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Production and Purification of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Degrading Enzyme from Streptomyces sp. AF-111

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Abstract

A poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) degrading bacterial strain designated as AF-111 was isolated from sewage sludge sample. The bacterium was identified by 16S rRNA gene sequencing. The results revealed that strain AF-111 showed 99 % similarity with Streptomyces althioticus strain NRRL B-3981 and designated as Streptomyces sp. strain AF-111. An extracellular PHBV depolymerase enzyme was produced under optimized conditions and purified through ammonium sulphate fractionation and column chromatography. The enzyme was purified to homogeneity, indicated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis and molecular weight was found to be approximately 51 kDa. Effect of temperature, pH, metal ions and inhibitors on the PHBV depolymerase activity was determined. The enzyme was stable at wide range of temperature (35–55 °C) and pH (6–8). PHBV depolymerase was stable in the presence of different metal ions except iron and zinc which had inhibitory effect on depolymerase activity. Both ethylenediamine teteracetic acid and phenylmethyl sulphonyl fluoride strongly inhibited enzyme activity which indicates that this enzyme belongs to the serine hydrolase family like other polyhydroxyalkanoate depolymerases. The results show that a depolymerase from strain AF-111 can effectively degrade PHBV, therefore, it can be applied in the process of biochemical monomer recycling.

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Acknowledgments

We are very grateful to Professor Toshiaki Nakajima-Kambe, Bioindustrial Sciences Division, Graduate School of Life and Environmental Sciences, University of Tsukuba, Japan, for providing the facility for identification of the actinomycetes strain through 16S rRNA sequencing.

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Authors and Affiliations

  1. Department of Microbiology, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, 45320, Pakistan

    Siddiq Akbar, Fariha Hasan, Akhtar Nadhman, Samiullah Khan & Aamer Ali Shah

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  1. Siddiq Akbar
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  2. Fariha Hasan
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  3. Akhtar Nadhman
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  4. Samiullah Khan
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  5. Aamer Ali Shah
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Correspondence to Aamer Ali Shah.

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Akbar, S., Hasan, F., Nadhman, A. et al. Production and Purification of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Degrading Enzyme from Streptomyces sp. AF-111. J Polym Environ 21, 1109–1116 (2013). https://doi.org/10.1007/s10924-013-0600-4

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  • DOI: https://doi.org/10.1007/s10924-013-0600-4

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