Drosophila lebanonensis ADH: analysis of recombinant wild‐type enzyme and site‐directed mutants

FEBS Letters

Atrian, Sílvia¹  Gonzàlez-Duarte, Roser¹  Albalat, Ricard¹ 

[1] Departament de Genètica, Facultat de Biologia, Universitat de Barcelona, Av. Diagonal 645, E-08028 Barcelona, Spain

关键词: Short-chain dehydrogenase;    Alcohol dehydrogenase;    Drosophila lebanonensis;    Site-directed mutagenesis;    Catalytic efficiency;   

DOI  :  10.1016/0014-5793(94)80451-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Unique amino acid substitutions occur in D. lebanonensis ADH. They are found within the putative NAD + -binding domain and affect residues that are otherwise highly conserved in all other species of the genus. To restore the consensus ainino acids, we have constructed an expression system for this enzyme in E. coli, and engineered two mutants, Ala¹³ Gly and Asn⁵⁶ Thr. The biochemical and kinetic features of these retromutants are consistent with increased catalytic efficiency and thermal stability. Thus, results show that wild-type D. lebanonensis ADH can be improved by site-directed mutagenesis.

【 授权许可】

Unknown