Enzymatic Transesterification Research Articles

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.

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.

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.

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.

Utilization of microalgae for agricultural runoff remediation and sustainable biofuel production through an integrated biorefinery approach

Generally wastewater such agricultural runoff is considered a nuisance; however, it could be harnessed as a potential source of nutrients like nitrates and phosphates in integrated biorefinery context. In the current study, microalgae Chlorella sp. S5 was used for bioremediation of agricultural runoff and the leftover algal biomass was used as a potential source for production of biofuels in an integrated biorefinery context. The microalgae Chlorella sp. S5 was cultivated on Blue Green (BG 11) medium and a comprehensive optimization of different parameters including phosphates, nitrates, and pH was carried out to acquire maximum algal biomass enriched with high lipids content. Dry biomass was quantified using the solvent extraction technique, while the identification of nitrates and phosphates in agricultural runoff was carried out using commercial kits. The algal extracted lipids (oils) were employed in enzymatic trans-esterification for biodiesel production using whole-cell biomass of Bacillus subtilis Q4 MZ841642. The resultant fatty acid methyl esters (FAMEs) were analyzed using Fourier transform infrared (FTIR) spectroscopy and gas chromatography coupled with mass spectrometry (GC–MS). Subsequently, both the intact algal biomass and its lipid-depleted algal biomass were used for biogas production within a batch anaerobic digestion setup. Interestingly, Chlorella sp. S5 demonstrated a substantial reduction of 95% in nitrate and 91% in phosphate from agricultural runoff. The biodiesel derived from algal biomass exhibited a noteworthy total FAME content of 98.2%, meeting the quality standards set by American Society for Testing and Materials (ASTM) and European union (EU) standards. Furthermore, the biomethane yields obtained from whole biomass and lipid-depleted biomass were 330.34 NmL/g VSadded and 364.34 NmL/g VSadded, respectively. In conclusion, the findings underscore the potent utility of Chlorella sp. S5 as a multi-faceted resource, proficiently employed in a sequential cascade for treating agricultural runoff, producing biodiesel, and generating biogas within the integrated biorefinery concept.Graphical

An overview on eco-friendly polyglycerol esters of fatty acid, synthesis and applications

Abstract Polyglycerol esters (PEGs) which are non-ionic surfactants acting as emulsifiers, wetting agents and viscosity modifying agents are used in the cosmetic, pharmaceutical and food industries. They have been proposed as an alternative to ethoxylated glycol-based non-ionic surfactants due to safety issues. PGEs are composed of fatty acid, which is a lipophilic moiety, and polyglycerol, which is a hydrophilic moiety. They are synthesized by several methods, such as direct esterification of fatty acids and polyglycerols, chemical transesterification of fatty acid methyl esters and polyglycerol, enzymatic transesterification using Lipozyme 435, using glycerol carbonate as raw material, using microwave irradiation, etc. PGEs (Polyglycerol esters) are claimed to be green alternatives to the existing emulsifiers used in the chemical industry as the raw material used for synthesis is obtained from vegetable oils which are renewable, and therefore, eco-friendly surfactants for use in a broad number of applications including food, cosmetics, textiles and personal care. The major challenges in the synthesis of polyglycerol fatty acid esters are to increase yield and control the esterification level while minimising side reactions.

Enzymatic transesterification of Phoenix dactylifera L. oil using lipase immobilized on activated carbon and optimization using response surface methodology

Enzymatic transesterification of Phoenix dactylifera L. oil using lipase immobilized on activated carbon and optimization using response surface methodology

Condition optimization and kinetic evaluation of Novozym 435-catalyzed synthesis of aroma pyridine esters

A new method for rapid synthesis of pyridine esters without stirring based on Novozym 435 catalysis had been reported, which was from pyridine methanols and vinyl esters. Based on single factor experiment, the optimization of the enzymatic transesterification was received by response surface methodology (RSM). The high yield of 82% was still reached when Novozym 435 had been recycled 12 times. Under optimal conditions, a total of 22 pyridine esters were synthesized. The kinetic model of Novozym 435-catalyzed transesterification of vinyl acetate with 3-pyridinemethanol was established by enzymatic assay. It was discovered that the reaction followed the Ping-Pong Bi-Bi mechanism, and 3-pyridinemethanol inhibited the reaction. Gas chromatography-mass spectrometry-olfactometry (GC–MS-O) was used to analyze the synthesized pyridine esters, of which pyridin-3-ylmethylhexanoate (3e), pyridin-3-ylmethylcinnamate (3i), and 2-(pyridin-2-yl)ethyl acetate (3 s) possessed strong and excellent aromas. These three flavor compounds (3e, 3i and 3 s) were found to have good stability at the specified temperature by thermogravimetric (TG) analysis.

