Lipase Lipozyme Research Articles

Ultrasound assisted enzymatic synthesis of OPO structured lipids by covalent organic framework immobilized lipase

In this study, the covalent organic framework immobilized Rhizomucor miehie lipase COF@RML as a novel biocatalyst was applied in the enzymatic synthesis of OPO structured lipids (1, 3-dioleoyl-2-palmitoylglycerol). The impact of reaction medium, substrate molar ratio, enzyme addition amount, reaction time and temperature on the enzymatic synthesis of OPO structured lipids were studied. Furthermore, the effects of ultrasonic power and ultrasonic time on the synthesis of OPO structural lipids were studied. The results showed that ultrasonication could increase the yeild of OPO structured lipids by improving substrate mass transfer and enzyme particle dispersion. The optimal process for the synthesis of OPO structured lipids was obtained. When the ultrasonic power was set at 90 W, ultrasonic time at 12 minutes, enzyme addition amount at 10 wt%, substrate molar ratio at 1:8, reaction temperature at 45 °C, and reaction time at 6 hours, the yield of OPO structured lipids reached a remarkable 51.27 %. Finally, the commercial lipase Lipozyme RM IM was compared with the COF@RML. The findings indicated that COF@RML immobilized enzyme had better application value in the synthesis of OPO structured lipids.

Sustainable one-pot solvent-free enzymatic synthesis of capric acid-rich structured lipids to enhance the nutritional value of grape seed oil

Grape seed oil (GSO) is a valuable resource for the pharmaceutical, cosmetic, and food industries due to its fatty acid composition, which provides anti-inflammatory, cardioprotective, antimicrobial, and anticancer properties. GSO is also recognized for its sensory attributes and serves as an essential component in our dietary intake, providing vital fatty acids for metabolic reactions. The enzymatic synthesis of structured lipids (SL) using GSO was investigated in this study. The SL was created by incorporating medium chain fatty acids (capric acid – C10) at the sn-1 and sn-3 positions and long chain fatty acids in the internal position of the triacylglycerols (TAG). GSO was selected based on its composition rich in unsaturated fatty acids, mainly linoleic acid. The acidolysis reactions were catalyzed by an immobilized commercial lipase Lipozyme TLIM®. The incorporation degree of C10 into GSO reached 36%. Health lipid indices were calculated to determine the C10 incorporation effect on the improvement of nutritional quality of oil. The SL showed lower oxidative stability index (OSI) values than the original oil and higher degree of fatty acid (DFA) values. The simpler, solvent-free method not only enhances the health benefits of oils but also gives valuable properties to GSO, making it an even more attractive option for food industries.

Kinetic modeling and optimization of the mono- and diglycerides synthesis mediated by the lipase Lipozyme® TL 100 L immobilized on clayey support.

Mono- and diglycerides play a crucial role in the food industry as multifunctional food additives and emulsifiers. Their importance stems from their unique properties, which allow them to improve the quality, texture, and stability of various food products. Here, results of the kinetic modeling of the mono- and diglycerides synthesis mediated by the lipase Lipozyme® TL 100 L immobilized on the clayey support Spectrogel® type C are reported. The support was characterized by TEM, SEM, and FTIR. Firstly, the influence of pH and lipase load on the immobilization process was analyzed, resulting in an enzymatic activity of 93.2 ± 0.7Ug-1 under optimized conditions (170.9Ug-1 of lipase and pH of 7.1). Afterward, the effects of reaction temperature and concentration of immobilized biocatalyst in the feedstock conversion were evaluated. At optimized parameters, a triglycerides conversion of 97% was obtained at 36.5°C, 7.9 vol.% of enzyme, a glycerol to feedstock molar ratio of 2:1, and 2h. The optimized conditions were used to determine the kinetic constants of the elementary reactions involved in the glycerolysis, where a fit superior to 0.99 was achieved between experimental values and predicted data.

Enzymatic In Situ Interesterification of Rapeseed Oil with Methyl Formate in Diesel Fuel Medium

The purpose of this research was to evaluate the process of enzymatic biodiesel synthesis by directly using rapeseed as a raw material, extracting the oil contained within and interesterifying with a mixture of methyl formate and mineral diesel, choosing the amount of mineral diesel so that the ratio between it and the rapeseed oil in the seeds was 9:1. As the final product of the interesterification process, a mixture of mineral diesel and biodiesel was obtained directly, which is conventionally produced by mixing the mineral diesel and biodiesel. The tests were performed using enzymatic catalysis using the lipase Lipozyme TL TIM. Process optimization was performed using the response surface methodology. A model describing the interaction of three independent variables and their influence on the yield of rapeseed oil methyl esters was developed. The physical and chemical indicators of the product obtained under optimal interesterification conditions were evaluated.

