Enzymatic Transesterification Research Articles

Insight into in situ enzymatic transesterification modification of polyethylene terephthalate fibers with polyethylene glycol

Polyethylene terephthalate (PET) fabric was enzymatically modified with polyethylene glycol (PEG). Lipase from Aspergillus oryzae was used as a catalyst. The wetting behavior of the modified PET fabric was characterized by the water contact angle, water adsorption dynamics and moisture regain. The initial water contact angle of the modified fabric decreased from 129.3° to 74.9°. The water absorption dynamics results indicated that the equilibrium water absorption amount of the modified fabric was approximately 10 times greater than that of the untreated sample. The initial absorption rate (Vinit) of the modified fabric was almost 30 times higher than that of the untreated fabric. The moisture regain reached 3.52% after modification. Scanning electron microscopy revealed granular and sheet coatings on the surface of the modified fabric. Chemical linkage of PEG onto the PET fibers was confirmed by attenuated total reflection–Fourier transform infrared spectroscopy. The modification had no significant effect on the thermal properties of the PET fabrics according to the thermogravimetric curve and differential scanning calorimetry results. The mechanical properties of the modified fabric were slightly improved. Therefore, in situ enzymatic transesterification between PET and PEG could be a potential novel modification approach for adding value to PET fiber textiles.

Enzymatic synthesis of aromatic biobased polymers in green, low-boiling solvents

Given the urge to accelerate the substitution of petrol-derived solvents not only in more traditional fields like pharmaceuticals, personal care, or electronics but also in innovative research processes, this work focuses on the utilisation of four biobased solvents as media for the enzymatic synthesis of aliphatic-aromatic polyesters. As building blocks, the lignin-derived diethyl-2,4-pyridinedicarboxylate was selected as the potentially biobased, aromatic component while more classical diols such as 1,4-butanediol and 1,8-octanediol were used as the aliphatic portion. Results show that among the tested green solvents (cyclohexanone, phenetole, anisole and eucalyptol), the most suitable medium for lipase B from Candida antarctica-catalysed polycondensation reactions was eucalyptol that allowed reach monomer conversions >95 % and number average molecular weights up to 3500 g·mol−1. On the other hand, cyclohexanone led to the lowest monomer conversions (<80 %) and molecular weights (Mn<500 g·mol−1) confirming once again the unsuitability of ketone-containing solvents for enzymatic esterification and transesterification reactions. The lipase could be used up to three times, in eucalyptol as a solvent, without a significant decrease in monomer conversion or molecular weight.

Zr-based MOF as a support for lipase immobilization to enhance enzymatic transesterification for biodiesel production

The development of novel biocatalysts is essential to promote the commercialization of biodiesel production by transesterification reaction. In this paper, Rhizopus oryzae lipase (ROL) was immobilized on an amino-functionalized zirconium-based metal organoskeleton by interfacial adsorption. The immobilization conditions were optimized and the enzymatic properties were tested, and the resulting novel biocatalysts exhibited higher stability and heat resistance. SEM, XRD and BET analyses were used to characterize the biocatalysts and carrier materials. The catalytic performance of ROL@UiO-66-NH2 in the production of biodiesel by transesterification reaction was explored, and the production process was optimized by response surface method. The results showed that the conversion rate of FAEE reached 82.05% at molar ratio of ethanol/oil of 15.43:1, reaction temperature of 50.28°C, reaction time of 120.9 min, DES addition of 48.08 wt%, biocatalyst addition of 3 wt%, and ultrasonic power of 90 W. In addition, ROL@UiO-66-NH2 demonstrated good recyclability, with the catalytic efficiency remaining at 71.87% after five cycles.

