||Cloning and Expression of a Lignin Peroxidase Gene from 'Streptomyces viridosporus' in 'Streptomyces lividans'.
Wang, Z. ;
Bleakley, B. H. ;
Crawford, D. L. ;
Hertel, G. ;
Rafii., F. ;
||Idaho Univ., Moscow. Dept. of Bacteriology and Biochemistry.;Corvallis Environmental Research Lab., OR.;Department of Energy, Washington, DC.;National Science Foundation, Washington, DC.
||EPA-R-815300-01-0 ;DE-FG786ER13586; EPA/600/J-92/173;
Molecular cloning ;
Gene expression ;
Recombinant DNA ;
Agar gel electrophoresis ;
Polyacrylamide gel electrophoresis ;
Substrate specificity ;
Southern blotting ;
Hydrogen peroxide ;
Streptomyces lividans ;
Streptomyces viridosporus ;
||Most EPA libraries have a fiche copy filed under the call number shown. Check with individual libraries about paper copy.
||A lignin peroxidase gene was cloned from Streptomyces viridosporus T7A into Streptomyces lividans TK64 in plasmid pIJ702. Bg/II-digested genomic DNA (4-10 kb) of S. viridosporus was shotgun-cloned into S. lividans after insertion into the melanin (mel+) gene of pIJ702. Transformants expressing pIJ702 with insert DNA were selected based upon the appearance of thiostrepton resistant tsr(supr)/mel(sup-) colonies on regeneration medium. Lignin peroxidase-expressing clones were isolated from this population by screening of transformants on a tsr-poly B-411 dye agar medium. In the presence of H2O2 excreted by S. lividans, colonies of lignin peroxidase-expressing clones decolorized the dye. Among 1000 transformants screened, 2 dye-decolorizing clones were found. One, pIJ702/TK64.1(TK64.1), was further characterized. TK64.1 expressed significant extracellular 2,4-dichlorophenol (2,4-DCP) peroxidase activity (= assay for S. viridosporus lignin peroxidase). Under the cultural conditions employed, plasmidless S. lividans TK64 had a low background level of 2,4-DCP oxidizing activity. (Copyright (c) 1990 Elsevier Science Publishers B.V. (Biomedical Division).
||Pub. in Jnl. of Biotechnology 13, p131-144 1990. See also PB90-108184. Sponsored by Corvallis Environmental Research Lab., OR., Department of Energy, Washington, DC., and National Science Foundation, Washington, DC.
|NTIS Title Notes
||Reprint: Cloning and Expression of a Lignin Peroxidase Gene from 'Streptomyces viridosporus' in 'Streptomyces lividans'.
||PC A03/MF A01