Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada.
Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada.
Biochim Biophys Acta Gen Subj. 2019 Feb;1863(2):304-312. doi: 10.1016/j.bbagen.2018.10.018. Epub 2018 Nov 1.
The camphor-degrading microorganism, Pseudomonas putida strain ATCC 17453, is an aerobic, gram-negative soil bacterium that uses camphor as its sole carbon and energy source. The genes responsible for the catabolic degradation of camphor are encoded on the extra-chromosomal CAM plasmid. A monooxygenase, cytochrome P450, mediates hydroxylation of camphor to 5-exo-hydroxycamphor as the first and committed step in the camphor degradation pathway, requiring a dioxygen molecule (O) from air. Under low O levels, P450 catalyzes the production of borneol via an unusual reduction reaction. We have previously shown that borneol downregulates the expression of P450. To understand the function of P450 and the consequences of down-regulation by borneol under low O conditions, we have studied chemotaxis of camphor induced and non-induced P. putida strain ATCC 17453. We have tested camphor, borneol, oxidized camphor metabolites and known bacterial attractants (d)-glucose, (d) - and (l)-glutamic acid for their elicitation chemotactic behavior. In addition, we have used 1-phenylimidazole, a P450 inhibitor, to investigate if P450 plays a role in the chemotactic ability of P. putida in the presence of camphor. We found that camphor, a chemoattractant, became toxic and chemorepellent when P450 was inhibited. We have also evaluated the effect of borneol on chemotaxis and found that the bacteria chemotaxed away from camphor in the presence of borneol. This is the first report of the chemotactic behaviour of P. putida ATCC 17453 and the essential role of P450 in this process.
具有降解樟脑能力的微生物 Pseudomonas putida 菌株 ATCC 17453 是一种需氧、革兰氏阴性土壤细菌,以樟脑作为其唯一的碳源和能源。负责樟脑分解代谢的基因编码在染色体外的 CAM 质粒上。一种单加氧酶,细胞色素 P450,介导樟脑的羟化作用,将其转化为 5-外消旋羟基樟脑,作为樟脑降解途径的第一步和关键步骤,需要来自空气的一个氧分子 (O)。在低 O 水平下,P450 通过异常还原反应催化龙脑的生成。我们之前已经表明,龙脑醇通过下调 P450 的表达。为了了解 P450 的功能以及在低 O 条件下龙脑醇下调的后果,我们研究了诱导和非诱导的 Pseudomonas putida 菌株 ATCC 17453 的樟脑诱导的趋化性。我们已经测试了樟脑、龙脑醇、氧化樟脑代谢物和已知的细菌趋化剂 (d)-葡萄糖、(d) -和 (l)-谷氨酸的趋化行为。此外,我们还使用了 1-苯基咪唑,一种 P450 抑制剂,来研究 P450 是否在存在樟脑的情况下发挥了 Pseudomonas putida 趋化能力的作用。我们发现,作为一种趋化剂的樟脑在 P450 被抑制时变得有毒和化学排斥。我们还评估了龙脑醇对趋化作用的影响,发现细菌在龙脑醇存在的情况下从樟脑中趋化。这是首次报道 Pseudomonas putida ATCC 17453 的趋化行为以及 P450 在这个过程中的重要作用。