From the N-terminal amino acid sequence of the Aspergillus niger F044 lipase

a potential homologous gene A84689 of the anl（the gene encoding the Aspergillus niger lipase） was found by means of bioinformatics.Based on the nucleotide sequence of the A84689

primers were designed to amplify anl.Nucleotide sequencing of the genomic DNA and cDNA revealed that the anl had an open reading frame comprising 1044 nucleotides

containing three introns of 54

45 and 51 bp.The deduced amino acid sequence of the anl encoded 297 amino acid residues including a signal sequence of 27 amino acid residues.The cDNA fragment coding the mature ANL（the protein of the Aspergillus niger lipase） was subcloned into the pPIC9K expression vector and the reconstructed expression plasmid pPIC9K-anl was then transformed into Pichia pastoris GS115.The anl was actively expressed in the Pichia pastoris GS115 and the recombinated lipase protein was secreted into the cultural broth.