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绿针假单胞菌P3中3-羟基苯甲酸降解基因的鉴定与分析

Identification and Analysis of 3-Hydroxybenzoic Acid Degradation Gene in Pseudomonas chlororaphis P3

  • 摘要: 3-羟基苯甲酸(3-hydroxybenzoic acid, 3-HBA)是一种具有工业应用价值的平台化合物,在微生物代谢中也起到重要的作用。绿针假单胞菌P3是一株稳定的吩嗪-1-甲酰胺高产突变株,具有良好生理生化特性,可以作为代谢工程改造的平台菌株。本研究通过探究P3菌株在不同浓度3-HBA中的生长状况,发现其对3-HBA的耐受性较高,能够在3-HBA过量积累的环境下生长。此外,绿针假单胞菌P3可以利用3-HBA为唯一碳源生长,表明P3菌株中存在3-HBA降解基因。通过序列比较分析,我们找到了3-HBA的降解基因;经外源表达和全细胞催化,鉴定3-HBA的降解产物为龙胆酸,证明了绿针假单胞菌P3通过龙胆酸途径降解3-HBA。本研究揭示了绿针假单胞菌P3中的3-HBA降解途径,为研究其他芳香族化合物的降解途径和假单胞菌异源合成并积累3-HBA奠定了基础。

     

    Abstract: 3-Hydroxybenzoic acid(3-HBA) is a platform chemical with high practical application value, and plays an important role in microbial metabolism. Pseudomonas chlororaphis P3 is a high yield of phenazine-1-carboxamide mu tant strain with outstanding physiological and biochemical characteristics, which can be developed as a platform organism via metabolic engineering. In this research, the growth of P3 strain under different concentrations of3-HBA was evaluated, finding that P3 strain showed tolerance of growing in the 3-HBA over-accumulated environment. In addition, P3 strain could be cultured when applying 3-HBA as the sole carbon source, indicating that 3-HBA can be degraded. After sequence alignment, a gene had been identified to catalyze the degradation of 3-HBA. It was certificated that 3-HBA was degraded by gentisate pathway in P3 strain through exogenous expression and whole cell catalysis. This research revealed a degradation pathway of 3-HBA in P. chlororaphis P3, which laid a foundation for biodegradation of other aromatic compounds as well as exogenous synthesis and accumulation of 3-HBA in Pseudomonas.

     

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