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Review
. 2022 Mar 28;7(14):11530-11543.
doi: 10.1021/acsomega.2c00357. eCollection 2022 Apr 12.

Emerging 3D Printing Strategies for Enzyme Immobilization: Materials, Methods, and Applications

Yun Shao  1 , Zhijun Liao  1 , Bingbing Gao  1 , Bingfang He  1
Affiliations
Review

Emerging 3D Printing Strategies for Enzyme Immobilization: Materials, Methods, and Applications

Yun Shao et al. ACS Omega. .

Abstract

As the strategies of enzyme immobilization possess attractive advantages that contribute to realizing recovery or reuse of enzymes and improving their stability, they have become one of the most desirable techniques in industrial catalysis, biosensing, and biomedicine. Among them, 3D printing is the emerging and most potential enzyme immobilization strategy. The main advantages of 3D printing strategies for enzyme immobilization are that they can directly produce complex channel structures at low cost, and the printed scaffolds with immobilized enzymes can be completely modified just by changing the original design graphics. In this review, a comprehensive set of developments in the fields of 3D printing techniques, materials, and strategies for enzyme immobilization and the potential applications in industry and biomedicine are summarized. In addition, we put forward some challenges and possible solutions for the development of this field and some possible development directions in the future.

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Conflict of interest statement

The authors declare no competing financial interest.

