Journal of Siberian Federal University. Biology / Cloning and Characterization of Polyhydroxybutyrate Synthase from Methylobacterium extorquens AM1

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Issue
Journal of Siberian Federal University. Biology. 2016 9 (2)
Authors
Zamakhaeva, Svetlana A.; Fedorov, Dmitry N.; Doronina, Nina V.; Trotsenko, Yuri A.
Contact information
Zamakhaeva, Svetlana A.:G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms RAS 5 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; Pushchino State Institute of Natural Sciences 3 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; Fedorov, Dmitry N.:G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms RAS 5 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; Pushchino State Institute of Natural Sciences 3 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; ; Doronina, Nina V.:G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms RAS 5 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; Pushchino State Institute of Natural Sciences 3 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; Trotsenko, Yuri A.:G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms RAS 5 Prospect Nauki, Pushchino, Moscow region, 142290, Russia; Pushchino State Institute of Natural Sciences 3 Prospect Nauki, Pushchino, Moscow region, 142290, Russia
Keywords
Methylobacterium extorquens; phaC; methylotrophs; polyhydroxybutyrate; PHB-synthase; Methylobacterium extorquens; phaC
Abstract

Multiple genes encoding putative PHB synthases (up to 5 in single strain) were found in Methylobacterium genomes. As a result of phylogenetic analysis proteins PhaC1, PhaC2, PhaC3 were identified as class I PHB synthases, PhaC4 proteins were identified as class III PHB synthases, while PhaC5 apparently belongs to uncharacterized class of PHB synthases. Firstly, the recombinant class I PBH synthase (EC 2.3.1.B2) encoded by phaC1 gene from Methylobacterium extorquens AM1 was purified and characterized. Molecular mass of enzyme monomer was 78 kDa. Michaelis constant (Km) for PhaC1 was 1,3 mM and maximal reaction rate (Vmax) was 0,1 μmol∙min–1∙mg–1. The deletion mutant of Methylobacterium extorquens in the phaC gene was obtained which is promising for further study of peculiarities of methylobacteria’s PHB biosynthesis

Pages
169-179
Paper at repository of SibFU
https://elib.sfu-kras.ru/handle/2311/20363

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