Next Article in Journal
Immobilized Palladium Nanoparticles on Zirconium Carboxy-Aminophosphonates Nanosheets as an Efficient Recoverable Heterogeneous Catalyst for Suzuki–Miyaura and Heck Coupling
Next Article in Special Issue
Direct Conversion of Propylene Oxide to 3-Hydroxy Butyric Acid Using a Cobalt Carbonyl Ionic Liquid Catalyst
Previous Article in Journal
Catalytic Acetalization: An Efficient Strategy for High-Value Utilization of Biodiesel-Derived Glycerol
Previous Article in Special Issue
Caprolactam-Based Brønsted Acidic Ionic Liquids for Biodiesel Production from Jatropha Oil
Article Menu
Issue 6 (June) cover image

Export Article

Catalysts 2017, 7(6), 185; doi:10.3390/catal7060185

Lipase-Catalyzed Synthesis of Indolyl 4H-Chromenes via a Multicomponent Reaction in Ionic Liquid
Key Laboratory of Molecular Enzymology and Engineering of Ministry of Education, School of Life Sciences, Jilin University, 2699 Qianjin Street, Changchun 130000, China
State Key Laboratory of Supramolecular Structure and Materials, Jilin University, 2699 Qianjin Street, Changchun 130000, China
Department of Urology, China-Japan Union Hospital of Jilin University, 126 Xiantai Street, Changchun 130000, China
Authors to whom correspondence should be addressed.
Academic Editor: Sandrine Bouquillon
Received: 25 April 2017 / Accepted: 31 May 2017 / Published: 8 June 2017


Synthesis of indolyl 4H-chromenes via a three-component reaction catalyzed by lipase in ionic liquidsis reported here for the first time. High yields (77–98%) were obtained when Mucor miehei lipase was used as the catalyst in [EMIM][BF4]. Furthermore, [EMIM][BF4] exhibited good reusability in this enzymatic reaction. This study affords a new example of lipase catalytic promiscuity and broadens the application range of ionic liquid in biocatalysis.
lipase; promiscuity; multicomponent; ionic liquid; indolyl-4H-chromene

1. Introduction

Chromene and its derivatives can be found in many natural products [1]. With their vital role in pharmacology, they have been utilized in many fields, such as medicinal, bioorganic, and pharmaceutical chemistry [2,3,4,5,6,7]. The substructure of indole is a key structural motif in a variety of bioactive compounds [8,9]. Therefore, indoyl 4H-chromene has the combined properties of both indole and chromene moieties that provide new promising biological activities [10,11]. Many methods have been presented for the synthesis of these derivatives and various catalysts have been used, such as ZnO nanoparticle, polystyrene-supported p-toluenesulfonic acid, 4-dimethylaminopyridine (DMAP), β-cyclodextrin, oleic acid, 1,4-diazabicyclo[2.2.2]octane (DABCO), microbial cyclosophoraose, etc. [12,13,14,15,16,17]. However, most of the reported methods have encountered drawbacks, such as the utilization of environmentally hazardous and expensive catalysts, long reaction times, high reaction temperatures, and complicated reaction processes. Thus, a new alternative synthetic method for the indolyl 4H-chromene is still highly desirable.
Enzyme catalytic promiscuity is the “hidden skill” of the enzyme to catalyze different type of organic reactions [18,19,20,21,22,23,24,25]. This useful enzyme property makes it possible to catalyze multistep reactions in a multicomponent reaction (MCR). Recently, a series of enzymatic MCRs have been described to produce complex skeletons, and these findings have significantly broadened the use of enzymes in organic synthesis [26,27,28,29]. However, most of these enzymatic promiscuous reactions used organic solvent as the reaction media, which are volatile and toxic to the environment. Moreover, the deactivation of enzymes could be observed in these organic solvents, particularly at high temperatures.
It is known that room temperature ionic liquids (ILs) have been broadly used as attractive media in enzymatic reactions for their distinct features, such as the negligible vapor pressure, high thermal stability, and excellent biocompatibility [30,31]. Another advantage of ILs is that they can be easily recovered as the reaction media in biocatalysis [32,33]. However, only a few studies have used ionic liquid as the reaction media for enzyme catalytic promiscuous reactions. Sharma et al. reported a novel combination of enzyme and ionic liquid [HMIM] Br for the oxidation of aryl alcohols/acetates [34]. Yu and co-workers exhibited the asymmetric cross aldol reactions of aromatic aldehydes with ketones catalyzed by lipase in [BMIM][PF6] [35]. As part of our investigation on the enzymatic synthesis of heterocyclic compounds, a mild and efficient method for the synthesis of indolyl 4H-chromenes via a MCR catalyzed by lipase in ionic liquid (Scheme 1) is herein reported for the first time.

