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A novel nicotine catabolic plasmid pMH1 in Pseudomonas sp. strain HF-1

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Attempts were made to acquire a plasmid-loss mutant via various methods (spontaneous mutation, SDS, and mitomycin C), among which the method involving mitomycin C (10 µg/mL) has been proven successful. Concomitant with the loss of the plasmid in Pseudomonas sp. strain HF-1, the cured derivative was identified as having a nicotine-negative (Nic-) phenotype, named mutant strain 6-13 (Nic-). After plasmids were transferred from strain HF-1 (named plasmid pMH1) to the mutant strain 6-13, the mutant strain acquired nicotine-degrading ability, called 6-13 transformant (Nic+). There were no differences in growth or nicotine-degrading efficiency between strain HF-1 (wild-type strain) and strain 6-13 transformant. After pMH1 was transferred to Escherichia coli strain Top10 (Nic-), a distant relative of Pseudomonas, it also gained nicotine-degrading ability, showing the highest nicotine degradation efficiency at pH 7.0, the optimal pH for growth of E. coli. The hsp gene, which encodes 6-hydroxy-3-succinoylpyridine hydroxylase, is involved in nicotine degradation in Pseudomonas putida strain S16 and was present in pMH1 but not in pAO1, the well-known nicotine degradation plasmid in Arthrobacter nicotinovorans. It was demonstrated that plasmid pMH1 is a novel nicotine-degrading plasmid.

Des essais visant à obtenir un mutant par perte plasmidique ont été réalisés à l’aide de différentes méthodes (mutation spontanée, SDS, mitomycine C), parmi lesquelles la méthode impliquant la mitomycine C (10 µg/mL) s’est révélée la plus fructueuse. De façon concomitante à la perte de plasmide chez la souche HF-1, la souche curée a été identifiée par le phénotype Nic-, et appelée souche mutante 6-13 (Nic-). Lors du transfert de plasmides de la souche HF-1 (plasmide pMH1) à la souche mutante 6-13, la dégradation de la nicotine a été rétablie chez le transformant 6-13 (Nic+). Il n’y avait pas de différence de croissance et d’efficacité de dégradation de la nicotine entre la souche HF-1 (sauvage) et le transformant 6-13. Lors du transfert de pMH1 à Escherichia coli Top10 (Nic-), un parent éloigné du genre Pseudomonas, la capacité de dégradation de la nicotine était aussi rétablie, l’efficacité de dégradation étant la plus élevée à pH 7, le pH optimal pour la croissance de E. coli. Le gène hsp codant la 6-hydroxy-3-succinlypyridine hydroxylase, impliquée dans la dégradation de la nicotine chez Pseudomonas putida S16, était présent dans le plasmide pMH-1, mais absent du plasmide pAO1, le plasmide bien connu impliqué dans la dégradation de la nicotine chez Arthrobacter nicotinovorans. Ceci démontre que le plasmide pMH1 est un nouveau plasmide impliqué dans le catabolisme de la nicotine.

Document Type: Research Article

Publication date: 2009-03-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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