Bobik T A, Xu Y, Jeter R M, Otto K E, Roth J R
Department of Microbiology and Cell Science, University of Florida, Gainesville 32611, USA.
J Bacteriol. 1997 Nov;179(21):6633-9. doi: 10.1128/jb.179.21.6633-6639.1997.
The propanediol utilization (pdu) operon of Salmonella typhimurium encodes proteins required for the catabolism of propanediol, including a coenzyme B12-dependent propanediol dehydratase. A clone that expresses propanediol dehydratase activity was isolated from a Salmonella genomic library. DNA sequence analysis showed that the clone included part of the pduF gene, the pduABCDE genes, and a long partial open reading frame (ORF1). The clone included 3.9 kbp of pdu DNA which had not been previously sequenced. Complementation and expression studies with subclones constructed via PCR showed that three genes (pduCDE) are necessary and sufficient for propanediol dehydratase activity. The function of ORF1 was not determined. Analyses showed that the S. typhimurium propanediol dehydratase was related to coenzyme B12-dependent glycerol dehydratases from Citrobacter freundii and Klebsiella pneumoniae. Unexpectedly, the S. typhimurium propanediol dehydratase was found to be 98% identical in amino acid sequence to the Klebsiella oxytoca propanediol dehydratase; this is a much higher identity than expected, given the relationship between these organisms. DNA sequence analyses also supported previous studies indicating that the pdu operon was inherited along with the adjacent cobalamin biosynthesis operon by a single horizontal gene transfer.
鼠伤寒沙门氏菌的丙二醇利用(pdu)操纵子编码丙二醇分解代谢所需的蛋白质,包括一种依赖辅酶B12的丙二醇脱水酶。从沙门氏菌基因组文库中分离出一个表达丙二醇脱水酶活性的克隆。DNA序列分析表明,该克隆包含pduF基因的一部分、pduABCDE基因以及一个长的部分开放阅读框(ORF1)。该克隆包含3.9 kbp以前未测序的pdu DNA。通过PCR构建的亚克隆的互补和表达研究表明,三个基因(pduCDE)对于丙二醇脱水酶活性是必需且足够的。ORF1的功能尚未确定。分析表明,鼠伤寒沙门氏菌的丙二醇脱水酶与弗氏柠檬酸杆菌和肺炎克雷伯菌中依赖辅酶B12的甘油脱水酶相关。出乎意料的是,发现鼠伤寒沙门氏菌的丙二醇脱水酶与产酸克雷伯菌的丙二醇脱水酶在氨基酸序列上有98%的同一性;考虑到这些生物体之间的关系,这一同一性比预期的要高得多。DNA序列分析也支持了先前的研究,表明pdu操纵子是通过一次水平基因转移与相邻的钴胺素生物合成操纵子一起遗传的。