Immobilisation of Candida rugosa lipase on a highly hydrophobic support: A stable immobilised lipase suitable for non-aqueous synthesis

Ivan Kurtovic; Tim D Nalder; Helen Cleaver; Susan N Marshall

doi:10.1016/j.btre.2020.e00535

Immobilisation of Candida rugosa lipase on a highly hydrophobic support: A stable immobilised lipase suitable for non-aqueous synthesis

Biotechnol Rep (Amst). 2020 Sep 29:28:e00535. doi: 10.1016/j.btre.2020.e00535. eCollection 2020 Dec.

Authors

Ivan Kurtovic¹, Tim D Nalder^{1

2}, Helen Cleaver¹, Susan N Marshall¹

Affiliations

¹ Nelson Research Centre, The New Zealand Institute for Plant and Food Research Limited, 293-297 Akersten Street, Nelson, 7010, New Zealand.
² School of Life and Environmental Sciences, Deakin University, 75 Pigdons Road, Waurn Ponds, 3216, Victoria, Australia.

Abstract

Lipase from Candida rugosa (CrL) was immobilised on highly hydrophobic, octadecyl methacrylate resin (Lifetech™ ECR8806M) via interfacial adsorption. The aim was to produce a stable biocatalyst suitable for use in a range of lipid-modifying reactions. Immobilisation was carried out in 10 mM phosphate buffer (pH 6.0) over 24 h at 21 °C. High protein binding of 58.7 ± 4.9 mg/g dry support accounted for ∼53 % of the applied protein. The activity recovery against tributyrin was 74.0 ± 1.1 %. The specific activity of immobilised CrL against tributyrin was considerably higher than that of Novozym® 435, at 1.79 ± 0.05 and 1.08 ± 0.04 U/mg bound protein, respectively. Incubation with high concentrations (10 % w/v) of both Triton X-100 and SDS resulted in only a small reduction in immobilised lipase activity. Solvent-free synthesis of glycerides by the FFA-saturated immobilised CrL was successful over 6 reaction cycles, with no apparent loss of activity.

Keywords: 4-MU, 4-methylumbelliferone; 4-MUHP, methyl 4-methylumbelliferyl hexylphosphonate; C18M, octadecyl methacrylate resin (Lifetech™ ECR8806M); CaLB, lipase B fromCandida antarctica; Candida rugosa lipase; CrL, Candida rugosa lipase; Enzyme stability; FFA, free fatty acid; Glyceride synthesis; Interfacial adsorption; Restructured lipids.