Production and separation of formate dehydrogenase from Candida boidinii

摘要: Abstract The production of FDH in a 40-l bioreactor was carried out with methanol as the inducer to reach final intracellular activity 35 U/g cell. Among different permeabilization methods studied, treatment toluene at relatively small amount resulted cells highest activity. Crude cell extract obtained by ultrasonically breaking down wall treated polyethyleneimine (PEI) separate from other proteins. By adding PEI low concentration 0.04 mg/ml extract, ∼50% proteins formed aggregates and precipitated; however, not these remained solution. After treatment, specific increased 1.6-fold. SDS-PAGE analysis showed that precipitation removed some impurity protein molecules cannot be separated affinity chromatography Sepharose-Procion Blue HERB separation ligand, thus improved efficiency. adsorbed column eluted KCl Adding 5 mM NAD + 0.2 M elution 1.38-fold compared 1 M KCl. Overall, dye chromatographic process high recovery yield 56% 5.5-fold increase crude extract.