Acidolysis Reaction Research Articles

Intensified synthesis of olive oil-based structured lipids based on enzymatic acidolysis using multi-frequency ultrasound

Intensified synthesis of structured lipids based on olive oil and capric acid through enzymatic acidolysis was investigated utilizing a multifrequency hexagonal ultrasonic reactor. The ultrasound-assisted process achieved a 59 % conversion of olive oil into structured lipids in just 4 h, compared to a 52 % conversion in 6 h with the conventional approach. The optimized parameters for the ultrasound-assisted process were a 4:1 reactant molar ratio, 4 % enzyme loading, 60 °C temperature, 30 W ultrasonic power, a 60 % duty cycle, and a combined frequency as 22 + 33 kHz. Sn-2 positional analysis confirmed the enzyme’s specificity for the glycerol 1,3 positions in the acidolysis reaction. Only 20 % loss in enzyme activity was observed after the sixth cycle, indicating the enzyme’s reusability within the process. Overall, the study effectively highlights the use of ultrasound as a powerful approach to intensify the enzymatic synthesis of designer lipids, while also elucidating the optimal operating conditions.

Enhancing the Thermostability and Catalytic Activity of the Lipase from Rhizopus oryzae via Introducing N-Glycosylation.

Lipase from Rhizopus oryzae (ROL) exhibits remarkable sn-1,3 stereoselectivity and catalytic activity, but its poor thermostability limits its applications in the production of 1,3-dioleoyl-2-palmitoyl glycerol (OPO, a high-quality substitute for human milk fat). In this work, a semirational method was proposed to engineer the thermostability and catalytic activity of 4M (ROL mutant in our previous study). First, a computer-aided design is performed using 4M as a template, and N-glycosylation mutants are then recombinantly expressed and screened in Pichia pastoris, the optimal mutant N227 exhibited a half-life of 298.8 h at 45 °C, which is 7.23-folds longer than that of 4M. Its catalytic activity also reached 1043.80 ± 61.98 U/mg, representing a 29.2% increase compared to 4M (808.02 ± 47.02 U/mg). Molecular dynamics simulations of N227 suggested that the introduction of glycan enhanced the protein rigidity, while the strong hydrogen bonds formed between the glycan and the protein stabilized the lipase structure, thereby improving its thermostability. The acidolysis reaction between oleic acid (OA) and glycerol tripalmitate (PPP) was successfully carried out using immobilized N227, achieving a molar conversion rate of 90.2% for PPP. This engineering strategy guides the modification of lipases, while the glycomutants obtained in this study have potential applications in the biosynthesis of OPO.

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.

Bio-imprinted lipase-catalyzed production of medium- and long-chain structured lipids rich in n-3 polyunsaturated fatty acids by acidolysis

Medium- and long-chain structured lipids (MLSLs) rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) were synthesized by acidolysis of fish oil (Omax 1812, a mixture of sardine and anchovy oils) with caprylic acid (CA) using a lipase bio-imprinted with fatty acids as a catalyst. A bio-imprinted lipase for the acidolysis reaction was first prepared under optimal conditions, and its activity was 1.43 times higher than that of the non-bioimprinted. The bio-imprinted lipase was used as a catalyst to produce such MLSLs, and the reaction conditions were optimized. Under the optimal conditions, equilibrium was achieved in 8 hr, and the total and the sn-1, 3 CA contents in the obtained product were 28.72% and 42.26%. Additionally, the total and the sn-2 DHA contents were 14.23% and 21.10%, respectively, and the total and the sn-2 EPA contents were 16.43% and 10.14%, respectively. These obtained MLSLs, rich in sn-1, 3 CA, and sn-2 DHA, EPA can deliver significant health benefits.

