Identification of nicotine biotransformation intermediates by Agrobacterium tumefaciens strain S33 suggests a novel nicotine degradation pathway

Wang, Shuning; Huang, Haiyan; Xie, Kebo; Xu, Ping

doi:10.1007/s00253-012-4007-2

Cited by 52 publications

(62 citation statements)

References 34 publications

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“…The 2,5-DHP is converted via a ring cleavage reaction to render N-formylmaleamic acid, maleamic acid, maleic acid, and fumaric acid (13). In addition to the pyridine and pyrrolidine pathways, some other nicotine degradation pathways were also proposed, including a variant of the pyridine and pyrrolidine pathways, which is designated the VPP pathway in bacteria (10,11). In the upper VPP pathway, nicotine is converted to 6HPON through 6HN and 6HMM, just like in the pyridine pathway.…”

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confidence: 99%

“…Many microorganisms that use various nicotine degradation pathways have been isolated from the environment, including Arthrobacter (6), Pseudomonas (7,8), Ochrobactrum (9), Agrobacterium (10), Shinella (11), and Aspergillus (12). The pathways of nicotine degradation have been widely investigated by researchers.…”

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confidence: 99%

“…Two strains, Agrobacterium tumefaciens strain S33 and Shinella sp. strain HZN7, with the VPP pathway, were isolated and characterized recently (10,11). However, the molecular mechanisms involved were rarely reported.…”

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Molecular Mechanism of Nicotine Degradation by a Newly Isolated Strain, Ochrobactrum sp. Strain SJY1

Tang

Zhu

et al. 2015

Appl Environ Microbiol

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A newly isolated strain, SJY1, identified as Ochrobactrum sp., utilizes nicotine as a sole source of carbon, nitrogen, and energy. Strain SJY1 could efficiently degrade nicotine via a variant of the pyridine and pyrrolidine pathways (the VPP pathway), which highlights bacterial metabolic diversity in relation to nicotine degradation. A 97-kbp DNA fragment containing six nicotine degradation-related genes was obtained by gap closing from the genome sequence of strain SJY1. Three genes, designated vppB, vppD, and vppE, in the VPP pathway were cloned and heterologously expressed, and the related proteins were characterized. The vppB gene encodes a flavin-containing amine oxidase converting 6-hydroxynicotine to 6-hydroxy-N-methylmyosmine. Although VppB specifically catalyzes the dehydrogenation of 6-hydroxynicotine rather than nicotine, it shares higher amino acid sequence identity with nicotine oxidase (38%) from the pyrrolidine pathway than with its isoenzyme (6-hydroxy-L-nicotine oxidase, 24%) from the pyridine pathway. The vppD gene encodes an NADH-dependent flavin-containing monooxygenase, which catalyzes the hydroxylation of 6-hydroxy-3-succinoylpyridine to 2,5-dihydroxypyridine. VppD shows 62% amino acid sequence identity with the hydroxylase (HspB) from Pseudomonas putida strain S16, whereas the specific activity of VppD is ϳ10-fold higher than that of HspB. VppE is responsible for the transformation of 2,5-dihydroxypyridine. Sequence alignment and phylogenetic analysis suggested that the VPP pathway, which evolved independently from nicotinic acid degradation, might have a closer relationship with the pyrrolidine pathway. The proteins and functional pathway identified here provide a sound basis for future studies aimed at a better understanding of molecular principles of nicotine degradation. N icotine, which easily passes biological membranes, is significantly harmful to humans and is considered a potentially addictive drug (1, 2). Large quantities of tobacco waste with high concentrations of nicotine are generated annually by cigarette and cigar manufacturing (3). Due to its water solubility, nicotine can easily spread in the environment, emerging as a threat to public health (3). As a result, the U.S. Environmental Protection Agency classified nicotine as a "toxic release inventory" chemical in 1994 (4). Microbes play significant roles in removing nicotine from the environment, and in the meantime, they serve as the catalysts for biotransformation. Nicotine could potentially be used for the production of value-added chemicals. 6-Hydroxy-3-succinoylpyridine (HSP), an important precursor for the synthesis of compounds with biological activities, was efficiently produced by an engineered nicotine-degrading strain (5). Therefore, the study of the mechanism of nicotine degradation provides useful information for both bioremediation and biocatalysis.Many microorganisms that use various nicotine degradation pathways have been isolated from the environment, including Arthrobacter (6), Pseudomonas (7, 8), Ochroba...

