Cloning of a laccase gene from a novel basidiomycete Trametes sp. 420 and its heterologous expression in Pichia pastoris.
The laccase gene lacD, cloned from a novel laccase-producing basidiomycete Trametes sp. 420, contained 2,052 base pairs (bp) interrupted by 8 introns. lacD displayed a relatively high homology with laccase genes from other white rot fungi, whereas the homology between lacD and laccase genes from plants, insects, or bacteria was less than 25%. A 498-amino acid peptide encoded by the lacD cDNA was heterologously expressed in the Pichia pastoris strain GS115, resulting in the highest yield of laccase (8.3 x 10(4) U/l) as determined with ABTS (2,2'-azinobis [3-ethylbenzothia-zoline-6-sulfonic acid]) as the substrate. Additionally, the enzyme activity of recombinant laccase on decolorization of some industrial dyes was assessed.
Hong
Yu-zhi
YZ
School of Life Sciences and Modern Experiment Technology Center, Anhui University, Hefei, 230039, P.R. China.
Zhou
Hong-min
HM
Tu
Xiao-ming
XM
Li
Jian-feng
JF
Xiao
Ya-zhong
YZ
D000880:Anthraquinones / Q000378:metabolism; D052160:Benzothiazoles / Q000378:metabolism; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D005070:Evans Blue / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic; D015966:Gene Expression Regulation, Fungal; D007203:Indigo Carmine; D007211:Indoles / Q000378:metabolism; D042845:Laccase / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010843:Pichia / Q000235:genetics*; D020072:Polyporales / Q000201:enzymology / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013451:Sulfonic Acids / Q000378:metabolism
D000880:Anthraquinones; D052160:Benzothiazoles; D004271:DNA, Fungal; D005656:Fungal Proteins; D007211:Indoles; D011994:Recombinant Proteins; D013451:Sulfonic Acids; C002502:2,2'-azino-di-(3-ethylbenzothiazoline)-6-sulfonic acid; D005070:Evans Blue; D007203:Indigo Carmine; D042845:Laccase