FEBS Letters | |
An attempt to transform class characteristics within the alcohol dehydrogenase family | |
Hedberg, Jesper J1  Höög, Jan-Olov1  Strömberg, Patrik1  | |
[1] Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden | |
关键词: Alcohol dehydrogenase; Recombinant protein; Mutagenesis; Enzyme kinetics; | |
DOI : 10.1016/S0014-5793(98)01100-4 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
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-hydroxymethylglutathione and 12-hydroxydodecanoate through increased K m values. Overall results show that it is not possible, by single point mutations, to completely transform enzyme characteristics between these two classes of alcohol dehydrogenase.
【 授权许可】
Unknown
【 预 览 】
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RO201912020306571ZK.pdf | 93KB | download |