Biodegradation and metabolic pathway of nicotine in Rhodococcus sp. Y22.
Nicotine in tobacco is harmful to health and the environment, so there is an environmental requirement to remove nicotine from tobacco and tobacco wastes. In this study, the biotransformation of nicotine by Rhodococcus sp. Y22 was investigated, and three metabolites (NIC1, NIC4 and NIC5) were isolated by column separation, preparative TLC and solid plate's method, respectively. NIC1 was identified as 6-hydoxynicotine based on the results of NMR, MS, HPLC-UV and HRESIMS analysis; NIC4 was a novel compound and identified as 5-(3-methyl-[1,3]oxazinan-2-ylidene)-5H-pyridin-2-one based on the results of NMR, MS and UV analysis; NIC5 was identified as nicotine blue based on the results of NMR and MS analysis. Meanwhile, two metabolites NIC2 and NIC3 were identified as 6-hydroxy-N-methylmyosmine and 6-hydroxypseudooxynicotine by HRESIMS analysis, respectively. According to these metabolites, the possible pathway of nicotine degradation by Rhodococcus sp. Y22 was proposed. The nicotine can be transformed to nicotine blue through two pathways (A and B), and 6-hydroxy-N-methylmyosmine is the key compound, which can be converted to 6-hydroxypseudooxynicotine (pathway A) and 5-(3-methyl-[1,3]oxazinan-2-ylidene)-5H-pyridin-2-one (pathway B), respectively. Moreover, the encoding gene of nicotine dehydrogenase, ndh, was amplified from Rhodococcus sp. Y22, and its transcriptional level could be up-regulated obviously under nicotine induction. Our studies reported the key metabolites and possible biotransformation pathway of nicotine in Rhodococcus sp. Y22, and provided new insights into the microbial metabolism of nicotine.
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan University, Kunming, 650091, People's Republic of China.|R & D Center of China Tobacco Yunnan Industrial Co., Ltd., Kunming, 650024, People's Republic of China.
Xiaowei
X
Gong
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan University, Kunming, 650091, People's Republic of China.|Yunnan Comtestor Co., Ltd., Kunming, 650106, People's Republic of China.
Guanghui
G
Ma
R & D Center of China Tobacco Yunnan Industrial Co., Ltd., Kunming, 650024, People's Republic of China.
Yanqing
Y
Duan
R & D Center of China Tobacco Yunnan Industrial Co., Ltd., Kunming, 650024, People's Republic of China.
Donglai
D
Zhu
R & D Center of China Tobacco Yunnan Industrial Co., Ltd., Kunming, 650024, People's Republic of China.
Yongkuan
Y
Chen
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan University, Kunming, 650091, People's Republic of China.
Ke-Qin
KQ
Zhang
State Key Laboratory for Conservation and Utilization of Bio-Resources in Yunnan, Yunnan University, Kunming, 650091, People's Republic of China. jinkui960@ynu.edu.cn.
Jinkui
J
Yang
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001673:Biodegradation, Environmental; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D055432:Metabolomics; D009538:Nicotine / Q000031:analogs & derivatives / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D012240:Rhodococcus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D014026:Nicotiana / Q000737:chemistry
C502317:6-hydroxynicotine; D001426:Bacterial Proteins; D009538:Nicotine
Biodegradation; Metabolites; Nicotine; Nicotine dehydrogenase; Rhodococcus sp. Y22
Biotechnol Lett. 2004 Oct;26(19):1493-6
15604785
Arch Microbiol. 2008 May;189(5):511-7
18071673
Mol Chem Neuropathol. 1996 May-Aug;28(1-3):3-11
8871936
Mol Microbiol. 1994 Sep;13(5):929-36
7815950
Biodegradation. 2014 Feb;25(1):95-107
23604517
J Biol Chem. 2011 Nov 11;286(45):39179-87
21949128
PLoS Genet. 2013 Oct;9(10):e1003923
24204321
Appl Environ Microbiol. 2009 Feb;75(3):772-8
19060159
Methods. 2001 Dec;25(4):402-8
11846609
Res Microbiol. 2011 Apr;162(3):285-91
21288482
Nucleic Acids Res. 2005 Jul 07;33(12):e107
16002786
J Biol Chem. 1960 Mar;235:795-9
14402193
ScientificWorldJournal. 2013 Dec 29;2013:125385
24470788
J Bacteriol. 2003 Mar;185(6):1976-86
12618462
Appl Environ Microbiol. 2015 Jan;81(1):272-81
25344232
Appl Microbiol Biotechnol. 1999 Jun;51(6):786-93
10422226
Arch Mikrobiol. 1973 Mar 26;90(2):167-9
4707595
J Mol Evol. 2013 Aug;77(1-2):22-30
23884627
J Mol Biol. 1998 Dec 18;284(5):1323-39
9878353
Res Microbiol. 2010 Sep;161(7):626-33
20600861
J Appl Microbiol. 2009 Sep;107(3):838-47
19486412
Res Microbiol. 2009 Apr;160(3):200-4
19298855
Appl Microbiol Biotechnol. 2012 Sep;95(6):1567-78
22466953
Appl Microbiol Biotechnol. 2015 May;99(9):3775-85
25805341
Proc Natl Acad Sci U S A. 2000 Nov 7;97(23 ):12493-7
11050152
Arch Mikrobiol. 1969;65(2):98-104
5384624
Appl Microbiol Biotechnol. 2006 Jan;69(5):493-8
16333621
Curr Opin Biotechnol. 2005 Jun;16(3):282-90
15961029
Microbiology. 2007 May;153(Pt 5):1556-65
17464070
Sci Rep. 2012;2:377
22530095
PLoS One. 2014 Jul 23;9(7):e103324
25054198
FEBS J. 2006 Apr;273(7):1528-36
16689938