2. Results and Discussion

Initially, we carried out the model MCR in [EMIM][BF4] with salicylaldehyde (1a), indole (2a), and cyclohexane-1,3-dione (3) as the substrates catalyzed by different enzymes. The results are presented in Table 1. It could be observed that the catalytic activity was affected dramatically by the organism from which the enzyme was obtained. Mucor miehei lipase (MML) was identified to be the most efficient catalyst for this MCR in [EMIM][BF4]. The denatured MML and BSA did not exhibit activity for this MCR (Entries 6 and 7), and no reaction was observed in the absence of enzyme (Entry 8), which suggests that a specific conformation of lipase is necessary for the catalytic activity.
In this study, eight ILs of imidazolium type were selected for this enzymatic synthesis of indolyl 4H-chromene. As shown in Table 2, the yields were dramatically affected by the anion of ILs. MML was active in [BMIM][OTf], [BMIM]N(Tf)2, [BMIM][BF4], and [BMIM][PF6], but exhibited low activities in [BMIM][Ac] and [BMIM][NO3]. The low nucleophilicity of anion (PF6, BF4, N(Tf)2, or OTf) may have helped the ILs maintain the enzyme performance [36,37]. Higher yields were obtained in hydrophilic ionic liquid ([BMIM][BF4] and [BMIM][OTf]), which could be attributed to the preferable solubility of substrates in ILs. The effect of the cation of ionic liquid has also been studied (Entries 6–8). An apparent decrease of yield could be found in the results when the alkyl chain of the cation was elongated. One plausible explanation is that the high viscosity of ILs is harmful to the mass transfer [38,39,40]. Compared to the solvents (ethanol and water), MML exhibited a higher enzyme performance in [EMIM][BF4]. Therefore, [EMIM][BF4] was chosen as the suitable IL for further study.
Lipase catalyzed synthesis of indolyl 4H-chromene was carried out at a temperature range of 30 to 80 °C in [EMIM][BF4]. The effect of temperature on the reaction is demonstrated in Figure 1. The yield increased with the enhancement of temperature from 30 °C to 60 °C, and the maximum yield was achieved at 60 °C. Further increase in the reaction temperature resulted in an appreciable loss of the reaction yield. Generally, high temperature is known to increase the colliding probability between enzyme and substrate, which is conducive to form the enzyme-substrate complexes and improve the reaction rate. However, excess temperature could destroy the conformation of enzyme and decrease enzyme catalytic performance.
The effect of enzyme dosage was also studied (Figure 2). It could be found in Figure 2 that the yield increased along with the increasing amount of MML from 20 to 60 mg. But the yield could not be enhanced by further increasing the dosage of MML. Therefore, 60 mg of MML turned out to be sufficient to catalyze the reaction.
To test the robustness of the reaction, various substituted salicylaldehydes or indoles have been used for the synthesis of indolyl 4H-chromenes. As shown in Table 3, all reactions provided indolyl 4H-chromenes in good to excellent isolated yields (77–98%). It is noteworthy that the salicylaldehydes or indoles with electron donating groups (OH, methyl or methoxyl) provided higher yields than those substrates with electron withdrawing groups (F, Cl, Br, or NO2).
Another significant property of ILs is their reusability, which is responsible for their environmental friendliness character and the industrial applications. In this study, the IL was recovered by filtration from the reaction system and washed with cold ethyl acetate to remove the residual substrates. It was then dried under vacuum and reused for the next cycle. The results shown in Figure 3 indicated that the recovered IL was recycled for ten runs without any negative effect on the yield of indolyl 4H-chromene.
Generally, the immobilization is a powerful tool to enhance the reusability and stability of enzymes in modern biotechnology [41,42,43]. In this study, MML was immobilized on SBA-15 mesoporous silica via physical adsorption according to the previous study in our Lab [44,45], and the immobilized MML was used in the synthesis of indolyl 4H-chromenes. Our results indicated that the immobilized MML showed a lower catalytic performance than free MML (Table 4), but exhibited a satisfactory reusability (Figure 4). When the immobilized enzyme was reused, a slight loss of the catalytic activity could be observed, which may be due to the leakage of protein from SBA-15. Considering the operational simplicity of the immobilized enzyme on magnetic nanoparticles [46,47,48], research on using immobilized MML attached onto the magnetic Fe3O4 nanoparticles by covalent attachment for recycling is currently undergoing and will be reported in the future.

