Article pubs.acs.org/JAFC
Cite This: J. Agric. Food Chem. 2018, 66, 12361−12367
Choline-Chloride-Based Eutectic Solvent for the Efficient Production of Docosahexaenoyl and Eicosapentaenoyl Ethanolamides via an Enzymatic Process Huipei Liang,† Xiaoli Qin,‡ Chin Ping Tan,§ Daoming Li,∥ and Yonghua Wang*,†
Downloaded via UNIV OF NORTH DAKOTA on November 21, 2018 at 09:24:42 (UTC). See https://pubs.acs.org/sharingguidelines for options on how to legitimately share published articles.
†
School of Food Science and Engineering, South China University of Technology, Guangzhou, Guangdong 510640, People’s Republic of China ‡ College of Food Science, Southwest University, Chongqing 400715, People’s Republic of China § Department of Food Technology, Faculty of Food Science and Technology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia ∥ School of Food and Biological Engineering, Shaanxi University of Science and Technology, Xi’an, Shaanxi 710021, People’s Republic of China S Supporting Information *
ABSTRACT: Docosahexaenoyl and eicosapentaenoyl ethanolamides (DHEA and EPEA) have physiological functions, including immunomodulation, brain development, and anti-inflammation, but their efficient production is still unresolved. In this study, choline-chloride-based natural deep eutectic solvents are used as media to improve the production of DHEA and EPEA. The water content showed a key effect on the reactant conversion. Adding water to choline chloride−glucose (CG, molar ratio of 5:2) led to a significant increase (13.03% for EPEA and 27.95% for DHEA) in the yields after 1 h. The high yields of EPEA (96.84%) and DHEA (90.06%) were obtained under the optimized conditions [fish oil ethyl esters/ethanolamine molar ratio of 1:2, temperature of 60 °C, 1 h, enzyme loading of 2195 units, and CG containing 8.50% water of 43.30% (w/w, relative to total reactants)]. The products could be easily separated using centrifugation. In summary, the research has the potential to produce fatty acyl ethanolamides. KEYWORDS: natural deep eutectic solvents, docosahexaenoyl ethanolamide, eicosapentaenoyl ethanolamide, enzymatic synthesis
■
INTRODUCTION Fatty acyl ethanolamides have recently attracted substantial attention as a result of their various chemical and physiological functions, which are associated with the length and structure of the alkyl chain. These compounds with an alkyl chain length of 8−18 are widely used as non-ionic surfactants in lubricants, cosmetics, and textiles.1−3 Very recently, fatty acyl ethanolamides with a long alkyl chain (16−22) act as a special lipid messenger and have biological functions.4−12 For example, docosahexaenoyl ethanolamide (DHEA) and eicosapentaenoyl ethanolamide (EPEA) have beneficial functions, such as antiinflammation,13−15 antitumor,16 and neurological17,18 functions. It has also been confirmed that DHEA and EPEA have greater antiproliferative activities than their parent fatty acids (i.e., docosahexaenoic and eicosapentaenoic acids).19 Therefore, it is of interest to produce DHEA and EPEA with high purity to understand their functions more deeply and broaden their applications. The production of fatty acyl ethanolamides is generally accomplished by the reaction of ethanolamine with an acyl donor, such as a fatty acid,20 fatty acid methyl ester,21 and fatty acid ethyl ester,22 catalyzed by lipases. However, there are some problems with the current approaches, such as low yield,23−27 time consuming, and slow reaction speed. It may be improved to some extent when organic solvents (hexane and acetonitrile) are used as reaction media, but this approach still © 2018 American Chemical Society
suffers from the low purity of the product, complicated separation of the product, and environmental hazards.21 The efficient enzymatic synthesis of polyunsaturated fatty acyl ethanolamides is still an unresolved issue. We hypothesized that a potential reason behind such issues is the poor affinity of lipases toward substrates as a result of the heterogeneous reaction. To overcome these issues, it is urgent to find an alternative green solvent with more lipase affinity to efficiently synthesize polyunsaturated fatty acyl ethanolamides enzymatically. Recently, natural deep eutectic solvents (NADES), as widely used green media, have been successfully employed in the production of various functional triglycerides,28 glycolipids,29 and epoxidated oil.30 Numerous studies have demonstrated that NADES can enhance the yields, facilitate the separation of products,31 and protect the enzymes from inactivation.32 In addition, NADES are easy to prepare at low cost and have ecofriendly and biodegradable properties. These advantages make NADES promising solvents for potential applications in the food and pharmaceutical fields. To the best of our knowledge, there is little information on the exploitation of NADES as Received: Revised: Accepted: Published: 12361
