title: Functional Identification of a Novel Gene, moaE, for 3-Succinoylpyridine Degradation in Pseudomonas putida S16.
abstract: Microbial degradation of N-heterocyclic compounds, including xanthine, quinoline, nicotinate, and nicotine, frequently requires molybdenum hydroxylases. The intramolecular electron transfer chain of molybdenum hydroxylases consists of a molybdenum cofactor, two distinct [2Fe-2S] clusters, and flavin adenine dinucleotide. 3-Succinoylpyridine monooxygenase (Spm), responsible for the transformation from 3-succinoylpyridine to 6-hydroxy-3-succinoylpyridine, is a crucial enzyme in the pyrrolidine pathway of nicotine degradation in Pseudomonas. Our previous work revealed that the heterotrimeric enzyme (SpmA, SpmB, and SpmC) requires molybdopterin cytosine dinucleotide as a cofactor for their activities. In this study, we knocked out four genes, including PPS_1556, PPS_2936, PPS_4063, and PPS_4397, and found that a novel gene, PPS_4397 encoding moaE, is necessary for molybdopterin cytosine dinucleotide biosynthesis. Resting cell reactions of the moaE deletion mutant incubated with 3 g l(-1) nicotine at 30 °C resulted in accumulation of 3-succinoylpyridine, and the strain complemented by the moaE gene regained the ability to convert 3-succinoylpyridine. In addition, reverse transcription-quantitative polymerase chain reaction analysis indicated that the transcriptional levels of the genes of moaE, spmA, and spmC of Pseudomonas putida S16 were distinctly higher when grown in nicotine medium than in glycerol medium.
authors: State Key Laboratory of Microbial Metabolism, and School of Life Sciences &Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.|Joint International Research Laboratory of Metabolic &Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
authors: Yi
authors: Y
authors: Jiang
authors: State Key Laboratory of Microbial Metabolism, and School of Life Sciences &Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.|Joint International Research Laboratory of Metabolic &Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
authors: Hongzhi
authors: H
authors: Tang
authors: State Key Laboratory of Microbial Metabolism, and School of Life Sciences &Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.|Joint International Research Laboratory of Metabolic &Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
authors: Geng
authors: G
authors: Wu
authors: State Key Laboratory of Microbial Metabolism, and School of Life Sciences &Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.|Joint International Research Laboratory of Metabolic &Developmental Sciences, Shanghai Jiao Tong University, Shanghai 200240, People's Republic of China.
authors: Ping
authors: P
authors: Xu
mesh_terms: D001673:Biodegradation, Environmental; D004789:Enzyme Activation; D009538:Nicotine / Q000378:metabolism*; D016958:Pseudomonas putida / Q000378:metabolism*; D011725:Pyridines / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D013386:Succinates / Q000378:metabolism*; D013466:Sulfurtransferases / Q000378:metabolism*
chemical_list: C507268:6-hydroxy-3-succinoyl-pyridine; D011725:Pyridines; D013386:Succinates; D009538:Nicotine; D013466:Sulfurtransferases; C085585:molybdopterin synthase