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首页> 外文期刊>Journal of bacteriology >PduL Is an Evolutionarily Distinct Phosphotransacylase Involved in B12-Dependent 1,2-Propanediol Degradation by Salmonella enterica Serovar Typhimurium LT2
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PduL Is an Evolutionarily Distinct Phosphotransacylase Involved in B12-Dependent 1,2-Propanediol Degradation by Salmonella enterica Serovar Typhimurium LT2

机译：PduL是一种进化上独特的磷酸转酰酶，参与由肠炎沙门氏菌血清型鼠伤寒沙门氏菌LT2依赖于B12的1,2-丙二醇降解。

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Salmonella enterica degrades 1,2-propanediol (1,2-PD) in a coenzyme B12-dependent manner. Previous enzymatic assays of crude cell extracts indicated that a phosphotransacylase (PTAC) was needed for this process, but the enzyme involved was not identified. Here, we show that the pduL gene encodes an evolutionarily distinct PTAC used for 1,2-PD degradation. Growth tests showed that pduL mutants were unable to ferment 1,2-PD and were also impaired for aerobic growth on this compound. Enzyme assays showed that cell extracts from a pduL mutant lacked measurable PTAC activity in a background that also carried a pta mutation (the pta gene was previously shown to encode a PTAC enzyme). Ectopic expression of pduL corrected the growth defects of a pta mutant. PduL fused to eight C-terminal histidine residues (PduL-His8) was purified, and its kinetic constants were determined: the Vmax was 51.7 ± 7.6 μmol min?1 mg?1, and the Km values for propionyl-PO42? and acetyl-PO42? were 0.61 and 0.97 mM, respectively. Sequence analyses showed that PduL is unrelated in amino acid sequence to known PTAC enzymes and that PduL homologues are distributed among at least 49 bacterial species but are absent from the Archaea and Eukarya.
机译：沙门氏菌以辅酶B 12 依赖性方式降解1,2-丙二醇（1,2-PD）。以前对粗细胞提取物进行的酶分析表明，该过程需要磷酸转酰酶（PTAC），但未鉴定到所涉及的酶。在这里，我们显示 pduL 基因编码用于1，2-PD降解的进化上独特的PTAC。生长测试表明， pduL 突变体无法发酵1,2-PD，并且也对该化合物的有氧生长不利。酶法检测表明， pduL 突变体的细胞提取物在可携带 pta 突变的背景中缺乏可测量的PTAC活性（ pta 基因以前是（显示编码PTAC酶）。 pduL 的异位表达纠正了 pta 突变体的生长缺陷。纯化融合到八个C末端组氨酸残基（PduL-His 8 ）的PduL，并确定其动力学常数： V max 为丙酰-PO 的51.7±7.6μmolmin ？1 mg ？1 和 K m 值> 4 2？和乙酰基-PO 4 2？分别为0.61和0.97 mM。序列分析表明，PduL的氨基酸序列与已知的PTAC酶无关，并且PduL的同源物分布在至少49种细菌中，而 Archaea 和 Eukarya 中却没有。

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