A novel nicotine catabolic plasmid pMH1 in Pseudomonas sp strain HF-1
A novel nicotine catabolic plasmid pMH1 in Pseudomonas sp strain HF-1
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DOI：
10.1139/w08-135
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发表时间：
2009-03-01
影响因子：
2.8
通讯作者：
Lv, Zhenmei
中科院分区：
文献类型：
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作者：
Wang, Meizhen;Yang, Guiqin;Lv, Zhenmei
我们尝试通过多种方法（自发突变、SDS、丝裂霉素C）获得质粒丢失突变体，其中丝裂霉素C （10 μ g/mL）的方法获得了成功。随着假单胞菌菌株HF-1的质粒丢失，固化衍生物被鉴定为尼古丁阴性（Nic(-)）表型，命名为突变菌株6-13 （Nic(-)）。将质粒从菌株HF-1（命名为质粒pMH1）转移到突变菌株6-13后，突变菌株获得了尼古丁降解能力，称为6-13转化（Nic(+)）。菌株HF-1（野生型）与菌株6-13的生长和尼古丁降解效率无显著差异。pMH1转移到假单胞菌的远亲大肠杆菌菌株Top10 （Nic(-)）中后，也获得了尼古丁降解能力，在pH 7.0时尼古丁降解效率最高，pH 7.0是大肠杆菌生长的最佳pH。编码6-羟基-3-琥珀酰吡啶羟化酶的hsp基因参与了腐烂假单胞菌菌株S16的尼古丁降解，该基因存在于pMH1中，而不存在于烟酸节杆菌中众所周知的尼古丁降解质粒pAO1中。结果表明，pMH1质粒是一种新型的尼古丁降解质粒。
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 mu g/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.