Recent Trends in Enzyme Immobilization—Concepts for Expanding the Biocatalysis Toolbox
Abstract
:1. Introduction
2. Discussion: Novel Support Technologies
2.1. Polysaccharides
2.2. DNA
2.3. Chitosan
2.4. Renewables
2.5. Metal–Organic Frameworks
2.6. Controlled Pore Glass
2.7. Magnetic Nanoparticles
3. Integrating Immobilization into Developing Biocatalytic Technology
3.1. Flow Biocatalysis
3.2. 3D-Printed Biocatalytic Scaffolds
3.3. Multi-Enzymatic Cascade Reactions
3.4. Integrating Enzyme Immobilization and Protein Engineering
4. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Conflicts of Interest
References
- Mix, S.; Moody, T.S.; Taylor, S.J. Biocatalysis—How Secret Should It Be? Chemical Knowledge Hub. Available online: https://www.chemicalsknowledgehub.com/article/14595/ (accessed on 16 August 2020).
- Mullin, R.; Moody, T.S. Ticking a new box in enzyme chemistry. Chem. Eng. News 2019, 97, 34–35. [Google Scholar]
- Moody, T.; Mix, S. Managing and redesigning chemical processes with enzymes. Spec. Chem. Mag. 2019, 22–25. Available online: https://www.specchemonline.com/ (accessed on 16 August 2020).
- Wu, S.; Snajdrova, R.; Moore, J.C.; Baldenius, K.; Bornscheuer, U.T. Biocatalysis: Enzymatic Synthesis for Industrial Applications. Angew. Chem. Int. Ed. 2021, 60, 88–119. [Google Scholar] [CrossRef] [PubMed]
- Federsel, H.-J.; Pesti, J.; Thompson, M.P. Immobilized Enzymes: Application in Organic Synthesis. In Catalyst Immobilization. Methods and Applications; Benaglia, M., Puglisi, A., Eds.; Wiley-VCH: Weinheim, Germany, 2020; Chapter 13; pp. 437–463. [Google Scholar] [CrossRef]
- Salvi, H.M.; Yadav, G.D. Process intensification using immobilized enzymes for the development of white biotechnology. Catal. Sci. Technol. 2021, 11, 1994–2020. [Google Scholar] [CrossRef]
- Bilal, M.; Iqbal, H.M. Naturally-derived biopolymers: Potential platforms for enzyme immobilization. Int. J. Biol. Macromol. 2019, 130, 462–482. [Google Scholar] [CrossRef] [PubMed]
- Reis, C.; Sousa, E.; Serpa, J.; Oliveira, R.; Santos, J. Design of immobilized enzyme biocatalysts: Drawbacks and opportunities. Química Nova 2019, 42, 768–783. [Google Scholar] [CrossRef]
- Rodriguez-Abetxuko, A.; Sánchez-Dealcázar, D.; Muñumer, P.; Beloqui, A. Tunable Polymeric Scaffolds for Enzyme Immobilization. Front. Bioeng. Biotechnol. 2020, 8, 830. [Google Scholar] [CrossRef]
- Kong, H.J. Designing alginate hydrogels to maintain viability of immobilized cells. Biomaterials 2003, 24, 4023–4029. [Google Scholar] [CrossRef]
- Singh, R.; Kennedy, J. Immobilization of yeast inulinase on chitosan beads for the hydrolysis of inulin in a batch system. Int. J. Biol. Macromol. 2017, 95, 87–93. [Google Scholar] [CrossRef] [PubMed]
- Lv, M.; Ma, X.; Anderson, D.P.; Chang, P.R. Immobilization of urease onto cellulose spheres for the selective removal of urea. Cellulose 2017, 25, 233–243. [Google Scholar] [CrossRef]
- Nawaz, M.A.; Karim, A.; Bibi, Z.; Rehman, H.U.; Aman, A.; Hussain, D.; Ullah, M.; Qader, S.A.U. Maltase entrapment approach as an efficient alternative to increase the stability and recycling efficiency of free enzyme within agarose matrix. J. Taiwan Inst. Chem. Eng. 2016, 64, 31–38. [Google Scholar] [CrossRef]
