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Main Title Cloning and Expression of a Lignin Peroxidase Gene from 'Streptomyces viridosporus' in 'Streptomyces lividans'.
Author Wang, Z. ; Bleakley, B. H. ; Crawford, D. L. ; Hertel, G. ; Rafii., F. ;
CORP Author Idaho Univ., Moscow. Dept. of Bacteriology and Biochemistry.;Corvallis Environmental Research Lab., OR.;Department of Energy, Washington, DC.;National Science Foundation, Washington, DC.
Publisher c1990
Year Published 1990
Report Number EPA-R-815300-01-0 ;DE-FG786ER13586; EPA/600/J-92/173;
Stock Number PB92-180389
Additional Subjects Molecular cloning ; Gene expression ; Lignocellulose ; Plasmids ; Recombinant DNA ; Agar gel electrophoresis ; Polyacrylamide gel electrophoresis ; Substrate specificity ; Southern blotting ; Hydrogen peroxide ; Reprints ; Streptomyces lividans ; Streptomyces viridosporus ; Lignin peroxidase
Library Call Number Additional Info Location Last
NTIS  PB92-180389 Some EPA libraries have a fiche copy filed under the call number shown. 07/26/2022
Collation 16p
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).