In industrial production, the use of enzyme-catalyzed reactions to prepare biochemical products often faces many difficulties. Traditional immobilization technology showed the non-ideal immobilization effect, the activity of the enzyme may be greatly affected, and the final production efficiency is still not ideal. For these dilemmas, this research takes aldo-keto reductase as the object, and tries to use the fast-developing 3D printing technology to optimize the immobilization rate of enzyme, increase the activity of the immobilized enzyme, and open up its potential for industrial applications. In order to ensure that the enzyme activity after immobilization is not affected, single factor experiments and orthogonal experiments were set up, and the optimal conditions for 3D printing were determined through response surface analysis. In addition, the repeated use performance of the printed aldo-keto reductase was also studied, and the results showed that it still retains about 50% of the enzyme activity after six times, reflecting the huge industrial application potential. Finally, the immobilized enzyme was further characterized by SEM to explore the loading of aldo-keto reductase on the printing material. The results are of great significance for the immobilization of catalysts, the production of chiral alcohols, and the application of 3D printing.
Get full access to this article
View all access options for this article.
References
1.
PeiR, FanL, ZhaoF, et al.3D-Printed metal-organic frameworks within biocompatible polymers as excellent adsorbents for organic dyes removal. J Hazard Mater, 2020; 384:121418; doi: 10.1016/j.jhazmat.2019.121418
2.
BettermannS, StuhrR, MoritzHU, et al.Customizable 3D-printed stirrers for investigation, optimization and scale-up processes of batch emulsion copolymerizations. Chemical Engineering Science, 2019; 206:50–62; doi: 10.1016/j.ces.2019.05.026
3.
ShenX, YangM, CuiC, et al.In situ immobilization of glucose oxidase and catalase in a hybrid interpenetrating polymer network by 3D bioprinting and its application. Colloids & Surfaces A Physicochemical & Engineering Aspects, 2019; 568:411–418; doi: 10.1016/j.colsurfa.2019.02.021
4.
GuoL, LuL, YinM, et al.Valorization of refractory keratinous waste using a new and sustainable bio-catalysis. Chemical Engineering Journal, 2020; 397:125420; doi: 10.1016/j.cej.2020.125420
5.
HuangZX, CaoSL, XuP, et al.Preparation of a novel nanobiocatalyst by immobilizing penicillin acylase onto magnetic nanocrystalline cellulose and its use for efficient synthesis of cefaclor. Chemical Engineering Journal, 2018; 346:361–368; doi: 10.1016/j.cej.2018.04.026
6.
ChenK, ArnoldFH. Engineering new catalytic activities in enzymes. Nat Catal, 2020; 3(3):203–213; doi: 10.1038/s41929-019-0385-5
7.
EganPF. Integrated design approaches for 3D printed tissue scaffolds: Review and outlook. Materials (Basel), 2019; 12(15); doi: 10.3390/ma12152355
8.
YangC, ZhengZ, YounisMR, et al.3D printed enzyme functionalized scaffold facilitates diabetic bone regeneration. Adv Funct Materials, 2021; 31(20):2101372; doi: 10.1002/adfm.202101372
9.
MohanA, SandhyaraniN. A 3D printed low volume hybrid enzyme fuel cell for low power applications. Applied Surface Science, 2021; 555:149720; doi: 10.1016/j.apsusc.2021.149720
10.
MaierM, RadtkeCP, HubbuchJ, et al.On-demand production of flow-reactor cartridges by 3D printing of thermostable enzymes. Angew Chem Int Ed Engl, 2018; 57(19):5539–5543; doi: 10.1002/anie.201711072
11.
ThA, AnB, AsA, et al.Bile duct reconstruction using scaffold-free tubular constructs created by Bio-3D printer-ScienceDirect. Regenerative Therapy, 2021; 16:81–89; doi: 10.1016/j.reth.2021.02.001
12.
NarupaiB, SmithPT, NelsonA. 4D printing of multi stimuli responsive protein based hydrogels for autonomous shape transformations. Adv Funct Materials, 2021; 31(23):2011012; doi: 10.1002/adfm.202011012
13.
ValottaA, MaierMC, SoritzS, et al.3D printed ceramics as solid supports for enzyme immobilization: An automated DoE approach for applications in continuous flow. J Flow Chem, 2021; 11(3):675–689; doi: 10.1007/s41981-021-00163-4
14.
ShenS, ShenJ, ShenH, et al.Dual-enzyme crosslinking and post-polymerization for printing of polysaccharide-polymer hydrogel. Front Chem, 2020; 8:36; doi: 10.3389/fchem.2020.00036
15.
BlanchetteCD, KnipeJM, StolaroffJK, et al.Printable enzyme-embedded materials for methane to methanol conversion. Nat Commun, 2016; 7:11900; doi: 10.1038/ncomms11900
16.
PalmaraG, FrascellaF, RoppoloI, et al.Functional 3D printing: Approaches and bioapplications. Biosens Bioelectron, 2021; 175(80):112849; doi: 10.1016/j.bios.2020.112849
17.
