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Esterification of polyunsaturated fatty acids by various forms of immobilized lipase from Rhizomucor miehei

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Lipids

Abstract

Ethyl esterification specificity of a lipase from Rhizomucor miehei for polyunsaturated fatty acids (PUFA) was compared at 1 and 100 mM to study molecular recognition of PUFA. The chemical shift of methylene adjacent to carboxyl groups in the nuclear magnetic resonance spectrum of docosahexaenoic acid (DHA) in ethanol moved to a lower magnetic field as the concentration of DHA increased, suggesting that the degree of dissociation of DHA decreased. Specificity constants or apparent second-order rate constants (V max/K m or catalytic power) for 1 mM esterification by immobilized lipases were higher than the native lipase. Immobilized hydrophobic carrier of low mass transfer resistance for the esterification substrate may improve maximal velocity and affinity for the substrate. Higher specificity constants for 1 mM substrates were observed using immobilized lipases fixed on an anion exchange resin with glutaraldehyde and on a cation exchange carrier with carbodiimide. Activity yields measured with 1 mM PUFA substrate were high. For the substrates at a concentration of 100 mM, higher specific constants with these bifunctional reagents were not observed but higher activity yields were found.

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Abbreviations

CMC:

critical micelle concentration

DHA:

docosahexaenoic acid

K m or K Ac1X, K Ac2X :

Michaelis constant

NMR:

nuclear magnetic resonance

PUFA:

polyunsaturated fatty acid

V max, V Ac2X, V Ac1X :

maximal velocity

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Author notes

  1. Prodyut Kumar Roy

    Present address: Central Drug Research Institute, Chattar Manzil, 226001, Lucknow, India

  2. Qinglong Chang

    Present address: Soil and Plant Analysis Lab, University of Wisconsin, 53705, Madison, WI

  3. Cao Shu-gui

    Present address: National Laboratory of Enzyme Engineering, Jilin University, 130023, Changchun, China

  4. Makoto Fukatsu

    Present address: College of Science and Technology, Nihon University, 1-8 Kanda, 101-8308, Surugadai, Japan

Authors and Affiliations

  1. Agency of Industrial Science and Technology, National Institute of Bioscience and Human-Technology, 1-1 Higashi, 305-8566, Tsukuba, Japan

    Yoshitsugu Kosugi, Prodyut Kumar Roy, Qinglong Chang, Cao Shu-gui, Makoto Fukatsu, Kenji Kanazawa & Hiroshi Nakanishi

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  1. Yoshitsugu Kosugi
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  2. Prodyut Kumar Roy
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  3. Qinglong Chang
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  4. Cao Shu-gui
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  5. Makoto Fukatsu
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  6. Kenji Kanazawa
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  7. Hiroshi Nakanishi
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Correspondence to Yoshitsugu Kosugi.

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Kosugi, Y., Kumar Roy, P., Chang, Q. et al. Esterification of polyunsaturated fatty acids by various forms of immobilized lipase from Rhizomucor miehei . Lipids 35, 461–466 (2000). https://doi.org/10.1007/s11745-000-545-7

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  • DOI: https://doi.org/10.1007/s11745-000-545-7

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