title: Geotrichum candidum produces several lipases with markedly different substrate specificities.
abstract: We have purified and examined the substrate specificity of four lipases from two strains of the mould Geotrichum candidum, ATCC 34614 and CMICC 335426. We have designated the lipases I and II (ATCC 34614), and A and B (CMICC 335426). The enzymes are monomeric and have similar molecular masses and pI. Thus, lipases I and II have native molecular masses of 50.1 kDa and 55.5 kDa, and pI of 4.61 and 4.47, respectively. Lipases A and B are very similar to lipases I and II with native molecular masses of 53.7 kDa and 48.9 kDa, and pI of 4.71 and 4.50, respectively. Treatment with endo-beta-N-acetylglucosaminidase caused a reduction in molecular mass of approximately 4.5 kDa for all four lipases, indicating that these enzymes are glycosylated. Western blotting shows that the lipases are related. However, lipase B from CMICC 335426 shows a remarkable specificity for unsaturated substrates with a double bond at position 9 (cis configuration), and this specificity is not exhibited by the other three lipases. No lipase of this unique specificity has previously been purified to homogeneity. Structural studies using these four lipases should allow insight into the molecular basis of this remarkable specificity.
authors.affiliation: Unilever Research, Colworth Laboratory, Sharnbrook, England.
authors.forename: C M
authors.initials: CM
authors.lastname: Sidebottom
authors.forename: E
authors.initials: E
authors.lastname: Charton
authors.forename: P P
authors.initials: PP
authors.lastname: Dunn
authors.forename: G
authors.initials: G
authors.lastname: Mycock
authors.forename: C
authors.initials: C
authors.lastname: Davies
authors.forename: J L
authors.initials: JL
authors.lastname: Sutton
authors.forename: A R
authors.initials: AR
authors.lastname: Macrae
authors.forename: A R
authors.initials: AR
authors.lastname: Slabas
mesh_terms: D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000378:metabolism; D005848:Geotrichum / Q000201:enzymology*; D006031:Glycosylation; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008049:Lipase / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D013379:Substrate Specificity
chemical_list: D005227:Fatty Acids; D007527:Isoenzymes; D008049:Lipase