Synthesis of dietary lipids from pumpkin (Cucurbita pepo. L) oil obtained by enzymatic extraction: a sustainable approach.

This study aimed to assess the nutritional properties of dietary lipids obtained through the modification of aqueous enzymatically extracted pumpkin seed (Cucurbita pepo. L) oil. The optimal growth conditions for producing pectinase using strain Aspergillus sp. 391 were determined, and partial characterization of pectinase and commercial cellulase was conducted. The enzymatic extraction was performed at pH 4.0, 50°C, for 24h, using a combination of pectinase and cellulase for optimum effectiveness. The crude oil obtained was analyzed for acid, peroxide, and fatty acid composition. The study found a high amount of unsaturated fatty acids, mainly linoleic acid (C18:2), and a 59% oil recovery rate. Subsequently, this oil was subjected to enzymatic acidolysis with capric acid in solvent-free media, catalyzed by lipase Lipozyme RM IM®, resulting in a product with a higher incorporation degree (48.39 ± 0.5mol%), observed after 24h at 60°C using molar ratio oil:acid capric of 1:9 (run 4). The nutritional properties of this oil were improved.

1,3-Selective hydrolysis of blackcurrant seed oil for the concentration of alpha- and gamma-linolenic acids

Blackcurrant seed oil contains alpha-linolenic acid (a-Ln) and gamma-linolenic acid (g-Ln), essential fatty acids of different therapeutic effects. Lipid classes of various a-Ln/g-Ln ratios were previously produced by partial hydrolysis catalyzed by 1,3-specific lipases Lipozyme RM IM and Lipozyme TL IM, conducted in supercritical CO2 (scCO2) in a continuous packed-bed reactor. In this work, free fatty acids (FFA) were separated from glycerides using preparative chromatography on silica gel with scCO2 as mobile phase in order to obtain two clean, natural products of different a-Ln/g-Ln ratios, and fatty acid profiles in hydrolysates obtained under different conditions were reviewed. Using Lipozyme RM IM and maintaining the FFA mass fraction in hydrolysate below that of partial glycerides, the a-Ln/g-Ln ratio increased from 1.3 in oil to 10.7 in FFA. Lipozyme RM IM-mediated hydrolysis showed steric hindrance of g-Ln, while silica support of Lipozyme TL IM promoted acyl migration.

Improvements in the Modeling and Kinetics Processes of the Enzymatic Synthesis of Pentyl Acetate

In this work, the enzymatic synthesis of pentyl acetate obtained from acetic acid and pentan-1-ol using the commercial immobilized lipase Lipozyme®435 was studied. Specifically, the effects of several variables of the process on the kinetics were shown, such as the initial concentration of the acetic acid, the alcohol/acid molar ratio, and the possible reuse of the enzyme, while other variables, such as temperature, agitation, and the enzyme/acid ratio were held constant. The kinetics were determined by assessing the acetic acid concentration throughout the reactive process. Experimental data were correlated with the rate equation consisting of a modified version of the Bi–Bi Ping-Pong mechanism. The results showed that when no hydrophobic solvents were used with the reagents in stoichiometric proportion, a high molar fraction of acetic acid (x0,acid ≈ 0.50) caused the loss of enzymatic activity, achieving a conversion of only 5%. However, when there was an excess of pentan-1-ol, the reaction occurred successfully. Under optimal conditions (solvent-free conditions, x0,alcohol/x0,acid = 2, and x0,acid = 0.33), it was found that the enzyme could be reused up to 10 times without a loss of activity, reaching conversions higher than 80% after 8 h. Therefore, those conditions are advantageous in terms of productivity.