- Singh, V.; Singh, D. Glucose Oxidase Immobilization on Guar Gum–Gelatin Dual-Templated Silica Hybrid Xerogel. Ind. Eng. Chem. Res. 2014, 53, 3854–3860. [Google Scholar] [CrossRef]
- Prakash, O.; Jaiswal, N. Immobilization of a Thermostable α-Amylase on Agarose and Agar Matrices and its Application in Starch Stain Removal. World Appl. Sci. J. 2011, 13, 572–577. [Google Scholar]
- Kara, F.; Demirel, G.; Tümtürk, H. Immobilization of urease by using chitosan–alginate and poly(acrylamide-co-acrylic acid)/κ-carrageenan supports. Bioprocess Biosyst. Eng. 2006, 29, 207–211. [Google Scholar] [CrossRef] [PubMed]
- Bilal, M.; Asgher, M.; Iqbal, H.M.N.; Hu, H.; Zhang, X. Gelatin-Immobilized Manganese Peroxidase with Novel Catalytic Characteristics and Its Industrial Exploitation for Fruit Juice Clarification Purposes. Catal. Lett. 2016, 146, 2221–2228. [Google Scholar] [CrossRef]
- Rocha-Martin, J.; Acosta, A.; Berenguer, J.; Guisan, J.M.; Lopez-Gallego, F. Selective oxidation of glycerol to 1,3-dihydroxyacetone by covalently immobilized glycerol dehydrogenases with higher stability and lower product inhibition. Bioresour. Technol. 2014, 170, 445–453. [Google Scholar] [CrossRef] [PubMed]
- Yovcheva, T.; Vasileva, T.; Viraneva, A.; Cholev, D.; Bodurov, I.; Marudova, M.; Bivolarski, V.; Iliev, I. Effect of immobilization conditions on the properties of β-galactosidase immobilized in xanthan/chitosan multilayers. J. Phys. Conf. Ser. 2017, 794, 12032. [Google Scholar] [CrossRef]
- Costas, L.; Bosio, V.E.; Pandey, A.; Castro, G.R. Effects of Organic Solvents on Immobilized Lipase in Pectin Microspheres. Appl. Biochem. Biotechnol. 2008, 151, 578–586. [Google Scholar] [CrossRef]
- Gür, S.D.; Idil, N.; Aksöz, N. Optimization of Enzyme Co-Immobilization with Sodium Alginate and Glutaraldehyde-Activated Chitosan Beads. Appl. Biochem. Biotechnol. 2017, 184, 538–552. [Google Scholar] [CrossRef]
- Klein, W.P.; Thomsen, R.P.; Turner, K.B.; Walper, S.A.; Vranish, J.; Kjems, J.; Ancona, M.G.; Medintz, I.L. Enhanced Catalysis from Multienzyme Cascades Assembled on a DNA Origami Triangle. ACS Nano 2019, 13, 13677–13689. [Google Scholar] [CrossRef] [PubMed]
- Verma, M.L.; Kumar, S.; Das, A.; Randhawa, J.S.; Chamundeeswari, M. Chitin and chitosan-based support materials for enzyme immobilization and biotechnological applications. Environ. Chem. Lett. 2020, 18, 315–323. [Google Scholar] [CrossRef]
- Verma, M.L.; Kumar, S.; Das, A.; Randhawa, J.S.; Chamundeeswari, M. Enzyme Immobilization on Chitin and Chitosan-Based Supports for Biotechnological Applications. In Sustainable Agriculture Reviews 35, 1st ed.; Crini, G., Lichtfouse, E., Eds.; Springer: Cham, Switzerland, 2019; pp. 147–173. [Google Scholar] [CrossRef]
- Bösiger, P.; Tegl, G.; Richard, I.M.; Le Gat, L.; Huber, L.; Stagl, V.; Mensah, A.; Guebitz, G.M.; Rossi, R.M.; Fortunato, G. Enzyme functionalized electrospun chitosan mats for antimicrobial treatment. Carbohydr. Polym. 2018, 181, 551–559. [Google Scholar] [CrossRef] [PubMed]
- Girelli, A.M.; Astolfi, M.L.; Scuto, F.R. Agro-industrial wastes as potential carriers for enzyme immobilization: A review. Chemosphere 2020, 244, 125368. [Google Scholar] [CrossRef] [PubMed]