3. Materials and Methods

3.1. Materials

C. rugosa lipase (CRL), Porcine pancreatic lipase (PPL), and Candida antarctica lipase B (CalB) were purchased from Sigma (Beijing, China). Mucor miehei lipase (MML) was purchased from Shanghai Dongfeng Biochemical Reagent Co., Ltd. (Shanghai, China). Lipase from Pseudomonas sp.(PSL) was purchased from Amano Pharmaceutical Co., Ltd. (Tokyo, Japan). All enzymes were lyophilized before being used in the reaction. Substituted salicylaldehydes, substituted indoles, and cyclohexane-1,3-dione were purchased from J&K Scientific (Beijing, China). Ionic liquids were purchased from Shanghai Chengjie Chemical Co. Ltd. (Shanghai, China). The ionic liquids used and their abbreviations are listed in the Supplementary Materials. SBA-15 was donated by Dr. Yazhuo Li from College of Chemistry, Jilin University. All the other chemical reagents were purchased from Shanghai Chemical Reagent Company (Shanghai, China). All the commercially available reagents and solvents were used without further purification. NMR spectra were recorded on an Inova 500 (500 MHz) spectrometer (Vernon Hills, IL, USA).

3.2. General Procedure of the Synthesis of Indolyl 4H-Chromenes Catalyzed by Lipase

A mixture of substituted salicylaldehyde (1 mmol), substituted indole (1 mmol), and cyclohexane-1,3-dione (1 mmol), MML (60 mg, protein content) in [EMIM][BF4] (5 mL) was stirred at 60 °C in a round-bottom flask for 3 h. The enzymatic reaction was monitored by TLC (0.5-mm silica gel plates, eluent: ethyl acetate/n-hexane: 1/4). Then, the mixture was filtered, and the filter cake was washed with cold ethanol and water. The resulting residue was dried under vacuum to provide the pure product. All the products were well characterized by their 1H-NMR spectral analysis. Each experiment was performed triplicate, and all the data were obtained based on the average values.

3.3. Reusability of [EMIM][BF4]s

To test the reusability of [EMIM][BF4] in repeated use, batch reaction was conducted under the optimal reaction conditions for 3 h. After the reaction had been completed, enzyme and the product were filtered, and the IL was washed with cold ethyl acetate in order to extract the residual substrates. Then, the recycled [EMIM][BF4] was dried under vacuum and reused for the next cycle under the same conditions.

3.4. Immobilization of MML

The commercial MML (1 g) was dissolved in phosphate buffer (0.1 L, pH 7.5, 0.1 M) at 4 °C for 120 min, and the insoluble residue was removed by centrifugation (8000 rpm, 5 min). Then, the entire supernatant was lyophilized. MML solution (10 mg/mL) was obtained by dispersing the lyophilized MML in phosphate buffer (pH 7.5, 0.1 M). MML solution (10 mL, 10 mg/mL) was put into the tube containing SBA-15 (0.5 g) at 4 °C for 120 min under stirring. After that, the immobilized MML was obtained from the supernatant by centrifugation and washed with the deionized water more than three times. The immobilized MML was dried for 24 h and the enzyme loading of the immobilized MML was determined according to Lowry method for protein concentration [49].

3.5. Systhesis of Indolyl 4H-Chromenes Catalyzed by the Immobilized MML

A mixture of 1a (1 mmol), 2a (1 mmol), and 3 (1 mmol), immobilized MML (60 mg, protein content) in [EMIM][BF4] (5 mL) was stirred at 60 °C in a round-bottom flask for 3 h. The reaction was monitored by TLC (0.5-mm silica gel plates, eluent: ethyl acetate/n-hexane: 1/4). Then, the mixture was isolated by filtration, and the obtained precipitation was washed with cold ethanol and water. The resulting residue was dissolved in CHCl3, and the insoluble immobilized MML was then recovered by centrifugation. The organic phase was dried under vacuum to provide the pure product.

