A second pectin lyase gene (pel2) from Aspergillus oryzae KBN616: Its sequence analysis and overexpression, and characterization of the gene products

A second pectin lyase gene, designated pel2, was isolated from a shoyu koji mold Aspergillus oryzae KBN616 and characterized. The structural gene comp rised 1306 hp with three introns. The ORF encoded 375 amino acids with a si gnal peptide of 19 amino acids. The deduced amino acid sequence showed high similarity to those of A. oryzae Pel1, Aspergillus niger pectin lyases and Glomerella cingulata PnlA. The pel2 gene was overexpressed under the contr ol of the promoter of the A. oryzae TEF1 gene for purification and enzymati c characterization of its gene product. The gene product exhibited two mole cular masses of 48 and 44 kDa due to different degrees of glycosylation. Bo th proteins had the Same pH optimum of 6.0 and temperature optimum of 50 de greesC.