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Appl. Environ. Microbiol. doi:10.1128/AEM.02145-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

The genomes of the non-clear zone forming and natural rubber-degrading species Gordonia polyisoprenivorans and Gordonia westfalica harbor genes expressing Lcp (latex clearing protein) activity in Streptomyces strains

Institut für Molekulare Mikrobiologie und Biotechnologie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 3, D-48149 Münster, Germany

^* To whom correspondence should be addressed. Email: steinbu{at}uni-muenster.de.

	Abstract

The latex clearing protein (Lcp_K30) from the rubber-degrading bacterium Streptomyces sp. strain K30 is involved in the cleavage of poly(cis-1,4-isoprene) yielding isoprenoid aldehydes and ketones. Lcp homologues were so far detected in all investigated clear zone forming rubber-degrading bacteria. Internal degenerated oligonucleotides derived from lcp genes of Streptomyces sp. strain K30 (lcp_K30), S. coelicolor strain A3(2), Nocardia farcinica strains IFM10152 and E1 were applied in PCR to investigate whether lcp homologues occur also in the non clear zone forming rubber-utilizing bacteria Gordonia polyisoprenivorans strains VH2 and Y2K, G. alkanivorans strain 44187 and G. westfalica strain Kb1 which grow adhesively on rubber. The 1230- and 1224-bp lcp homologous genes from G. polyisoprenivorans strain VH2 (lcp_VH2) and G. westfalica strain Kb1 (lcp_Kb1) were obtained after screening genomic libraries by degenerated PCR amplification, and their translational products exhibited 50 and 52% amino acid identity, respectively, to Lcp_K30. Recombinant lcp_VH2 and lcp_Kb1 harboring cells of the non-rubber degrading S. lividans strain TK23 were able to form clear zones and aldehydes on latex overlay-agar plates, thus indicating that lcp_VH2 and lcp_Kb1 encode functionally active proteins. Analysis by gel permeation chromatography demonstrated lower polymer concentrations and molecular weights of the remaining polyisoprenoid molecules after incubation with these recombinant S. lividans strains. RT-PCR analysis demonstrated that lcp_VH2 was transcribed in cells of G. polyisoprenivorans strain VH2 cultivated in presence of poly(cis-1,4-isoprene) but not in presence of sodium acetate. Anti-Lcp_K30 IgGs, which were raised in this study, were rather specific for Lcp_K30 and did not cross react with Lcp_VH2 and Lcp_Kb1. A lcp_VH2 disruption mutant was still able to grow with poly(cis-1,4-isoprene) as sole carbon source; therefore, lcp_VH2 seems not to be essential for rubber degradation in G. polyisoprenivorans.

This article has been cited by other articles:

• Yikmis, M., Arenskotter, M., Rose, K., Lange, N., Wernsmann, H., Wiefel, L., Steinbuchel, A. (2008). Secretion and Transcriptional Regulation of the Latex-Clearing Protein, Lcp, by the Rubber-Degrading Bacterium Streptomyces sp. Strain K30. Appl. Environ. Microbiol. 74: 5373-5382 [Abstract] [Full Text]