Lipase-Mediated Synthesis of Oleoyl Ethanolamide Starting from High-Oleic Sunflower Oil Soapstock

This work describes the lipase-mediated synthesis of oleoyl ethanolamide, a dietary supplement for body weight loss recently approved by FDA. The target compound is prepared by conversion of the oleic acid contained in a mixture of fatty acids recovered by enzymatic hydrolysis of soapstock, a side-product of high oleic sunflower oil refinement. The use of a packed-bed reactor (a glass column loaded with the commercial lipase Lipozyme 435) in continuous flow mode improves the space-time yield of the reaction and the catalyst productivity. The nontoxic, bioderived, and renewable solvent limonene is used in the reaction medium. The process has been run for more than 157 h of continuous operation, demonstrating the stability and efficiency of the biocatalyst. Additionally, at the end of the reaction, only oleoyl ethanolamide crystallizes from the reaction mixture, thus, it is collected by simple filtration of the outlet solution in 53% isolation yield, showing 99% chemical purity, while all the byproducts of the reaction are left behind in the mother liquors.

Immobilization of Rhizomucor miehei lipase on magnetic multiwalled carbon nanotubes towards the synthesis of structured lipids rich in sn-2 palmitic acid and sn-1,3 oleic acid (OPO) for infant formula use

Nowadays, breast milk is considered as the ideal food for infants owing to the most common oleic acid-palmitic acid-oleic acid (OA-PA-OA) fatty acid distribution of the human milk fat (HMF). This study reports the synthesis of 1,3-dioleoyl-2-palmotoylglycerol (OPO)-rich human milk fat substitutes in a two-step enzymatic acidolysis reaction with Rhizomucor miehei lipase (RML) immobilized on magnetic multi-walled carbon nanotubes(mMWCNTs). The immobilized RML (RML-mMWCNTs) showed better thermal and pH stability, convenient recovery and reusability than the free soluble form. Under optimized reaction conditions (1:8 tripalmitin (PPP)/OA, 10%wt. enzyme, 50 °C, 5 h), PA content at the sn-2 position and OA incorporation at the sn-1,3 positions reached 93.46% and 59.54%, respectively. Comparison tests have also showed that RML-mMWCNTs has better catalytic activity and reusability than the commercial lipase Lipozyme RM IM. The results suggest that RML-mMWCNTs is a promising biocatalyst for the synthesis of OPO-rich TAGs with potential use in infant formulas.

A parameter study of ultrasound assisted enzymatic esterification

This work is focused on the study of the esterification parameters for the ultrasound assisted synthesis of isoamyl acetate catalyzed by lipase Lipozyme 435 in a continuous loop reactor. Investigating the influence of different parameters shows that a higher concentration of ester (462 mg/g mixture) can be obtained at a temperature of 50 °C, flow rate 0.16 mL/min. The best ultrasonication conditions are: sonication applied continuously for a short time (20 min), ultrasound power 32 mW and amplitude 20%. The enzyme can be successfully reused tree times without loss of enzyme activity. Reaction kinetics for isoamyl acetate ultrasound assisted production showed that satisfactory reaction concentration (close to the equilibrium concentrations) could be reached in short reaction times (2 h). Ultrasound assisted enzymatic esterification is consequently a cleaner and a faster process.

Regioselective enzymatic hydrolysis of blackcurrant seed oil in supercritical CO2

Continuous flow hydrolysis of blackcurrant seed oil dissolved in supercritical carbon dioxide (scCO2) was catalyzed by immobilized lipase Lipozyme TL IM. HPLC-MS and acid-base titration were used to determine composition of oil and hydrolysis products comprised of tri-, di- and monoglycerides, free fatty acids and glycerol. The oil contained 10 % linolenic and 2 % stearidonic fatty acids (omega-3), 51 % linoleic and 19 % gamma-linolenic fatty acids (omega-6), 12 % oleic acid, 5 % palmitic acid, and 1% stearic acid. Effects of enzyme load (17−500 mg), pressure (15−30 MPa), temperature (30−50 °C), and CO2 flow rate (0.2–4 g/min) on the conversion were evaluated. The reaction was reversible and the time to reach steady state was enzyme load-dependent and took up to 200 min. A slight drop of enzyme activity after four experimental runs and total reaction time 17 h was indicated by the decrease of free fatty acid content in the product from 0.72 % to 0.69 %. Due to the regioselectivity of lipase-catalyzed hydrolysis, the omega-3/omega-6 ratio was increased from 17:83 in oil to 23:77 in produced diglycerides.