Effect of geometrical modifications on the mixing and enzymatic conversion of structured lipids in internal-loop airlift bioreactors

Airlift bioreactors present a compelling alternative for reactions utilizing immobilized enzymes, offering advantages over stirred tank and packed-bed reactors. This study proposed a new configuration for an internal-loop airlift bioreactor (1 L working volume) to produce structured triglycerides using immobilized lipases in a viscous fluid. Different internal tube configurations were studied, which varied in diameter (17.5–78.6 mm), angle (-2,0,5,10 and 12°), and the addition of a rounded bottom to the bioreactor, which improved the three-phase mixture flow. The hydrodynamic effect caused by these modifications was evaluated by studying the mixing time. The best internal tube configuration, featuring a truncated cone-shaped, exhibited a 5° angle and a 64.5-mm diameter, resulting in a mixing time of 54 s. This value was about three times less than the mixing time obtained by the standard tube (171 s) at 2 vvm. Sequentially, grape seed oil acidolysis reactions with capric acid (C10) catalyzed by the Lipozyme RM IM were performed, obtaining structured MLM triglycerides with an incorporation degree of 32.32% ± 1.01%. These results indicated good biocatalyst operational stability, with a half-life of 14.13 days. Thus, this work unveils a new configuration for an airlift bioreactor collectively driving promising advancements in structured lipid production.

Lipase-Catalyzed Preparation and Optimization of Structured Phosphatidylcholine Containing Nervonic Acid.

This study investigated the incorporation of nervonic acid into the chemical structure of phosphatidylcholine via a lipase-catalyzed acidolysis reaction to obtain a functional phospholipid. Lipase immobilization was conducted, and Amberlite XAD7-HP was selected as a carrier to immobilize phospholipase A1 (PLA1) for subsequent experiments. The main acidolysis reaction parameters, including enzyme load, substrate ratio, temperature, and water content, were studied against the reaction time. The optimum reaction conditions obtained were enzyme load, 20%; reaction temperature, 55 °C; water content, 1%; and reaction time, 9 h. The maximum incorporation of nervonic acid into phosphatidylcholine was 48 mol%, with PC recovery at 61.6 mol%. The positional distribution of structured phosphatidylcholine shows that nervonic acid was found in the sn-1 position due to enzyme specificity and in the sn-2 position, possibly due to acyl migration.

Enzymatic synthesis of structured lipids from sacha inchi (Plukenetia volubilis) oil with capric acid via acidolysis reaction in stirred tank and packed bed mini reactors

The present study describes an efficient and sustainable synthesis method for producing functional triglycerides (FTs) by conducting an acidolysis reaction of sacha inchi (Plukenetia volubilis) oil with capric acid (C10), catalyzed by Rhizopus oryzae (RO) lipase immobilized in low-cost corn cob powder, to obtain medium-large-medium chain (MLM) triglycerides. Initially, acidolysis reactions were performed between sacha inchi oil, C10, and the immobilized enzyme in a stirred tank at various temperatures (35 °C, 45 °C, 55 °C, and 59 °C), different molar ratios (1:2, 1:3, 1:4 oil:fatty acid), and various hydrolytic enzyme activities (83.6, 124, and 217 IU/mg). Subsequently, to optimize the degree of incorporation, the best condition from the stirred tank reactions (45 °C, 1:3 M ratio, and 217 U of hydrolytic activity) was selected, considering the highest nutritional quality of the lipid in terms of the migration of the sn2 position, and was tested in a packed bed reactor with a spatial time of 112 min, residence time of 107 min, and a flow rate of 0.5 mL/min. The highest incorporation degree achieved in packed bed reactor was 36% at 45 °C. Afterward, it was developed for the best condition an integrative platform for the recycling of the RO lipase immobilized in low-cost corn cob powder (up to four successive cycles without any make up). Subsequent assays on the functional triglycerides revealed atherogenic and thrombogenicity lipid index values of 0.04 ± 0.10 and 4.15 ± 0.05, respectively, which were lower than those of pure sacha inchi oil, indicating the potential to enhance the nutritional quality of the oil through C10 incorporation.