Techno-economic assessment of co-production of biodiesel and epoxy fatty acid methyl ester as renewable fuel and plasticizer from waste cooking oil

Diversifying waste cooking oil (WCO)-based biodiesel production towards epoxy fatty acid methyl esters (EFAMEs) offers a double gain, boosting revenue channels in the biodiesel sector and driving the development of sustainable phthalate plasticizer alternatives. This study explored the techno-economic feasibility of co-derived biodiesel and EFAMEs from WCO using a three-step process involving enzymatic transesterification, urea complexation, and performic acid epoxidation. The result shows that a plant processing 61,300 tonnes/year of WCO, may produce ∼27,770 tonnes/year of biodiesel and ∼30,180 tonnes/year of EFAMEs, with a total heating duty of 1,124 kW and annual electricity consumption of 34.9 million kWh. The initial capital investment of the plant in China amounts to 136.8 MM CNY, with an annual manufacturing cost of 460 MM CNY in 2022. Techno-economic evaluation demonstrats a net present value (NPV) of 421.9 MM CNY at a 10 % discount rate, an internal rate of return (IRR) of 37.69 %, and discounted payback period of 2.37 years. The break-even point (BEP) is estimated at an annual sales volume of total products of 27,101 tonnes/year, including 12,790, 11,769, 2,542, and 2,390 tonnes/year of epoxy plasticizers, biodiesel, glycerol, and polymeric grease, respectively, collectively utilizing 42.38 % of the processing capacity. Monte Carlo simulation indicates minimal risk of the project, with probabilities of 98.5 % for IRR > 10 % and 97.8 % for a positive NPV. This study affirms the economic viability of co-producing biodiesel and EFAMEs from WCO for industrial application.

Modeling of enzymatic transesterification for omega-3 fatty acids enrichment in fish oil

Mathematical models of transesterification commonly assume that oil is a mixture of triacylglycerols, where each component has only one type of acid attached. This article aims to show how a different assumption on acid distribution affects the results of acylglycerols fraction composition. Experiments of fish oil ethanolysis have been performed at different enzyme loadings and ethanol concentrations, leading to enrichments from 35 % to 52 % of ω3 mass fraction in acylglycerols, by losing 12.1 % of ω3 as ethyl esters. A kinetic model is developed assuming both all acids of the same type on each acylglycerol and all acids randomly distributed on the available positions. The two different assumptions showed strong discrepancies on the acylglycerols fraction compositions predictions, demonstrating how the initial fatty acids distribution is important when an accurate description of the acylglycerols fraction is desired.

Modeling the Effects of Temperature in Enzymatic Biodiesel Synthesis of Jatropha Oil: An Optimal Control Approach.

Biodiesel, an alternative to diesel, is produced through the enzymatic transesterification of vegetable oil or animal fats. Enzymatic transesterification of oil is gaining more importance, as this method possesses no such disadvantages that are associated with the chemical process. Temperature is the most important factor in enzymatic transesterification for biodiesel synthesis. In this study, a mathematical model is developed to understand the effects of temperature on the enzymatic transesterification of Jatropha curcas oil. Reaction rates are expressed as a function of temperature using an appropriate function, and the effects of temperature on the overall enzymatic system are investigated using the mathematical model. A suitable temperature is determined using the mathematical model, keeping the other reaction conditions unchanged that gives maximum yield. Furthermore, an optimal control problem (OCP) is formulated to identify the temperature control strategy that maximizes the biodiesel yield while minimizing production costs. Simulated outcomes obtained from the mathematical model are analogized with the experimental results to make them acceptable. The optimal profile of temperature is determined from the developed OCP, which maximizes the biodiesel yield.

Enzymatic Transesterification Using Different Immobilized Lipases and its Biodiesel Effect on Gas Emission

Biodiesel, a third-generation bio-fuels, offering several advantages over regular diesel fuel. Waste cooking oil (WCO) emerges as an ideal feedstock due to its availability and easy accessibility. In this work, biodiesel is utilized from two different types of immobilized lipases: Rhizomucor miehei lipase (RMIM) and Candida antarctica lipase B (CALB). The impact of the molar ratio of oil to methyl acetate (1:3-1:12) was evaluated for both lipases, and the resultant biodiesel was tested in diesel engine. The enzymatic transesterification was carried out in ultrasonic assistance and the results showed that the greatest yield of 81.20% at 45℃, using CALB as a biocatalyst, 1.8% (w/v) lipase and oil to methyl acetate molar ratio of 1:12 within 3 hours. Triacetin, by-product was determined their concentration for each molar ratio and analyzed using FTIR range of 500cm-1 to 4000cm-1, revealing a significant absorption peak at 1238.90cm-1. Biodiesel was blended with commercial diesel fuel in varying quantities of 7, 10, and 20% by volume (B20). The results were compared to Industrial Diesel Fuel 7% (B7) and Commercial Diesel Fuel 10% (B10). NOx and CO2 emission drops as the percentage of diesel/biodiesel blends increases, supporting WCO as a cost-effective biodiesel feedstock with low petrol pollution.

