Efficient expression of laccase gene from white-rot fungus Schizophyllum commune in a transgenic tobacco plant

• Published in: FEMS microbiology letters Vol. 286; no. 1; pp. 130 - 135
• Main Authors: Hirai, Hirofumi, Kashima, Yoshiyuki, Hayashi, Katsuma, Sugiura, Tatsuki, Yamagishi, Kenji, Kawagishi, Hirokazu, Nishida, Tomoaki
• Format: Journal Article
• Language: English
• Published: Oxford, UK Oxford, UK : Blackwell Publishing Ltd 01.09.2008; Blackwell Publishing Ltd; Wiley-Blackwell; Oxford University Press
• Subjects: Amino Acid Sequence; Base Sequence; Biological and medical sciences; Complementary DNA; CpG-dinucleotide motif; Electrophoresis; Enzymes; Fundamental and applied biological sciences. Psychology; Fungal Proteins - chemistry; Fungal Proteins - genetics; Fungal Proteins - metabolism; Fungi; Gel electrophoresis; Gene Expression; Gene sequencing; Genes; Growth, nutrition, metabolism, transports, enzymes. Molecular biology; Laccase; Laccase - chemistry; Laccase - genetics; Laccase - metabolism; Ligninolytic enzymes; Microbiology; Molecular Sequence Data; Mycology; Nicotiana - genetics; Nicotiana - metabolism; Phytoremediation; Plants, Genetically Modified - genetics; Plants, Genetically Modified - metabolism; Pollutants; Polyacrylamide; Protein Engineering; RNA-directed DNA polymerase; Rot; Schizophyllum - enzymology; Schizophyllum commune; Tobacco; transgenic plant; Transgenic plants; Trichlorophenol; Trichlorophenols; White rot fungi; white-rot fungi; CpG-dinucleotide motif; trichlorophenol; laccase; transgenic plant; Enzyme; Stimulant plant; Gene expression; Nicotiana tabacum; Basidiomycota; Fungi; Schizophyllum commune; Dicotyledones; Angiospermae; Wood destroying fungi; Spermatophyta; Transgenic plant; Oxidoreductases; Solanaceae; Laccase
• ISSN: 0378-1097; 1574-6968
• DOI: 10.1111/j.1574-6968.2008.01267.x

Ligninolytic enzymes produced by white-rot fungi are effective degraders of recalcitrant aromatic environmental pollutants. However, gene sequences of these enzymes are rich in CpG dinucleotides, which are particularly unfavorable to efficient expression in plants. In order to develop a phytoremediation technique with a ligninolytic enzyme-producing transgenic plant, laccase cDNA (scL) from white-rot fungus Schizophyllum commune was used as a model ligninolytic enzyme, and we attempted to obtain the efficient expression of scL in a transgenic tobacco plant by decreasing the CpG-dinucleotide motif content. We constructed a mutagenized scL sequence, scL12, decreasing the CpG-dinucleotide motif content by 12%, and scL12 was introduced into the tobacco plant. Much higher laccase activity was detected in transgenic scL12 plants than in transgenic scL plants and wild-type plants. Using reverse transcriptase-PCR analysis, scL12 was translated in transgenic scL12 plants whereas mRNA of scL was not detected in the transgenic scL plants, and scL, which is the product of the scL12 gene, was produced in the transgenic scL12 plants using native-polyacrylamide gel electrophoresis analysis. Moreover, transgenic scL12 plants were able to remove trichlorophenol more effectively than transgenic scL plants and wild-type plants. These results suggest that decreasing CpG-dinucleotide motif content in fungal target genes is a useful method for efficient expression of these genes in transgenic plants.