Molecular cloning and expression of a laccase from Ganoderma lucidum, and its antioxidative properties.
Laccases are multicopper-containing oxidases that catalyze the oxidation of many aromatic compounds with concomitant reduction of oxygen to water. Interest in this enzyme has arisen in many fields of industry, including detoxification, wine stabilization, paper processing, and enzymatic conversion of chemical intermediates. In this study, we cloned a laccase gene (GLlac1) from the white-rot fungus Ganoderma lucidum. The cloned gene consists of 4,357 bp, with its coding region interrupted by nine introns, and the upstream region has putative CAAT and TATA boxes as well as several metal responsive elements (MREs). We also cloned a full-length cDNA of GLlac1, which contains an uninterrupted open reading frame (ORF) of 1,560 bp coding for 520 amino acids with a putative 21-residue signal sequence. The DNA and deduced amino acid sequences of GLlac1 were similar but not identical to those of other fungal laccases. GLlac1 was released from the cells when expressed in P. pastoris, and had high laccase activity. In addition, GLlac1 conferred antioxidative protection from protein degradation, and thus may be useful in bio-medical applications.
Joo
Seong Soo
SS
Division and Institute of Life Sciences, Kangwon National University, Chuncheon 200-701, Korea.
Ryu
In Wang
IW
Park
Ji-Kook
JK
Yoo
Yeong Min
YM
Lee
Dong-Hyun
DH
Hwang
Kwang Woo
KW
Choi
Hyoung-Tae
HT
Lim
Chang-Jin
CJ
Lee
Do Ik
DI
Kim
Kyunghoon
K
D000595:Amino Acid Sequence; D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D042845:Laccase / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D020940:Reishi / Q000201:enzymology*
D000975:Antioxidants; D005656:Fungal Proteins; D042845:Laccase