In-situ stabilized lipase in calcium carbonate microparticles for activation in solvent-free transesterification for biodiesel production

Biodiesel serves as a crucial biofuel alternative to petroleum-based diesel fuels, achieved through enzymatic transesterification of oil substrates. This study aims to investigate stabilized lipase (LP) within calcium carbonate (CaCO3) microparticles as a catalyst for solvent-free transesterification in biodiesel synthesis. The specific hydrolysis activity of the in-situ immobilized LP was 66% of that of free LP. However, the specific transesterification activity of immobilized LP in the solvent-free phase for biodiesel production was 2.29 times higher than that of free LP. These results suggest that the interfacial activation of LP molecules is facilitated by the inorganic CaCO3 environment. The immobilized LP demonstrated higher biodiesel production levels with superior stability compared to free LP, particularly regarding methanol molar ratio and temperature. To the best of our knowledge, there are no previous reports on the in-situ immobilization of LP in a CaCO3 carrier without any crosslinker as an interfacial-activated biocatalyst for biodiesel production.

An Exploratory Study of the Enzymatic Hydroxycinnamoylation of Sucrose and Its Derivatives.

Phenylpropanoid sucrose esters are a large and important group of natural substances with significant therapeutic potential. This work describes a pilot study of the enzymatic hydroxycinnamoylation of sucrose and its derivatives which was carried out with the aim of obtaining precursors of natural phenylpropanoid sucrose esters, e.g., vanicoside B. In addition to sucrose, some chemically prepared sucrose acetonides and substituted 3'-O-cinnamates were subjected to enzymatic transesterification with vinyl esters of coumaric, ferulic and 3,4,5-trimethoxycinnamic acid. Commercial enzyme preparations of Lipozyme TL IM lipase and Pentopan 500 BG exhibiting feruloyl esterase activity were tested as biocatalysts in these reactions. The substrate specificity of the used biocatalysts for the donor and acceptor as well as the regioselectivity of the reactions were evaluated and discussed. Surprisingly, Lipozyme TL IM catalyzed the cinnamoylation of sucrose derivatives more to the 1'-OH and 4'-OH positions than to the 6'-OH when the 3'-OH was free and the 6-OH was blocked by isopropylidene. In this case, Pentopan reacted comparably to 1'-OH and 6'-OH positions. If sucrose 3'-O-coumarate was used as an acceptor, in the case of feruloylation with Lipozyme in CH3CN, 6-O-ferulate was the main product (63%). Pentopan feruloylated sucrose 3'-O-coumarate comparably well at the 6-OH and 6'-OH positions (77%). When a proton-donor solvent was used, migration of the 3'-O-cinnamoyl group from fructose to the 2-OH position of glucose was observed. The enzyme hydroxycinnamoylations studied can shorten the targeted syntheses of various phenylpropanoid sucrose esters.

A multi reaction kinetic model to describe the enzymatic transesterification reaction of jatropha oil using a fermented solid containing lipases

The advancement of more precise tools for sustainable process design in enzymatic biodiesel synthesis from renewable sources is crucial. Kinetics of solvent-free transesterification reactions were conducted across a temperature spectrum from 30 °C to 60 °C, utilizing Jatropha curcas oil (TG) and ethanol as substrates, alongside a fermented solid by Rhizopus homothallicus as the biocatalyst. The dynamics of chemical species concentrations were monitored through High-performance Thin-Layer Chromatography. Maximum productivities were achieved at 35 °C and 60 °C for biodiesel (293.24 and 299.02 g kg biocat−1 h−1, respectively), at 40 °C for diglycerides (1018.36 g kg biocat−1 h−1), and at 35 °C for monoglycerides (560.75 g kg biocat−1 h−1). Maximum yields were determined at 30 °C for fatty acid ethyl esters (0.56 g gTG−1), and at 40 °C for diglycerides (0.53 g gTG−1) and monoglycerides (0.30 g gTG−1). Based on the experimental findings, a kinetic model was formulated encompassing three reversible transesterification reactions. Individual reactions were structured following classical biochemical kinetics, inclusive of ethanol inhibition. Model fitting was executed through non-linear multivariable regression techniques, with the minimum of the average coefficient of variation of the residuals (ACVR) serving as the objective function. The resulting fit of the kinetic model to the experimental data proved satisfactory, with an ACVR of less than 5 % across all instances. Notably, the maximum biodiesel productivity, obtained in this work, represented the highest value, compared to other related studies, using a fermented solid as a biocatalyst.