LIPASE FROM Rhizopus oryzae IMMOBILIZED ON CORN COB POWDER: A NEW APPROACH FOR DIETETIC TRIGLYCERIDE SYNTHESIS IN A FIXED BED REACTOR

This work explores the possibility of using corn cob powder as a carrier for immobilization by physical adsorption of lipase from Rhizopus oryzae used in acidolysis reaction of grape seed oil with capric acid (C10) to synthesize dietary triglycerides (TAGs) formed from medium-long-medium chain (MLM) fatty acids in a fixed bed reactor. First, enzymatic loading was investigated to improve hydrolytic activity and the immobilized biocatalyst was characterized in terms of biochemical parameters and thermal stability. An enzyme load of 30 mg mL-1 was quite adequate, as about 90% of the immobilization yield was achieved. The central composite rotatable design (CCRD) experimental design revealed that, for the immobilized enzyme, the optimal hydrolytic activity was obtained at pH 7.75 and 41 °C. Stirred tank reactions were optimized in CCRD statistical analysis revealing the most desirable conditions for molar ratio of 1:9.36 (oil:acid), 45 °C and 15% biocatalyst, obtaining a maximum ID value of 86.93 ± 2.25%. In a fixed bed reactor, the synthesis showed an average ID of 51.13 ± 0.77, demonstrating the potential of this configuration. It is concluded that corn cob powder as an immobilization carrier is a promising alternative for the enzymatic synthesis of MLM triacylglycerols.

Enzymatic production process of capric acid-rich structured lipids: Development of formulation as a new therapeutic approach

The present work reports an optimization of the synthesis of MLM-type (medium, long, medium) structured lipids (SL) through an acidolysis reaction of grape seed oil with capric acid catalyzed by Rhizopus oryzae lipase immobilized. At first, tests were carried out by preparing the biocatalysts using enzyme loadings (0.15 to 1 g of enzymatic powder) for each gram of support. Enzyme loading was used 0.3 g of enzymatic powder, and hydrolytic activity of 1860 ± 23.4 IU/g was reached. Optimized conditions determined by the Central Composite Rotatable Design (CCRD) revealed that the acidolysis reaction reached approximately 59 % incorporation degree (%ID) after 24 h, in addition to the fact that the biocatalyst could maintain the incorporation degree in five consecutive cycles. From this high incorporation degree, cell viability assays were performed with murine fibroblast cell lines and human cervical adenocarcinoma cell lines. Concerning the cytotoxicity assays, the concentration of MLM-SL to 1.75 and 2 % v/v were able to induce cell death in 56 % and 64 % of adenocarcinoma cells, respectively. Human cervical adenocarcinoma cells showed greater sensitivity to the induction of cell death when using emulsions with MLM-SL > 1.75 % v/v compared to emulsions with lower content indicating a potential for combating carcinogenic cells.

Bioimprinted lipase-catalyzed synthesis of medium- and long-chain structured lipids rich in docosahexaenoic acid for infant formula.

Medium- and long-chain structured lipids (MLSLs) rich in docosahexaenoic acid (DHA) were obtained in shorter reaction time by acidolysis of single-cell oil (DHASCO) from Schizochytrium sp. with caprylic acid (CA) using a lipase bioimprinted with fatty acids as a catalyst. The conditions for preparation of the bioimprinted lipase for the acidolysis reaction were firstly optimized and the activity of the obtained lipase was 2.17 times higher than that of the non-bioimprinted. The bioimprinted lipase was then used as a catalyst and the reaction conditions were optimized. Under the optimal conditions, the equilibrium could be achieved in 4h, and the total and sn-1,3 CA contents in the product were 29.18% and 42.34%, respectively, and the total and sn-2 DHA contents were 46.26% and 70.12%, respectively. Such MLSLs rich in sn-1,3 CA and sn-2 DHA are beneficial for DHA absorption, and thus have potential for use in infant formula.

Investigation on the catalysis performance of metal-doped zeolites on ring-opening acidolysis reaction

Metal-incorporated zeolites as heterogeneous Lewis acid were applied in the ring-opening acidolysis of ethylene oxide with acrylic acid to prepare 2-hydroxyethyl acrylate (HEA). The catalysis characteristics, including Lewis acid strength and internal mass transfer resistance, were systematically investigated to facilitate the rational design of zeolite catalysts. Through modifying the Lewis acid strength of zeolites via different metals (Sn, Zr and Ti) incorporation and the internal mass transfer resistance of zeolites from the particle size, pore diameter or pore structure, it was found that the increased Lewis acidity can accelerate both the main and side reactions, resulting in the high activity but low selectivity of zeolite catalysts; the enhanced internal diffusion was the key to limiting subsequent cascade alcoholysis side reactions as well as the decline of catalytic performance. Based on these findings, Zr-containing zeolites featured with moderate Lewis acidity and uniform intracrystalline mesoporosity were designed and prepared, presenting excellent catalysis and recycling performance. These results demonstrated the great potential of the developed zeolite catalysts for the green synthesis of HEA as well as the similar ring-opening acidolysis of epoxides.