Ultra-small magnetic Candida antarctica lipase B nanoreactors for enzyme synthesis of bixin-maltitol ester.

Bixin has desirable bioactivities but poor water solubility, which limits its practical applications. Enzymatic transesterification of methyl to alditol groups in bixin by Candida antarctica lipase B (CALB) improves bixin water solubility. Herein, magnetic CALB nanoreactors with diameter of 11.7nm and CALB layer thickness of 3.5nm were developed by covalently linking CALB onto silicon covered Fe3O4 nanoparticles. The CALB loading capacity in nanoreactors achieved 30%. The Michaelis constant (Km) and maximum reaction rate of magnetic CALB nanoreactors were 56.1mmol/L and 0.2mmol/(L·min). Magnetic CALB nanoreactors could circularly catalyze bixin-maltitol ester synthesis and keep catalytic efficiency of 62.6% after eight repetitive enzymatic reactions. Additionally, the optimal bixin-maltitol ester synthesis procedure was heating bixin-maltitol mixture at molar ratio of 1:7 in anhydrous 2-methyl-2-butanol-dimethylsulfoxide (8:2, v/v) at 50°C for 24h. Bixin-maltitol ester showed improved water solubility at pH 5.5 and 7.0.

Combined immobilized lipases for effective biodiesel production from spent coffee grounds

This work describes the enzymatic transesterification of the oil extracted from SCGs for synthesis of biodiesel as a promising alternative to diesel fuels based on petroleum. Biocatalysts from various sources were tested for biodiesel synthesis using coffee oil among which CaCO3- immobilized Staphylococcus aureus and Bacillus stearothermophilus showed the highest conversion yields (61 ± 2.64% and 64.3 ± 1.53%, respectively) in 4 h. In further optimizing reaction parameters, methanol to oil molar ratio, biocatalyst quantity, water content, as well as incubation time and temperature markedly improved oil-to-biodiesel conversion up to 99.33 ± 0.57 % in a solvent free reaction after 12 h at 55 °C. A mixture of inexpensive CaCO3-immobilized bacterial lipases at a 1:1 ratio was the best environment-friendly catalyst for biofuel synthesis as well as the ideal trade-off between conversion and cost. Obtained coffee biodiesel remained stable beyond 40 days at ambient storage conditions and its chemical characteristics were comparable to those of other known biodiesels according to the European requirements (EN14214). Collectively, SCGs, after oil extraction, could be an ideal substrate for the production of an environment-friendly biodiesel by using appropriate mixture of CaCO3-immobilized lipases.

Kinetic study on the production of biodegradable lubricant by enzymatic transesterification of high oleic palm oil

Kinetic studies are necessary to identify inhibitors of lipase enzyme deactivation in reaction systems because varying substrate concentrations can affect the enzyme's catalytic activity. The present study aimed to analyze the reaction kinetics of palm-based polyol ester production catalyzed by commercial lipase Novozyme 435 (N435). The enzymatic transesterification reaction was performed in a solvent-free medium. The effect of substrates concentration, specifically high oleic palm methyl ester (HO-PME) and trimethylolpropane (TMP), on the kinetic constant was studied at the initial reaction rate. The study was conducted based on the Ping-pong Bi-bi model, assuming that both substrates could inhibit the reaction. The reaction was carried out at 70 °C and 15.25 mbar with a 3 % (w/w) N435 enzyme load to investigate the effect of various HO-PME and TMP concentrations. The kinetic constants obtained are as follows: Km(HOPME) = 61.112 mol/L, Km(TMP) = 0.336 mol/L, Ki(HOPME)= 0.002 mol/L, Ki(TMP) = 2.415 mol/L and Vmax = 17.24 mol/L.hr. The results implied that N435 has higher affinity towards TMP than HO-PME. Inhibition constant indicated a lower inhibitory function of the TMP than HO-PME (Ki(TMP) >Ki(HOPME)). The reaction kinetics obtained in this study agreed well with the model used with TMP and HO-PME as competitive inhibitor during enzymatic transesterification.

