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Lipase-catalyzed fractionation of conjugated linoleic acid isomers

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Lipids

Abstract

The abilities of lipases produced by the fungus Geotrichum candidum to selectively fractionate mixtures of conjugated linoleic acid (CLA) isomers during esterification of mixed CLA free fatty acids and during hydrolysis of mixed CLA methyl esters were examined. The enzymes were highly selective for cis-9,trans-11–18∶2. A commercial CLA methyl ester preparation, containing at least 12 species representing four positional CLA isomers, was incubated in aqueous solution with either a commercial G. candidum lipase preparation (Amano GC-4) or lipase produced from a cloned high-selectivity G. candidum lipase B gene. In both instances selective hydrolysis of the cis-9,trans-11–18∶2 methyl ester occurred, with negligible hydrolysis of other CLA isomers. The content of cis-9,trans-11–18∶2 in the resulting free fatty acid fraction was between 94 (lipase B reaction) and 77% (GC-4 reaction). The commercial CLA mixture contained only trace amounts of trans-9,cis-11–18∶2, and there was no evidence that this isomer was hydrolyzed by the enzyme. Analogous results were obtained with these enzymes in the esterification in organic solvent of a commercial preparation of CLA free fatty acids containing at least 12 CLA isomers. In this case, G. candidum lipase B generated a methyl ester fraction that contained >98% cis-9,trans-11–18∶2. Geotrichum candidum lipases B and GC-4 also demonstrated high selectivity in the esterification of CLA with ethanol, generating ethyl ester fractions containing 96 and 80%, respectively, of the cis-9,trans-11 isomer. In a second set of experiments, CLA synthesized from pure linoleic acid, composed essentially of two isomers, cis-9,trans-11 and trans-10,cis-12, was utilized. This was subjected to esterification with octanol in an aqueous reaction system using Amano GC-4 lipase as catalyst. The resulting ester fraction contained up to 97% of the cis-9,trans-11 isomer. After adjustment of the reaction conditions, a concentration of 85% trans-10,cis-12–18∶2 could be obtained in the unreacted free fatty acid fraction. These lipase-catalyzed reactions provide a means for the preparative-scale production of high-purity cis-9,trans-11–18∶2, and a corresponding CLA fraction depleted of this isomer.

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Abbreviations

Ag+-HPLC:

silver-ion-high-performance liquid chromatography

CLA:

conjugated linoleic acid

DPG:

diphosphatidylglycerol

FAME:

fatty acid methyl ester

FFA:

free fatty acid

GC:

gas chromatography

HPLC:

high-performance liquid chromatography

TLC:

thin-layer chromatography

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Author information

Authors and Affiliations

  1. Southern Crop Frotection, Food Research Center, Agriculture and Agri-Food Canada, N1G 2W1, Guelph, Ontario, Canada

    John K. G. Kramer

  2. Loders Croklaan, 60137, Glen Hlyn, Illinois

    Gerald McNeill

  3. U.S. Center for Food Safety and Applied Nutrition, Food and Drug Administration, DC 20204, Washington

    Najibulla Sehat, Jan Fritsche, Magdi M. Mossoba & Martin P. Yurawecz

  4. Eastern Regional Research Center, ARS, USDA, 600 E. Mermaid Lane, 19038, Wyndmoor, PA

    Michael J. Haas, Karen Scott & Thomas A. Foglia

Authors
  1. Michael J. Haas
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  2. John K. G. Kramer
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  4. Karen Scott
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  5. Thomas A. Foglia
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  6. Najibulla Sehat
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  9. Martin P. Yurawecz
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Correspondence to Michael J. Haas.

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Haas, M.J., Kramer, J.K.G., McNeill, G. et al. Lipase-catalyzed fractionation of conjugated linoleic acid isomers. Lipids 34, 979–987 (1999). https://doi.org/10.1007/s11745-999-0448-7

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  • DOI: https://doi.org/10.1007/s11745-999-0448-7

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