a recombinant <i>cupriavidus</i><i>necator</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/9b205fed16704397af1b10a1e215cbb6
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Biosynthesis of Poly(3-hydroxybutyrate-<i>co</i>-3-hydroxyhexanoate) from CO<sub>2</sub> by a Recombinant <i>Cupriavidus</i><i>necator</i>

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