- Brígida, A.I.S.; Pinheiro, Á.D.T.; Ferreira, A.L.O.; Gonçalves, L.R.B. Immobilization of Candida antarctica Lipase B by Adsorption to Green Coconut Fiber. Appl. Biochem. Biotechnol. 2008, 146, 173–187. [Google Scholar] [CrossRef] [PubMed]
- Cristóvão, R.O.; Silvério, S.C.; Tavares, A.P.M.; Brígida, A.I.S.; Loureiro, J.M.; Boaventura, R.A.R.; Macedo, E.A.; Coelho, M.A.Z. Green coconut fiber: A novel carrier for the immobilization of commercial laccase by covalent attachment for textile dyes decolourization. World J. Microbiol. Biotechnol. 2012, 28, 2827–2838. [Google Scholar] [CrossRef] [PubMed]
- Borgio, J.F. Immobilization of Microbial (Wild and Mutant Strains) Amylase on Coconut Fiber and Alginate Matrix for Enhanced Activity. Am. J. Biochem. Mol. Biol. 2011, 1, 255–264. [Google Scholar] [CrossRef]
- Bonet-Ragel, K.; López-Pou, L.; Tutusaus, G.; Benaiges, M.D.; Valero, F. Rice husk ash as a potential carrier for the immobilization of lipases applied in the enzymatic production of biodiesel. Biocatal. Biotransform. 2018, 36, 151–158. [Google Scholar] [CrossRef]
- Kessi, E.; Arias, J.L. Using Natural Waste Material as a Matrix for the Immobilization of Enzymes: Chicken Eggshell Membrane Powder for β-Galactosidase Immobilization. Appl. Biochem. Biotechnol. 2018, 187, 101–115. [Google Scholar] [CrossRef]
- Bassan, J.C.; Bezerra, T.M.D.S.; Peixoto, G.; Da Cruz, C.Z.P.; Galán, J.P.M.; Vaz, A.B.D.S.; Garrido, S.S.; Filice, M.; Monti, R. Immobilization of Trypsin in Lignocellulosic Waste Material to Produce Peptides with Bioactive Potential from Whey Protein. Materials 2016, 9, 357. [Google Scholar] [CrossRef] [Green Version]
- Pandey, D.; Daverey, A.; Arunachalam, K. Biochar: Production, properties and emerging role as a support for enzyme immobilization. J. Clean. Prod. 2020, 255, 120267. [Google Scholar] [CrossRef]
- Ye, N.; Kou, X.; Shen, J.; Huang, S.; Chen, G.; Ouyang, G. Metal-Organic Frameworks: A New Platform for Enzyme Immobilization. ChemBioChem 2020, 21, 2585–2590. [Google Scholar] [CrossRef]
- Hu, C.; Bai, Y.; Hou, M.; Wang, Y.; Wang, L.; Cao, X.; Chan, C.-W.; Sun, H.; Li, W.; Ge, J.; et al. Defect-induced activity enhancement of enzyme-encapsulated metal-organic frameworks revealed in microfluidic gradient mixing synthesis. Sci. Adv. 2020, 6, eaax5785. [Google Scholar] [CrossRef] [Green Version]
- Cassimjee, K.E.; Kadow, M.; Wikmark, Y.; Humble, M.S.; Rothstein, M.L.; Rothstein, D.M.; Bäckvall, J.-E. A general protein purification and immobilization method on controlled porosity glass: Biocatalytic applications. Chem. Commun. 2014, 50, 9134–9137. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Engelmark Cassimjee, K.; Federsel, H.-J. EziG: A Universal Platform for Enzyme Immobilization. In Biocatalysis: An Industrial Perspective; de Gonzalo, G., de Maria, P.D., Eds.; RSC Catalysis Series No. 29; The Royal Society of Chemistry: London, UK, 2018; Chapter 13; pp. 345–362. [Google Scholar] [CrossRef]
- Cassimjee, K.E.; Hendil-Forssell, P.; Volkov, A.; Krog, A.; Malmo, J.; Aune, T.E.V.; Knecht, W.; Miskelly, I.R.; Moody, T.S.; Humble, M.S. Streamlined Preparation of Immobilized Candida antarctica Lipase B. ACS Omega 2017, 2, 8674–8677. [Google Scholar] [CrossRef] [PubMed]
- Bilal, M.; Zhao, Y.; Rasheed, T.; Iqbal, H.M. Magnetic nanoparticles as versatile carriers for enzymes immobilization: A review. Int. J. Biol. Macromol. 2018, 120, 2530–2544. [Google Scholar] [CrossRef]