September 3, 2018 November 1, 2018 November 5, 2018 November 5, 2018 DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
Article
Journal of Agricultural and Food Chemistry Table 1. NADES Were Formed by Mixing Different Components group number 1 2 3 4 5 6 7 8 9 10 11
component 1 betaine betaine choline choline choline choline choline choline choline choline choline
chloride chloride chloride chloride chloride chloride chloride chloride chloride
component 2
component 3
molar ratio
abbreviation
urea glycerol urea malonic acid glycerol glucose glucose glucose glucose glucose glucose
water
1:2:1 1:2 1:2 1:1 1:1:1 5:2 5:2:3 5:2:5 5:2:7 5:2:9 5:2:15
BUH BGly CU CM CGlyU CG CG−5.10% water CG−8.50% water CG−11.90% water CG−15.30% water CG−20.30% water
urea water water water water water
a 25 mL reaction vial and diluted with the NADES (43.3% of the total weight of the reactants). After the addition of 2195 units of Novozym 435, the reaction mixture was incubated at 60 °C under magnetic stirring (500 revolutions/min) for 1 h. At the end of the reaction, the lipase was filtered. The samples obtained were diluted with ethanol and quantified using high-performance liquid chromatography (HPLC). To investigate the influence of the best NADES on the aminolysis, factors including the fish oil ethyl ester/ethanolamine molar ratio (1:1, 1:1.50, 1:2, 1:2.50, and 1:3), water content of the NADES (detail composition listed in Table 1), reaction temperature (30, 40, 50, 60, and 70 °C), and reaction time (10, 20, 30, 40, 60, 120, and 180 min) were studied. Determination of the Fatty Acyl Ethanolamides Using HPLC. The reaction samples obtained were analyzed using a Dionex HPLC system equipped with a U3000 pump on an XTerra RP18 column (250 × 4.6 mm, 5 μm, Waters) at 215 nm with an ultraviolet (UV) detector. The column was maintained at 35 °C. Methanol and water were used for gradient elution. The concentration of methanol was 80% in the first 15 min, increased linearly from 80 to 90% during 20 min, and decreased linearly from 90 to 80% during 10 min. The flow rate was 1 mL/min. Standard curves of EPEA and DHEA were established to calculate the concentration of EPEA and DHEA that were produced in the reaction. The yields of EPEA and DHEA that were produced in the reaction were calculated using eqs 1 and 2
solvents for the enzymatic production of fatty acyl ethanolamides. The influence of NADES on the production performance and substrate selectivity of lipases is expected to be studied. Therefore, the aim of this study was to develop an efficient and green approach to produce fatty acyl ethanolamines from polyunsaturated fatty acid ethyl esters and ethanolamine, to favor improved product yields, and simplify the separation and purification processes. First, the effect of six types of NADES that are used as reaction media on the product yields and substrate selectivity of a lipase was investigated. Second, the influence of selected NADES in the presence of various water levels, reaction temperature, and polyunsaturated fatty acid ethyl ester/ethanolamine molar ratio on the fatty acyl ethanolamide yields was examined.
■
MATERIALS AND METHODS
Chemicals and Reagents. Fish oil ethyl esters (an average molecular weight of 340 g/mol) containing 39.20% eicosapentaenoic acid ethyl ester (EPA-EE) and 43.60% docosahexaenoic acid ethyl ester (DHA-EE) were obtained from Zhoushan Sinomega Biotech Engineering Co., Ltd. (Zhejiang, China). Docosahexaenoyl ethanolamide (≥98%) was obtained from Cayman Chemical. Eicosapentaenoyl ethanolamide (≥98%) was purchased from Sigma-Aldrich. Eicosapentaenoic acid ethyl ester (98%) was purchased from the Shanghai Yuan Ye Biological Technology Co., Ltd. (Shanghai, China). Docosahexaenoic acid ethyl ester (GC, >97%) was obtained from the Tokyo Chemical Industry Co., Ltd. (Shanghai, China). Choline chloride (98%), D-(+)-glucose (≥99.50%), urea (99%), glycerol (99%), and malonic acid (99.50%) were purchased from Aladdin (Shanghai, China). Betaine (98%, anhydrous) was purchased from Macklin (Shanghai, China). Ethanolamine (≥99%) was obtained from the Guangzhou Chemical Reagent Factory (Guangzhou, China). Novozym 435 (immobilized Candida antarctica lipase B) was purchased from Novozymes A/S (Bagsvaerd, Denmark). Methanol of chromatographic grade was purchased from Shanghai Ling Feng Chemical Reagent Co., Ltd. (Shanghai, China). All of the other reagents were obtained commercially and were of analytical grade. Preparation of Different NADES. The detailed composition of the various NADES is listed in Table 1. CG (group number 6) was prepared by mixing choline chloride and glucose using vacuum evaporation (500 revolutions/min) at 80 °C. Other NADES were prepared using direct heating to 80 °C under magnetic stirring. Finally, homogeneous and colorless liquids were formed, and the resultant NADES were sealed and stored in a silica gel dryer until use. Preparation of Fatty Acyl Ethanolamides. Fatty acyl ethanolamides (EPEA and DHEA) were prepared using the aminolysis of fish oil ethyl esters with ethanolamine in the presence of different NADES as reaction solvents. To evaluate the influence of various NADES on the aminolysis, the reactants containing fish oil ethyl ester and ethanolamine at a molar ratio of 1:2 were weighed into
