A unique class I polyhydroxyalkanoate synthase (PhaC) from Brevundimonas sp. KH11J01 exists as a functional trimer: A comparative study with PhaC from Cupriavidus necator H16

Abstract

Polyhydroxyalkanoates (PHAs) are natural biodegradable polyesters that are produced by numerous prokaryotic microorganisms primarily as a carbon- and energy reserve. The PhaC enzyme catalyzes the last step in the PHA biosynthesis pathway and synthesizes PHA polymers from hydroxyalkanoic acids. A type I PhaC from a PHAproducing marine bacterium Brevundimonas sp. KH11J01 (BrPhaC) was identified, produced recombinantly and characterized. Its properties were compared with its homolog from C. necator H16 (RePhaC). Unlike other PhaCs, it was found that BrPhaC is a lag-phase free enzyme organized as a trimer, even without the presence of a substrate. The enzymatic reaction is initiated instantly irrespective of temperature, in contrast to RePhaC in which the duration of the lag-phase was highly affected by temperature. At 10 ◦C BrPhaC was 40% active whereas RePhaC was barely active. The significance of using marine microorganisms, harboring cold-active PHA biosynthesis enzymes, for energy efficient PHA production, is also discussed briefly. The unique trimeric organization of BrPhaC challenges our understanding of the PhaC reaction mechanisms, which is mainly based on the crystal structures of the inactive forms of the enzyme.