Solimonas fluminis has an active latex-clearing protein
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The utilization of rubber (poly (cis-1,4-isoprene)) by rubber-degrading bacteria depends on the synthesis of rubber oxygenases that cleave the polymer extracellularly to low molecular weight products that can be taken up and used as a carbon source. All so far described Gram-negative rubber-degrading species use two related ≈ 70 kDa rubber oxygenases (RoxA and RoxB) for the primary attack of rubber while all described Gram-positive rubber-degrading strains use RoxA/RoxB-unrelated latex-clearing proteins (Lcps, ≈ 40 kDa) as rubber oxygenase(s). In this study, we identified an lcp orthologue in a Gram-negative species (Solimonas fluminis). We cloned and heterologously expressed the lcp gene of S. fluminis HR-BB, purified the corresponding Lcp protein (LcpHR-BB) from recombinant Escherichia coli, and biochemically characterised the LcpHR-BB activity. LcpHR-BB cleaved polyisoprene to a mixture of C20 and higher oligoisoprenoids at a specific activity of 1.5 U/mg. Furthermore, spectroscopic investigation identified LcpHR-BB as a b-haem-containing protein with an oxidised, fivefold coordinated (open) haem centre. To the best of our knowledge, this is the first report that Gram-negative bacteria can have an active rubber oxygenase of the Lcp type.
KeywordsRubber oxygenase Latex-clearing protein Polyisoprene Biodegradation Haem dioxygenase
We thank the Weber and Schaer Company (Hamburg) for providing polyisoprene.
This work was supported by a grant of the Deutsche Forschungsgemeinschaft to D. J.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Arenskötter M, Baumeister D, Berekaa MM, Pötter G, Kroppenstedt RM, Linos A, Steinbüchel A (2001) Taxonomic characterization of two rubber degrading bacteria belonging to the species Gordonia polyisoprenivorans and analysis of hyper variable regions of 16S rDNA sequences. FEMS Microbiol Lett 205:277–282CrossRefGoogle Scholar
- Chia K-H, Nanthini J, Thottathil GP, Najimudin N, Haris MRHM, Sudesh K (2014) Identification of new rubber-degrading bacterial strains from aged latex. Polym Degrad Stab 109:354–361. https://doi.org/10.1016/j.polymdegradstab.2014.07.027 CrossRefGoogle Scholar
- Coenen A, Oetermann S, Steinbüchel A (2019) Identification of LcpRBA3(2), a novel regulator of lcp expression in Streptomyces coelicolor A3(2). Appl Microbiol Biotechnol 7:e50562–e50512. https://doi.org/10.1007/s00253-019-09896-8
- Heisey RM, Papadatos S (1995) Isolation of microorganisms able to metabolize purified natural rubber. Appl Environ Microbiol 61:3092–3097Google Scholar
- Hiessl S, Schuldes J, Thuermer A, Halbsguth T, Broeker D, Angelov A, Liebl W, Daniel R, Steinbüchel A (2012) Involvement of two latex-clearing proteins during rubber degradation and insights into the subsequent degradation pathway revealed by the genome sequence of Gordonia polyisoprenivorans strain VH2. Appl Environ Microbiol 78:2874–2887. https://doi.org/10.1128/AEM.07969-11 CrossRefGoogle Scholar
- Ibrahim E, Arenskötter M, Luftmann H, Steinbüchel A (2006) Identification of poly (cis-1,4-isoprene) degradation intermediates during growth of moderately thermophilic actinomycetes on rubber and cloning of a functional lcp homologue from Nocardia farcinica strain E1. Appl Environ Microbiol 72:3375–3382. https://doi.org/10.1128/AEM.72.5.3375-3382.2006
- Imai S, Ichikawa K, Muramatsu Y, Kasai D, Masai E, Fukuda M (2011) Isolation and characterization of Streptomyces, Actinoplanes, and Methylibium strains that are involved in degradation of natural rubber and synthetic poly (cis-1,4-isoprene). Enzym Microb Technol 49:526–531. https://doi.org/10.1016/j.enzmictec.2011.05.014 CrossRefGoogle Scholar
- Linh DV, Huong NL, Tabata M, Imai S, Iijima S, Kasai D, Anh TK, Fukuda M (2017) Characterization and functional expression of a rubber degradation gene of a Nocardia degrader from a rubber-processing factory. J Biosci Bioeng 123:412–418. https://doi.org/10.1016/j.jbiosc.2016.11.012 CrossRefGoogle Scholar
- Nanthini J, Ong SY, Sudesh K (2017) Identification of three homologous latex-clearing protein (lcp) genes from the genome of Streptomyces sp. strain CFMR 7. Gene 628:146–155. https://doi.org/10.1016/j.gene.2017.07.039
- Rose K, Tenberge KB, Steinbüchel A (2005) Identification and characterization of genes from Streptomyces sp. strain K30 responsible for clear zone formation on natural rubber latex and poly (cis-1,4-isoprene) rubber degradation. Biomacromolecules 6:180–188. https://doi.org/10.1021/bm0496110 CrossRefGoogle Scholar
- Sharma V, Siedenburg G, Birke J, Mobeen F, Jendrossek D, Prakash T (2018) Metabolic and taxonomic insights into the Gram-negative natural rubber degrading bacterium Steroidobacter cummioxidans sp. nov., strain 35Y. PloS One 13(5):e0197448. https://doi.org/10.1371/journal.pone.0197448 CrossRefGoogle Scholar
- Tsuchii A, Takeda K (1990) Rubber-degrading enzyme from a bacterial culture. Appl Environ Microbiol 56:269–274Google Scholar
- Warneke S, Arenskötter M, Tenberge KB, Steinbüchel A (2007) Bacterial degradation of poly (trans-1,4-isoprene) (gutta percha). Microbiology (Reading, Engl) 153:347–356. https://doi.org/10.1099/mic.0.2006/000109-0
- Watcharakul S, Röther W, Birke J, Umsakul K, Hodgson B, Jendrossek D (2016) Biochemical and spectroscopic characterization of purified latex clearing protein (Lcp) from newly isolated rubber degrading Rhodococcus rhodochrous strain RPK1 reveals novel properties of Lcp. BMC Microbiol 16:92. https://doi.org/10.1186/s12866-016-0703-x CrossRefGoogle Scholar