Lipase catalysed transesterification of palm stearin with ferulic acid in solvent-free media

Recently, several scientific groups investigated the suitable enzymatic route for synthesis of structured phenolic lipids for cosmetic and nutritive purposes. The feruloylated palm stearin (FPS) was obtained from enzymatic transesterification reactions of palm stearin with ferulic acid in solvent-free media with 1 mmol of glycerol or without it, using non-specific lipases, NovoCor ADL (Candida antarctica lipase A); Novozyme435® (Candida antarctica lipase B), and 1,3-specific lipase Lipozyme TL 100L from Thermomyces lanuginosus. The individual lipases and their combinational systems were examined. ESI+-MS/MS confirmed the formation of 19 feruloylated compounds. FPSs were produced by all lipases and their combinations. The positive synergetic effect occurred for 1,3-specific lipase and unspecified lipases with mass ratio 8:2, respectively. The antioxidant activities of all FPSs were examined. FPS produced by NovoCor ADL lipase in presence of free glycerol showed an antioxidant activity 10.3-times higher. In conclusion, the two low-cost lipases NovoCor ADL and Lipozyme TL 100L and its combi-lipase system could be considered suitable low-cost alternative to the expensive wide use N435 lipase in the solvent free production process of FPS for food and cosmetic applications.

Enzymatic synthesis optimization of isoamyl butyrate from fusel oil

Many food, cosmetic and pharmaceutical industries have increased their interest in short-chain esters due to their flavor properties. From the industrial standpoint, enzyme reactions are the most economical strategy to reach green products with neither toxicity nor damage to human health. Isoamyl butyrate (pear flavor) was synthesized by isoamyl alcohol (a byproduct of alcohol production) and butyric acid with the use of the immobilized lipase Lipozyme TL IM and hexane as solvents. Reaction variables (temperature, butyric acid concentration, isoamyl alcohol:butyric acid molar ratio and enzyme concentration) were investigated in ester conversion (%), concentration (mol L-1) and productivity (mmol ester g-1 mixture . h), by applying a sequential strategy of the Fractional Factorial Design (FFD) and the Central Composite Rotatable Design (CCRD). High isoamyl butyrate conversion of 95.8% was achieved at 24 hours. At 3 hours, the highest isoamyl butyrate concentration (1.64 mol L-1) and productivity (0.19 mmol ester g-1 mixture . h) were obtained under different reaction conditions. Due to high specificity and selectivity of lipases, process parameters of this study and their interaction with the Lipozyme TL IM are fundamental to understand and optimize the system so as to achieve maximum yield to scale up. Results show that fusel oil may be recycled by the green chemistry process proposed by this study.

Antioxidant activities of chlorogenic acid derivatives with different acyl donor chain lengths and their stabilities during in vitro simulated gastrointestinal digestion

In this study, chlorogenic acid (CA) was acylated with vinyl esters of different carbon chain lengths under the action of the lipase Lipozyme RM. Five CA derivatives (C2-CA, C4-CA, C6-CA, C8-CA, and C12-CA) with different lipophilicities were obtained, and their digestive stabilities and antioxidant activities were evaluated. The lipophilicities were positively correlated with the digestive stabilities of CA derivatives. The antioxidant activities of CA derivatives did not change with the reduction of phenolic hydroxyl groups, and their capacity to scavenge 2,2′-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+•) and 1,1-diphenyl-2-picrylhydrazyl (DPPH•) were similar to those of CA. In cellular antioxidant activity (CAA) tests, it was found that the capacity of these derivates to cross cell membranes were enhanced upon enhancing lipophilicity, and their antioxidant activities were improved. C12-CA showed the best antioxidant activity with a median effective dose (EC50) of 9.40 μg/mL, which was significantly lower than that of CA (i.e., 29.08 μg/mL).