EPEA yield (%) =
DHEA yield (%) =
n(EPEA) × 100% m(EPA‐EE)/M(EPA‐EE) n(DHEA) × 100% m(DHA‐EE)/M(DHA‐EE)
(1)
(2)
where n(EPEA) is the moles of EPA-EE converted to EPEA, n(DHEA) is the mole sof DHA-EE converted to DHEA, m(EPA-EE) and m(DHA-EE) are the masses of EPA-EE and DHA-EE, respectively, and M(EPA-EE) and M(DHA-EE) represent the molecular weights of EPA-EE and DHA-EE, respectively. Determination of the Viscosity of CG−Water. The viscosity of CG with different water contents was tested at room temperature using a rotational viscometer (NDJ-1, Shanghai Changji Geological Instrument Co., Ltd., China). The CG sample was placed in a 100 mL special beaker. An appropriate rotor was selected so that the pointer read more than 10 grid. The pointer reading can be recorded until the pointer stabilized at a position. Determination of the Water Activity of CG−Water. The water content of CG was measured at room temperature using a water activity meter (Aqualab 3TE series, Pullman, WA, U.S.A.). Statistical Analysis. All of the experiments were performed in triplicate. The data were analyzed in Origin 9.0 software and were expressed as the mean values ± standard deviations. The differences between the mean values were analyzed by an analysis of variance using the SPSS software (version 14.0 demo, SPSS, Inc., Chicago, IL, U.S.A.). 12362
DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
Article
Journal of Agricultural and Food Chemistry
■
RESULTS AND DISCUSSION Influence of the NADES on Aminolysis. As shown in Figure 1, the use of betaine-based NADES as reaction solvents
Figure 1. Effect of the NADES on fatty acyl ethanolamide yields and the preference of Novozym 435 for fatty acid ethyl esters. Reaction conditions: fish oil ethyl ester/ethanolamine molar ratio of 1:2, enzyme loading of 2195 units, NADES of 43.30% (w/w, relative to total reactants), and reaction temperature of 60 °C. The equilibrium time of the reaction was 5 h for the solvent-free system and CM, 1 h for BUH and BGly, 2 h for CU and CGlyU, and 3 h for CG.
resulted in a lower or similar performance compared to a solvent-free system. However, choline chloride-based NADES (CU, CM, CGlyU, and CG) significantly improved the yields of EPEA and DHEA. In particular, when CG was used as the media, the highest yields of EPEA and DHEA reached approximately 90 and 70% after 3 h, respectively. One of the possible explanations for the high yields of EPEA and DHEA in CG is that glucose in the choline chloride-based NADES may protect the lipase from inactivation. It has been reported that choline chloride NADES could enhance the catalytic activity and stability of the enzyme.32,33 Alternatively, previous studies suggested that choline chloride−sugar-based NADES could decrease the surface tension of the oil/droplets in an aqueous environment, lower surface tension, and favor the retention of the activity of a lipase.34 Thus, the enzyme catalytic efficiency was improved. Interestingly, choline chloride-based NADES showed a different influence on the yields and the EPEA/DHEA ratio. For example, Novozym 435 had a stronger preference for EPA-EE than DHA-EE in CM, CU, and CGlyU. In particular, Novozym 435 had the strongest preference with an EPEA/DHEA ratio of 2.09 in the presence of CM, but the yields of EPEA and DHEA were not the highest (approximately 70 and 30% after 5 h, respectively). These results indicate that choline chloride-based NADES consisting of different components not only enhance the efficiency of the Novozym-435-catalyzed aminolysis reaction but also affect the selectivity of the lipase for EPA-EE and DHA-EE. The selectivity of the lipases may be related to the nature of the NADES themselves. The interaction between the choline chloride-based NADES and enzyme or water is affected by hydrogen bonding, which may alter the catalytic properties of the enzyme.28 Effect of CG with Different Water Contents on Aminolysis. The yields were remarkably improved in the initial stage of the reaction by adding water (Figure 2). For
Figure 2. Effect of the water content of CG on the (A) EPEA yield and (B) DHEA yield. Reaction conditions: fish oil ethyl ester/ ethanolamine molar ratio of 1:2, enzyme loading of 2195 units, CG− water of 43.30% (w/w, relative to total reactants), and temperature of 60 °C.