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Molecular Mechanism of Nicotine Degradation by a Newly Isolated Strain, Ochrobactrum sp. Strain SJY1

Tang

Zhu

et al. 2015

Appl Environ Microbiol

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“…The enzyme was purified 17.6-fold to give a yield of 13.4% and a specific activity of 5.1 U/mg. The molecular mass of the purified enzyme was estimated to be 45 kDa by SDS-PAGE (Figure 1), similar to that of HSP hydroxylases from Pseudomonas putida S16 and Agrobacterium tumefaciens S33 [23,24]. Table 1.…”

Section: Purification and Identification Of Hsphzzmentioning

confidence: 97%

“…The optimum temperature and pH of HSPH ZZ were determined to be 30 • C and 8.5, respectively (Figure 3). The pH optimum was higher than that of reported HSP hydroxylases from P. putida S16 and A. tumefaciens S33 (pH 8.0) [23,24]. The thermostability of HSPH ZZ was evaluated at three different temperatures (30 • C, 35 • C, and 40 • C) with increasing incubation times up to 120 min.…”

Section: Effect Of Temperature and Ph On Activity Of The Recombinant mentioning

confidence: 99%

Characterization of a Novel Nicotine Hydroxylase from Pseudomonas sp. ZZ-5 That Catalyzes the Conversion of 6-Hydroxy-3-Succinoylpyridine into 2,5-Dihydroxypyridine

et al. 2017

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A novel nicotine hydroxylase was isolated from Pseudomonas sp. ZZ-5 (HSPH ZZ ). The sequence encoding the enzyme was 1206 nucleotides long, and encoded a protein of 401 amino acids. Recombinant HSPH ZZ was functionally overexpressed in Escherichia coli BL21-Codon Plus (DE3)-RIL cells and purified to homogeneity after Ni-NTA affinity chromatography. Liquid chromatography-mass spectrometry (LC-MS) analyses indicated that the enzyme could efficiently catalyze the conversion of 6-hydroxy-3-succinoylpyridine (HSP) into 2,5-dihydroxypyridine (2,5-DHP) and succinic acid in the presence of nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FAD). The kinetic constants (K m , k cat , and k cat /K m ) of HSPH ZZ toward HSP were 0.18 mM, 2.1 s −1 , and 11.7 s −1 mM −1 , respectively. The optimum temperature, pH, and optimum concentrations of substrate and enzyme for 2,5-DHP production were 30 • C, 8.5, 1.0 mM, and 1.0 µM, respectively. Under optimum conditions, 85.3 mg/L 2,5-DHP was produced in 40 min with a conversion of 74.9%. These results demonstrated that HSPH ZZ could be used for the enzymatic production of 2,5-DHP in biotechnology applications.

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Isolation and characterization of a novel nicotinophilic bacterium, Arthrobacter sp. aRF‐1 and its metabolic pathway

Ruan

Gao

Fang

et al. 2018

Biotech and App Biochem

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Nicotine is a potent parasympathomimetic stimulant and an important natural alkaloid mainly found in the Nicotiana genus of plants. It can directly threaten ecological security and human health in tobacco waste, wastewater, and other forms of tobacco production. Therefore, it is the basis of nicotine pollution prevention and of great application value to explore efficient and a wide range of nicotinophilic bacteria for tobacco industry and environmental protection. In this study, one nicotinophilic bacterium was isolated from the soil, which accumulated tobacco waste over 50 years at a Hefei cigarette factory. The strain was named aRF-1, which was identified as Arthrobacter sp. by analysis. The nicotine degradation tests showed that the optimum temperature for cell growth and metabolism of nicotine of Arthrobacter sp. aRF-1 was 30 °C, and the optimum initial pH value was about 7.0. Under the optimum experimented conditions, it can tolerance nicotine concentration as high as 8 g·L . The highest removal rate of nicotine was 93.8% in 72 H in nonsterilization contaminated soil by Arthrobacter sp. aRF-1. LC-MS/MS was used to analyze the nicotine metabolic intermediates of strain Arthrobacter sp. aRF-1. A total of nine major metabolites that were detected were able to metabolize nicotine along a variant pathway of pyridine and pyrrolidine, and there may be more than two nicotine metabolic pathways for Arthrobacter sp. aRF-1 through the analysis of the main intermediate products.

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Identification of nicotine biotransformation intermediates by Agrobacterium tumefaciens strain S33 suggests a novel nicotine degradation pathway

Cited by 52 publications

References 34 publications

Molecular Mechanism of Nicotine Degradation by a Newly Isolated Strain, Ochrobactrum sp. Strain SJY1

Molecular Mechanism of Nicotine Degradation by a Newly Isolated Strain, Ochrobactrum sp. Strain SJY1

Characterization of a Novel Nicotine Hydroxylase from Pseudomonas sp. ZZ-5 That Catalyzes the Conversion of 6-Hydroxy-3-Succinoylpyridine into 2,5-Dihydroxypyridine

Isolation and characterization of a novel nicotinophilic bacterium, Arthrobacter sp. aRF‐1 and its metabolic pathway

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