Figures

Figure 1
Figure 1
(A) Arrangement of two micromixers in series with carbonic anhydrase (CA) and formate dehydrogenase (FDH) enzymes immobilized in separate micromixers (domino immobilization) of SLA 3D printing for the enzymatic cascade reduction of CO2. Reprinted with permission from ref (11c) Copyright 2021 Elsevier. (B) Digital images of FDM 3D-printed scaffolds for enzymes immobilization. Reprinted from ref (12b). Copyright 2019 American Chemical Society. (C) Photograph of the FDM printed reactor. Reprinted from ref (12c). Copyright 2016 American Chemical Society. (D) Photograph of two FDM printed enzyme/substrate-incorporated 48-well plates (side-by-side/layer-by-layer design) glued onto a transparent polystyrene microplate, with glucose oxidase immobilized near the center of each well. Reprinted with permission from ref (12d). Copyright 2018 Elsevier. (E) Photograph of an extrusion-based printer: Cure-on-dispense setup used in combination with a Gesim BioScaffolder 3.1. Reprinted with permission from ref (13b). Copyright 2020 Frontiers Media S.A. (F) Side view of 3D-printed lattice structures. Reprinted with permission from ref (13a). Copyright 2021 Institute of Physics Publishing. (G) The image of three-dimensional printed grid pattern using preheated gelatin 10 represents proper gelation. Reprinted with permission from ref (13c). Copyright 2020 Whioce Publishing Pte. Ltd.
Figure 2
Figure 2
Various 3D printing materials for enzyme immobilization. (A) SEM image of ZIF-8/carbonic anhydrase (CA) and formate dehydrogenase (FDH) in situ thin film. Reprinted with permission from ref (11c). Copyright 2021 Elsevier. (B) Image of in situ metal–organic framework growth and encapsulation process. Reprinted with permission from ref (15c). Copyright 2020 Elsevier. (C) Digital images of three types of 3D-printed polylactic acid scaffolds with adjustable aperture structures: cube (I), sphere (II), and semicircle (III). Reprinted with permission from ref (16d). Copyright 2021 Elsevier. (D) Agarose hydrogel scaffolds of different shapes and sizes can be printed. Reprinted with permission from ref (17b). Copyright 2018 John Wiley and Sons Ltd. (E) Picture of the 3D bioprinted constructs with the ratio of acrylamide: hydroxyapatite/sodium alginate = 4:1.2:1. Reprinted with permission from ref (17d). Copyright 2020 Elsevier. (F) 3D printing of the graphene-polylactic acid electrode. Reprinted with permission from ref (16a). Copyright 2020 Elsevier. (G) SEM micrographs of the carbon black-polylactic acid electrode surface after electrolysis in 1 M NaOH (−1.4 to +1.2 V vs the Ag|AgCl polarization range). Reprinted with permission from ref (16b). Copyright 2022 Elsevier. (H) Hydrogel structure is inserted in the reactor housing with connection to the fluidic system. Reprinted with permission from ref (17a). Copyright 2018 Frontiers Media S.A. (I) Top view image of the printed hydrogels using the additives Deuteron XG. Reprinted with permission from ref (17c). Copyright 2018 John Wiley and Sons Ltd.
Figure 3
Figure 3
(A) Production of immobilized glucose oxidase/catalase by 3D bioprinted hybrid interpenetrating polymer network hydrogel. Reprinted with permission from ref (18c). Copyright 2019 Elsevier. (B) Picture of 3D-printed lattices for the physical entrapment of enzymes. Reprinted with permission from ref (18b). Copyright 2020 Frontiers Media S.A. (C) Manufacturing process of agarose-based, compartmentalized biocatalytic flow reactors. Reprinted with permission from ref (18a). Copyright 2019 John Wiley and Sons Ltd. (D) 3D jet writing of hydrogel fibers allows yielding precisely oriented hydrogel fibers loaded with enzymes. Reprinted with permission from ref (18d). Copyright 2020 John Wiley and Sons Ltd.
Figure 4
Figure 4
(A) Strategy for acetyltransferase p300/CBP associated factor (PCAF) and peptidylarginine deiminase type 1 (PAD) covalent immobilization. Reprinted with permission from ref (19a). Copyright 2018 John Wiley and Sons Ltd. (B) 3D-printed nylon part undergoes several modifications to covalently immobilize enzymes. Reprinted with permission from ref (19c). Copyright 2017 Royal Society of Chemistry. (C) Schematic of the silver nanoparticles coated on the polypropylene/polydopamine/polyethylenimine membrane used for covalent immobilization of the glucose oxidase and horseradish peroxidase enzymes. Reprinted with permission from ref (19b). Copyright 2020 John Wiley and Sons Ltd. (D) Sequential stages for covalent immobilization of Candida rugosa lipase on lattice geopolymers. Reprinted with permission from ref (19d). Copyright 2021 Elsevier.
Figure 5
Figure 5
(A) Experimental setup of pencil graphite electrodes based on 3D printed enzymatic biofuel cell. Reprinted with permission from ref (20b). Copyright 2019 Elsevier Ltd. (B) Final structure optimized 3D printed bioelectrodes of glucose oxidase or laccase immobilization. Reprinted with permission from ref (20c). Copyright 2020 Institute of Electrical and Electronics Engineers Inc. (C) Preparation routes of 3D-printed xylanase which is applied to digest the corn cob reaction system enzymatically. Reprinted with permission from ref (21c). Copyright 2020 Elsevier. (D) Preparation routes of 3D-printed aldo-keto reductases-IA. Reprinted with permission from ref (21d). Copyright 2022 Elsevier. (E) Image of 3D-printed labware. Reprinted from ref (21a). Copyright 2020 American Chemical Society. (F) Photograph of the polycaprolactone-modified paper immobilized α-glucosidase after cutting. Reprinted with permission from ref (21b). Copyright 2019 BioMed Central Ltd.
Figure 6
Figure 6
(A) Representation of the 3D-printed glucose oxidase biosensor. Reprinted with permission from ref (22b). Copyright 2020 Elsevier. (B) Schematic illustration of the preparation process for the biosensor. Reprinted with permission from ref (22c). Copyright 2021 Nature Publishing Group. (C) Hemostatic photo images within 110 s: a negative control group without treatment (upper), the hemostatic effect of hybrid hydrogel (Hgel) (lower). Reprinted with permission from ref (22g). Copyright 2016 Royal Society of Chemistry. (D) Schematic illustration of 3D printed alginate/glucose oxidase/catalase-assisted biomineralized calcium phosphate nanosheets scaffolds. Reprinted with permission from ref (22f). Copyright 2021 Wiley-VCH Verlag.
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