Lundgren Benjamin R, Harris Joshua R, Sarwar Zaara, Scheel Ryan A, Nomura Christopher T
1Department of Chemistry, State University of New York - College of Environmental Science and Forestry, 1 Forestry Drive, Syracuse, New York, 13210, USA.
1Department of Chemistry, State University of New York - College of Environmental Science and Forestry, 1 Forestry Drive, Syracuse, New York, 13210, USA 2Center for Applied Microbiology, State University of New York - College of Environmental Science and Forestry, 1 Forestry Drive, Syracuse, New York, 13210, USA.
Microbiology (Reading). 2015 Nov;161(Pt 11):2232-42. doi: 10.1099/mic.0.000163. Epub 2015 Aug 25.
A variety of soil-dwelling bacteria produce polyhydroxybutyrate (PHB), which serves as a source of energy and carbon under nutrient deprivation. Bacteria belonging to the genus Pseudomonas do not generally produce PHB but are capable of using the PHB degradation product (R)-3-hydroxybutyrate [(R)-3-HB] as a growth substrate. Essential to this utilization is the NAD+-dependent dehydrogenase BdhA that converts (R)-3-HB into acetoacetate, a molecule that readily enters central metabolism. Apart from the numerous studies that had focused on the biochemical characterization of BdhA, there was nothing known about the assimilation of (R)-3-HB in Pseudomonas, including the genetic regulation of bdhA expression. This study aimed to define the regulatory factors that govern or dictate the expression of the bdhA gene and (R)-3-HB assimilation in Pseudomonas aeruginosa PAO1. Importantly, expression of the bdhA gene was found to be specifically induced by (R)-3-HB in a manner dependent on the alternative sigma factor RpoN and the enhancer-binding protein PA2005.This mode of regulation was essential for the utilization of (R)-3-HB as a sole source of energy and carbon. However, non-induced levels of bdhA expression were sufficient for P. aeruginosa PAO1 to grow on ( ± )-1,3-butanediol, which is catabolized through an (R)-3-HB intermediate. Because this is, we believe, the first report of an enhancer-binding protein that responds to (R)-3-HB, PA2005 was named HbcR for (R)-3-hydroxybutyrate catabolism regulator.
多种土壤细菌可产生聚羟基丁酸酯(PHB),在营养缺乏时作为能量和碳源。假单胞菌属的细菌一般不产生PHB,但能够利用PHB降解产物(R)-3-羟基丁酸酯[(R)-3-HB]作为生长底物。这种利用的关键是NAD⁺依赖性脱氢酶BdhA,它将(R)-3-HB转化为乙酰乙酸,乙酰乙酸可轻易进入中心代谢。除了众多关注BdhA生化特性的研究外,关于假单胞菌中(R)-3-HB的同化作用,包括bdhA表达的基因调控,一无所知。本研究旨在确定调控铜绿假单胞菌PAO1中bdhA基因表达和(R)-3-HB同化作用的调控因子。重要的是,发现bdhA基因的表达由(R)-3-HB特异性诱导,其方式依赖于替代σ因子RpoN和增强子结合蛋白PA2005。这种调控模式对于将(R)-3-HB用作唯一能量和碳源至关重要。然而,bdhA的非诱导表达水平足以使铜绿假单胞菌PAO1在(±)-1,3-丁二醇上生长,(±)-1,3-丁二醇通过(R)-3-HB中间体进行分解代谢。因为我们认为这是首个关于对(R)-3-HB作出反应的增强子结合蛋白的报道,所以将PA2005命名为HbcR,即(R)-3-羟基丁酸酯分解代谢调节因子。