Thermomyces lanuginosus lipase immobilized on magnetic nanoparticles and its application in the hydrolysis of fish oil

Abstract

This article describes the immobilization and stabilization of lipase from Thermomyces lanuginosus (TLL) onto magnetic nanoparticles, which led to a markedly improved performance of the enzyme. The optimal immobilization conditions were found to be a pH of 7.5, 40°C, 4 hr, and 4.4 mg TLL protein/g of support of lipase. These results were used to enrich the docosahexaenoic acid (DHA) which is used in the fields of medicine and food. When the immobilized lipase was utilized to enrich DHA, the DHA content increased with the increasing amount of enzyme up to 4.4 mg TLL protein/g of support of lipase. The water content also influenced the DHA content, with the highest DHA content at 50% water, 40°C, and a pH of 7 after 14 hr that is, 4.4‐fold the initial content of DHA fish oil for TLL‐GMNPs. After six cycles of successive usage, it still remained over 82% relative hydrolysis. Practical applications One of the most important applications in use Omega 3, it is important for human health, industrial applications in the fields of food and medicine.

Novozym 435: the “perfect” lipase immobilized biocatalyst?

“…42 The heterogeneity of the natural substrate [43][44][45][46][47][48] has converted lipases in enzymes with a very broad specificity, accepting substrates very different from glycerides (even amides). Thus, lipases are used in vitro to catalyze reactions different from those of the natural hydrolase function, [49][50][51][52][53][54] such as esterification, [55][56][57][58][59][60][61] acidolysis, [62][63][64][65][66][67] interesterificaton, [68][69][70][71] transesterification, [72][73][74][75][76] aminolysis, [77][78][79][80][81] perhydrolysis, 82,83 etc., together with a collection of the so-called promiscuous reactions. [84][85][86][87][88]...…”

“…As previously stated, the natural substrates of lipases are glycerides. [49][50][51][52][53][54] These molecules have low solubility in water, therefore they form insoluble drops, where the lipase must act. For this purpose, lipases have a peculiar mechanism of action, called interfacial activation, which permits lipases to become adsorbed on the hydrophobic surface of the glyceride drops and act in the interface (this is why lipases are called "interfacial enzymes").…”

“…However, the in vitro oil hydrolysis capability of lipases has some applications such as in the production of free fatty acids. 42,[49][50][51][52][53][54] Thus, the hydrolysis of waste cooking oil under solventfree conditions was performed using N435 under ultrasound irradiation to produce free fatty acids. 401 After 2 h, a yield of 75.19% was obtained.…”

Suitability of Recombinant Lipase Immobilised on Functionalised Magnetic Nanoparticles for Fish Oil Hydrolysis

“…Carbon nanomaterial immobilised lipase B of Candida antarctica also retained higher activity (≥50%) up to five reuse cycles at 60 • C [20]. Magnetic nanoparticle immobilised lipase sourced from Thermomyces lanuginosus retained 82% relative hydrolysis after the six reuse cycles [30]. Superior biocatalyst recyclability was reported due to nanoparticle-enzyme covalent binding and efficient magnetic separation of superparamagnetic nanoparticle without any mechanical shearing, thus presenting a cost-effective method with potential use for economically immobilised biocatalyst processes.…”

Enhanced biological activity of Candida rugosa lipase via immobilization on magnetic nanoparticles

“…The activity for immobilized and free lipases was tested via hydrolysis process according to [23]. 5 mL 0.1 M phosphate buffer, 4 mL olive oil emulsion, a specific amount was added to the reaction system from the immobilized and free enzyme.…”