4. Conclusions

In conclusion, an efficient synthesis of indolyl 4H-chromenes via a three-component reaction catalyzed by lipase in ionic liquid has been developed for the first time. This novel strategy offers several advantages including a simple workup procedure, environmental friendliness, shorter reaction time with excellent yields, as well as the ability to convert a variety of substrates. Excitingly, ionic liquid exhibited a good reusability in the lipase catalyzed reaction. This green method provides not only a new case of lipase promiscuous reaction in organic synthesis, but also expands the utility of ionic liquid in enzyme promiscuous reaction.

Supplementary Materials

The following are available online at, Figure S1: Data of products, Figure S2: Spectra of products, Table S1: Ionic liquids and the abbreviations.


We gratefully acknowledge the financial support from the Fund of Basic Scientific Research from Jilin University (for Constructing the Scientific and Technological Platform, 2017) and the Foundation of Changchun BC&HC Pharmaceutical Technology Co., Ltd. (No. 3R116V401465).

Author Contributions

Weian Zhang, Ziyuan Zhao, Zhi Wang, and Chao Guo performed the experiments; Chunyu Wang and Weian Zhang analyzed the data; Rui Zhao and Lei Wang wrote the paper.

Conflicts of Interest

The authors declare no conflict of interest.


  1. Limsuwan, S.; Trip, E.N.; Kouwen, T.R.H.M.; Piersma, S.; Hiranrat, A.; Mahabusarakam, W.; Voravuthikunchai, S.P.; Dijl, J.M.; Kayser, O. Rhodomyrtone: A new candidate as natural antibacterial drug from Rhodomyrtus tomentosa. Phytomedicine 2009, 16, 645–651. [Google Scholar] [CrossRef] [PubMed]
  2. Hardcastle, I.R.; Cockcroft, X.L.; Curtin, N.J.; El-Murr, M.D.; Leahy, J.J.; Stockley, M.; Golding, B.T.; Rigoreau, L.; Richardson, C.; Smith, G.C.M.; et al. Discovery of Potent Chromen-4-one Inhibitors of the DNA-Dependent Protein Kinase (DNA-PK) Using a Small-Molecule Library Approach. J. Med. Chem. 2005, 48, 7829–7846. [Google Scholar] [CrossRef] [PubMed]
  3. Melander, R.J.; Minvielle, M.J.; Melander, C. Controlling bacterial behavior with indole-containing natural products and derivatives. Tetrahedron 2014, 70, 6363–6372. [Google Scholar] [CrossRef] [PubMed]
  4. Ambrus, J.I.; Kelso, M.J.; Bremner, J.B.; Ball, A.R.; Casadei, G.; Lewis, K. Structure-activity relationships of 2-aryl-1H-indole inhibitors of the NorA efflux pump in Staphylococcus aureus. Bioorg. Med. Chem. Lett. 2008, 18, 4294–4297. [Google Scholar] [CrossRef] [PubMed]
  5. Kidwai, M.; Saxena, S.; Khan, M.K.R.; Thukral, S.S. Aqua mediated synthesis of substituted 2-amino-4H-chromenes and in vitro study as antibacterial agents. Bioorg. Med. Chem. Lett. 2005, 15, 4295–4298. [Google Scholar] [CrossRef] [PubMed]
  6. Li, Q.; Guo, Y.; Xu, J.; Shao, S. Novel indole based colorimetric and “turn on” fluorescent sensors for biologically important fluoride anion sensing. J. Photochem. Photobiol. B 2011, 103, 140–144. [Google Scholar] [CrossRef] [PubMed]
  7. Kniess, T.; Laube, M.; Bergmann, R.; Sehn, F.; Graf, F.; Steinbach, J.; Wuest, F.; Pietzsch, J. Radiosynthesis of a 18 F-labeled 2,3-diarylsubstituted indole via McMurry coupling for functional characterization of cyclooxygenase-2 (COX-2) in vitro and in vivo. Bioorg. Med. Chem. 2012, 20, 3410–3421. [Google Scholar] [CrossRef] [PubMed]
