An attempt to transform class characteristics within the alcohol dehydrogenase family

作者：Hedberg JJ; Stromberg P; Hoog JO^*

来源：FEBS Letters, 1998, 436(1): 67-70.

DOI：10.1016/S0014-5793(98)01100-4

• alcohol dehydrogenase
• recombinant protein
• mutagenesis
• enzyme kinetics

Human class I alcohol dehydrogenase was mutated at positions 57 and 115, exchanging for Asp and Arg respectively, in an attempt to introduce glutathione-dependent formaldehyde dehydrogenase characteristics. In addition, class III alcohol dehydrogenase, identical to glutathione-dependent formaldehyde dehydrogenase, was mutated at position 115, introducing Ser or Lys. The attempted class transformation was partly successful considering a higher affinity for 12-hydroxydodecanoate and a lower affinity for ethanol that was monitored for the class I mutant. However, the class I mutant displayed neither glutathione-dependent formaldehyde dehydrogenase activity nor fatty acid activation of alcohol oxidation, Interestingly, both class III mutants showed reduced activities for S-hydroxymethyl-glutathione and 12-hydroxydodecanoate through increased K-m values. Overall results show that

• 出版日期1998-9-25