example, when the water content of CG increased from 0 to 5.10%, the yields of EPEA and DHEA increased remarkably from 82.42 to 95.45% and from 57.98 to 85.93% after 1 h, respectively. The increase in the yields with the addition of water could be explained by the following possibilities: (1) The addition of water led to an increase in the catalytic activity of the lipase, because an appropriate water activity is needed to maintain the enzymatic conformation and favors enzymatic activity,35 thus increasing the reactivity of substrates. (2) A reduction in the viscosity of the system (CG with water added) may improve the mass transfer of the substrates (Figure S1 of the Supporting Information). (3) Ethanol, one of reaction products, may interact with CG by hydrogen bonding. It has been reported that compounds that can donate electrons or protons, including ethylene glycol and glycerol, could be associated with deep eutectic solvents by hydrogen bonding.35,36 In this case, ethanol as a protic solvent is presumably associated with CG, which may attenuate the inhibition of lipase by ethanol, thus protecting the activity and stability of lipase. Alternatively, bonded ethanol may drive the reaction to the right, achieving high yields of products. 12363
DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
Article
Journal of Agricultural and Food Chemistry
and DHEA (94.31%) at 2 h were slightly higher than those (96.84 and 90.06%, respectively) at 1 h, a longer reaction time may result in the production of some byproducts and a deepening in the color of the products. Therefore, 1 h of reaction was chosen in the subsequent experiments. Effect of the Temperature on Aminolysis. Generally, the reaction temperature affects the lipase activity, reaction rate, and physical properties of the reaction system. The effect of the reaction temperature on the aminolysis was investigated. As shown in Figure 4, the yield of EPEA gradually increased
Although adding water led to high product yields in the initial stage of reaction, the water content ranging from 5.10 to 20.30% showed a less pronounced effect on the product yields compared to water contents below 5.10% (Figure 2). However, a previous study has reported that the water content above 5% led to the production of a low content of oleoylethanolamide.37 The possible explanation was that water was produced from the amidation between oleic acid and ethanolamine. As the amidation proceeded, the water was continually accumulated; therefore, the above 5% water was added to the reaction, presumably because the excess water in the reaction system weakened the reaction toward the right, thereby lowering the oleoylethanolamide content. While in our study, choline chloride, glucose, and water were the components of NADES and the water activity value of CG with a water content ranging from 8.50 to 20.30% was from 0.2 to 0.3 (Figure S2 of the Supporting Information); therefore, the minor changes in water activity of CG with different water contents have less influence on the product yields. The results imply that the water content of the substrates or CG does not need to be exactly controlled to a specific value or a narrow range in industrial applications. Effect of the Reaction Time on Aminolysis. The effect of the reaction time on the yields of fatty acyl ethanolamides and fatty acid ethyl ester preference of Novozym 435 was studied. Although the moles of EPA-EE (2.02) and DHA-EE (2.08) were near equal in the fish oil ethyl esters, the molar yield of EPEA was linearly increased to 93.94% within 30 min and was higher than that of DHEA (77.95%) (Figure 3). The
Figure 4. Effect of the reaction temperature on fatty acyl ethanolamide yields and the preference of Novozym 435 for fatty acid ethyl esters. Reaction conditions: fish oil ethyl ester/ethanolamine molar ratio of 1:2, enzyme loading of 2195 units, CG−8.50% water of 43.30% (w/w, relative to total reactants), and reaction time of 1 h.
