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Biochemical characterization of a new type of intracellular PHB depolymerase from Rhodospirillum rubrum with high hydrolytic activity on native PHB granules

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Abstract

A Rhodospirillum rubrum gene that is predicted to code for an extracellular poly(3-hydroxybutyrate) (PHB) depolymerase by the recently published polyhydroxyalkanoates (PHA) depolymerase engineering database was cloned. The gene product (PhaZ3 Rru ) was expressed in recombinant E. coli, purified and biochemically characterized. PhaZ3 Rru turned out, however, to share characteristics of intracellular PHB depolymerases and revealed a combination of properties that have not yet been described for other PHB depolymerases. A fusion of PhaZ3 Rru with the enhanced cyan fluorescent protein was able to bind to PHB granules in vivo and supported the function as an intracellular PHB depolymerase. Purified PhaZ3 Rru was specific for short-chain-length polyhydroxyalkanoates (PHASCL) and hydrolysed both untreated native PHB granules as well as trypsin-activated native PHB granules to a mixture of mono- and dimeric 3-hydroxybutyrate. Crystalline (denatured) PHB granules were not hydrolysed by PhayZ3 Rru . Low concentrations of calcium or magnesium ions (1–5 mM) reversibly (EDTA) inhibited the enzyme. Our data suggest that PhaZ3 Rru is the representative of a new type of the growing number of intracellular PHB depolymerases.

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Acknowledgements

This work was supported by the Deutsche Forschungsgemeinschaft (DFG).

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  1. Institut für Mikrobiologie, Universität Stuttgart, Allmandring 31, 70550, Stuttgart, Germany

    Anna Sznajder & Dieter Jendrossek

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  1. Anna Sznajder
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  2. Dieter Jendrossek
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Correspondence to Dieter Jendrossek.

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Sznajder, A., Jendrossek, D. Biochemical characterization of a new type of intracellular PHB depolymerase from Rhodospirillum rubrum with high hydrolytic activity on native PHB granules. Appl Microbiol Biotechnol 89, 1487–1495 (2011). https://doi.org/10.1007/s00253-011-3096-7

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  • DOI: https://doi.org/10.1007/s00253-011-3096-7

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