Suitability ofGeotrichum candidum lipase for the stereospecific analysis of some triglycerides (original) (raw)

Abstract

A procedure is described for determining the stereospecific structure of triacid triglycerides containing oleic acid. The method utilizes the unique specificity of the lipase system from_Geotricum candidum_ for hydrolyzing fatty acids which contain_cis_-9-unsaturation.

The procedure involves a pancreatic lipase hydrolysis of 10–20 mg of substrate to determine the fatty acids in the β-position and the incubation of another 50 mg of triglyceride with_G. candidum_ lipase to obtain diglyceride for further treatment. The α,α- and α,β-diglycerides are collected separately, converted to phenyl phosphatides and treated with phospholipase A. The analysis of the monoglycerides, produced with pancreatic lipase and the analysis of the β-lysophosphatides allows the determination of the ratios of the original 2 position oleic acid_sn_-triglycerides while the analysis of the α-lysophosphatides aids in the determination of the original_sn_-triglycerides which contained oleic acid in the 3 position. The 1 position oleic acid_sn_-triglycerides are calculated by difference.

Racemic and enantiomeric triglycerides containing palmitic, stearic and oleic acids were synthesized and used to establish the limits of the new method. The good agreement between the actual and observed values for a mixture of isomers indicates that the procedure will be useful in the analysis of triacid triglycerides which contain oleic acid. The application of the procedure to triacid triglycerides which contain other unsaturated fatty acids is discussed.

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Author notes

1. J. Sampugna
   Present address: Chemistry Department, University of Maryland, College Park, Md.

Authors and Affiliations

1. Department of Animal Industries, University of Connecticut, 06268, Storrs, Connecticut
   J. Sampugna & R. G. Jensen

Authors

1. J. Sampugna
2. R. G. Jensen

Additional information

Scientific contribution No. 323, Agricultural Experiment Station, University of Connecticut, Storrs.

The numbering used in this manuscript is according to the rules for “stereospecific numbering” as outlined in the IUPAC-IUB Commission on Biochemical Nomenclature (CBN), The Nomenclature of Lipids, European J. Biochem. 2 (1967) 127–131. In this system, the glycerol molecule is viewed in a Fischer projection formula with the secondary hydroxyl (or derived group) to the left and the hydrogen on the asymmetric carbon to the right. Then, the carbinol group at the top is position 1 and the group at the bottom is position 3. When stereospecific numbering is indicated the numbering is prefixed with_sn_; where the racemic glyceride is intended by the formula, the numbering is prefixed with_rac_. Also in this paper, an abbreviated representation of triglyceride nomenclature is used and when stereospecific numbering is intended, the first fatty acid depicted is in position 1 while the last fatty acid mentioned is in position 3. For example,_sn_-PSO is used as an abbreviation for_sn_-glyceryl-1-palmitate-2-stearate-3-oleate. In the abbreviated formula,_rac_-PSO, palmitic and oleic acids are considered to occupy the 1 and 3 positions equally. Where the Greek letters α and β are employed, no knowledge of optical isomerism is intended; thus, in these instances racemates, enantiomers or partial racemates could be indicated.

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Sampugna, J., Jensen, R.G. Suitability of_Geotrichum candidum_ lipase for the stereospecific analysis of some triglycerides.Lipids 3, 519–529 (1968). https://doi.org/10.1007/BF02530896

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• Received: 31 May 1968
• Issue date: November 1968
• DOI: https://doi.org/10.1007/BF02530896

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