Publication Date
7-1976
Advisor(s) - Committee Chair
Martin Houston, Lewis, Lockwood, David Hartman
Degree Program
Department of Biology
Degree Type
Master of Science
Abstract
NADP+-dependent 5-ketogluconate reductase (D-Gluconate:NAD(P)+ oxidoreductase E.C. 1.1.1.69) isozymes were detected in cell-free extracts of Gluconobacter suboxydans (American Type Cultures Collection, Bethesda, Maryland, strain 621). Cell-free enzyme extracts were prepared from cells grown in media which contained glucose, gluconate, glycerol, sorbitol, or mannitol as the sole carbon source in 2% concentration. The extracts were fractionated using analytical polyacrylamide disc-gel electrophoresis. Gels were stained specifically for 5-ketogluconate reductase. Three isozymes were present in each extract from cells grown on the five different carbon sources. Gels stained in a reaction mixture which contained glycerol exhibited two of the isozymes.
An attempt was made to separate the three isozymes using anion-exchange chromatography. A stepwise anion-exchange procedure on DEAE Sephadex resulted in the separation of one of the isozymes from the other two and indicated that this isozyme might actually be two isozymes with the same electrophoretic mobility. Results fro linear gradient anion-exchange chromatography disproved this possibility.
A comparison of results obtained from tracticelation by linear gradient anion-exchange of extracts which had been previously fractionated on Sephadex Gel-100 gel and those which had not, indicated that gel filtration did not improve separation of the isozymes.
Disciplines
Biology | Life Sciences
Recommended Citation
Dianis, Sharon, "Electrophoretic Heterogeneity of Nicotinamide Adenine Dinucleotide Phosphate-Dependent 5-Ketogluconate Reductase of Gluconobacter Suboxydans" (1976). Masters Theses & Specialist Projects. Paper 2257.
https://digitalcommons.wku.edu/theses/2257