Biosynthesis of polyhydroxyalkanoates from styrene by <i>Enterobacter</i> spp. isolated from polluted environment

doi:10.1007/s11515-017-1446-2

Front. Biol.

2017, Vol. 12

Issue (3) : 210-218 https://doi.org/10.1007/s11515-017-1446-2

RESEARCH ARTICLE

Biosynthesis of polyhydroxyalkanoates from styrene by Enterobacter spp. isolated from polluted environment

Arooj Arshad, Bisma Ashraf, Iftikhar Ali(

), Nazia Jamil

Department of Microbiology and Molecular Genetics, University of the Punjab, Quaid-e-Azam Campus, Lahore-54590, Pakistan

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Abstract

BACKGROUND: Styrene and its metabolites are known to have serious adverse effects on human health and hence, strategies to prevent its release, eradicate it from the environment, and understand its route of degradation are being considered.

METHODS: A total of 18 strains were isolated from 4 samples of diesel contaminated soils. Among them 5 strains were selected for their ability to degrade styrene and use it as a sole carbon source to produce PHA. These strains were identified as Enterobacter spp. on the basis of 16S rRNA gene sequencing. Bacteria were screened for their ability to produce PHA by utilizing glucose and styrene as a carbon sources. Screening for PHA production was done by Nile blue A, Sudan black B, and phase contrast microscopy and the selected 3 strains showed positive results. Growth kinetics along with time profiling of PHA was performed for glucose and styrene as carbon sources.

RESULTS:PHA extraction was done at equal intervals of 12 h by sodium hypochlorite method which showed that these strains accumulate maximum amount of PHA after 48 h in glucose (30.60%). FTIR analysis of PHA was done which revealed homopolymer PHB and copolymer (PHB-co-PHV) production in strains by utilizing glucose and styrene. Gas chromatography mass spectrometry was carried out to identify the metabolites produced by bacterial strains grown on styrene. Metabolites of styrene degradation included propyne and phenylalanine. Genomic DNA isolation was carried out to amplify phaC gene which encodes PHA synthase enzyme.

CONCLUSIONS: The conversion of styrene to polyhydroxyalkanoates (PHA) provides a new and unique link between an aromatic environmental pollutant and aliphatic PHA accumulation.

Keywords biodegradable polymers environmental pollutants PHA FTIR recycling bacteria

Corresponding Author(s): Iftikhar Ali

Online First Date: 04 May 2017 Issue Date: 19 June 2017

Cite this article:

Arooj Arshad,Bisma Ashraf,Iftikhar Ali, et al. Biosynthesis of polyhydroxyalkanoates from styrene by Enterobacter spp. isolated from polluted environment[J]. Front. Biol., 2017, 12(3): 210-218.

URL:

https://academic.hep.com.cn/fib/EN/10.1007/s11515-017-1446-2
https://academic.hep.com.cn/fib/EN/Y2017/V12/I3/210

Fig.1 Selection of PHA producers.

Fig.2 Growth curve of PHA producing strains.

Fig.3 (A) FTIR spectra of PHA extracted from Enterobacter strain 3A with glucose as a carbon source. Sharp absorption peaks at 618 cm⁻¹, 972 cm⁻¹, 1403 cm⁻¹, 1658 cm⁻¹, and 2899 cm⁻¹ corresponds to ketones, esters, aromatics, CH₃ and CH₂ functional groups respectively. (B) FTIR spectra of PHA extracted from Enterobacter strain 3A with styrene as a carbon source. Sharp absorption peaks at 601 cm⁻¹, 1110 cm⁻¹, 1411 cm⁻¹, 1750 cm⁻¹ and 2911 cm⁻¹ which corresponds to ketones, esters, aromatics and aliphatic compounds. (C) FTIR spectra of PHA extracted from Enterobacter strain 11 with glucose as a carbon source. Sharp absorption peaks at 624 cm⁻¹, 1095 cm⁻¹, 1408 cm⁻¹, 1630 cm⁻¹, 1735 cm⁻¹ and 2935 cm⁻¹ which corresponds to ketones, esters, aromatics and aliphatic compounds. (D) FTIR spectra of PHA extracted from Enterobacter strain 11 with styrene as a carbon source. Sharp absorption peaks at 615 cm⁻¹, 975 cm⁻¹, 1399 cm⁻¹, 1732 cm⁻¹and 2850 cm⁻¹ which corresponds to ketones, esters, aromatics, primary and secondary amines and aliphatic compounds.

Fig.4 Gas chromatographs of sample from Enterobacter strain 3A (A) and from Enterobacter strain 11 (B). The control sample reported only styrene peak (C).

Tab.1 Identification of isolated bacteria on the basis of 16S rRNA gene sequencing

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