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Purification and characterization of diacetyl reductase from Kluyveromyces marxianus
Author(s) -
Schwarz J.G.,
Hang Y.D.
Publication year - 1994
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.1994.tb00867.x
Subject(s) - chromatofocusing , diacetyl , kluyveromyces marxianus , chemistry , isoelectric point , chromatography , methylglyoxal , biochemistry , size exclusion chromatography , enzyme , acetoin , ammonium , ion chromatography , column chromatography , yeast , fermentation , organic chemistry , saccharomyces cerevisiae
Diacetyl reductase from Kluyveromyces marxianus NRRL Y‐1196 was purified 27.5‐fold with a yield of 13% by ammonium sulphate fractionation, DEAE‐anion exchange chromatography, hydroxyapatite chromatography and chromatofocusing. The purified enzyme was most active at pH 7.0 and exhibited optimal activity at 40°C. The K m and V max values for diacetyl were 2.5 mmol 1 ‐1 and 0.026 mmol 1 ‐1 min ‐1 , respectively. The enzyme did not react with monoaldehydes or monoketones, but reduced acetoin, diacetyl and methylglyoxal with NADH as a cofactor. The enzyme had an isoelectric point (pl) of pH 5.8, and its molecular weight was 50 kDa.