  8. Kumar, D.; Reddy, V.B.; Sharad, S.; Dube, U.; Kapur, S. A facile one-pot green synthesis and antibacterial activity of 2-amino-4H-pyrans and 2-amino-5-oxo-5, 6, 7, 8-tetrahydro-4H-chromenes. Eur. J. Med. Chem. 2009, 44, 3805–3809. [Google Scholar] [CrossRef] [PubMed]
  9. Casapullo, A.; Bifulco, G.; Bruno, I.; Riccio, R. New Bisindole Alkaloids of the Topsentin and Hamacanthin Classes from the Mediterranean Marine Sponge Rhaphisia lacazei. J. Nat. Prod. 2000, 63, 447–451. [Google Scholar] [CrossRef] [PubMed]
  10. Kathrotiya, H.G.; Patel, M.P. Microwave-assisted synthesis of 3’-indolyl substituted 4H-chromenes catalyzed by DMAP and their antimicrobial activity. Med. Chem. Res. 2012, 21, 3406–3416. [Google Scholar] [CrossRef]
  11. Kasralikar, H.M.; Jadhavar, S.C.; Bhusare, S.R. Synthesis and molecular docking studies of oxochromenyl xanthenone and indolyl xanthenone derivatives as anti-HIV-1 RT inhibitors. Bioorg. Med. Chem. Lett. 2015, 25, 3882–3886. [Google Scholar] [CrossRef] [PubMed]
  12. Ghosh, P.P.; Das, A.R. Nanocrystalline and reusable ZnO catalyst for the assembly of densely functionalized 4H-chromenes in aqueous medium via one-pot three component reactions: A greener “NOSE” approach. J. Org. Chem. 2013, 78, 6170–6181. [Google Scholar] [CrossRef] [PubMed]
  13. Vijay, V.S.; Yong, S.J.; Yeon, T.J. A metal-free tandem C–C/C–O bond formation approach to densely functionalized indolyl 4H-chromenes catalyzed by polystyrene-supported p-toluenesulfonic acid under solvent-free conditions. Mol. Divers. 2015, 19, 367–383. [Google Scholar]
  14. Ganesan, A.; Kothandapani, J.; Nanubolu, J.B.; Ganesan, S.S. Oleic acid: A benign Brønsted acidic catalyst for densely substituted indole derivative synthesis. RSC Adv. 2015, 5, 28597–28600. [Google Scholar] [CrossRef]
  15. Pratibha, R.; Madhulika, S.; Snehlata, Y.; Jaya, S.; Jagdamba, S. β-Cyclodextrin: A Biomimetic Catalyst used for the Synthesis of 4H-chromene-3-carbonitrile and Tetrahydro-1H-xanthen-1-one Derivatives. Catal. Lett. 2015, 145, 2020–2028. [Google Scholar]
  16. Rajesh, U.C.; Kholiya, R.; Thakur, A.; Rawat, D.S. [TBA][Gly] ionic liquid promoted multi-component synthesis of 3-substituted indoles and indolyl-4H-chromenes. Tetrahedron Lett. 2015, 56, 1790–1793. [Google Scholar] [CrossRef]
  17. Dindulkar, S.D.; Jeong, D.; Cho, E.; Kim, D.; Jung, S. Microbial cyclosophoraose as a catalyst for the synthesis of diversified indolyl 4H-chromenes via one-pot three component reactions in water. Green Chem. 2016, 18, 3620–3627. [Google Scholar] [CrossRef]
  18. Kapoor, M.; Gupta, M.N. Lipase promiscuity and its biochemical applications. Process Biochem. 2012, 47, 555–569. [Google Scholar] [CrossRef]
  19. Guan, Z.; Fu, J.P.; He, Y.H. Biocatalytic promiscuity: Lipase-catalyzed asymmetric aldol reaction of heterocyclic ketones with aldehydes. Tetrahedron Lett. 2012, 53, 4959–4961. [Google Scholar] [CrossRef]
  20. Rivera-Ramírez, J.D.; Escalante, J.; López-Munguía, A.; Marty, A.; Castillo, E. Thermodynamically controlled chemoselectivity in lipase-catalyzed aza-Michael additions. J. Mol. Catal. B Enzym. 2015, 112, 76–82. [Google Scholar] [CrossRef]
