Abstract
In this study, regenerated cellulose-based macrogels with abundant carboxyl groups and interconnected microporous structures were synthesized from cellulose fibers (CFs)/cellulose nanofibers (CNFs) and utilized as stable, efficient, and recyclable platforms for enzyme immobilization. CNFs with different carboxyl contents were prepared by a TEMPO (2, 2, 6, 6 -tetramethylpiperidine-1-oxyl radical)-medium oxidation system. The cellulose-based macrogels were prepared by dissolving CFs and CNFs in a NaOH/urea aqueous solution and a followed extrusion process. The carboxyl groups within the macrogels were then activated by an N-(3-Dimethylaminopropyl)-N’-ethylcarbodiimide hydrochloride (EDC·HCl)/Nhydroxysuccinimide (NHS) system, which provides these macrogels with efficient binding sites for enzyme immobilization. The three-dimensional network structures of the macrogels facilitate the enzymes to infiltrate into the core of the macrogels and be fixed on the abundant pore walls. The immobilized enzymes maintain activity at a higher pH than free enzymes. Moreover, the immobilized enzymes have higher temperature tolerance, and their optimal reaction temperature is about 10 °C higher than that of free enzymes. The immobilized enzymes retain 24% of the initial activity after repeated use for 7 times. The microporous regenerated cellulose-based macrogels show potential in the fields of enzyme immobilization and biocatalysis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abouhmad A, Dishisha T, Amin MA, Hatti-Kaul R (2017) Immobilization to positively charged cellulose nanocrystals enhances the antibacterial activity and stability of hen egg white and T4 lysozyme. Biomacromol 18:1600–1608. https://doi.org/10.1016/S0141-0229(98)00084-2
Arcot LR, Lundahl M, Rojas OJ, Laine J (2014) Asymmetric cellulose nanocrystals: thiolation of reducing end groups via NHS–EDC coupling. Cellulose 21:4209–4218. https://doi.org/10.1007/s10570-014-0426-9
Arola S, Tammelin T, Setala H, Tullila A, Linder MB (2012) Immobilization-stabilization of proteins on nanofibrillated cellulose derivatives and their bioactive film formation. Biomacromol 13:594–603. https://doi.org/10.1021/bm201676q
Chen H, Hsieh YL (2005) Enzyme immobilization on ultrafine cellulose fibers via poly(acrylic acid) electrolyte grafts. Biotechnol Bioeng 90:405–413. https://doi.org/10.1002/bit.20324
Chi MC, Lyu RC, Lin LL, Huang HB (2008) Characterization of Bacillus kaustophilus leucine aminopeptidase immobilized in Ca-alginate/k-carrageenan beads. Biochem Eng J 39:376–382. https://doi.org/10.1016/j.bej.2007.10.008
Erramuspe VIB, Fazeli E, Nareoja T, Trygg J, Hanninen P, Heinze T, Fardim P (2016) Advanced cellulose fibers for efficient immobilization of enzymes. Biomacromol 17(10):3188–3197. https://doi.org/10.1021/acs.biomac.6b00865
Franssen MCR, Steunenberg P, Scott EL, Zuilhof H, Sanders JP (2013) Immobilised enzymes in biorenewables production. Chem Soc Rev 42:6491–6533. https://doi.org/10.1039/C3CS00004D
Gupta PK, Uniyal V, Naithani S (2013) Polymorphic transformation of cellulose I to cellulose II by alkali pretreatment and urea as an additive. Carbohydr Polym 94:843–849. https://doi.org/10.1016/j.carbpol.2013.02.012
Hartmann M, Kostrov X (2013) Immobilization of enzymes on porous silicas-benefits and challenges. Chem Soc Rev 42:6277–6289. https://doi.org/10.1039/C3CS60021A