- Cui, J.; Cui, L.; Jia, S.; Su, Z.; Zhang, S. Hybrid Cross-Linked Lipase Aggregates with Magnetic Nanoparticles: A Robust and Recyclable Biocatalysis for the Epoxidation of Oleic Acid. J. Agric. Food Chem. 2016, 64, 7179–7187. [Google Scholar] [CrossRef]
- Gao, J.; Yu, H.; Zhou, L.; He, Y.; Ma, L.; Jiang, Y. Formation of cross-linked nitrile hydratase aggregates in the pores of tannic-acid-templated magnetic mesoporous silica: Characterization and catalytic application. Biochem. Eng. J. 2017, 117, 92–101. [Google Scholar] [CrossRef]
- Tural, B.; Tarhan, T.; Tural, S. Covalent immobilization of benzoylformate decarboxylase from Pseudomonas putida on magnetic epoxy support and its carboligation reactivity. J. Mol. Catal. B Enzym. 2014, 102, 188–194. [Google Scholar] [CrossRef]
- Zlateski, V.; Fuhrer, R.; Koehler, F.M.; Wharry, S.; Zeltner, M.; Stark, W.J.; Moody, T.S.; Grass, R.N. Efficient Magnetic Recycling of Covalently Attached Enzymes on Carbon-Coated Metallic Nanomagnets. Bioconj. Chem. 2014, 25, 677–684. [Google Scholar] [CrossRef]
- Romero-Fernández, M.; Paradisi, F. Protein immobilization technology for flow biocatalysis. Curr. Opin. Chem. Biol. 2020, 55, 1–8. [Google Scholar] [CrossRef] [PubMed]
- Britton, J.; Majumdar, S.; Weiss, G.A. Continuous flow biocatalysis. Chem. Soc. Rev. 2018, 47, 5891–5918. [Google Scholar] [CrossRef]
- Thompson, M.P.; Peñafiel, I.; Cosgrove, S.C.; Turner, N.J. Biocatalysis Using Immobilized Enzymes in Continuous Flow for the Synthesis of Fine Chemicals. Org. Process Res. Dev. 2019, 23, 9–18. [Google Scholar] [CrossRef]
- Bolivar, J.M.; Mannsberger, A.; Thomsen, M.S.; Tekautz, G.; Nidetzky, B. Process intensification for O2-dependent enzymatic transformations in continuous single-phase pressurized flow. Biotechnol. Bioeng. 2019, 116, 503–514. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bolivar, J.M.; Luley-Goedl, C.; Leitner, E.; Sawangwan, T.; Nidetzky, B. Production of glucosyl glycerol by immobilized sucrose phosphorylase: Options for enzyme fixation on a solid support and application in microscale flow format. J. Biotechnol. 2017, 257, 131–138. [Google Scholar] [CrossRef]
- Valikhani, D.; Bolivar, J.M.; Pfeiffer, M.; Nidetzky, B. Multivalency Effects on the Immobilization of Sucrose Phosphorylase in Flow Microchannels and Their Use in the Development of a High-Performance Biocatalytic Microreactor. ChemCatChem 2017, 9, 161–166. [Google Scholar] [CrossRef]
- Ye, J.; Chu, T.; Chu, J.; Gao, B.; He, B. A Versatile Approach for Enzyme Immobilization Using Chemically Modified 3D-Printed Scaffolds. ACS Sustain. Chem. Eng. 2019, 7, 18048–18054. [Google Scholar] [CrossRef]
- Mayer, S.F.; Kroutil, W.; Faber, K. Enzyme-Initiated Domino (Cascade) Reactions. Chem. Soc. Rev. 2001, 30, 332–339. [Google Scholar] [CrossRef]
- Schoffelen, S.; Van Hest, J.C.M. Multi-enzyme systems: Bringing enzymes together in vitro. Soft Matter 2011, 8, 1736–1746. [Google Scholar] [CrossRef]
- Ricca, E.; Brucher, B.; Schrittwieser, J.H. Multi-Enzymatic Cascade Reactions: Overview and Perspectives. Adv. Synth. Catal. 2011, 353, 2239–2262. [Google Scholar] [CrossRef]