from 90.55 to 96.84%, while the yield of DHEA increased remarkably from 57.03 to 90.06%, as the temperature increased from 30 to 60 °C. A high lipase activity and a reduced viscosity of the reaction mixture at higher temperatures may be responsible for the improvement of the yields of EPEA and DHEA. Interestingly, the EPEA/DHEA ratio was higher at lower temperatures (≤50 °C) but stayed almost constant at higher temperatures (≥60 °C). The low catalytic performance of lipase and the high viscosity of the reaction mixture caused by low temperatures (≤50 °C) could lead to the apparent preference of Novozym 435 for EPA-EE. The yields began to decrease when the temperature was raised to 70 °C, possibly as a result of the inactivation of the lipase. Therefore, 60 °C was chosen for the remaining experiments. Effect of the Molar Ratio of Fish Oil Ethyl Ester/ Ethanolamine on Aminolysis. The yields of EPEA and DHEA were increased dramatically when the fish oil ethyl ester/ethanolamine molar ratio was increased from 1:1 to 1:2 (Figure 5). It is suggested that a molar ratio of less than 1:1 resulted in the low yield of fatty acyl ethanolamide formation. This could be due to the formation of the byproduct (fatty acyl esteramide) at the fatty acid ester (or free fatty acid)/ ethanolamine molar ratio of less than 1:1.38 Alternatively, the EPEA/DHEA ratio was rapidly decreased from 1.73 to 1.08 with the increase in the molar ratio from 1:1 to 1:2, suggesting that Novozym 435 preferred EPA-EE compared to DHA-EE at a higher level of fish oil ethyl esters (fish oil ethyl ester/ ethanolamine molar ratio of 1:1) (Figure 5). An additional increase in the ethanolamine level (fish oil ethyl ester/
Figure 3. Effect of the reaction time on fatty acyl ethanolamide yields and the preference of Novozym 435 for fatty acid ethyl esters. Reaction conditions: fish oil ethyl ester/ethanolamine molar ratio of 1:2, enzyme loading of 2195 units, CG−8.50% water of 43.30% (w/w, relative to total reactants), and temperature of 60 °C.
results indicate that Novozym 435 preferred EPA-EE to DHAEE in the very initial stage of the reaction. As the reaction proceeded for 30 min, the EPEA/DHEA ratio was sharply reduced to 1.21. This could be explained because most EPAEE has been consumed to form EPEA and the decreased EPAEE concentration in the reaction system may attenuate the reaction of EPA-EE with ethanolamine. The yield of DHEA gradually increased with increasing reaction times from 30 to 60 min, after which the yields of EPEA and DHEA were kept almost constant, indicating that the reaction had almost reached equilibrium. Although the yields of EPEA (98.32%) 12364
DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
Article
Journal of Agricultural and Food Chemistry
Figure 5. Effect of the molar ratio of fish oil ethyl ester/ethanolamine on fatty acyl ethanolamide yields and the preference of Novozym 435 for fatty acid ethyl esters. Reaction conditions: enzyme loading of 2195 units, CG−8.50% water of 43.30% (w/w, relative to total reactants), temperature of 60 °C, and reaction time of 1 h.
ethanolamine molar ratio from 1:2 to 1:3) resulted in a reduction in the yields (from 96.84 to 94.36% for EPEA and from 90.06 to 87.37% for DHEA). It has been reported that excess ethanolamine lowered the activity and stability of Novozym 435, and the content of fatty acyl ethanolamide was decreased from 21.60 to 15.00% with the increase in the fatty acid ethyl ester/ethanolamine molar ratio from 1:1 to 1:3 in a solvent-free system.27 However, when CG−8.50% water was used as a reaction solvent in this study, the highest yields of EPEA (96.84%) and DHEA (90.06%) were obtained at the molar ratio of 1:2, after which the yields of EPEA and DHEA slightly decreased by 2.46 and 2.69%, respectively. One of the possible explanations is that CG−water not only improved the mass transfer between lipase and the reaction mixtures but also attenuated the decrease in the activity and stability of lipase.35,36,39 Thus, Novozym 435 has a high tolerance to polar ethanolamine in CG−water. Separation of the Products. The mixture after the aminolysis reaction was subjected to centrifugation, resulting in two layers (Figure 6A). The fatty acyl ethanolamide products were almost all (99.41%) present in the upper layer, while almost all CG−8.50% water and ethanolamine remained in the lower phase (panels B and C of Figure 6). The peak areas of EPEA and DHEA in the upper layer were 165- and 171-fold higher than those in the lower layer, respectively, suggesting that the centrifugation separated the products (EPEA and DHEA) from the NADES and ethanolamine. The separation was simple and effective. In conclusion, using NADES as a reaction medium to enzymatically synthesize EPEA and DHEA provided significant advantages over the use of organic solvents. First, the separation and purification of products is simplified. Unreacted ethanolamine could be removed from the reaction mixtures using centrifugation. However, when organic solvent is used as the reaction solvent, the organic solvent and unreacted ethanolamine require removal from the reaction mixtures using vacuum evaporation. Second, the NADES could be more environmentally friendly and safer than organic solvents. In summary, highly efficient production (approximately 96.84% for EPEA and 90.06% for DHEA at 1 h) was achieved via the lipase-catalyzed aminolysis of fish oil ethyl esters and
Figure 6. (A) Separation of the fatty acyl ethanolamides from the NADES and the HPLC chromatogram of crude products of the (B) upper phase and (C) lower phase.
ethanolamine in the presence of CG−water. The approach proposed is an efficient and green technology, which shows potential for industrial applications.