HeWJ, ZhangL, YiSY, et al.An aldo-keto reductase is responsible for Fusarium toxin-degrading activity in a soil Sphingomonas strain. Sci Rep, 2017; 7(1):9549; doi: 10.1038/s41598-017-08799-w
ZhangW, ZhuT, LiH, et al.Key sites insight on the stereoselectivity of four mined aldo-keto reductases toward α-keto esters and halogen-substituted acetophenones. Appl Microbiol Biotechnol, 2019; 103(15):6119–6128; doi: 10.1007/s00253-019-09932-7
20.
LiuH, HoffBH, AnthonsenT. Chemo-enzymatic synthesis of the antidepressant duloxetine and its enantiomer. Chirality, 2000; 12(1):26–29.
21.
PeiR, WuW, ZhangY, et al.Characterization and catalytic-site-analysis of an Aldo-Keto reductase with excellent solvent tolerance. Catalysts, 2020; 10(10):1121; doi: 10.3390/catal10101121
22.
JampalaP, PreethiM, RamanujamS, et al.Immobilization of levan-xylanase nanohybrid on an alginate bead improves xylanase stability at wide pH and temperature. Int J Biol Macromol, 2017; 95:843–849; doi: 10.1016/j.ijbiomac.2016.12.012
23.
DaiXY, KongLM, WangXL, et al.Preparation, characterization and catalytic behavior of pectinase covalently immobilized onto sodium alginate/graphene oxide composite beads. Food Chem, 2018; 253:185–193; doi: 10.1016/j.foodchem.2018.01.157
24.
SuryawanshiRK, JanaUK, PrajapatiBP, et al.Immobilization of Aspergillus quadrilineatus RSNK-1 multi-enzymatic system for fruit juice treatment and mannooligosaccharide generation. Food Chem, 2019; 289(AUG.15):95–102; doi: 10.1016/j.foodchem.2019.03.035
25.
LiuS, HuY, ZhangJ, et al.Bioactive and biocompatible macroporous scaffolds with tunable performances prepared based on 3D printing of the pre-crosslinked sodium alginate/hydroxyapatite hydrogel ink. Macro Materials & Eng, 2019; 304(4):1800698; doi: 10.1002/mame.201800698
26.
FadnavisNW, SheeluG, KumarBM, et al.Gelatin Blends with Alginate: Gels for Lipase Immobilization and Purification. Biotechnol Prog, 2003; 19(2):557–564; doi: 10.1021/bp010172f
27.
NguyenLT, LauYS, YangKL. Entrapment of cross-linked cellulase colloids in alginate beads for hydrolysis of cellulose. Colloids Surf B Biointerfaces, 2016; 145:862–869; doi: 10.1016/j.colsurfb.2016.06.008
28.
KumarL, NagarS, MittalA, et al.Immobilization of xylanase purified from Bacillus pumilus VLK-1 and its application in enrichment of orange and grape juices. J Food Sci Technol, 2014; 51(9):1737–1749; doi: 10.1007/s13197-014-1268-z
29.
SahaK, VermaP, SikderJ, et al.Synthesis of chitosan-cellulase nanohybrid and immobilization on alginate beads for hydrolysis of ionic liquid pretreated sugarcane bagasse. Renewable Energy, 2019; 133:66–76; doi: 10.1016/j.renene.2018.10.014
30.
AhmedIN, ChangR, TsaiWB. Poly(acrylic acid) nanogel as a substrate for cellulase immobilization for hydrolysis of cellulose. Colloids Surf B Biointerfaces, 2017; 152(Complete):339–343; doi: 10.1016/j.colsurfb.2017.01.040
31.
LiC, YoshimotoM, FukunagaK, et al.Characterization and immobilization of liposome-bound cellulase for hydrolysis of insoluble cellulose. Bioresour Technol, 2007; 98(7):1366–1372; doi: 10.1016/j.biortech.2006.05.028
32.
WonK, KimS, KimKJ, et al.Optimization of lipase entrapment in Ca-alginate gel beads. Process Biochemistry, 2005; 40(6):2149–2154; doi: 10.1016/j.procbio.2004.08.014
33.
KnezevicZ, BobicS, MilutinovicA, et al.Alginate-immobilized lipase by electrostatic extrusion for the purpose of palm oil hydrolysis in lecithin/isooctane system. Process Biochemistry, 2002; 38(3):313–318; doi: 10.1016/S0032-9592(02)00085-7
34.
Shuang, ZhangWenting. Immobilization of lipase with alginate hydrogel beads and the lipase-catalyzed kinetic resolution of -phenyl ethanol. Journal of Applied Polymer Science, 2014; 131(8):40178; doi: 10.1016/S0142-9612(02)00095-9
35.
ZhouX, LiuCHANG‐JUN. Three-dimensional printing for catalytic applications: Current status and perspectives. Adv Funct Materials, 2017; 27(30):1701134; doi: 10.1002/adfm.201701134
36.
JiandongC, ZhaohuiC, ZhiguoS, et al.Bioactive coating prepared by bio-3D printing of castor oil-based waterborne polyurethane mixed with carbonic anhydrase. CIESC Journal, 2018; 69(8):3577–3584; doi: 10.11949/j.issn.0438-1157.20180054
