Abstract
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, involving Pseudomonas, Exiguobacterium, Marinobacter, Citreicella, and Novosphingobium were 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 in Citreicella 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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Acknowledgments
This work was supported by the National Natural Science Foundation of China (41176111), and the Natural Science Foundation of Fujian Province of China (no. 2012 J01049). We thank Professor Zongze Shao for kindly providing strains. We are grateful to Mr. Cao for his kind assistance in phylogenetic analysis.
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The first two authors contributed equally to this work.
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Wen, W., Wang, S., Zhou, X. et al. Central Carbon Metabolism in Marine Bacteria Examined with a Simplified Assay for Dehydrogenases. Appl Biochem Biotechnol 170, 473–482 (2013). https://doi.org/10.1007/s12010-013-0200-8
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DOI: https://doi.org/10.1007/s12010-013-0200-8