  21. Le, Z.G.; Guo, L.T.; Jiang, G.F.; Yang, X.B.; Liu, H.Q. Henry reaction catalyzed by Lipase A from Aspergillus niger. Green Chem. Lett. Rev. 2013, 6, 277–281. [Google Scholar] [CrossRef]
  22. Wu, L.L.; Xiang, Y.; Yang, D.C.; Guan, Z.; He, Y.H. Biocatalytic asymmetric Mannich reaction of ketimines using wheat germ lipase. Catal. Sci. Technol. 2016, 6, 3963–3970. [Google Scholar] [CrossRef]
  23. Wang, Z.; Wang, C.Y.; Wang, H.R.; Zhang, H.; Su, Y.L.; Ji, T.F.; Wang, L. Lipase-catalyzed Knoevenagel condensation between α, β-unsaturated aldehydes and active methylene compounds. Chin. Chem. Lett. 2014, 25, 802–804. [Google Scholar] [CrossRef]
  24. Reetz, M.T.; Mondière, R.; Carballeira, J.D. Enzyme promiscuity: First protein-catalyzed Morita-Baylis-Hillman reaction. Tetrahedron Lett. 2007, 48, 1679–1681. [Google Scholar] [CrossRef]
  25. Izquierdo, D.F.; Barbosa, O.; Burguete, M.I.; Lozano, P.; Luis, S.V.; Fernandez-Lafuente, R.; García-Verdugo, E. Tuning lipase B from Candida antarctica C-C bond promiscuous activity by immobilization on poly-styrene-divinylbenzene beads. RSC Adv. 2014, 4, 6219–6225. [Google Scholar] [CrossRef]
  26. Chen, X.; Zhang, W.A.; Yang, F.J.; Guo, C.; Zhao, Z.Y.; Ji, D.; Zhou, F.; Wang, Z.; Zhao, R.; Wang, L. Synthesis of dihydropyrano[4,3-b]pyranes via a multicomponent reaction catalyzed by lipase. Green Chem. Lett. Rev. 2017, 10, 54–58. [Google Scholar] [CrossRef]
  27. Bora, P.P.; Bihani, M.; Bez, G. Multicomponent synthesis of dihydropyrano [2, 3-c] pyrazoles catalyzed by lipase from Aspergillus niger. J. Mol. Catal. B Enzym. 2013, 92, 24–33. [Google Scholar] [CrossRef]
  28. Kłossowski, S.; Wiraszka, B.; Berłożecki, S.; Ostaszewski, R. Model studies on the first enzyme-catalyzed Ugi reaction. Org. Lett. 2013, 15, 566–569. [Google Scholar] [CrossRef] [PubMed]
  29. Yang, F.J.; Wang, Z.; Wang, H.R.; Zhang, H.; Yue, H.; Wang, L. Enzyme catalytic promiscuity: Lipase catalyzed synthesis of substituted 2H-chromenes by a three-component reaction. RSC Adv. 2014, 4, 25633–25636. [Google Scholar] [CrossRef]
  30. Plechkova, N.V.; Seddon, K.R. Applications of ionic liquids in the chemical industry. Chem. Soc. Rev. 2008, 37, 123–150. [Google Scholar] [CrossRef] [PubMed]
  31. Shimomura, K.; Harami, H.; Matsubara, Y.; Nokami, T.; Katada, N.; Itoh, T. Lipase-mediated dynamic kinetic resolution (DKR) of secondary alcohols in the presence of zeolite using an ionic liquid solvent system. Catal. Today 2015, 255, 41–48. [Google Scholar] [CrossRef]
  32. Zhao, R.H.; Zhang, X.W.; Zheng, L.; Xu, H.; Li, M. Enantioselective esterification of (R,S)-flurbiprofen catalyzed by lipase in ionic liquid. Green Chem. Lett. Rev. 2017, 10, 23–28. [Google Scholar] [CrossRef]
  33. Liu, Y.; Guo, C.; Liu, C.Z. Efficient kinetic resolution of (R,S)-2-octanol catalyzed by magnetite-immobilized Yarrowia lipolytica lipase in mixed ionic liquids. Catal. Lett. 2014, 144, 1552–1556. [Google Scholar] [CrossRef]
  34. Sharma, U.K.; Sharma, N.; Kumar, R.; Kumar, R.; Sinha, A.K. Biocatalytic promiscuity of lipase in chemoselective oxidation of aryl alcohols/acetates: A unique synergism of CAL-B and [hmim] Br for the metal-free H2O2 activation. Org. Lett. 2009, 11, 4846–4848. [Google Scholar] [CrossRef] [PubMed]
