Isolation, Purification and Partial Characterization of Alcohol Dehydrogenase from Pig Liver

doi:10.7506/spkx1002-6630-201517034

Abstract

Abstract:

Electrophoresis-purity alcohol dehydrogenase (ADH) from pig liver was obtained through homogenization, buffer
extraction, ammonium sulfate fractionation, DEAE-Sepharose ion-exchange chromatography and Superdex-200 gel filtration
chromatography. Results showed that the specific activity of the purified ADH was 1 622.33 U/mg with an activity recovery
of 29.05% and a purification fold of 34.58. The relative molecular weight of the ADH was approximately 171.79 kD, in
which the subunit molecular mass was roughly 43.68 kD. The enzymatic properties showed that the optimum temperature and pH
for the ADH were 45 ℃ and 10.0, respectively. The enzyme was stable at 25–45 ℃ and pH 7.5–9.0, and its apparent Km towards
ethanol was 19 mmol/L. The enzyme activity of ADH could be strongly inhibited by n-butanol, chloroform, isopropanol, sodium
dodecyl sulfate, oxalic acid, Zn2+, Cu2+, and Ag+, and activated by Mg2+. EDTA had a dual effect on this enzyme.

Key words: pig liver, alcohol dehydrogenase, isolation and purification, enzymatic properties

CLC Number:

Q946.5

FU Ting, WANG Dan, WAN Ji, TANG Yunming. Isolation, Purification and Partial Characterization of Alcohol Dehydrogenase from Pig Liver[J]. FOOD SCIENCE, doi: 10.7506/spkx1002-6630-201517034.

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Isolation, Purification and Partial Characterization of Alcohol Dehydrogenase from Pig Liver

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