■
ASSOCIATED CONTENT
S Supporting Information *
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jafc.8b04804. Calculation of product yields as well as viscosity and water activity of CG−water, with the effect of the water content of CG on viscosity (Figure S1) and the effect of 12365
DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
Article
Journal of Agricultural and Food Chemistry
■
(10) Melis, M.; Pillolla, G.; Luchicchi, A.; Muntoni, A. L.; Yasar, S.; Goldberg, S. R.; Pistis, M. Endogenous fatty acid ethanolamides suppress nicotine-induced activation of mesolimbic dopamine neurons through nuclear receptors. J. Neurosci. 2008, 28, 13985− 13994. (11) Dennedy, M. C.; Friel, A. M.; Houlihan, D. D.; Broderick, V. M.; Smith, T.; Morrison, J. J. Cannabinoids and the human uterus during pregnancy. Am. J. Obstet. Gynecol. 2004, 190, 2−9. (12) Quintana, P. G.; García Liñares, G.; Chanquia, S. N.; Gorojod, R. M.; Kotler, M. L.; Baldessari, A. Improved enzymatic procedure for the synthesis of anandamide and N-fatty acylalkanolamine analogues: A combination strategy to antitumor activity. Eur. J. Org. Chem. 2016, 2016, 518−528. (13) Balvers, M. G. J.; Verhoeckx, K. C. M.; Plastina, P.; Wortelboer, H. M.; Meijerink, J.; Witkamp, R. F. Docosahexaenoic acid and eicosapentaenoic acid are converted by 3T3-L1 adipocytes to N-acyl ethanolamines with anti-inflammatory properties. Biochim. Biophys. Acta, Mol. Cell Biol. Lipids 2010, 1801, 1107−1114. (14) Meijerink, J.; Plastina, P.; Vincken, J.; Poland, M.; Attya, M.; Balvers, M.; Gruppen, H.; Gabriele, B.; Witkamp, R. F. The ethanolamide metabolite of DHA, docosahexaenoylethanolamine, shows immunomodulating effects in mouse peritoneal and RAW264.7 macrophages: Evidence for a new link between fish oil and inflammation. Br. J. Nutr. 2011, 105, 1798−1807. (15) Meijerink, J.; Poland, M.; Balvers, M. G. J.; Plastina, P.; Lute, C.; Dwarkasing, J.; van Norren, K.; Witkamp, R. F. Inhibition of COX-2-mediated eicosanoid production plays a major role in the antiinflammatory effects of the endocannabinoid N- docosahexaenoylethanolamine (DHEA) in macrophages. Br. J. Pharmacol. 2015, 172, 24−37. (16) Rovito, D.; Giordano, C.; Vizza, D.; Plastina, P.; Barone, I.; Casaburi, I.; Lanzino, M.; De Amicis, F.; Sisci, D.; Mauro, L.; Aquila, S.; Catalano, S.; Bonofiglio, D.; Andò, S. Omega-3 PUFA ethanolamides DHEA and EPEA induce autophagy through PPARγ activation in MCF-7 breast cancer cells. J. Cell. Physiol. 2013, 228, 1314−1322. (17) Rashid, M. A.; Katakura, M.; Kharebava, G.; Kevala, K.; Kim, H. N-docosahexaenoylethanolamine is a potent neurogenic factor for neural stem cell differentiation. J. Neurochem. 2013, 125, 869−884. (18) Lee, J.; Huang, B. X.; Kwon, H.; Rashid, M. A.; Kharebava, G.; Desai, A.; Patnaik, S.; Marugan, J.; Kim, H. Orphan GPR110 (ADGRF1) targeted by N-docosahexaenoylethanolamine in development of neurons and cognitive function. Nat. Commun. 2016, 7, 13123. (19) Brown, I.; Cascio, M. G.; Wahle, K. W. J.; Smoum, R.; Mechoulam, R.; Ross, R. A.; Pertwee, R. G.; Heys, S. D. Cannabinoid receptor-dependent and -independent anti-proliferative effects of omega-3 ethanolamides in androgen receptor-positive and -negative prostate cancer cell lines. Carcinogenesis 2010, 31, 1584−1591. (20) Chen, Y.; Wang, Y.; Jin, Q.; Wang, X.; Hu, J.; Wang, X. Preparation of arachidonoyl ethanolamide by enzymatic amidation of arachidonic acid purified from microbial oil. Process Biochem. 