  35. Zhang, Y.; Wang, N.; Xie, Z.B.; Zhou, L.H.; Yu, X.Q. Ionic liquid as a recyclable and efficient medium for lipase-catalyzed asymmetric cross aldol reaction. J. Mol. Catal. B Enzym. 2014, 110, 100–110. [Google Scholar] [CrossRef]
  36. Park, S.; Kazlauskas, R.J. Biocatalysis in ionic liquids-advantages beyond green technology. Curr. Opin. Biotechnol. 2003, 14, 432–437. [Google Scholar] [CrossRef]
  37. Sheldon, R.A.; Laua, R.M.; Sorgedragera, M.J.; Rantwijka, F.V.; Seddon, K.R. Biocatalysis in ionic liquids. Green Chem. 2002, 4, 147–151. [Google Scholar] [CrossRef]
  38. Hallett, J.P.; Welton, T. Room-temperature ionic liquids: Solvents for synthesis and catalysis. Chem. Rev. 2011, 111, 3508–3576. [Google Scholar] [CrossRef] [PubMed]
  39. Sanmamed, Y.A.; González-Salgado, D.; Troncoso, J.; Cerdeiriña, C.A.; Romaní, L. Viscosity-induced errors in the density determination of room temperature ionic liquids using vibrating tube densitometry. Fluid Phase Equilibr. 2007, 252, 96–102. [Google Scholar] [CrossRef]
  40. Han, D.D.; Row, K.H. Recent applications of ionic liquids in separation technology. Molecules 2010, 15, 2405–2426. [Google Scholar] [CrossRef] [PubMed]
  41. Rodrigues, R.C.; Ortiz, C.; Berenguer-Murcia, Á.; Torres, R.; Fernández-Lafuente, R. Modifying enzyme activity and selectivity by immobilization. Chem. Soc. Rev. 2013, 42, 6290–6307. [Google Scholar] [CrossRef] [PubMed]
  42. Manoel, E.A.; Santos, J.C.S.; Freire, D.M.G.; Rueda, N.; Fernandez-Lafuente, R. Immobilization of lipases on hydrophobic supports involves the open form of the enzyme. Enzyme Microb. Technol. 2015, 71, 53–57. [Google Scholar] [CrossRef] [PubMed]
  43. Rueda, N.; Santos, J.C.S.; Ortiz, C.; Torres, R.; Barbosa, O.; Rodrigues, R.C.; Berenguer-Murcia, Á.; Fernandez-Lafuente, R. Chemical Modification in the Design of Immobilized Enzyme Biocatalysts: Drawbacks and Opportunities. Chem. Rec. 2016, 16, 1436–1455. [Google Scholar] [CrossRef] [PubMed]
  44. Yu, D.H.; Wang, Z.; Zhao, L.F.; Cheng, Y.M.; Cao, S.G. Resolution of 2-octanol by SBA-15 immobilized Pseudomonas sp. lipase. J. Mol. Catal. B Enzym. 2007, 48, 64–69. [Google Scholar] [CrossRef]
  45. Zhang, H.; Xun, E.N.; Wang, J.X.; Chen, G.; Cheng, T.X.; Wang, Z.; Ji, T.F.; Wang, L. Immobilization of laccase for oxidative coupling of trans-resveratrol and its derivatives. Int. J. Mol. Sci. 2012, 13, 5998–6008. [Google Scholar] [CrossRef] [PubMed]
  46. Xun, E.N.; Lv, X.L.; Kang, W.; Wang, J.X.; Zhang, H.; Wang, L.; Wang, Z. Immobilization of Pseudomonas fluorescens lipase onto magnetic nanoparticles for resolution of 2-octanol. Appl. Biochem. Biotechnol. 2012, 168, 697–707. [Google Scholar] [CrossRef] [PubMed]
  47. Cipolatti, E.P.; Valério, A.; Henriques, R.O.; Moritz, D.E.; Ninow, J.L.; Freire, D.M.G.; Manoel, E.A.; Fernandez-Lafuente, R.; Oliveira, D. Nanomaterials for biocatalyst immobilization-state of the art and future trends. RSC Adv. 2016, 6, 104675–104692. [Google Scholar] [CrossRef]
  48. Zang, L.M.; Qiu, J.H.; Wu, X.L.; Zhang, W.J.; Sakai, E.; Wei, Y. Preparation of magnetic chitosan nanoparticles as support for cellulase immobilization. Ind. Eng. Chem. Res. 2014, 53, 3448–3454. [Google Scholar] [CrossRef]
  49. Lowry, O.H.; Rosebrough, N.J.; Farr, A.L.; Randall, R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 1951, 193, 265–275. [Google Scholar] [PubMed]
