Cloning and sequence analysis of cDNA encoding Rhizopus niveus lipase. |
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Authors: | W Kugimiya Y Otani M Kohno Y Hashimoto |
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Institution: | Central Research Institute, Fuji Oil Co., Ltd., Ibaraki, Japan. |
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Abstract: | Complementary DNA encoding Rhizopus niveus lipase (RNL) was isolated from the R. niveus IF04759 cDNA library using a synthetic oligonucleotide corresponding to the amino acid sequence of the enzyme. A clone, which had an insert of 1.0 kilobase pairs, was found to contain the coding region of the enzyme. The lipase gene was expressed in Escherichia coli as a lacZ fusion protein. The mature RNL consisted of 297 amino acid residues with a molecular mass of 32 kDa. The RNL sequence showed significant overall homology to Rhizomucor miehei lipase and the putative active site residues were strictly conserved. |
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