Isolation of carboxylester lipase (CEL) isoenzymes from Candida rugosa and identification of the corresponding genes.
The yeast Candida rugosa produces extracellular lipases which are widely used for industrial purposes. A commercial lipase preparation from this yeast can be separated into several isoenzymes which differ in carbohydrate content, isolelectric point, substrate specificity, and primary sequence. We have here purified and characterized three lipases, which also hydrolyze p-nitrophenyl esters, from a commercial preparation of this yeast. These three carboxylester lipases (CELs) elute differently on hydrophobic interaction chromatography, and have different carbohydrate contents and substrate specificities. Sequence analysis of their amino termini and peptides generated by LysC treatment showed that CEL-1 and CEL-3 probably have identical primary structure while CEL-2 was proven to be a different enzyme. Sequence comparison showed that both CEL-1 and CEL-3 are products of the LIP1 gene and that CEL-2 is the gene product of LIP2, cloned by Longhi et al. (Biochim. Biophys. Acta 1131, 227-232, 1992).
Diczfalusy
M A
MA
Department of Medical Laboratory Sciences and Technology, Karolinska Institutet, Huddinge University Hospital, Sweden.
Hellman
U
U
Alexson
S E
SE
D000595:Amino Acid Sequence; D002175:Candida / Q000201:enzymology* / Q000235:genetics*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005800:Genes, Fungal; D007527:Isoenzymes / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008049:Lipase / Q000235:genetics* / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D013379:Substrate Specificity
D007527:Isoenzymes; D010446:Peptide Fragments; D008049:Lipase