Stabilization of the lipase of Hypocrea pseudokoningii by multipoint covalent immobilization after chemical modification and application of the biocatalyst in oil hydrolysis

• Lipase produced by Hypocrea pseudokoningii was immobilized on covalent support.
• The lipase was activated during immobilization on hydrophobic support and it maintained its activation during amination process.
• The lipase was chemical modified.
• Glyoxyl derivative was more stable than the CNBr derivative.
• The multipoint immobilization increased the enzyme activity in all tested oils.

Lipase from Hypocrea pseudokoningii was purified using the support Octyl–Sepharose. This adsorption resulted in a 3-fold increase in activity of the immobilized enzyme. Following, still on this support, the lipase was enriched in surface amino groups (by reaction of carboxy groups with ethylendiamine). After amination, the lipase was desorbed from Octyl–Sepharose, while the 2-fold hyper-activation was maintained. The aminated lipase was also successfully immobilized on Glyoxyl–Agarose. The derivative was 45-fold more stable than was the free enzyme at 50 and 60 °C. The derivative was also stable in 50% of organic solvents such as methanol, ethanol, propanol and cyclohexane. The multipoint immobilization also increased the enzyme stability in relation to the free enzyme in the presence of ethanol, methanol and cyclohexane for up to 72 h. For example, the stabilized derivative was 9-fold more stable than the free enzyme in presence of methanol. The derivatives hydrolyzed fish, cupuaçu (Theobroma grandiflorum), bacuri (Latonia insignis) and murumuru (Astrocaryum murumuru) oils. The multipoint immobilization process increased the hydrolysis of oils up to 15-fold compared with the control, what makes these derivatives attractive for industrial application.

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