- Monti, D.; Ferrandi, E.E.; Zanellato, I.; Hua, L.; Polentini, F.; Carrea, G.; Riva, S. One-Pot Multienzymatic Synthesis of 12-Ketoursodeoxycholic Acid: Subtle Cofactor Specificities Rule the Reaction Equilibria of Five Biocatalysts Working in a Row. Adv. Synth. Catal. 2009, 351, 1303–1311. [Google Scholar] [CrossRef]
- Lopez-Gallego, F.; Schmidt-Dannert, C. Multi-enzymatic synthesis. Curr. Opin. Chem. Biol. 2010, 14, 174–183. [Google Scholar] [CrossRef] [PubMed]
- Zhang, Y.; Hess, H. Toward Rational Design of High-efficiency Enzyme Cascades. ACS Catal. 2017, 7, 6018–6027. [Google Scholar] [CrossRef] [Green Version]
- Bruggink, A.; Schoevaart, R.; Kieboom, T. Concepts of Nature in Organic Synthesis: Cascade Catalysis and Multistep Conversions in Concert. Org. Process Res. Dev. 2003, 7, 622–640. [Google Scholar] [CrossRef]
- Ji, Q.; Wang, B.; Tan, J.; Zhu, L.; Li, L. Immobilized multienzymatic systems for catalysis of cascade reactions. Process Biochem. 2016, 51, 1193–1203. [Google Scholar] [CrossRef]
- Mateo, C.; Chmura, A.; Rustler, S.; Van Rantwijk, F.; Stolz, A.; Sheldon, R.A. Synthesis of enantiomerically pure (S)-mandelic acid using an oxynitrilase–nitrilase bienzymatic cascade: A nitrilase surprisingly shows nitrile hydratase activity. Tetrahedron Asymmetry 2006, 17, 320–323. [Google Scholar] [CrossRef]
- Zhang, Y.; Wang, Q.; Hess, H. Increasing Enzyme Cascade Throughput by pH-Engineering the Microenvironment of Individual Enzymes. ACS Catal. 2017, 7, 2047–2051. [Google Scholar] [CrossRef]
- Findrik, Z.; Vasić-Rački, Đ. Biotransformation of d-methionine into l-methionine in the cascade of four enzymes. Biotechnol. Bioeng. 2007, 98, 956–967. [Google Scholar] [CrossRef] [PubMed]
- Hwang, E.T.; Lee, S. Multienzymatic Cascade Reactions via Enzyme Complex by Immobilization. ACS Catal. 2019, 9, 4402–4425. [Google Scholar] [CrossRef]
- Velasco-Lozano, S.; Benítez-Mateos, A.I.; López-Gallego, F. Co-immobilized Phosphorylated Cofactors and Enzymes as Self-Sufficient Heterogeneous Biocatalysts for Chemical Processes. Angew. Chem. Int. Ed. 2017, 56, 771–775. [Google Scholar] [CrossRef] [PubMed]
- Wu, M.; He, Q.; Shao, Q.; Zuo, Y.; Wang, F.; Ni, H. Preparation and Characterization of Monodispersed Microfloccules of TiO2 Nanoparticles with Immobilized Multienzymes. ACS Appl. Mater. Interfaces 2011, 3, 3300–3307. [Google Scholar] [CrossRef]
- Cao, L.; Van Rantwijk, F.; Sheldon, R.A. Cross-Linked Enzyme Aggregates: A Simple and Effective Method for the Immobilization of Penicillin Acylase. Org. Lett. 2000, 2, 1361–1364. [Google Scholar] [CrossRef]
- Sheldon, R.A. Cross-linked enzyme aggregates (CLEA®s): Stable and recyclable biocatalysts. Biochem. Soc. Trans. 2007, 35, 1583–1587. [Google Scholar] [CrossRef] [Green Version]
- Sheldon, R.A. Cross-Linked Enzyme Aggregates as Industrial Biocatalysts. Org. Process Res. Dev. 2011, 15, 213–223. [Google Scholar] [CrossRef]
- Sheldon, R.A. Characteristic features and biotechnological applications of cross-linked enzyme aggregates (CLEAs). Appl. Microbiol. Biotechnol. 2011, 92, 467–477. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Yamaguchi, H.; Kiyota, Y.; Miyazaki, M. Techniques for Preparation of Cross-Linked Enzyme Aggregates and Their Applications in Bioconversions. Catalysts 2018, 8, 174. [Google Scholar] [CrossRef] [Green Version]
