Direct assay for phosphotransacetylase and acetyl-coenzyme A carboxylase by high-performance liquid chromatography |
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| Authors: | B De Spiegeleer G Mannens G Slegers W Van den Bossche A Claeys |
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| Affiliation: | 1. Department of Pharmaceutical Chemistry and Drug Quality Control, State University of Ghent, Harelbekestraat 72, B-9000 Ghent, Belgium;2. Laboratory of Analytical Chemistry, Faculty of Pharmaceutical Sciences, State University of Ghent, Harelbekestraat 72, B-9000 Ghent, Belgium;1. Department of Chemical Engineering, University of Ulsan, 93 Daehak-Ro, Namgu, Ulsan 44610, Republic of Korea;2. Research Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), 45 Jongga-ro, Jung-gu, Ulsan 44429, Republic of Korea;3. Department of Green Chemistry and Environmental Biotechnology, Korea University of Science and Technology (UST), 217 Gajeong-ro, Yuseong-gu, Daejeon 34144, Republic of Korea;1. Department of Chemical and Biomolecular Engineering, Yonsei University, Seoul 03722, Republic of Korea;2. Integrated Science and Engineering Division, Yonsei University, Seoul 03722, Republic of Korea;1. Institute of Nanotechnology – CNR NANOTEC, Campus Ecotekne, Via Monteroni, 73100 Lecce, Italy;2. Clinical and Experimental Medicine PhD Program, University of Modena and Reggio Emilia, 41125 Modena, Italy;3. Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy;4. Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Genetic Resources for Vegetables, Medicinal and Special Plants, Crop Research Institute, Šlechtitelů 29, 78371 Olomouc, Czech Republic;5. Department of Biochemistry, Faculty of Science, Palacký University, Šlechtitelů 27, 78371 Olomouc, Czech Republic;6. Department of Life Sciences, University of Modena and Reggio Emilia, Via Campi 103, 41125 Modena, Italy;7. Department of Chemistry, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185 Rome, Italy;8. CREA – Research Center for Cereal and Industrial Crops, Via di Corticella 133, 40128 Bologna, Italy;1. Department of Chemistry, Faculty of Basic Science, University of Mohaghegh Ardabili, 56199-11367, Ardabil, Iran;2. Department of Chemistry, Faculty of Science, University of Kurdistan, 66177-15175, Sanandaj, Iran;1. School of Electrical and Computer Engineering, Institute of Information Technology, University of Seoul, 163 Seoulsiripdaero, Dongdaemun-gu, Seoul 02504, Republic of Korea;2. Université de Paris, CNRS, ITODYS, F-75006 Paris, France |
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| Abstract: | ![]()
A simple and specific assay to measure the activity of two coenzyme A derivative-processing enzymes, i.e., phosphotransacetylase (EC 2.3.1.8) and acetyl-coenzyme A carboxylase (EC 6.4.1.2), is described. The assay is based on the HPLC analysis of the short-chain coenzyme A derivatives formed by the enzymatic reaction, viz., acetyl-CoA and malonyl-CoA. For this purpose, ion-pair reversed-phase HPLC conditions are optimized. Furthermore, the influence of several variables on the enzyme reaction is studied in order to get maximum activity. Due to its short analysis time, good selectivity, and chromatogram information, HPLC proves to be an excellent method for the assay of these enzymes. |
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