Abstract: The laccase gene 1 had been cloned by RT-PCR from Coprinopsis cinerea okayama7#130, through analyzing of SignalP3.0 Server, a new primer had been designed that it didn’t have signal peptide sequence for laccase1(lac1) and construction of Pichia pastoris expression plasmid pPIC9K-lac1, then transformed into the Pichia pastoris GS115 and induction expression, studied properties of recombinant enzyme at last. The full length of laccase 1was 1593bp, which coded 530 amino acids and included 18 amino acids of Signal peptide sequence. SDS-PAGE show that obvious protein band was 65 kDa. The optimal fermentature was 45℃ and pH value was 4.3. The activity of restructed laccase remain unchanged basically after it staied in 45℃ for 3 hours, the stability of avticity was good in the scope from 4.0 to 10.0 of pH.The enzyme activity of secretion expression achieve to 1.108 U/mL.

Key words: RT-PCR, laccase gene, Pichia pastoris, secretion expression