Construction of a Pickering interfacial biocatalysis system in skim milk and enzymatic transesterification for enhancement of flavor and quality

Construction of a Pickering interfacial biocatalysis system in skim milk and enzymatic transesterification for enhancement of flavor and quality

Ultrasound-assisted intensification of Pickering interfacial biocatalysis preparation of vitamin A aliphatic esters

A novel approach to ultrasound-assisted Pickering interfacial biocatalysis (PIB) has been proposed and implemented for the efficient enzymatic transesterification production of vitamin A fatty acid esters. This is the first instance of exploiting the synergistic effect of ultrasound and the bifunctional modification of enzyme supports to accelerate biocatalytic performance in PIB systems. The optimal conditions were determined to be ultrasound power of 70 W, on/off time of 5 s/5 s, substrate molar ratio of 1:1, enzyme addition of 2 %, and a volume ratio of n-hexane to PBS of 3:1, a temperature of 40 °C, and a time of 30 min. The application of ultrasound technology not only improved lipase activity but also allowed for a reduction in emulsion droplet size to enhance interfacial mass transfer.Bifunctional modification of silica-based supports enhanced stability of immobilized enzymes by increasing hydrogen bonding while maintaining the active interface microenvironment. Compared with a non-ultrasound-assisted PIB system stabilized by mono-modified immobilized enzyme particles, the catalytic efficacy (CE) of the novel system reached 8.18 mmol g−1 min−1, which was enhanced by 3.33-fold, while the interfacial area was found to have increased by 17.5-fold. The results facilitated the conversion of vitamin A palmitate (VAP), vitamin A oleate (VAO), vitamin A linoleate (VAL), and vitamin A linolenate (VALn), with conversion rates of approximately 98.2 %, 97.4 %, 96.1 %, and 94.7 %, respectively. This represents the most efficient example that has been reported to our knowledge. Furthermore, the system demonstrated improved reusability, with a conversion rate of 62.1 % maintained even after 10 cycles. The findings presented in this paper provide valuable insights into an efficient and conveniently promising protocol for the development of PIB systems.

Combining lipase enzymatic techniques and antioxidants on the flavor of structured lipids (SLs) prepared from goat butter and coconut oil

Enzymatic transesterification can improve the quality of goat butter, however, the high temperature may lead to oil oxidation and produce a bad taste. In this study, goat butter (GB) and coconut oil (CO) were combined to synthesize structured lipids (SLs) by enzymatic transesterification, and antioxidants were added to inhibit oxidation to improve the oils’ flavor. The Lipozyme TLIM and tert-butylhydroquinone (TBHQ) were screened for SLs synthesis. The contents of SLs were 56.24% under the optimal conditions of 8 h reaction time, 5°C reaction temperature, 8% enzyme dosage, and 3:1 (w/w) substrate ratio, and the addition of antioxidants had no significant effect on the contents of SLs. It was demonstrated that short-chain fatty acids in the SLs were significantly reduced by 59.2%, and the medium-chain fatty acids were increased by 54.9% compared to GB. One hundred volatile compounds were identified in the lipid samples and classified as (1) long-chain alkane aromatic components and aromatic components, which remarkably decreased by 58.9%, and (2) alcohols, aldehydes, and ketones, which increased by 12.9%, through using solid-phase microextraction coupled with gas chromatography-mass spectrometry (SPME-GC-MS) and electronic nose. The relative aroma activity value (ROAV) of 9 key volatile aroma components was higher than 1 in the SLs, which greatly contributed to the reduction of goaty flavor and increasing cream flavor. The results suggested that the goaty flavor of GB could be significantly reduced by decreasing the short-chain fatty acids contents in SLs through enzymatic transesterification with CO, while the addition of antioxidants efficiently inhibited excessive lipids oxidation and increased the content of long-chain aldehydes, ketones, and lactones, thus enhancing the cream flavor, which had great potential to be utilized as natural food flavor additives.