Transesterification of phosphatidylcholine with DHA-rich algal oil using immobilized Candida antarctica lipase B to produce DHA-phosphatidylcholine

Docosahexaenoic acid (DHA) enriched with phospholipids (PLs) (DHA-PLs) is a type of structured PL with good physicochemical and nutritional properties. Compared to PLs and DHA, DHA-PLs has higher bioavailability and structural stability and many nutritional benefits. To improve the enzymatic synthesis of DHA-PLs, this study investigated the preparation of phosphatidylcholine (PC) enriched with DHA (DHA-PC) via enzymatic transesterification of algal oil, which is rich in DHA-triglycerides, using immobilized Candida antarctica lipase B (CALB). The optimized reaction system incorporated 31.2% DHA into the acyl chain of PC and converted 43.6% PC to DHA-PC within 72 h at 50 °C, 1:8 PC: algal oil mass ratio, 25% enzyme load (based on total substrate mass), and 0.02 g/mL molecular sieve concentration. Consequently, the side reactions of PC hydrolysis were effectively suppressed and products with high PC content (74.8%) were produced. Molecular structure analysis showed that exogenous DHA was specifically incorporated into the sn-1 site of the PC by immobilized CALB. Furthermore, the evaluation of reusability with eight cycles showed that the immobilized CALB had good operational stability in the present reaction system. Collectively, this study demonstrated the applicability of immobilized CALB as a biocatalyst for synthesizing DHA-PC and provided an improved enzyme-catalyzed method for future DHA-PL synthesis.

BIODIESEL PRODUCTION: POTENTIAL AND FUTURE TRENDS – A REVIEW

Biodiesel is a potential renewable energy that can reduce greenhouse gas (GHG) emissions and increase energy security. Biodiesel has been shown to have lower carbon emissions compared to petroleum diesel, and it can reduce GHG by as much as 86%. Governments around the world have set targets for renewable energy, with a specific focus on the use of biofuels like biodiesel. Biodiesel can be derived from various feedstocks such as animal lipids, vegetable oils, and waste oils. It can be made through the transesterification of triglyceride with ethanol or methanol. This reaction requires strong base catalysts, such as sodium hydroxide or potassium hydroxide, in order to produce methyl esters. The potential of biodiesel has led to advancements in its production, such as the use of enzymatic transesterification, novel feedstocks, and the optimization of production parameters. Additionally, various companies have ventured into biodiesel production with a range of business models and approaches.

Lipase-catalysed changes in essential oils revealed by comprehensive two-dimensional gas chromatography

Candida antarctica lipase A (CALA) was applied for the chemo-selective enzymatic transesterification of terpene and phenyl alcohols in 35 different essential oil samples. Comprehensive two-dimensional gas chromatography with mass spectrometry (GC×GC‒MS) analysis enabled the separation and tentative identification of a cohort of 125 compounds, allowing the instant visualisation of the reaction process changes, amid the complex chemical background of the samples. The results indicate that 42 out of 79 alcohols so-identified were fully or partially esterified within 48 h of reaction, with primary alcohols being the substrates of preference of the enzyme (90–100% conversion), followed by secondary alcohols (mostly ~ 80–100% conversion). No significant conversion of tertiary alcohols and phenols was observed using the tested conditions. Overall, the enzyme’s performance was consistent for primary alcohol substrates identified in multiple samples of different compositions. The observed selectivity, efficiency, robustness, scalability (enzyme/substrate working concentration ratio > 1:160), potential reusability, mild reaction conditions, and other factors make this process a greener and more sustainable alternative for industry applications, particularly for the manufacture of novel flavours and fragrances.Graphical

Flow enzymatic esterification of 5-hydroxymethylfurfural and liquid–liquid extraction of 5-acetyl-hydroxymethylfurfural using deep eutectic solvents in semicontinuous mode

This study explored for the first time the use of a flow-packed bed bioreactor for producing 5-acetyl-hydroxymethylfurfural (5-acetyl-HMF) and semicontinuous liquid–liquid extraction with a deep eutectic solvent (DES): choline chloride: glycerol= 1:2 (ChCl: GLY=1:2). The flow reaction system provided a specific productivity that was considerably higher than that observed in the batch reactor (1 gProduct/min·gBiocat vs. 0.07 gProduct/min·gBiocat). The biocatalyst Novozym 435® showed high operational stabilities in continuous mode and sequential batch mode without losses of activity. In continuous mode, an 80 % conversion was achieved with 100 mM 5-hydroxymethylfurfural (HMF) and a flow rate of 0.05 mL/min. The separation in the semicontinuous mode was more efficient than that for the extraction batch system (70 % vs. 38 %). Finally, in the future, a combination of continuous enzymatic transesterification and semicontinuous separation of 5-acetyl-HMF will provide an efficient, sustainable biocatalytic process that can be industrially scaled.