Purification and properties of histidinol dehydrogenase from Escherichia coli B.
Histidinol dehydrogenase has been purified from a derepressed mutant of Escherichia coli B. A molecular weight of about 91,000 was estimated by gel filtration. The native enzyme seems to be composed of two similar subunits which have a molecular weight of 52,000 as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The pI of the enzyme as determined by isoelectric focusing is 4.75. The enzyme is maximally active at pH 9.5. It is highly specific for NAD+ and histidinol, with a Km (NAD+) of 0.57 mM and a Km (histidinol) of 14 microM. Mn2+ is required for maximal activity. The enzyme is completely inactivated by 8 M-urea but regains its activity very quickly upon removal of the urea. Mn2+ and histidinol protect the enzyme from heat inactivation.
D000429:Alcohol Oxidoreductases / Q000302:isolation & purification* / Q000378:metabolism; D002413:Cations, Divalent / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D006641:Histidinol / Q000302:isolation & purification / Q000378:metabolism; D006358:Hot Temperature; D007526:Isoelectric Point; D007700:Kinetics; D008970:Molecular Weight; D009243:NAD / Q000378:metabolism; D013379:Substrate Specificity; D014508:Urea / Q000494:pharmacology
D002413:Cations, Divalent; D009243:NAD; D006641:Histidinol; D014508:Urea; D000429:Alcohol Oxidoreductases; C021358:histidinol dehydrogenase