Optimization of Enzymatic Transesterification of Acid Oil for Biodiesel Production Using a Low-Cost Lipase: The Effect of Transesterification Conditions and the Synergy of Lipases with Different Regioselectivity.

This study describes the enzymatic production of second-generation biodiesel using low-quality acid oil as a substrate. Biolipasa-R, a commercially available and low-cost lipase, was employed for enzymatic transesterification. Response surface methodology was applied to optimize the enzymatic transesterification process. The optimal conditions for biodiesel production, which comprised 42% lipase concentration (per weight of oil), 32% water content (per weight of oil), a methanol to oil molar ratio of 3:1, pH 7.0 and reaction temperature 30°C, resulted in the highest fatty acid methyl ester (FAME) content (71.3%). Subsequently, the synergistic effect of two lipases with different regioselectivities under the optimum transesterification conditions was studied, aiming at the enhancement of process efficiency. The transesterification efficiency of immobilized Biolipasa-R was determined and compared to that of Biolipasa-R in its free form. The results revealed a good performance on FAME content (66.5%), while the recycling of immobilized lipase resulted in a decrease in transesterification efficiency after three consecutive uses.

Efektifitas Reaksi Transesterifikasi Minyak Kelapa dan Sereh untuk Pembuatan Perisa Alami dengan Biokatalisator Lipase serta Penggunaan Ultrasonik

In general, flavor are synthesis through enzymatic hydrolysis and esterification reactions (up to 20 hours) using commercial ingredients based on free fatty acids and alcohol. Efficient endeavors utilizing enzymatic transesterification to expedite reactions are necessary to acquire flavored products. Substrates may originate from commercial or natural sources abundant in fatty acids and alcohol. Commercial fatty acids and alcohol (geraniol) are readily available in pure forms. Alternatively, fatty acids can be sourced from coconut oil, while geraniol can be derived from lemongrass oil, which is more cost-effective compared to commercial ingredients. Ultrasonics have emerged as a means to expedite enzymatic. The objective of this study is to investigate the impact of ultrasonic power and transesterification reaction time between coconut oil and geraniol on geraniol conversion. Utilizing lipase (415.99 U/g), coconut oil was subjected to transesterification with commercially obtained pure geraniol in a 1:3 weight ratio, conducted in an ultrasonic tank filled with water. Reaction durations spanned 30, 60, 90, 120, and 150 minutes, with ultrasonic powers set at 50, 70, and 90 watts. The study findings elucidated that the highest geraniol conversion rate of 50.59% was achieved with 70 watts of ultrasonic power over a 90-minute period. These optimal conditions were subsequently applied to transesterify coconut oil with geraniol from lemongrass oil, yielding a conversion rate of 90.29%. This finding demonsttrate the posibility of employing coconut oil and lemongrass oil in a one-stage transesterification process to produce natural flavors via enzymatic lipase catalysis expedited by ultrasonic technology, facilitating swift reaction times.

Preparation of functional oils rich in phytosterol esters and diacylglycerols by enzymatic transesterification

Phytosterol esters (PEs) and diacylglycerols (DAGs) have various health benefits in humans. In this study, PEs and DAGs were synthesized by lipase-catalyzed transesterification between a natural oil and phytosterols. First, commercial lipases were screened for transesterification and were further verified using multiple-ligand molecular docking. AYS “Amano” (a lipase from Candida rugosa) was found to be the optimum lipase. Subsequently, the enzymatic transesterification conditions were optimized. The optimized conditions were determined to be a 1:2 M ratio of phytosterols to oil, 100 mmol/L phytosterols, and 9 % AYS “Amano”, and 50 °C for 24 h in 20 mL n-hexane. Under these conditions, over 70 % of phytosterols were converted to PEs. In this study, an efficient enzymatic process was developed to produce value-added functional oils rich in PEs and DAGs, with PEs content ≥ 31.6 %, DAGs content ≥ 11.2 %, acid value ≤ 0.91 mg KOH/g, and peroxide value ≤ 2.38 mmol/kg.