Biodiesel production from non-edible high acid value phoenix seed oil using a cheap immobilized lipase

Fatty acid methyl ester (FAME) is one kind of renewable and biodegradable biodiesel energy. In this work, in order to decrease FAME production cost, a cheap immobilized lipase Lipozyme TLIM (Thermomyces lanuginosus lipase, 100$/kg) and waste phoenix seed (Firmiana platanifolia (L. f.) Marsili) oil with high free fatty acids (FFA) content (30.1 ± 0.4%) were used as catalyst and new feedstock, respectively. The effects of reaction variables were evaluated and optimized by response surface methodology. Results showed that Lipozyme TLIM can simultaneously catalyze the esterification and transesterification of high FFA phoenix seed oil to produce FAME. The conditions were optimized as follows: 11.8% water load added, 3.1:1 molar ratio of CH3OH to oil, and 7% lipase load at 33 °C for 6.47 h. High FAME yield (93.3 ± 1.6%) was achieved under the optimized conditions. Kinetic values (Vm and K’m) of FAME production were 2.66 × 10−2 mol/(L·min) and 7.4 mol/L, respectively, and the activation energy (Ea) was 42.62 kJ/mol. Therefore, a cheap immobilized lipase Lipozyme TLIM with a noted high level of tolerance of water could be useful in the industrial FAME production from phoenix seed oil with high FFA. After purified by molecular distillation, the contents of FAME, FFA, and water content of the final biodiesel product were 97.2%, 0.3%, and 0.04%, which were in accord with the quality standard of ASTM D6751 (>96.5%, <0.4%, and < 0.05%).

Simultaneous esterification and transesterification of waste phoenix seed oil with a high free fatty acid content using a free lipase catalyst to prepare biodiesel

Biodiesel is a typical renewable and green energy, that is usually produced by the transesterification of edible vegetable oils. However, the cost of edible vegetable oils as feedstock accounts for 60%–80% of the total cost of biodiesel production. To overcome these issues, in this work, a low-cost feedstock, waste phoenix seed oil (PSO) with a high free fatty acid (FFA) content (30.1 ± 0.6%), consisting of the main fatty acid, linoleic acid (30.2%), oleic acid (22.2%), and palmitic acid (17.4%), was developed as a new feedstock to prepare biodiesel. A novel one-step method including simultaneous esterification of FFAs and transesterification of the PSO to prepare biodiesel was developed. Free lipase was used as catalyst. The influences of the reaction parameters were explored and optimized by response surface methodology. The results showed that the free lipase Lipozyme TL100L from Thermomyces lanuginosus can simultaneously catalyze esterification and transesterification. Under the optimum conditions (enzyme load 9.7%, reaction time 6.9 h, reaction temperature 31 °C and substrate ratio (methanol/PSO) 4.3: 1 (mol/mol)), the maximum biodiesel yield (93.8 ± 0.5%) was achieved. The activation energy from biodiesel preparation using PSO with a high FFA content was 39.7 kJ/mol. Furthermore, the kinetic values for K'm and Vmax were 0.18 mol/L and 0.043 mol/(L·min), respectively. The reaction mechanism of the simultaneous esterification and transesterification was also proposed. These results indicate that PSO with a high FFA content can be used as a potential feedstock for biodiesel production.

Study of acetylated EGCG synthesis by enzymatic transesterification in organic media

(-)-Epigallocatechin-3-O-gallate (EGCG), the most abundant polyphenolic compound in catechins, exerts excellent physiological effects including antioxidant. However, with its high hydrophilicity and poor lipophilicity, the application of EGCG in oil products is limited. In this study, EGCG acetylated derivatives were prepared by transesterification of EGCG with vinyl ester in acetonitrile/isopropanol (1:1 v/v). Lipase Lipozyme RM IM was found to be the optimum catalyst at concentration of 12 U/g EGCG, with a molar ratio of 1:5 of EGCG to vinyl acetate as the substrates. And 83.2% conversion was obtained after 10 h reaction at 50 °C. Based on the ping-pong model, the kinetic equation was constructed to determine the reaction kinetic parameters. The analysis of the initial rate and progress curve indicated that the transesterification of EGCG and vinyl acetate was kinetically regulated. Two major acetylated derivatives were identified as 5″-O-acetyl-EGCG and 3″, 5″-di-O-acetyl-EGCG by LC-MS/MS and NMR. Their enhanced lipophilicity was confirmed by transmittance test and octanol–water partition coefficient. The antioxidant activity of di-acetylated EGCG was superior to mono-acetylated EGCG and EGCG, but slightly lower than tert-butyl hydroquinone (TBHQ) as determined by peroxide values (POV) and Rancimat test. Acetylated EGCG might be used as a potent antioxidant for controlling oxidation of oil.

