Arica, MYHalicigil, CAlaeddinoglu, GDenizli, A2020-06-252020-06-251999closedAccess1359-5113https://hdl.handle.net/20.500.12587/2823A methylotrophic hydroxypyruvate reductase was partially purified and characterized from Methylophilus spp. using the biomimetic dye, Cibacron Blue F3FA attached to poly(HEMA-EGDMA) microspheres. The absorption capacities of the dye-affinity microspheres were determined by changing pH and the concentration of the proteins in the adsorption medium. Hydroxypyruvate reductase was desorbed from the dye-affinity support specifically with 2 mM NADH solution. The enzyme was purified 10.4-fold with 47% yield. The molecular mass and subunit molecular mass of the enzyme was estimated to be 75 kDa and 37 kDa on the basis of its mobility in polyacrylamide and SDS-polyacrylamide gels, respectively. This suggested a homogeneous dimer structure. The optimal pH was between 5.0 and 7.0, and the maximum enzyme activity was obtained at 50 degrees C. The K-m values of hydroxpyruvate reductase were 0.222 mM for hydroxpyruvate and 0.067 mM for NADH. (C) 1999 Elsevier Science Ltd. All rights reserved.eninfo:eu-repo/semantics/closedAccesshydroxypyruvate reductaseNADH dependent enzymeMethylophilus spp.affinity adsorptionpoly(HEMA-EGDMA) microspheresCibacron Blue F3GAArticle3443753812-s2.0-0344224217Q1WOS:000081089900008Q1