Abstract
Publisher Summary This chapter focuses on the assay method, purification procedure, and properties of D-galactitol-6-phosphate dehydrogenase. This inducible enzyme functions in the catabolism of galactitol in Klebsiella pneumonia . The rate of D-galactitol 6-phosphate-dependent nicotinamide adenine dinucleotide dehydrogenase (NADH) formation is determined by measuring the rate of absorbance increase at 340 nm. The bacterial strain used is Klebsiella pneumoniae PRL-R3. The cells are harvested by centrifugation and then washed with 0.85% NaCl. The steps involved in the purification of D-galactitol-6-phosphate dehydrogenase are: (1) the preparation of cell extracts, (2) diethylaminoethyl (DEAE)-cellulose chromatography, (3) ammonium sulfate fractionation, and (4) Sephadex g-150 chromatography. D-Galactitol 6-phosphate and D-tagatose 6-phosphate are the only known substrates. No activity is detected when nicotinamide adenine dinucleotide kinase (NADP + ) and nicotinamide adenine dinucleotide phosphate dehydrogenase (NADPH) are substituted for NAD + and NADH, respectively, with any of the sugar phosphates tested. D-galactitol-6-phosphate dehydrogenase in the Sephadex G-150 fractions retains most of its activity for at least 2 weeks when is stored at 0–20°C.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