2018, 66, 120−125. (21) Whitten, K. M.; Makriyannis, A.; Vadivel, S. K. Enzymatic synthesis of N-acylethanolamines: Direct method for the aminolysis of esters. Tetrahedron Lett. 2012, 53, 5753−5755. (22) Couturier, L.; Taupin, D.; Yvergnaux, F. Lipase-catalyzed chemoselective aminolysis of various aminoalcohols with fatty acids. J. Mol. Catal. B: Enzym. 2009, 56, 29−33. (23) Fernandez-Perez, M.; Otero, C. Enzymatic synthesis of amide surfactants from ethanolamine. Enzyme Microb. Technol. 2001, 28, 527−536. (24) Plastina, P.; Meijerink, J.; Vincken, J. -P.; Gruppen, H.; Witkamp, R.; Gabriele, B. Selective synthesis of unsaturated Nacylethanolamines by lipase-catalyzed N-acylation of ethanolamine with unsaturated fatty acids. Lett. Org. Chem. 2009, 6, 444−447. (25) Wang, X.; Wang, T.; Wang, X. An improved method for synthesis of N-stearoyl and N-palmitoylethanolamine. J. Am. Oil Chem. Soc. 2012, 89, 1305−1313.
the water content of CG on water activity (Figure S2) (PDF)
AUTHOR INFORMATION
Corresponding Author
*E-mail:
[email protected]. ORCID
Chin Ping Tan: 0000-0003-4177-4072 Yonghua Wang: 0000-0002-3255-752X Funding
This work was supported by the National Outstanding Youth Science Foundation of China (31725022), the Molecular Enzyme and Engineering International Cooperation Base of South China University of Technology (2017A050503001), the Special Program of Guangdong Province for Leader Project in Science and Technology Innovation: Development of New Partial Glycerin Lipase (2015TX01N207), and the Science and Technology Planning Project of Guangdong Province (2016B090920082). Notes
The authors declare no competing financial interest.
■
ABBREVIATIONS USED EPEA, eicosapentaenoyl ethanolamide; DHEA, docosahexaenoyl ethanolamide; EPA-EE, eicosapentaenoic acid ethyl ester; DHA-EE, docosahexaenoic acid ethyl ester; NADES, natural deep eutectic solvents; HPLC, high-performance liquid chromatography; BUH, betaine−urea−water; BGly, betaine− glycerol; CU, choline chloride−urea; CM, choline chloride− malonic acid; CG, choline chloride−glucose; CGlyU, choline chloride−glycerol−urea.
■
REFERENCES
(1) Liu, K. J.; Nag, A.; Shaw, J. Lipase-catalyzed synthesis of fatty acid diethanolamides. J. Agric. Food Chem. 2001, 49, 5761−5764. (2) Mäki-Arvela, P.; Tkacheva, A.; Dosmagambetova, I.; Chapelliere, Y.; Hachemi, I.; Kumar, N.; Aho, A.; Murzin, D. Y. Thermal and catalytic amidation of stearic acid with ethanolamine for production of pharmaceuticals and surfactants. Top. Catal. 2016, 59, 1151−1164. (3) Mouad, A. M.; Taupin, D.; Lehr, L.; Yvergnaux, F.; Porto, A. L. M. Aminolysis of linoleic and salicylic acid derivatives with Candida antarctica lipase B: A solvent-free process to obtain amphiphilic amides for cosmetic application. J. Mol. Catal. B: Enzym. 2016, 126, 64−68. (4) Verme, J. L. The nuclear receptor peroxisome proliferatoractivated receptor-α mediates the anti-inflammatory actions of palmitoylethanolamide. Mol. Pharmacol. 2005, 67, 15−19. (5) Petrosino, S.; Iuvone, T.; Di Marzo, V. N-palmitoyl-ethanolamine: Biochemistry and new therapeutic opportunities. Biochimie 2010, 92, 724−727. (6) Ameri, A. The effects of cannabinoids on the brain. Prog. Neurobiol. 