Screening and Characterization of Metagenome-Derived PHA Synthase Gene (phaC) from Limestone Soil Sample of Gunung Lang, Ipoh, Perak.
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Date
2016
Authors
Kanapathi Pillai, K. Sudesh Kumar
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Abstract
In this study, metagenomic approach was applied to identify novel PHA synthase genes (phaCs). The presence of phaCs in limestone soil was screened. Phylogenetic analysis has revealed that limestone area harbors a great diversity of phaCs. A total of 87 partial phaCs were obtained. Among these, 7 partial phaC genes were selected for full length amplification through metagenome walking. One of the clones (SC8) encoded a new gene (phaCsca), with an 81 % identity to the phaC of Chromobacterium violaceum ATCC 12472 based on BLASTn results. It able to express successfully and produce up to 55 wt. % of PHA when it was cloned into a PHA-negative mutant of Cupriavidus necator PHB-4. Besides that, the PHA synthase activity of this transformant was 55 U/g protein. At the same time, the biodiversity of this limestone area was studied. Based on the 16S rDNA sequencing result, it can be seen that this location is rich in biodiversity with the presence of potentially novel organisms. In conclusion, this area is a suitable and conducive environment for the living of organisms especially for the PHA producers. This study presents a pioneering study on the discovery of phaC in a limestone area using metagenomic approach.
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Keywords
Metagenome-Derived , Synthase Gene , Limestone Soil