Extreme UV Resist Exhibiting Synergism between Chemical and Physical Crosslinking Mechanisms.

Carbon-fluorine bonds in fluorinated molecules can undergo homolytic cleavage reactions when electrons are injected, and the resulting radicals combine to form network structures characterized by reduced solubility. This crosslinking chemistry suggests a new category of patterning materials that function under electron beam (e-beam) and extreme ultraviolet (EUV) lithographic conditions. Although this chemistry enables the production of 50 nm or smaller-sized features of simple fluoroalkylated polymers, it is limited by the need for relatively large amounts of irradiation energy to achieve required solubility changes. Therefore, this study was undertaken to devise a sensitivity-enhancing strategy based on a synergistic combination of radical crosslinking and hydrogen-bonding interactions between highly fluoroalkylated copolymers. An alternating copolymer was synthesized using tert-butoxystyrene and a fluoroalkylated maleimide, the former of which produces active hydrogens through catalytic acidolysis reactions. When the polymer was blended with a catalytic amount of a photoacid generator and subjected to lithographic patterning tests under e-beam and EUV irradiation, the deprotection reactions of tert-butoxy moieties proceeded at room temperature and led to a solubility decrease. We presume the small number of hydroxyl moieties produced formed an intermolecular hydrogen-bonding network, which acted synergistically with the covalent crosslinks generated by C-F bonds. When 30 nm features of copolymer thin films were fabricated by EUV lithography, sensitivity was improved by 25-34% without significant deterioration of pattern quality, especially line-edge roughness. These results demonstrate that EUV resists with improved patterning capabilities can be achieved by combining catalytic acidolysis reactions and noncatalytic crosslinking chemistry.

Isolation of Fatty Acids from the Enzymatic Hydrolysis of Capsaicinoids and Their Use in Enzymatic Acidolysis of Coconut Oil.

Herein we report the optimization of enzymatic hydrolysis of a mixture of capsaicinoids, capsaicin and dihydrocapsaicin obtained from chili peppers, and the utilization of the isolated fatty acids for the modification of coconut oil using enzyme catalyzed acidolysis. This work was carried out as the fatty acids that can be isolated from capsaicinoid hydrolysis have been shown to possess interesting biological properties. These biological properties could be better exploited by incorporating the fatty acids into a suitable delivery vehicle. The enzymatic hydrolysis of the mixture of capsaicin and dihydrocapsaicin was carried out using Novozym® 435 in phosphate buffer (pH 7.0) at 50℃. The enzyme catalyst could be reused in multiple cycles of the hydrolysis reaction. The desired 8-methyl-6-trans-nonenoic acid and 8-methylnonanoic acid were isolated from the hydrolysis reaction mixture using a simple extraction procedure with a 47.8% yield. This was carried out by first extracting the reaction mixture at pH 10 with ethyl acetate to remove any dissolved capsaicinoids and vanillyl amine side product. The fatty acids were isolated after adjustment of the pH of the reaction mixture to 5 and second extraction with ethyl acetate. The acidolysis of coconut oil with the obtained fatty acids was performed using Lipozyme® TL IM. The performance of the acidolysis reaction was evaluated using 1H-NMR spectroscopy and verified in selected cases using gas chromatography. The best performing conditions involved carrying out the acidolysis reaction at 60℃ with a 1.2 w/w ratio of the fatty acids to coconut oil and 10% enzyme loading for 72 h. This resulted in the incorporation of 26.61% and 9.86% of 8-methyl-6-trans-nonenoic acid and 8-methylnonanoic acid, respectively, into the modified coconut oil product. This product can act as a potential delivery vehicle for these interesting compounds.

