Cascading Beta-oxidation Intermediates for the Polyhydroxyalkanoate Copolymer Biosynthesis by Metabolic Flux using Co-substrates and Inhibitors

Abstract

Polyhydroxyalkanoates (PHAs) are biopolymers that are produced within the microbial cells in the presence of excess carbon and nutrient limitation. Different strategies have been studied to increase the quality and quantity of this biopolymer which in turn can be utilized as biodegradable polymers replacing conventional petrochemical plastics. In the present study, Bacillus endophyticus, a gram-positive PHA-producing bacterium, was cultivated in the presence of fatty acids along with beta-oxidation inhibitor acrylic acid. A novel approach for incorporating different hydroxyacyl groups provided using fatty acids as co-substrate and beta-oxidation inhibitors to direct the intermediates to co-polymer synthesis was experimented. It was observed that higher fatty acids and inhibitors had a greater influence on PHA production. The addition of acrylic acid along with propionic acid had a positive impact, giving 56.49% of PHA along with sucrose which was 1.2-fold more than the control devoid of fatty acids and inhibitors. Along with the copolymer production, the possible PHA pathway functional leading to the copolymer biosynthesis was hypothetically interpreted in this study. The obtained PHA was analyzed by FTIR and 1H NMR to confirm the copolymer production, which indicated the presence of poly3hydroxybutyrate-co-hydroxyvalerate (PHB-co-PHV), poly3hydroxybutyrate-co-hydroxyhexanoate (PHB-co-PHx).

Keywords

• bacillus endophyticus
• fatty acids
• inhibitors
• polyhydroxyalkanoate
• shake flask cultivation