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Heterologous expression of Cupriavidus sp. USMAA2-4 PHA synthase gene in PHB−4 mutant for the production of poly(3-hydroxybutyrate) and its copolymers

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Abstract

Ecological deterioration and human health concerns arising from the usage of non-biodegradable plastics have prompted mankind to search for greener alternatives which are biodegradable, biocompatible and easily produced from renewable sources. Polyhydroxyalkanoates (PHA), among other biopolymers, are emerging as a viable replacement for fossil fuel-based synthetic plastics. A PHA-producing strain, identified as Cupriavidus sp. (designated Cupriavidus sp. USMAA2-4) was isolated from a soil sample from western peninsular Malaysia. Heterologous expression of the PHA synthase gene (phaC USMAA2-4) in mutant C. necator PHB−4 complemented its PHA-producing ability. More than 60 wt% of P(3HB) was synthesized from various plant oils. The highest P(3HB) production of 2.38 g/l at 68 wt% was attained when crude palm kernel oil was fed as the sole carbon source. The 3HV molar fraction in poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV)] was significantly affected by the type of the precursor used and their respective feeding time. The 3HV molar fraction ranged from 4 to 31 mol% when sodium propionate/valerate was fed at different cultivation times. In addition, with the supplementation of 4HB-monomer precursors, approximately 67 wt% P(3HB-co-4HB) with 4–5 mol% of 4-hydroxybutyrate monomer was synthesized, regardless of the precursor feeding time used. Variation in the molar fraction of the second monomer along with its biodegradability and biocompatibility characteristics promotes the potential of these copolymers as replacements for traditional commodity plastics.

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Acknowledgments

We thank Dr. M. E. Kovach for the pBBR1MCS-2 vector used in this study. We are grateful to Professor Dr. John Leslie for editing this manuscript. This work was supported by a SAGA grant 304/PBIOLOGI/653009/A118 from the Academy of Sciences Malaysia.

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  1. School of Biological Sciences, Universiti Sains Malaysia, 11800, Penang, Malaysia

    Yik-Kang Kek, Choy-Wan Chang, Al-Ashraf Amirul & Kumar Sudesh

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  1. Yik-Kang Kek
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  4. Kumar Sudesh
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Correspondence to Kumar Sudesh.

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Kek, YK., Chang, CW., Amirul, AA. et al. Heterologous expression of Cupriavidus sp. USMAA2-4 PHA synthase gene in PHB−4 mutant for the production of poly(3-hydroxybutyrate) and its copolymers. World J Microbiol Biotechnol 26, 1595–1603 (2010). https://doi.org/10.1007/s11274-010-0335-5

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