Huang XJ, Chen PC, Huang F, Ou Y, Chen MR, Xu ZK (2011) Immobilization of Candida rugosa lipase on electrospun cellulose nanofiber membrane. J Mol Catal B-Enzym 70:95–100. https://doi.org/10.1016/j.molcatb.2011.02.010
Incani V, Danumah C, Boluk Y (2013) Nanocomposites of nanocrystalline cellulose for enzyme immobilization. Cellulose 20:191–200. https://doi.org/10.1007/s10570-012-9805-2
Je HH, Noh S, Hong SG, Ju Y, Kim J, Hwang DS (2017) Cellulose nanofibers for magnetically-separable and highly loaded enzyme immobilization. Chem Eng J 323:425–433. https://doi.org/10.1016/j.cej.2017.04.110
Laurell T, Drott J, Rosengren L (1995) Silicon wafer intefrated enzyme reactors. Biosens Bioelectron 10:289–299. https://doi.org/10.1016/0956-5663(95)96848-S
Liu EY, Jung S, Yi H (2016) Improved protein conjugation with uniform, Macroporous Poly(acrylamide-co-acrylic acid) hydrogel microspheres via EDC/NHS chemistry. Langmuir 32:11043–11054. https://doi.org/10.1021/acs.langmuir.6b02591
Liu Z, Wang HS, Li B, Liu C, Jiang YJ, Yu G, Mu XD (2012) Biocompatible magnetic cellulose–chitosan hybrid gel microspheres reconstituted from ionic liquids for enzyme immobilization. J Mater Sci 22:15085–15091. https://doi.org/10.1039/C2JM33033D
Luan Q, Zhou WJ, Zhang H, Bao YP, Zheng MM, Shi J, Tang H, Huang FH (2018) Cellulose-based composite macrogels from cellulose fiber and cellulose nanofiber as intestine delivery vehicles for probiotics. J Agric Food Chem 66:339–345. https://doi.org/10.1021/acs.jafc.7b04754
Luo XG, Lei XJ, Cai N, Xie XP, Xue YN, Yu FQ (2016) Removal of heavy metal ions from water by magnetic cellulose-based beads with embedded chemically modified magnetite nanoparticles and activated carbon. ACS Sustain Chem Eng 4:3960–3969. https://doi.org/10.1021/acssuschemeng.6b00790
Mohamad NR, Marzuki NHC, Buang NA, Huyop F, Wahab RA (2015) An overview of technologies for immobilization of enzymes and surface analysis techniques for immobilized enzymes. Biotechnol Biotec Eq 29:205–220. https://doi.org/10.1080/13102818.2015.1008192
Patel SK, Choi SH, Kang YC, Lee JK (2017) Eco-friendly composite of Fe3O4-reduced graphene oxide particles for efficient enzyme immobilization. ACS Appl Mater Inter 9:2213–2222. https://doi.org/10.1021/acsami.6b05165
Saito T, Isogai A (2004) TEMPO-mediated oxidation of native cellulose the effect of oxidation conditions on chemical and crystal structure of the water insoluble fractions. Biomacromol 5:1983–1989. https://doi.org/10.1021/bm0497769
Saito T, Kimura S, Nishiyama Y, Isogai A (2007) Cellulose nanofibers prepared by TEMPO-mediated oxidation of native cellulose. Biomacromol 8:2485–2491. https://doi.org/10.1021/bm0703970
Saito T, Hirota M, Tamura N, Isogai A (2010) Oxidation of bleached wood pulp by TEMPO/NaClO/NaClO2 system: effect of the oxidation conditions on carboxylate content and degree of polymerization. J Wood Sci 56:227–232. https://doi.org/10.1007/s10086-009-1092-7
Sathishkumar P, Kamala-Kannan S, Cho M, Kim JS, Hadibarata T, Salim MR, Oh BT (2014) Laccase immobilization on cellulose nanofiber: The catalytic efficiency and recyclic application for simulated dye effluent treatment. J Mol Catal B-Enzym 100:111–120. https://doi.org/10.1016/j.molcatb.2013.12.008
Sheelu G, Kavitha G, Fadnavis NW (2008) Efficient immobilization of lecitase in gelatin hydrogel and degumming of rice bran oil using a spinning basket reactor. J Am Oil Chem Soc 85:739–748. https://doi.org/10.1007/s11746-008-1261-7
