Abstract
Lipases due to their ecological nature and catalytic versatility, are ideal for their application in the fish oil hydrolysis industry due to their selective property, which allows the preservation of polyunsaturated fatty acids (PUFAs) in the lipid structure. The objective of this research was to determine the activity and kinetic parameters of a commercial AY AMANO "30SD" lipase, as well as the temperature and time values to achieve an optimal degree of hydrolysis in semi-refined anchovy oil. The experiments were carried out in a jacketed minireactor with a working volume of 400 mL (oil-water-enzyme) with temperature control and pH 7.00, enzyme concentration 350 U/mL and stirring 160 rpm. A 3x3 factorial design and the response surface methodology were used. The results obtained from the study of the enzyme were: activity = 37 384.55 ± 395.07 U/g and kinetic parameters: Km = 7.98 g/L and Vmax. = 0.038887 g/Lxmin. Correspondingly, the following optimal parameters were obtained: Degree of hydrolysis 4.01%, temperature 46.86 °C and hydrolysis time 90 minutes, with a confidence level of 95% (p <0.05). Conclusions: The study allowed us to kinetically characterize the commercial lipase and determine the optimum degree of hydrolysis of the semi-refined anchovy oil.
Highlights
The fish oil industry in Peru has gained relevance since the discovery of its beneficial properties that its consumption provides due to its high content of polyunsaturated fatty acids (PUFAs) of the ω – 3 chain. (Valenzuela B. et al, 2012), In recent years the consumption of PUFA's in the human diet has increased but in higher concentrations, for which the way of consuming these essential fatty acids has been innovated in the form of concentrated supplements for people lacking in these components (Lopez H., 2016 ), this has developed over time techniques that allow obtaining high concentrations of polyunsaturated fatty acids (PUFAs) of the ω-3 chain, which are predominant eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Quian, et al, 2020)
Al., 2017) where the preference of lipase for fatty acid components is of order decreasing: palmitoleic acid> oleic acid> = palmitic acid> stearic acid> EPA> DHA, allowing the PUFA's to be preserved in the lipid structure (Hamed, H. et al, 2018), this hydrolysis process is complemented with other methods to achieve high concentrations of PUFA (Bonilla-Méndez & Hoyos-Concha, 2018), these have been raised in various investigations
3.1 Determination of enzyme activity The activities of the enzyme AY "AMANO" 30SD at dilutions 1: 6000, 1: 7000 and 1: 8000, the linearity range was found graphically at the times of 35, 30 and 25 minutes respectively (Figure 1), which resulted with activities of 36,972.79 U/g, 37,420.38 U/g and 37,760.48 U/g, U being the amount of micromole equivalents of fatty acid released per minute, working with a 12.5% olive oil emulsion substrate, the three experiences were found at the same reaction conditions with pH 7.00 and constant stirring of 160 RPM
Summary
(Valenzuela B. et al, 2012), In recent years the consumption of PUFA's in the human diet has increased but in higher concentrations, for which the way of consuming these essential fatty acids has been innovated in the form of concentrated supplements for people lacking in these components (Lopez H., 2016 ), this has developed over time techniques that allow obtaining high concentrations of polyunsaturated fatty acids (PUFAs) of the ω-3 chain, which are predominant eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (Quian, et al, 2020) These techniques that allow obtaining high concentrations of PUFA's have been investigated by various authors, for example molecular distillation (Cerón, Cardona, & Toro, 2012), formation of urea complexes (Gamez-Meza et al, 2003; Nagachinta, and Akoh, 2012), separation by chromatography (Dillon, Aponte, Tarozo, & Huang, 2013), extraction by supercritical fluids (Sahena et al, 2009), among others. Kahveci and Xu (2011) report in their research the use of salmon oil substrate, achieving the concentration of EPA that went from 5.46% to 10.00% and DHA increasing 2.8 times using lipase Candida rugosa, Miranda K. et al (2013) reported the use of a Candida antarctica Lipase for the production of structured diacylglycerols with 89.37% PUFA of the omega-3 chain (EPA, DHA and DPA) using fish oil as substrate, Valverde L. et al (2014) reported an increase in EPA that went from 19% to 61% and a DHA that went from 22% to 69% using Lipozyme TL-IM and QLG lipase with sardine and tuna oil substrate, Aarthy M. et al (2016) achieved an increase in EPA from 16.9% to 30.4% and DHA from 6.5% to 9.6% using cod, sardine, salmon and shark oil as substrate catalyzing the reaction with a Cryptococcus sp. lipase
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