Scheme 1. Synthesis of indolyl 4H-chromenes catalyzed by lipase in ionic liquid.
Scheme 1. Synthesis of indolyl 4H-chromenes catalyzed by lipase in ionic liquid.
Catalysts 07 00185 sch001
Figure 1. The effect of temperature on the synthesis of indolyl 4H-chromene. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML (60 mg, protein content), 3 h.
Figure 1. The effect of temperature on the synthesis of indolyl 4H-chromene. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML (60 mg, protein content), 3 h.
Catalysts 07 00185 g001
Figure 2. The effect of enzyme dosage on the synthesis of indolyl 4H-chromene. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML, 60 °C, 3 h.
Figure 2. The effect of enzyme dosage on the synthesis of indolyl 4H-chromene. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML, 60 °C, 3 h.
Catalysts 07 00185 g002
Figure 3. The reusability of [EMIM][BF4]. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML (60 mg, protein content), 60 °C, 3 h.
Figure 3. The reusability of [EMIM][BF4]. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML (60 mg, protein content), 60 °C, 3 h.
Catalysts 07 00185 g003
Figure 4. The reusability of immobilized MML in the synthesis of indolyl 4H-chromenes. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), enzyme (60 mg, protein content), 60 °C, 3 h.
Figure 4. The reusability of immobilized MML in the synthesis of indolyl 4H-chromenes. Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), enzyme (60 mg, protein content), 60 °C, 3 h.
Catalysts 07 00185 g004
Table 1. The effect of enzyme origin on the synthesis of indolyl 4H-chromene a.
Table 1. The effect of enzyme origin on the synthesis of indolyl 4H-chromene a.
EntryEnzymeIsolated Yield (%) b
1MML (Mucor miehei lipase)93
2PPL (Porcine pancreatic lipase)81
3PSL (Lipase from Pseudomonas sp.)62
4CRL (C. rugosa lipase)48
5CalB (C. antarctica lipase B)68
6denatured MML cN.D. d
7Albumin from bovine serum (BSA)N.D. d
8ControlN.D. d
a Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), enzyme (60 mg, protein content), 60 °C, 3 h. b Isolation procedure was described in Section 3.2. c The denatured MML was obtained by heating MML to 100 °C for 12 h in water before lyophilization. d N.D.: Not detected.
Table 2. The effect of ionic liquid on the synthesis of indolyl 4H-chromene.
Table 2. The effect of ionic liquid on the synthesis of indolyl 4H-chromene.
EntryIonic LiquidIsolated Yield (%)
Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), solvent (5 mL), Mucor miehei lipase (MML) (60 mg, protein content), 60 °C, 3 h.
Table 3. Synthesis of functionalized indolyl 4H-chromene derivatives.
Table 3. Synthesis of functionalized indolyl 4H-chromene derivatives.
Catalysts 07 00185 i001
Catalysts 07 00185 i002
Reaction condition: 1 (1 mmol), 2 (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), MML (60 mg, protein content), 60 °C, 3 h.
Table 4. Comparison of free MML and the immobilized MML.
Table 4. Comparison of free MML and the immobilized MML.
CatalystBound Protein (mg/g)Isolated Yield (%)
Free MML-93
Immobilized MML18086
Reaction condition: 1a (1 mmol), 2a (1 mmol), 3 (1 mmol), [EMIM][BF4] (5 mL), enzyme (60 mg, protein content), 60 °C, 3 h.
Catalysts EISSN 2073-4344 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top