|
Davydova Ye. Ye., Tishkov V. I.
Cloning of D-Amino Acid Oxidase Gene from the Yeast Trigonopsis
Variabilis
Abstract
D-amino-acid oxidase (DAAO) is used as one of the two biocatalysts for
two-step enzymatic conversion of cephalosporin C to 7-aminocephalosporanic
acid, a starting compound for the production of new generation of (3-lactam
antibiotics. The daao gene from T. variabilis was cloned and
expressed in E. coli cells. The cloning procedure included total mRNA
isolation, cDNA preparation by reverse transcriptase and polymerase chain
reaction with spesific primers. cDNA was cloned into E. coli expression
vector, pET23d ("Novagen") under the control of T7 promoter (final
plasmid pKDAOl). It is shown that recombinant E. coli BL21(DE3)/pLysS/pKDA01 strain
can express DAAO protein.
Copyright (C) Chemistry Dept., Moscow State University, 2002
|
|
