Aspergillus niger F044 Lipase:Gene Cloning,Overexpression and in vitro Refolding / 微生物学通报

ABSTRACT

From the N-terminal amino acid sequence of the Aspergillus niger F044 lipase,a potential homologous gene A84689 to 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 anl gene revealed an open reading frame of 1 044 nucleotides,containing three introns(54,45 and 51 nucleotides).The deduced amino acid sequence of the anl gene corresponds to 297 amino acid residues including a signal sequence of 27 amino acid residues.The cloned cDNA coding for mature Anl(the protein of the Aspergillus niger lipase)was overexpressed in Escherichia coli BL21(De3),and the recombinant Anl was purified.The denatured recombinant Anl by 8mol/L urea was refolded in vitro by dilution and DEAE Sepharose Fast Flow chromatography.