Central Carbon Metabolism in Marine Bacteria Examined with a Simplified Assay for Dehydrogenases
Central Carbon Metabolism in Marine Bacteria Examined with a Simplified Assay for Dehydrogenases
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使用简化的脱氢酶检测方法检查海洋细菌的中心碳代谢
DOI：
10.1007/s12010-013-0200-8
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发表时间：
2013-04
影响因子：
3
通讯作者：
Fang, Baishan
中科院分区：
文献类型：
--
作者：
Wen, Weiwei;Wang, Shizhen;Zhou, Xiaofen;Fang, Baishan
建立了一种简易的海洋细菌中心碳代谢关键酶活性测定平台。该平台在微孔板分析的基础上，通过统一酶的反应条件（包括温度、缓冲液和离子强度），降低了成本，简化了操作。研究了假单胞菌属、微小杆菌属、海杆菌属、柠檬菌属和新鞘氨醇菌属等16种海洋细菌的中心碳代谢。通过测定葡萄糖-6-磷酸脱氢酶、丙酮酸脱氢酶、2-酮戊二酸脱氢酶、苹果酸脱氢酶、苹果酸酶和异柠檬酸脱氢酶等6种中心碳代谢关键氧化还原酶在该途径中的活性，对它们进行了研究。结果表明，海洋柠檬藻（Citreicella marina）具有较高的苹果酸脱氢酶活性，其粗提液中苹果酸脱氢酶的比活力可达22 U/mg。结果还表明，进化关系较近的菌株在适应其所处生态位的同时，其中心碳代谢关键酶的活性也存在较大的变异性。
A simplified assay platform was developed to measure the activities of the key oxidoreductases in central carbon metabolism of various marine bacteria. Based on microplate assay, the platform was low-cost and simplified by unifying the reaction conditions of enzymes including temperature, buffers, and ionic strength. The central carbon metabolism of 16 marine bacteria, involvingPseudomonas,Exiguobacterium,Marinobacter,Citreicella, andNovosphingobiumwere studied. Six key oxidoreductases of central carbon metabolism, glucose-6-phosphate dehydrogenase, pyruvate dehydrogenase, 2-ketoglutarate dehydrogenase, malate dehydrogenase, malic enzyme, and isocitrate dehydrogenase were investigated by testing their activities in the pathway. High activity of malate dehydrogenase was found inCitreicella marina, and the specific activity achieved 22 U/mg in cell crude extract. The results also suggested that there was a considerable variability on key enzymes’ activities of central carbon metabolism in some strains which have close evolutionary relationship while they adapted to the requirements of the niche they (try to) occupy.
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影响因子：
11.6
作者：
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通讯作者：
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