Skip to main content

Table 2 Purification of the major NADPH-dependent alcohol dehydrogenase AdhB of Thermoanaerobacter sp. strain X514

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 512 2.5 1293 100 1.0
Cytoplasm 449 2.6 1150 89 1.0
Q-Sepharose 337 4.8 1599 123 1.9
(NH4)2SO4 precipitation 268 6.6 1772 137 2.6
Phenyl-Sepharose 86.8 14.8 1284 99 5.8
Superose 6 9.2 13.3 122 9.4 5.2
Blue-Sepharose 2.5 35.2 87.3 6.8 13.8
  1. NADPH-dependent ADH activity was measured in 50 mM Tris buffer (pH 7.5) supplemented with 2 mM DTE and 4 µM as NADPH (0.5 mM)-dependent reduction of acetaldehyde (10 mM) at 340 nm. The decrease in absorption of NADPH was followed at 340 nm. One unit represents one µmol of acetaldehyde reduced per minute