Formation rate of benzyl cations in various aqueous solutions containing different concentrations of acid but with a specific proton activity in lignin acidolysis

Abstract This study aimed to clarify the effect of different solvents, aqueous 1,4-dioxane, ethanol, or ethylene glycol at various mol%, on the formation rate of the benzyl cation, which forms via two steps, the protonation of the benzyl hydroxy or alkoxy group as a pre-equilibrium step and subsequent liberation of the water or corresponding alcohol molecule, respectively, in a model system of the lignin acidolysis, focusing on distinguishing the effects on both steps. For this aim, the acidolysis reactions were conducted in various aqueous solutions and organic solvents that showed a specific proton activity to establish the pre-equilibrium at a specific position. The formation rate of the benzyl cation showed minima in the intermediate mol% range. However, these rate differences were much smaller than those observed in the previous report where the aqueous solutions were prepared to contain a specific acid concentration and show different proton activities and thus the net effect of the solvent type on both steps was indistinguishable. These results clearly showed that the solvent type affects the first step (the proton activity) much more than the last step (the solvent-dissociating power). This paper also describes the preparation of the aqueous solutions to demonstrate a specific proton activity.

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.

Acidolysis of phospholipids with medium-chain fatty acids over M-SBA-15 (M = Zn, Al) silicas as efficient solid catalysts.

Efficient and sustainable production of structured phospholipids (SPLs) enriched in medium-chain fatty acids (MCFAs) in a heterogeneous manner is crucial for their potential applications in functional foods and drugs. Herein, for the first time, Zn- and Al-incorporated SBA-15 silicas were prepared by the coprecipitation method and further researched for catalytic synthesis of MCFA-enriched SPLs through acidolysis reaction of natural phospholipids with capric or caprylic acid. The as-prepared Zn- and Al-incorporated SBA-15 samples exhibited superior catalytic activities under mild experimental conditions (50 °C, 6h) to commercial homogeneous Lewis acids and benchmark enzymes. Correspondingly, the capric acid and caprylic acid incorporations were respectively achieved up to ~40.25 ± 0.40% (or 35.08 ± 0.09%) and 37.26 ± 0.38% (or 33.02 ± 0.13%) for Zn- (or Al-) incorporated SBA-15 catalyst. Moreover, various methods such as scanning electron microscopy with energy-dispersive X-ray spectrometry, ultraviolet-visible diffuse reflectance spectroscopy and pyridine-Fourier transform infrared spectroscopy were utilized to characterize the two catalysts in order to elucidate the possible structure-performance relationship. Accordingly, the above-mentioned satisfactory results are most probably due to the well-ordered mesostructures and large amounts of active Lewis acid sites existing in the investigated materials. Noticeably, the two catalysts featured good separation and excellent recyclability as well. The Zn- and Al-incorporated SBA-15 catalysts studied in this work might shed light on novel, sustainable and economic alternatives for effective SPL production to diminish the applications of conventional homogeneous catalysts and biocatalysts in food industries. © 2022 Society of Chemical Industry.

Advanced oxidation processes for wet-process phosphoric acid: Enhanced phosphorus recovery and removal of organic matters

Green and efficient phosphorus leaching from phosphate rock (PR) is the key to producing high-quality products and reducing energy consumption. Organic matters in the PR affect the efficiency of wet-process phosphoric acid production. In this study, advanced oxidation processes (AOPs) were employed to remove organic matters, thereby intensifying the acidolysis reaction of the PR. The characteristics, effects of reaction conditions on the leaching efficiency, and leaching reaction were systematically studied. Under the optimal reaction conditions with the H2O2 concentration of 0.04 mL/mL, reaction temperature of 70 °C, reaction time of 120 min, and particle size of −150 μm, a high leaching efficiency of 88.4% was achieved in the O3-H2O2 reaction (OHR) case. In addition, the total organic carbon content in the leach liquor is 12.2 mg/L, which is significantly decreased by 79.9%. This demonstrated the advantages of AOP systems in strengthening the efficient leaching of phosphorus as well as beneficial environmental impact.

Argan Oil as a Rich Source of Linoleic Fatty Acid for Dietetic Structured Lipids Production.

