Qiu Jiguo, Zhang Junjie, Zhang Yanting, Wang Yuhong, Tong Lu, Hong Qing, He Jian
Key Laboratory of Agricultural Environmental Microbiology, Ministry of Agriculture, College of Life Sciences, Nanjing Agricultural University, Nanjing, 210095, China.
Nanjing Yuanheng Institute for Environmental Studies Co., Ltd., Nanjing, 210049, China.
Curr Microbiol. 2017 Apr;74(4):508-514. doi: 10.1007/s00284-017-1205-2. Epub 2017 Feb 27.
We isolated a bacterial strain JQ135 from municipal wastewater, which was capable of efficiently degrading picolinic acid (PA). Based on the physico-biochemical characteristics and 16S rDNA analysis, strain JQ135 was identified as Alcaligenes faecalis. In addition, strain JQ135 produced an orange pigment when cultured in the Luria-Bertani medium, which is different from the previously reported strains of A. faecalis. During the degradation of PA by the resting strain JQ135 cells, only one intermediate, 6-hydroxypicolinic acid (6HPA), was detected by ultraviolet spectrophotometry, high-pressure liquid chromatography, and liquid chromatography-mass spectrometry. A random transposon mutagenesis library of strain JQ135 was constructed. One mutant, Mut-G31, could convert PA into 6HPA without further degradation. The disrupted gene (orf2) was amplified from Mut-G31, and its product showed 32% identity to the 3-deoxy-D-manno-octulosonic acid kinase (KdkA) from Haemophilus influenzae. Results from complementation analysis confirmed that GTG was the initiation codon of the kdkA-like orf2, and that it was essential for PA biodegradation by strain JQ135. This study provides the first genetic evidence for the bacterial degradation of PA.
我们从城市污水中分离出一株细菌菌株JQ135,它能够高效降解吡啶甲酸(PA)。基于生理生化特征和16S rDNA分析,菌株JQ135被鉴定为粪产碱菌。此外,菌株JQ135在Luria-Bertani培养基中培养时会产生橙色色素,这与先前报道的粪产碱菌菌株不同。在静止的菌株JQ135细胞降解PA的过程中,通过紫外分光光度法、高压液相色谱法和液相色谱-质谱法仅检测到一种中间产物6-羟基吡啶甲酸(6HPA)。构建了菌株JQ135的随机转座子诱变文库。一个突变体Mut-G31能够将PA转化为6HPA而不再进一步降解。从Mut-G31中扩增出中断的基因(orf2),其产物与流感嗜血杆菌的3-脱氧-D-甘露糖辛酸激酶(KdkA)具有32%的同源性。互补分析结果证实GTG是类kdkA的orf2的起始密码子,并且它对于菌株JQ135对PA的生物降解至关重要。本研究为细菌降解PA提供了首个遗传学证据。