Grochowski Laura L, Xu Huimin, White Robert H
Department of Biochemistry (0308), Virginia Polytechnic Institute and State University, Blacksburg, Virginia 24061.
J Bacteriol. 2005 Nov;187(21):7382-9. doi: 10.1128/JB.187.21.7382-7389.2005.
Recent work has raised a question as to the involvement of erythrose-4-phosphate, a product of the pentose phosphate pathway, in the metabolism of the methanogenic archaea (R. H. White, Biochemistry 43:7618-7627, 2004). To address the possible absence of erythrose-4-phosphate in Methanocaldococcus jannaschii, we have assayed cell extracts of this methanogen for the presence of this and other intermediates in the pentose phosphate pathway and have determined and compared the labeling patterns of sugar phosphates derived metabolically from [6,6-2H2]- and [U-13C]-labeled glucose-6-phosphate incubated with cell extracts. The results of this work have established the absence of pentose phosphate pathway intermediates erythrose-4-phosphate, xylose-5-phosphate, and sedoheptulose-7-phosphate in these cells and the presence of D-arabino-3-hexulose-6-phosphate, an intermediate in the ribulose monophosphate pathway. The labeling of the D-ara-bino-3-hexulose-6-phosphate, as well as the other sugar-Ps, indicates that this hexose-6-phosphate was the precursor to ribulose-5-phosphate that in turn was converted into ribose-5-phosphate by ribose-5-phosphate isomerase. Additional work has demonstrated that ribulose-5-phosphate is derived by the loss of formaldehyde from D-arabino-3-hexulose-6-phosphate, catalyzed by the protein product of the MJ1447 gene.
最近的研究提出了一个问题,即磷酸戊糖途径的产物4-磷酸赤藓糖是否参与产甲烷古菌的代谢(R. H. 怀特,《生物化学》43:7618 - 7627,2004)。为了探究詹氏产甲烷球菌中可能不存在4-磷酸赤藓糖的情况,我们检测了这种产甲烷菌的细胞提取物中是否存在该物质以及磷酸戊糖途径中的其他中间产物,并确定并比较了与细胞提取物一起孵育的[6,6-2H2]-和[U-13C]-标记的6-磷酸葡萄糖代谢产生的磷酸糖的标记模式。这项工作的结果表明,这些细胞中不存在磷酸戊糖途径的中间产物4-磷酸赤藓糖、5-磷酸木糖和7-磷酸景天庚酮糖,而存在磷酸核糖单磷酸途径的中间产物6-磷酸-D-阿拉伯-3-己酮糖。6-磷酸-D-阿拉伯-3-己酮糖以及其他磷酸糖的标记表明,这种6-磷酸己糖是5-磷酸核酮糖的前体,5-磷酸核酮糖又通过5-磷酸核糖异构酶转化为5-磷酸核糖。进一步的研究表明,5-磷酸核酮糖是由MJ1447基因的蛋白质产物催化6-磷酸-D-阿拉伯-3-己酮糖失去甲醛而产生的。