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Table 1 Purification of the major NADH-dependent alcohol dehydrogenase AdhE of Thermoanaerobacter sp. strain X514. NADH-dependent ADH activity was measured in 50 mM Tris buffer (pH 7.5) supplemented with 2 mM DTE and 4 µM resazurin as NADH (0.5 mM)-dependent reduction of acetaldehyde (10 mM) at 65 °C

From: Alcohol dehydrogenases AdhE and AdhB with broad substrate ranges are important enzymes for organic acid reduction in Thermoanaerobacter sp. strain X514

Purification step Total protein
[mg]
Specific activity
[U mg−1]
Total activity
[U]
Yield
[%]
Purification
[fold]
Cell-free extract 1423 1.2 1734 100.0 1.0
Cytoplasm 1196 1.8 2183 125.9 1.5
Q-Sepharose 877 2.7 2323 134.0 2.2
(NH4)2SO4 precipitation 511 4.7 2413 139.2 3.9
Phenyl-Sepharose 220 7.3 1602 92.4 6.0
Superose 6 2.8 61.2 173 9.9 50.2
  1. The decrease in absorption of NADH was followed at 340 nm. One unit represents one µmol of acetaldehyde reduced per minute. The major ALDH activity (CoA and NAD+-dependent oxidation of acetaldehyde) co-purified due to the bifunctionality of AdhE (Additional file 1: Table S1)