Expression of laccase gene lcc1 in Coprinopsis cinerea under control of various basidiomycetous promoters.
Coprinopsis cinerea laccase gene lcc1 was expressed in this basidiomycete under naturally non-inductive conditions using various homologous and heterologous promoters. Laccase expression was achieved in solid and liquid media with promoter sequences from the C. cinerea tub1 gene, the Agaricus bisporus gpdII gene, the Lentinus edodes priA gene and the Schizophyllum commune Sc3 gene. As measured by enzyme activity in liquid cultures, a 277-bp gpdII promoter fragment, followed by a 423-bp priA fragment, was most efficient. A shorter priA sequence of 372 bp was inactive. tub1 promoter fragments were reasonably active, whereas the S. commune Sc3 promoter sequence was less active, in comparison. Irrespective of the promoter used, addition of copper to the medium increased enzymatic activities for highly active transformants by 10- to 50-fold and for less active transformants for 2- to 7-fold. The highest enzymatic activities (3 U/ml) were reached with the gpdII promoter in the presence of 0.1 mM CuSO(4).
Kilaru
Sreedhar
S
Molecular Wood Biotechnology, Institute of Forest Botany, Georg-August-University Göttingen, Germany.
Hoegger
Patrik J
PJ
Majcherczyk
Andrzej
A
Burns
Claire
C
Shishido
Kazuo
K
Bailey
Andy
A
Foster
Gary D
GD
Kües
Ursula
U
D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics*; D001709:Biotechnology / Q000379:methods; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D042845:Laccase / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D014170:Transformation, Genetic
D004271:DNA, Fungal; D042845:Laccase