- Mateo, C.; Van Langen, L.M.; Van Rantwijk, F.; Sheldon, R.A. A new, mild cross-linking methodology to prepare cross-linked enzyme aggregates. Biotechnol. Bioeng. 2004, 86, 273–276. [Google Scholar] [CrossRef] [PubMed]
- Vaidya, B.K.; Kuwar, S.S.; Golegaonkar, S.B.; Nene, S.N. Preparation of cross-linked enzyme aggregates of l-aminoacylase via co-aggregation with polyethyleneimine. J. Mol. Catal. B Enzym. 2012, 74, 184–191. [Google Scholar] [CrossRef]
- Perez, D.I.; Van Rantwijk, F.; Sheldon, R.A. Cross-Linked Enzyme Aggregates of Chloroperoxidase: Synthesis, Optimization and Characterization. Adv. Synth. Catal. 2009, 351, 2133–2139. [Google Scholar] [CrossRef]
- Sheldon, R.A. Industrial Applications of Asymmetric Synthesis using Cross-Linked Enzyme Aggregates. In Comprehensive Chirality, 1st ed.; Carreira, E.M., Yamamoto, H., Eds.; Elsevier: Amsterdam, The Netherlands, 2012; Volume 9, pp. 353–366. [Google Scholar]
- Mahmod, S.S.; Yusof, F.; Jami, M.S.; Khanahmadi, S. Optimizing the preparation conditions and characterization of a stable and recyclable cross-linked enzyme aggregate (CLEA)-protease. Bioresour. Bioprocess. 2016, 3, 3. [Google Scholar] [CrossRef] [Green Version]
- Sheldon, R.A. CLEAs, Combi-CLEAs and ‘Smart’ Magnetic CLEAs: Biocatalysis in a Bio-Based Economy. Catalysts 2019, 9, 261. [Google Scholar] [CrossRef] [Green Version]
- Ahumada, K.; Martínez-Gil, A.; Moreno-Simunovic, Y.; Illanes, A.; Wilson, L. Aroma Release in Wine Using Co-Immobilized Enzyme Aggregates. Molecules 2016, 21, 1485. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Bernal, C.; Rodríguez, K.; Martínez, R. Integrating enzyme immobilization and protein engineering: An alternative path for the development of novel and improved industrial biocatalysts. Biotechnol. Adv. 2018, 36, 1470–1480. [Google Scholar] [CrossRef] [PubMed]
- Singh, R.K.; Tiwari, M.K.; Singh, R.; Lee, J.-K. From Protein Engineering to Immobilization: Promising Strategies for the Upgrade of Industrial Enzymes. Int. J. Mol. Sci. 2013, 14, 1232–1277. [Google Scholar] [CrossRef] [PubMed]
- Redeker, E.S.; Ta, D.T.; Cortens, D.; Billen, B.; Guedens, W.; Adriaensens, P. Protein Engineering For Directed Immobilization. Bioconj. Chem. 2013, 24, 1761–1777. [Google Scholar] [CrossRef]
- Bilal, M.; Iqbal, H.M.; Guo, S.; Hu, H.; Wang, W.; Zhang, X. State-of-the-art protein engineering approaches using biological macromolecules: A review from immobilization to implementation view point. Int. J. Biol. Macromol. 2018, 108, 893–901. [Google Scholar] [CrossRef]
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Federsel, H.-J.; Moody, T.S.; Taylor, S.J.C. Recent Trends in Enzyme Immobilization—Concepts for Expanding the Biocatalysis Toolbox. Molecules 2021, 26, 2822. https://doi.org/10.3390/molecules26092822
Federsel H-J, Moody TS, Taylor SJC. Recent Trends in Enzyme Immobilization—Concepts for Expanding the Biocatalysis Toolbox. Molecules. 2021; 26(9):2822. https://doi.org/10.3390/molecules26092822
Chicago/Turabian StyleFedersel, Hans-Jürgen, Thomas S. Moody, and Steve J.C. Taylor. 2021. "Recent Trends in Enzyme Immobilization—Concepts for Expanding the Biocatalysis Toolbox" Molecules 26, no. 9: 2822. https://doi.org/10.3390/molecules26092822