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Applied Microbiology and Biotechnology

, Volume 102, Issue 23, pp 10245–10257 | Cite as

Rhizobacter gummiphilus NS21 has two rubber oxygenases (RoxA and RoxB) acting synergistically in rubber utilisation

  • Jakob Birke
  • Wolf Röther
  • Dieter JendrossekEmail author
Environmental biotechnology
  • 191 Downloads

Abstract

Biodegradation of poly(cis-1,4-isoprene) (rubber) by Gram-negative bacteria has been investigated on the enzymatic level only in Steroidobacter cummioxidans 35Y (previously Xanthomonas sp. 35Y). This species produces two kinds of rubber oxygenases, RoxA35Y and RoxB35Y, one of which (RoxB35Y) cleaves polyisoprene to a mixture of C20- and higher oligoisoprenoids while the other (RoxA35Y) cleaves polyisoprene and RoxB35Y-derived oligoisoprenoids to the C15-oligoisoprenoid 12-oxo-4,8-dimethyltrideca-4,8-diene-1-al (ODTD). ODTD can be taken up by S. cummioxidans and used as a carbon source. Gram-positive rubber-degrading bacteria employ another type of rubber oxygenase, latex clearing protein (Lcp), for the initial oxidative attack of the polyisoprene molecule. In this contribution, we examined which type of rubber oxygenase is present in the only other well-documented Gram-negative rubber-degrading species, Rhizobacter gummiphilus NS21. No homologue for an Lcp protein but homologues for a putative RoxA and a RoxB protein (the latter identical to a previously postulated LatA-denominated rubber cleaving enzyme) were identified in the genome of strain NS21. The roxANS21 and roxBNS21 genes were separately expressed in a ∆roxA35Y/∆roxB35Y background of S. cummioxidans 35Y and restored the ability of the mutant to produce oligoisoprenoids. The RoxANS21 and RoxBNS21 proteins were each purified and biochemically characterised. The results—in combination with in silico analysis of databases—indicate that Gram-negative rubber-degrading bacteria generally utilise two synergistically acting rubber oxygenases (RoxA/RoxB) for efficient cleavage of polyisoprene to ODTD.

Keywords

Rubber oxygenase Latex clearing protein Polyisoprene Biodegradation Haem dioxygenase 

Notes

Acknowledgements

We thank Weber and Schaer company (Hamburg) for providing polyisoprene and PreSens (Regensburg) for sensor spots.

Funding

This work was supported by a grant of the Deutsche Forschungsgemeinschaft to D J. (JE 152 18-1).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethic approval

This article does not contain any studies with human participants or animals performed by any of the authors.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Institute of MicrobiologyUniversity of StuttgartStuttgartGermany
  2. 2.Novartis Pharma Stein AGSteinSwitzerland

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