Novel concepts for the biocatalytic synthesis of second-generation biodiesel

Biodiesel is synthesized by the transesterification of triglycerides of oils with short-chain alcohols, such as methanol and ethanol. According to the Renewable Energy Directive guidelines (RED II 2018/2001/EU) the contribution of advanced biofuels, which do not include edible oils, towards the overall EU target, is at 1% in 2025 and at least 3.5% in 2030. Bioprocesses that valorize non-edible oils for the production of second-generation biodiesel could play a critical role in achieving this goal. Immobilized lipases, as well as other enzyme classes, such as cutinases and acyltransferases, are utilized as biocatalysts for this process. For the sustainability of the process, renewable materials can be used as immobilization matrices, or even enzymes anchored on the cells as whole-cell biocatalysts. Membrane reactors can also be employed to facilitate the enzymatic transesterification by conducting a continuous enzymatic reaction and simultaneously separate the products in a single operation. The advances on the aforementioned fast-pacing fields are presented in this work.

Optimization and synthesis process of biodiesel production from coconut oil using central composite rotatable design of response surface methodology

In the transportation and power production industries, the use of renewable and environmentally friendly fuels has grown in importance. Biodiesel derived from coconut oil contains over 90% saturated fatty acids. Biodiesel was made using alkaline transesterification since coconut oil has a free fatty acid content of less than 2.5%. Enzymatic or chemical transesterification are both possible. For the synthesis of coconut biodiesel, the optimal processing conditions are 60 °C for 1 h, a 6:1 ratio, 1% potassium hydroxide and a 95% yield. According to the experiment, 55 °C was the ideal reaction temperature for using coconut oil to produce biodiesel. Sixty minutes was the ideal amount of time to extract biodiesel from coconut oil. The methanol-to-oil molar ratio raised yield from 6:1 to 8:1, a 95% increase. Significant amounts of an alkaline catalyst, which allows soap to develop under the influence of fatty acids, are responsible for the high yield response; it is concluded that 1 wt% would be an appropriate catalyst concentration for the present investigation. The central composite rotatable design (CCRD) of the response surface methodology method is used to optimize several process parameters, including temperature, reaction duration, methanol-to-oil ratio and catalyst concentration. The CCRD optimization approach produced better results. The following are the final, optimized results: coconut oil methyl ester ratio: 96.69%, temperature: 55 °C; duration: 59.2 min; catalyst concentration: 0.7; molar ratio: 6.4.

Potential Application of Bacterial Lipase from Bacillus subtilis in the Production of Biodisel

This work aims to evaluate the potential of bacterial lipase of Bacillus subtilis in biodiesel production. Assays were performed to obtain, purify, and immobilize the enzyme and compare chemical and enzymatic transesterification tests. The Tryptic Soy Broth (TSB) culture medium without supplementation showed the highest enzymatic activity and purification yields when using 60% of (NH4)2SO4. Citral was the most efficient agent for lipase immobilization. Even though the free enzymes showed a higher activity rate, the immobilized enzymes are associated with a better-quality product. In conclusion, the lipase enzyme showed positive potential in biodiesel production.

Enzymatic synthesis of tyrosol esters in organic solvents and ionic liquids: Correlation between enzyme activity and solvent properties

Tyrosol is a principle phenolic compound present in olive oil and wine with great pharmacological potentials due to its strong antioxidant property. Its application can be highly extended by increasing its lipophilicity through esterification. In this study, tyrosol esters were synthesized by enzymatic transesterification of tyrosol with vinyl esters, with a commercial lipase from Novozymes (Novozym 435, Candida antarctica lipase B (CALB) immobilized on macroporous acrylic resin beads) as the catalyst, both in organic solvents and in ionic liquids. Methyl tert-butyl ether (MTBE) was the best solvent, offering complete conversions towards the synthesis of tyrosol esters with varying chain lengths (C2-C18) within 1 h. Taking the enzymatic transesterification between tyrosol and vinyl acetate as the model reaction, 18 organic solvents and 24 ionic liquids were screened, and their solvent properties (i.e. hydrophobicity, polarity, viscosity, water activity, and partition coefficients of substrate and product) were correlated to the enzyme activity. Upon enzyme screening, four new lipases were found to be highly efficient, relative to the best-performed Novozym 435 and Lipozyme TLIM (Thermomyces lanuginosus lipase (TLL) immobilized on silica gel), in tyrosol ester synthesis. This work provides enzymatic methods of esterifying tyrosol that are much more efficient than those reported in literature, and will shed light on re-consideration of the roles of solvents in nonaqueous enzymology.