Argan oil is rich in long-chain unsaturated fatty acids (FA), mostly oleic and linoleic, and natural antioxidants. This study addresses the production of low-calorie structured lipids by acidolysis reaction, in a solvent-free system, between caprylic (C8:0; system I) or capric (C10:0; system II) acids and argan oil, used as triacylglycerol (TAG) source. Three commercial immobilized lipases were tested: Novozym® 435, Lipozyme® TL IM, and Lipozyme® RM IM. Higher incorporation degree (ID) was achieved when C10:0 was used as acyl donor, for all the lipases tested. Lipozyme® RM IM yielded the highest ID for both systems (28.9 ± 0.05 mol.% C10:0, and 11.4 ± 2.2 mol.% C8:0), being the only catalyst able to incorporate C8:0 under the reaction conditions for biocatalyst screening (molar ratio 2:1 FA/TAG and 55 °C). The optimal conditions for Lipozyme® RM IM in system II were found by response surface methodology (66 °C; molar ratio FA/TAG of 4:1), enabling to reach an ID of 40.9 mol.% of C10:0. Operational stability of Lipozyme® RM IM in system II was also evaluated under optimal conditions, after eight consecutive 24 h-batches, with biocatalyst rehydration between cycles. The biocatalyst presented a half-life time of 103 h.

Process intensification of acidolysis reaction catalysed by enzymes for synthesis of designer lipids using sonication

Designer lipids find a wide range of applications in food, pharmaceuticals and nutraceutical industries. The current work focuses on intensified production of designer lipids using ultrasound assisted enzymatic acidolysis reaction between long chain triglycerides from Linseed oil (LO) and MCFAs i.e. octanoic acid or decanoic acid. The synthesised designer lipids contain sn-1,3 position on glycerol backbone occupied by MCFAs and sn-2 position occupied by PUFAs, thus having the potential of numerous applications in food and pharma sector. Studies into effect of ultrasound parameters and reaction conditions on the yield demonstrated that the optimum operating conditions as molar ratio of LO: MCFAs of 1:5, temperature of 40 °C and Novozyme 435 dosage of 5% (based on substrate weight) as well as ultrasound conditions of 50% duty cycle and 90 min of irradiation time resulted in maximum yield as 96%. Comparison of ultrasound assisted synthesis with the conventional synthesis (less than 20% yield in 90 min and requirement of 26 h to reach equilibrium yield of 82.6%) demonstrated the intensification benefits as higher yield and lower reaction time for the ultrasound assisted approach. It was also elucidated that Novozyme 435 was effectively recycled for 12 cycles without much effect on the yield. Overall the work clearly demonstrated the efficacy of the ultrasound intensified approach for synthesis of designer lipids, also elucidating key design information for the optimum operating conditions.

Synthesis of lipase/silica biocatalysts through the immobilization of CALB on porous SBA-15 and their application on the resolution of pharmaceutical derivatives and on nutraceutical enrichment of natural oil

• A recombinant lipase was produced in a low-cost medium. • The immobilization process was studied. • The use of ammonium fluoride, APTES and GA allowed a most active biocatalyst. • Resolution of racemic myo -inositol derivatives was done with eep>99 %. • It was possible to obtain lipids containing 181.6 mg of EPA and DHA in coconut oil. A recombinant lipase from Candida antarctica , LIPB, was obtained through fermentation using an alternative low-cost medium and immobilized on the distinct mesoporous silica particles. The performance and versatility of the new biocatalysts was evaluated, for the first time, on two biotechnological interest reactions: resolution of racemic myo-inositol derivative and acidolysis reaction of coconut oil. For the racemate, the covalent biocatalyst SBA-15-F-APTES-GA-LIPB exhibited the highest conversions and enantiomeric excess of up to 99 %. It was the first time that covalent biocatalyst was used for resolution of racemic myo-inositol derivative. The most active biocatalyst (SBA-15-F-APTES-GA-LIPB) was used on ten different reaction cycles maintaining its activity, highlighting the operational stability of the developed biocatalyst. Moreover, it was possible to produce food oils enriched with polyunsaturated fatty acids (PUFA), obtaining lipids containing 181.6 mg of EPA and DHA in coconut oil. The preliminary economic analyses also indicate that the enzymatic production of such nutraceuticals and pharmaceutics compounds may be economically feasible, principally exploring low-cost strategies for enzyme production.