Kinetic modeling, recovery, and molecular characterization of poly-beta-hydroxybutyrate polymer in Acinetobacter baumannii isolate P39

Abstract

To control the poly-β-hydroxybutyrate (PHB) biopolymer production by Acinetobacter baumannii isolate P39 kinetic modeling of the fermentation process, polymer downstream processing, enzymological analysis, and molecular characterization of PHA synthase, key biosynthetic enzyme, should be addressed. A. baumannii isolate P39 produced 0.15g/L PHB after 24h of incubation with a polymer content of 28% per dry weight. Logistic and Leudeking-Piret models were used for describing cell growth and PHB production, respectively. They showed good agreement with the experimental data describing both cell growth and PHB production (average regression coefficient r2:0.999). The growth-associated production of PHB biopolymer as an electron acceptor was confirmed using Leudeking-Piret model and victim substrate experiment. The best method of recovery of PHB biopolymer was chemical digestion using sodium hypochlorite, since it produced the largest amount of polymer and highest molecular weight (16,000g/mole) in comparison to other recovery methods. DTNB assay showed high activity of PHA synthase enzyme, 600U activity, and 153.8U/mg specific activity. Molecular analysis of PHA synthase enzyme confirmed class III identity. Taken together, micelle model was proposed for polyhydroxybutyrate formation in A. baumannii isolate P39.