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嗜热栖热菌P2丙醇代谢的蛋白质组分析

Proteome analysis of Sulfolobus solfataricus P2 propanol metabolism.

作者信息

Chong Poh Kuan, Burja Adam M, Radianingtyas Helia, Fazeli Alireza, Wright Phillip C

机构信息

Biological and Environmental Systems Group, Department of Chemical and Process Engineering, University of Sheffield, Mappin Street, Sheffield S1 3JD, UK.

出版信息

J Proteome Res. 2007 Apr;6(4):1430-9. doi: 10.1021/pr060575g. Epub 2007 Feb 22.

Abstract

Sulfolobus solfataricus P2 is able to metabolize n-propanol as the sole carbon source. An average n-propanol consumption rate of 9.7 and 3.3 mg/L/hr was detected using GC-MS analysis from S. solfataricus cultures grown in 0.40 and 0.16% w/v n-propanol, respectively. The detection of propionaldehyde, the key intermediate of n-propanol degradation, produced at a rate of 1.3 and 1.0 mg/L/hr in 0.40 and 0.16% w/v n-propanol cultures, further validated the ability of S. solfataricus to utilize n-propanol. The translational and transcriptional responses of S. solfataricus grown on n-propanol versus glucose were also investigated using quantitative RT-PCR and iTRAQ approaches. Approximately 257 proteins with > or =2 MS/MS spectra were identified and quantified via iTRAQ. The global quantitative proteome overview obtained showed significant up-regulation of acetyl-CoA synthetases, propionyl-CoA carboxylase, and methylmalonyl-CoA mutase enzymes. This led to the proposition that the propionyl-CoA formed from n-propanol degradation is catabolised into the citrate cycle (central metabolism) via succinyl-CoA intermediates. In contrast, evidence obtained from these analysis approaches and in vivo stable isotope labeling experiments, suggests that S. solfataricus is only capable of converting isopropyl alcohol to acetone (and vice versa) but lacks the ability to further metabolize these compounds.

摘要

嗜热栖热菌P2能够将正丙醇作为唯一碳源进行代谢。使用气相色谱 - 质谱联用(GC - MS)分析,分别在0.40%和0.16%(w/v)正丙醇中培养的嗜热栖热菌培养物中,检测到的平均正丙醇消耗速率分别为9.7和3.3毫克/升/小时。正丙醇降解的关键中间体丙醛在0.40%和0.16%(w/v)正丙醇培养物中的生成速率分别为1.3和1.0毫克/升/小时,这进一步证实了嗜热栖热菌利用正丙醇的能力。还使用定量逆转录 - 聚合酶链反应(RT - PCR)和同位素标记相对和绝对定量(iTRAQ)方法研究了嗜热栖热菌在正丙醇与葡萄糖上生长时的翻译和转录反应。通过iTRAQ鉴定并定量了约257种具有≥2个串联质谱(MS/MS)谱图的蛋白质。获得的全球定量蛋白质组概况显示,乙酰辅酶A合成酶、丙酰辅酶A羧化酶和甲基丙二酰辅酶A变位酶显著上调。这表明由正丙醇降解形成的丙酰辅酶A通过琥珀酰辅酶A中间体分解代谢进入柠檬酸循环(中心代谢)。相比之下,从这些分析方法和体内稳定同位素标记实验获得的证据表明,嗜热栖热菌仅能够将异丙醇转化为丙酮(反之亦然),但缺乏进一步代谢这些化合物的能力。

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