A purple non-sulfur bacterium producing polyhydroxybutyrate and the conserved region of pha synthase gene

Somporn Tanskul,Aekkaraj Nualla-Ong,Supaporn Srisai

doi:10.14393/bj-v32n1a2016-33801

A purple non-sulfur bacterium producing polyhydroxybutyrate and the conserved region of pha synthase gene

Somporn Tanskul, Aekkaraj Nualla-Ong + Show 1 more

Open Access

https://doi.org/10.14393/bj-v32n1a2016-33801

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Journal: Bioscience Journal	Publication Date: Jan 1, 2016
Citations: 4	License type: cc-by

Affiliation: Prince of Songkla University

Abstract

This study aimed to screen purple non-sulfur bacteria capable of accumulating granules or polyhydroxybutyrate (PHB) inside the cells, identify the potent strain, assay the enzyme or PHA synthase, and compare the PHB synthase gene with that of related strains. A total of 58 strains of purple non-sulfur bacteria were isolated from 108 samples of chicken feces in the chicken-egg farm of the Department of Animal Science, Faculty of Natural Resources at Prince of Songkla University, Hat Yai, Thailand. After cultivating the bacteria in glutamate malate (GM) medium without added glutamic acid under light (3,000 Lux) at 35 o C for 5 days, the intracellular biopolymer granules of the bacteria were observed by using a Confocal Laser Scanning Microscope (CLSM) with excitation and emission wavelength of 530 and 605 nm, respectively. Gas chromatography (GC) was carried out for quantitative analysis of PHB. There were five strains, CH12, CH52, CH72, CH90 and CH92, showed biopolymer granules under CLSM, and accumulated PHB 5, 1.7, 1.5, 1.4 and 1.8% (w w -1 ) of the cell dry weight (CDW), respectively. The 16S rDNA sequence analysis of CH12 strain showed a high homology of 100% correlation to that of Rhodopseudomonas palustris strain NCIB8288. Regarding the taxonomic characteristics and 16S rDNA sequence analysis, CH12 strain was identified as Rps. palustris NCIB8288. The PHA synthase activity of the crude extract from CH12 strain was 25 units/mL. The conserved regions could be aligned and selected among 5 strains of Rhodopseudomonas palustris (strains BisA53, TIE-1, CGA009, HaA2 and BisB18). The purified PCR product was obtained for further studies.

Highlights

Plastics are not environmentally friendly materials to the circumstances due to the long-term degradation
A total of 108 samples of chicken feces in the chicken-egg farm of the Department of Animal Science, Faculty of Natural Resources at Prince of Songkla University, Hat Yai were used for isolation of purple non-sulfur bacteria
Glutamate-malate (GM) medium was prepared for growing the purple non-sulfur bacteria. 12.5 mL of glutamate malate (GM) medium without added glutamic acid in a 15-mL screw-cap tube used for cultivation of the bacteria

Summary

Introduction

Plastics are not environmentally friendly materials to the circumstances due to the long-term degradation. PHAs are found in various kinds of microorganisms as energy and carbon reserve (SALEHIZADEH; LOOSDRECHT, 2004) under aerobic condition (GOBI; VADIVELU, 2014) They can be found in natural environments , for example, lake (ESTEVE et al, 1996), river (LOPEZ et al, 1995), knots of plant rhizoids (NDOYE et al., 1994), deep sea under high temperature (GUEZENNEC et al, 1998), and activated sludge (MINO et al, 1998). They can be observed in bacteria under unusual growth conditions or depletion of an essential nutrient other than the carbon source (ALDERETE et al, 1993)

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