Map of the IncP1β plasmid pTSA encoding the widespread genes (tsa) for p-toluenesulfonate degradation in Comamonas testosteroni T-2

• Published in: Applied and environmental microbiology 2001-04, Vol.67 (4), p.1508-1516
• Main Authors: TRALAU, Tewes, COOK, Alasdair M, RUFF, Juergen
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Biology of microorganisms of confirmed or potential industrial interest; Biotechnology; Comamonas testosteroni; Fundamental and applied biological sciences. Psychology; Genetics; Genetics and Molecular Biology; insertion sequence IS1071; Mission oriented research; p-toluenesulfonate; p-Toluenesulfonic acid; tsa gene; tsa operon
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.67.4.1508-1516.2001

The catabolic IncP1β plasmid pTSA from Comamonas testosteroni T-2 was mapped by subtractive analysis of restriction digests, by sequencing outwards from the tsa operon (toluenesulfonate degradation), and by generating overlapping, long-distance-PCR amplification products. The plasmid was estimated to comprise 72 ± 4 kb. The tsa region was found to be a composite transposon flanked by two IS 1071 elements. A cryptic tsa operon was also present in the tsa transposon. Those backbone genes and regions which we sequenced were in the same order as the corresponding genes in resistance plasmid R751, and identities of about 99% were observed. Enrichment cultures with samples from four continents were done to obtain organisms able to utilize p -toluenesulfonate as the sole source of carbon and energy for aerobic growth. Most (15) of the 16 cultures (13 of them isolates) were obtained from contaminated sites and were attributed to three metabolic groups, depending on their metabolism of p -toluenesulfonate. The largest group contained the tsa transposon, usually (six of seven isolates) with negligible differences in sequence from strain T-2.