Farzaneh Barati
1 
, Ezat Asgarani
1* , Sara Gharavi
1, Mohammad Reza Soudi
21 Department of Biotechnology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran
2 2 Department of Microbiology, Faculty of Biological Sciences, Alzahra University, Tehran, Iran
Abstract
Introduction: Poly(3-hydroxybutyrate) (PHB) is a well-known biodegradable polymer produced by some microorganisms and can be a suitable alternative for petrochemical plastics. PHB synthase encoded by phbC gene is the main enzyme in PHB biosynthesis pathway in Ralstonia eutropha. The aim of current study was the transformation of R. eutropha PTCC 1615 with its own phbC gene and evaluation of the overexpression effect on PHB accumulation.
Methods: DNA fragment including phbC gene and its promoter and terminator regions, was isolated from R. eutropha PTCC 1615, inserted into pET28a(+) vector, and transferred to the competent bacteria using calcium chloride and heat shock method. The effect of the cloned gene expression on PHB production was investigated with absorption of crotonic acid produced through PHB dehydration. Statistical analyses were carried out by SPSS software.
Results: PHB content of cells of the engineered strain was 1.4 times more than that of the native bacteria. This significant difference can be an important finding for improvement of biopolymer production.
Conclusion: Overexpression of phbC, the critical gene in PHB biosynthesis pathway, in R. eutropha PTCC 1615 had considerable effect on PHB accumulation.