Mechanism of Microbial Degradation of Nicotine

Wada, Einosuke; Yamasaki, Kyoji

doi:10.1126/science.117.3033.152

Cited by 30 publications

(14 citation statements)

References 4 publications

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“…The initial steps of nicotine metabolism (nicotine, PN, and SP) by Pseudomonas strains were confirmed 60 years ago (42). The degrading mechanism at the gene and enzyme levels had not been demonstrated until NicA was found in P. putida strain S16 (18).…”

Section: Discussionmentioning

confidence: 88%

Cloning of a Novel Nicotine Oxidase Gene from Pseudomonas sp. Strain HZN6 Whose Product Nonenantioselectively Degrades Nicotine to Pseudooxynicotine

Qiu

Zhang

et al. 2013

Appl Environ Microbiol

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bPseudomonas sp. strain HZN6 utilizes nicotine as its sole source of carbon, nitrogen, and energy. However, its catabolic mechanism has not been elucidated. In this study, self-formed adaptor PCR was performed to amplify the upstream sequence of the pseudooxynicotine amine oxidase gene. A 1,437-bp open reading frame (designated nox) was found to encode a nicotine oxidase (NOX) that shows 30% amino acid sequence identity with 6-hydroxy-L-nicotine oxidase from Arthrobacter nicotinovorans. The nox gene was cloned into a broad-host-range cloning vector and transferred into the non-nicotine-degrading bacteria Escherichia coli DH5␣ (DH-nox) and Pseudomonas putida KT2440 (KT-nox). The transconjugant KT-nox obtained nicotine degradation ability and yielded an equimolar amount of pseudooxynicotine, while DH-nox did not. Reverse transcription-PCR showed that the nox gene is expressed in both DH5␣ and KT2440, suggesting that additional factors required for nicotine degradation are present in a Pseudomonas strain(s), but not in E. coli. The mutant of strain HZN6 with nox disrupted lost the ability to degrade nicotine, but not pseudooxynicotine. These results suggested that the nox gene is responsible for the first step of nicotine degradation. The (RS)-nicotine degradation results showed that the two enantiomers were degraded at approximately the same rate, indicating that NOX does not show chiral selectivity. Site-directed mutagenesis revealed that both the conserved flavin adenine dinucleotide (FAD)-binding GXGXXG motif and His456 are essential for nicotine degradation activity.

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Section: Discussionmentioning

confidence: 88%

Cloning of a Novel Nicotine Oxidase Gene from Pseudomonas sp. Strain HZN6 Whose Product Nonenantioselectively Degrades Nicotine to Pseudooxynicotine

Qiu

Zhang

et al. 2013

Appl Environ Microbiol

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“…HF-1 (Shao et al 2009). And nicotine could be rapidly catabolized by strain HF-1 into maleamic acid and fumaric acid to supply the energy for its growth via the pyridine pathway (Wada and Yamasaki 1953;Tang et al 2008). Thus, rapid nicotine degradation by strain HF-1 in tobacco wastewater may have alleviated the potential oxidative damage and contributed to shorten the exposure time of other microbes in the SBRs to nicotine.…”

Section: Discussionmentioning

confidence: 98%

Reduction of oxidative stress by bioaugmented strain Pseudomonas sp. HF-1 and selection of potential biomarkers in sequencing batch reactor treating tobacco wastewater

et al. 2010

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Oxidative stress induced by toxic pollutants is generally responsible for the poor performance of many sequencing batch reactors (SBRs) treating organic wastewater. In this study, the oxidative stress in two SBR systems that dealt with tobacco wastewater was monitored by measuring four popular biomarkers (superoxide dismutase, SOD; catalase, CAT; glutathione, GSH; and malondialdehyde, MDA). In the non-BA (non-bio-augmented) system, more intense oxidative stress was induced by a higher concentration of nicotine in tobacco wastewater, and excessive oxidative stress was induced by 250 mg/l of nicotine at the final stage. However, when a nicotine-degrading bacterial strain Pseudomonas sp. HF-1 was added to the BA (bio-augmented) system, the oxidative stress was significantly reduced compared to the non-BA system (p < 0.01).These results suggested that the oxidative stress was mainly induced by nicotine in the SBR treatment of tobacco wastewater, and that bioaugmentation with strain HF-1 would be a potential strategy to reduce the oxidative stress and thereby improve the performance in SBRs. Additionally, the positive correlation between the nicotine content and CAT, GSH and MDA activity in both systems implied that these parameters can be used as biomarkers for reflecting the performance of SBR treatment of tobacco wastewater, and in monitoring nicotine environmental pollution.

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“…The degradation mechanism of nicotine in Pseudomonas has been studied for more than 50 years (21,28). However, there have been few studies at the genetic level reported so far.…”

Section: Discussionmentioning

confidence: 99%

Functional Identification of Two Novel Genes from Pseudomonas sp. Strain HZN6 Involved in the Catabolism of Nicotine

Qiu

Wen

et al. 2012

Appl Environ Microbiol

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Nicotine is a natural alkaloid produced by tobacco plants, and the mechanisms of its catabolism by microorganisms are diverse. In the present study, we reported the mutation, cloning, and identification of two novel genes involved in nicotine degradation from the newly isolated Pseudomonas sp. strain HZN6. Transposon mutagenesis identified a HZN6 mutant in which the nicotine-degrading pathway was blocked at pseudooxynicotine. A 3,874-bp DNA fragment flanking the transposon insertion site was obtained through self-formed adaptor PCR. Two open reading frames (designated pao and sap) were analyzed, and the deduced amino acid sequences shared 29% identity with 6-hydroxy-L-nicotine oxidase from Arthrobacter nicotinovorans and 49% identity with an aldehyde dehydrogenase from Bartonella henselae. Both pao and sap were cloned and functionally expressed in recombinant Escherichia coli BL21. The pao gene encoded a novel pseudooxynicotine amine oxidase with noncovalently bound flavin adenine dinucleotide (FAD) and exhibited substrate specificity removing the methylamine from pseudooxynicotine with the formation of 3-succinoylsemialdehyde-pyridine and hydrogen dioxide. The sap gene encoded a NADP ؉ -dependent 3-succinoylsemialdehyde-pyridine dehydrogenase that catalyzed the dehydrogenation of 3-succinoylsemialdehyde-pyridine to 3-succinoyl-pyridine. Genetic analyses indicated that the pao gene played an essential role in nicotine or pseudooxynicotine mineralization in strain HZN6, whereas the sap gene did not. This study provides novel insight into the nicotine-degrading mechanism at the genetic level in Pseudomonas spp.

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Mechanism of Microbial Degradation of Nicotine

Cited by 30 publications

References 4 publications

Cloning of a Novel Nicotine Oxidase Gene from Pseudomonas sp. Strain HZN6 Whose Product Nonenantioselectively Degrades Nicotine to Pseudooxynicotine

Cloning of a Novel Nicotine Oxidase Gene from Pseudomonas sp. Strain HZN6 Whose Product Nonenantioselectively Degrades Nicotine to Pseudooxynicotine

Reduction of oxidative stress by bioaugmented strain Pseudomonas sp. HF-1 and selection of potential biomarkers in sequencing batch reactor treating tobacco wastewater

Functional Identification of Two Novel Genes from Pseudomonas sp. Strain HZN6 Involved in the Catabolism of Nicotine

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