1999, 58, 315−348. (7) Fu, J.; Astarita, G.; Gaetani, S.; Kim, J.; Cravatt, B. F.; Mackie, K.; Piomelli, D. Food intake regulates oleoylethanolamide formation and degradation in the proximal small intestine. J. Biol. Chem. 2007, 282, 1518−1528. (8) Schwartz, G. J.; Fu, J.; Astarita, G.; Li, X.; Gaetani, S.; Campolongo, P.; Cuomo, V.; Piomelli, D. The lipid messenger OEA links dietary fat intake to satiety. Cell Metab. 2008, 8, 281−288. (9) Fu, J.; Gaetani, S.; Oveisi, F.; Lo Verme, J.; Serrano, A.; Rodríguez de Fonseca, F.; Rosengarth, A.; Luecke, H.; Di Giacomo, B.; Tarzia, G.; Piomelli, D. Oleylethanolamide regulates feeding and body weight through activation of the nuclear receptor PPAR-α. Nature 2003, 425, 90−93. 12366
DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
Article
Journal of Agricultural and Food Chemistry (26) Wang, X.; Chen, Y.; Jin, Q.; Huang, J.; Wang, X. Synthesis of linoleoyl ethanolamide. J. Oleo Sci. 2013, 62, 427−33. (27) Wang, X.; Chen, Y.; Ma, Y.; Jin, Q.; Wang, X. Lipozyme 435catalyzed synthesis of eicosapentaenoyl ethanolamide in a solvent-free system. J. Mol. Catal. B: Enzym. 2015, 122, 233−239. (28) Zeng, C.; Qi, S.; Xin, R.; Yang, B.; Wang, Y. Enzymatic selective synthesis of 1,3-DAG based on deep eutectic solvent acting as substrate and solvent. Bioprocess Biosyst. Eng. 2015, 38, 2053−2061. (29) Pöhnlein, M.; Ulrich, J.; Kirschhöfer, F.; Nusser, M.; MuhleGoll, C.; Kannengiesser, B.; Brenner-Weiß, G.; Luy, B.; Liese, A.; Syldatk, C.; Hausmann, R. Lipase-catalyzed synthesis of glucose-6-Ohexanoate in deep eutectic solvents. Eur. J. Lipid Sci. Technol. 2015, 117, 161−166. (30) Zhou, P.; Wang, X.; Zeng, C.; Wang, W.; Yang, B.; Hollmann, F.; Wang, Y. Deep eutectic solvents enable more robust chemoenzymatic epoxidation reactions. ChemCatChem 2017, 9, 934−936. (31) Gu, L.; Huang, W.; Tang, S.; Tian, S.; Zhang, X. A novel deep eutectic solvent for biodiesel preparation using a homogeneous base catalyst. Chem. Eng. J. 2015, 259, 647−652. (32) Kim, S. H.; Park, S.; Yu, H.; Kim, J. H.; Kim, H. J.; Yang, Y.; Kim, Y. H.; Kim, K. J.; Kan, E.; Lee, S. H. Effect of deep eutectic solvent mixtures on lipase activity and stability. J. Mol. Catal. B: Enzym. 2016, 128, 65−72. (33) Huang, Z.; Wu, B.; Wen, Q.; Yang, T.; Yang, Z. Deep eutectic solvents can be viable enzyme activators and stabilizers. J. Chem. Technol. Biotechnol. 2014, 89, 1975−1981. (34) Lan, D.; Wang, X.; Zhou, P.; Hollmann, F.; Wang, Y. Deep eutectic solvents as performance additives in biphasic reactions. RSC Adv. 2017, 7, 40367−40370. (35) Durand, E.; Lecomte, J.; Baréa, B.; Dubreucq, E.; Lortie, R.; Villeneuve, P. Evaluation of deep eutectic solvent-water binary mixtures for lipase-catalyzed lipophilization of phenolic acids. Green Chem. 2013, 15, 2275−2282. (36) Durand, E.; Lecomte, J.; Baréa, B.; Piombo, G.; Dubreucq, E.; Villeneuve, P. Evaluation of deep eutectic solvents as new media for Candida antarctica B lipase catalyzed reactions. Process Biochem. 2012, 47, 2081−2089. (37) Wang, X.; Wang, X.; Wang, T. Synthesis of oleoylethanolamide using lipase. J. Agric. Food Chem. 2012, 60, 451−457. (38) Tufvesson, P.; Annerling, A.; Hatti-Kaul, R.; Adlercreutz, D. Solvent-free enzymatic synthesis of fatty alkanolamides. Biotechnol. Bioeng. 2007, 97, 447−453. (39) Xin, R.; Qi, S.; Zeng, C.; Khan, F. I.; Yang, B.; Wang, Y. A functional natural deep eutectic solvent based on trehalose: Structural and physicochemical properties. Food Chem. 2017, 217, 560−567.
12367
DOI: 10.1021/acs.jafc.8b04804 J. Agric. Food Chem. 2018, 66, 12361−12367
