Enhanced production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) with modulated 3-hydroxyvalerate fraction by overexpressing acetolactate synthase in Cupriavidus necator H16

• Published in: International journal of biological macromolecules Vol. 242; no. Pt 4; p. 125166
• Main Authors: Jo, Young Yun, Park, Soyoung, Gong, Gyeongtaek, Roh, Soonjong, Yoo, Jin, Ahn, Jung Ho, Lee, Sun-Mi, Um, Youngsoon, Kim, Kyoung Heon, Ko, Ja Kyong
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.07.2023
• Subjects: Acetolactate synthase; Cupriavidus necator H16; poly(3-hydroxybutyrate-co-3-hydroxyvalerate); Acetolactate synthase; Cupriavidus necator H16; poly(3-hydroxybutyrate-co-3-hydroxyvalerate)
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2023.125166

The elastomeric properties of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), a biodegradable copolymer, strongly depend on the molar composition of 3-hydroxyvalerate (3HV). This paper reports an improved artificial pathway for enhancing the 3HV component during PHBV biosynthesis from a structurally unrelated carbon source by Cupriavidus necator H16. To increase the intracellular accumulation of propionyl-CoA, a key precursor of the 3HV monomer, we developed a recombinant strain by genetically manipulating the branched-chain amino acid (e.g., valine, isoleucine) pathways. Overexpression of the heterologous feedback-resistant acetolactate synthase (alsS), (R)-citramalate synthase (leuA), homologous 3-ketothiolase (bktB), and the deletion of 2-methylcitrate synthase (prpC) resulted in biosynthesis of 42.5 % (g PHBV/g dry cell weight) PHBV with 64.9 mol% 3HV monomer from fructose as the sole carbon source. This recombinant strain also accumulated the highest PHBV content of 54.5 % dry cell weight (DCW) with 24 mol% 3HV monomer from CO2 ever reported. The lithoautotrophic cell growth and PHBV production by the recombinant C. necator were promoted by oxygen stress. The thermal properties of PHBV showed a decreasing trend of the glass transition and melting temperatures with increasing 3HV fraction. The average molecular weights of PHBV with modulated 3HV fractions were between 20 and 26 × 104 g/mol. [Display omitted] •The properties PHBV copolymer depend on the molar composition of 3-hydroxyvalerate.•C. necator was engineered to produce PHBV from structurally unrelated carbon source.•The CY8 strain was developed by engineering branched chain amino acid pathway.•The CY8 strain produced PHBV with the highest 3HV fraction from CO2 ever reported.•The lithoautotrophic PHBV production by CY8 strain was enhanced by oxygen stress.