Lipase-Catalysed In Situ Transesterification of Waste Rapeseed Oil to Produce Diesel-Biodiesel Blends

Rapeseed oil of high acidity, an agricultural industry by-product unsuitable for food, was used as an inexpensive raw material for the production of biodiesel fuel. The use of rapeseed oil that is unsuitable for food and lipase as a catalyst makes the biodiesel production process environmentally friendly. Simultaneous oil extraction and in situ transesterification using diesel as an extraction solvent was investigated to obtain a diesel-biodiesel blend. The diesel and rapeseed oil blend ratio was 9:1 (w/w). The enzymatic production of biodiesel from rapeseed oil with high acidity and methanol using eleven different lipases as biocatalysts was studied. The most effective biocatalyst, lipase—Lipozyme TL IM (Thermomyces lanuginosus), which is suitable for in situ transesterification—was selected, and the conversion of rapeseed oil into fatty acid methyl ester was evaluated. The influence of the amount of methanol and lipase, the reaction temperature and the reaction time were investigated to achieve the highest degree of transesterification. The optimal reaction conditions, when the methanol to oil molar ratio was 5:1, were found to be a reaction time of 5 h, a reaction temperature of 25 °C and a lipase (Lipozyme TL IM) concentration of 5% (based on oil weight). Under these optimal conditions, 99.90% (w/w) of the rapeseed oil was extracted from the seed and transesterified. The degree of transesterification obtained was 98.76% (w/w). Additionally, the glyceride content in the biodiesel fuel was investigated and met the requirements perfectly.

Preparation of phosphatidylcholine containing capric acid at the sn-1 position via Novozym® 435-catalyzed acidolysis

Abstract PC from soybean and C10:0 were subjected to lipase-mediated acidolysis/interesterification for the preparation of PC containing C10:0 (C10:0-PC). Free fatty acid form of C10:0 was selected as the best acyl donor in comparison to triacylglycerol from of C10:0 and ethyl ester from of C10:0 because higher C10:0-PC yield (46.2 ± 1.8 mol%) was obtained when free fatty acid form of C10:0 was used as acyl donor while that of triacylglycerol and ethyl ester were 24.9 ± 3.1 mol% and N.D., respectively in a preliminary reaction condition. In addition, Novozym® 435 was selected as the best catalyst compared to the commercial lipases Lipozyme® TL IM, Lipozyme® RM IM and Lipase OF® because the highest C10:0-PC yield (46.2 ± 1.8 mol%) was obtained when Novozym® 435 was used as a catalyst in a preliminary reaction condition. The reaction was then optimized based on the evaluation of acyl ratio (C10:0/PC), solvents, and reaction time, with mainly pursing C10:0-PC yield. It was demonstrated that high levels of acyl ratio decrease C10:0-incorporation, that solvents having low polarity are more effective in this reaction, and that the hydrolysis reaction was superior to the acidolysis reaction at the early stage of the process. Finally, C10:0-PC yield of 61.8 ± 3.0 mol% was obtained in the optimum reaction conditions, In this reaction system, Novozym® 435 showed regiospecificity at the sn-1 position of the PC molecule.

Incorporation of Caprylic Acid into a Single Cell Oil Rich in Docosahexaenoic Acid for the Production of Specialty Lipids.

SUMMARYResearch backgroundNew sources of docosahexaenoic acid have recently been investigated aiming at infant formula fortification and dietary supplementation, among which the single cell oil with 40-50% of this acid.Experimental approachFor this purpose, such an oil was blended with caprylic acid in amount substance ratio ranging from 1:1 to 5:1 and the blends were interesterified using either Novozym 435 or Lipozyme TL IM as the catalyst. The influence of the amount of excess free caprylic acid in the substrate, as well as the type of enzyme on the triacylglycerol rearrangement resulting from the synthesis of the structured lipids were evaluated.Results and conclusionsThe regiospecific lipase Lipozyme TL IM seemed to induce transesterification among single cell oil triacylglycerols preferably by acidolysis with caprylic acid, which was directly proportional to the ratio of this acid in the substrate. In reactions catalyzed by the non-regiospecific lipase Novozym 435, a higher incorporation of caprylic acid into single cell oil triacylglycerols was observed than when using Lipozyme TL IM, independently of the oil/caprylic acid molar ratio.Novelty and scientific contributionThese results revealed the importance of combining the choice of the type of lipase, either regiospecific or not, with the amount ratios of free fatty acids and the substrate in acidolysis when aiming to produce structured lipids as a source of docosahexaenoic acid.