Characterization of alcohol dehydrogenase from permeabilized brewer's yeast cells immobilized on the derived attapulgite nanofibers.
Alcohol dehydrogenase (ADH) from permeabilized brewer's yeast was immobilized on derived attapulgite nanofibers via glutaraldehyde covalent binding. The effect of immobilization on ADH activity, optimum temperature and pH, thermal, pH and operational stability, reusability of immobilized ADH, and bioreduction of ethyl 3-oxobutyrate (EOB) to ethyl(S)-3-hydroxybutyrate ((S)-EHB) by the immobilized ADH were investigated. The results show the immobilized ADH retained higher activity over wider ranges of pH and temperature than those of the free. The optimum temperature and pH were 7.5 and 35 degrees C, respectively, and 58% of the original activity was retented after incubation at 35 degrees C for 32 h. More importantly, in bioreduction of EOB mediated by immobilized ADH, the conversion of substrate and enantiomeric excess (ee) of product reached 88% and 99.2%, respectively, within 2 h and retained about 42% of the initial activity after eight cycles.
Zhao
Quan
Q
College of Pharmaceutical Sciences, Chongqing Medical University, Chongqing, 400016, China.
Hou
Yi
Y
Gong
Geng-Hao
GH
Yu
Ming-An
MA
Jiang
Lan
L
Liao
Fei
F
D000426:Alcohol Dehydrogenase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D004795:Enzyme Stability; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005976:Glutaral / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D017616:Magnesium Compounds / Q000737:chemistry*; D015394:Molecular Structure; D057139:Nanofibers / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D017655:Silicon Compounds / Q000737:chemistry*; D013696:Temperature
D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D017616:Magnesium Compounds; D017655:Silicon Compounds; D000426:Alcohol Dehydrogenase; D005976:Glutaral; C026325:attapulgite