Subramanian A, Kennel SJ, Oden PI, Jacobson KB, Woodward J, Doktycz MJ (1999) Comparison of techniques for enzyme immobilization on silicon supports. Enzyme Microb Tech 4:26–34. https://doi.org/10.1016/S0141-0229(98)00084-2
Sulaiman S, Mokhtar MN, Naim MN, Baharuddin AS, Sulaiman A (2015) A review: potential usage of cellulose nanofibers (CNF) for enzyme immobilization via covalent interactions. Appl Biochem Biotechnol 175:1817–1842. https://doi.org/10.1007/s12010-014-1417-x
Sun Q, Aguila B, Lan PC, Ma S (2019) Tuning Pore heterogeneity in covalent organic frameworks for enhanced enzyme accessibility and resistance against denaturants. Adv Mater 31:e1900008. https://doi.org/10.1002/adma.201900008
Takashi N, Ikuyo M, Koichi H (2004) All-cellulose composite. Macromolecules 37:7683–7687
Tang H, Chen H, Duan B, Lu A, Zhang L (2013) Swelling behaviors of superabsorbent chitin/carboxymethylcellulose hydrogels. J Mater Sci 49:2235–2242. https://doi.org/10.1007/s10853-013-7918-0
Wang C, Yan Q, Liu HB, Zhou XH, Xiao SJ (2011) Different EDC/NHS activation mechanisms between PAA and PMAA brushes and the following amidation reactions. Langmuir 27:12058–12068. https://doi.org/10.1021/la202267p
Wang Y, Zhao M, Song K, Wang L, Tang S, Riley WW (2010) Partial hydrolysis of soybean oil by phospholipase A1 (Lecitase Ultra). Food Chem 121:1066–1072. https://doi.org/10.1016/j.foodchem.2010.01.051
Wang YJ, Jian X, Luo GS, Dai YY (2008) Immobilization of lipase by ultrafiltration and cross-linking onto the polysulfone membrane surface. Bioresource Technol 99:2299–2303. https://doi.org/10.1016/j.biortech.2007.05.014
Wang ZG, Wan LS, Liu ZM, Huang XJ, Xu ZK (2009) Enzyme immobilization on electrospun polymer nanofibers: an overview. J Mol Catal B- Enzym 56:189–195. https://doi.org/10.1016/j.molcatb.2008.05.005
Weishaupt R, Siqueira G, Schubert M, Tingaut P, Maniura-Weber K, Zimmermann T, Thony-Meyer L, Faccio G, Ihssen J (2015) TEMPO-oxidized nanofibrillated cellulose as a high density carrier for bioactive molecules. Biomacromol 16:3640–3650. https://doi.org/10.1021/acs.biomac.5b01100
Yang D, Zhong LX, Yuan TQ, Peng XW, Sun RC (2013) Studies on the structural characterization of lignin, hemicelluloses and cellulose fractionated by ionic liquid followed by alkaline extraction from bamboo. Ind Crop Prod 43:141–149. https://doi.org/10.1016/j.indcrop.2012.07.024
Ye P, Xu ZK, Che AF, Wu J, Seta P (2005) Chitosan-tethered poly(acrylonitrile-co-maleic acid) hollow fiber membrane for lipase immobilization. Biomaterials 26:6394–6403. https://doi.org/10.1016/j.biomaterials.2005.04.019
Yu D, Jiang L, Li Z, Shi J, Xue J, Kakuda Y (2011) Immobilization of Phospholipase A1 and its application in soybean oil degumming. J Am Oil Chem Soc 89:649–656. https://doi.org/10.1007/s11746-011-1943-4
Yu DY, Ma Y, Xue SJ, Jiang LZ, Shi J (2013) Characterization of immobilized phospholipase A1 on magnetic nanoparticles for oil degumming application. LWT - Food Sci Technol 50:519–525. https://doi.org/10.1016/j.lwt.2012.08.014
Zhan JF, Jiang ST, Pan LJ (2013) Immobilization of Phospholipase A1 Using a Polyvinyl Alcohol-Alginate matrix and evaluation of the effects of immobilization. Braz J Chem Eng 30:721–728. https://doi.org/10.1590/S0104-66322013000400004
Zhang H, Yang C, Zhou WJ, Luan Q, Li WL, Deng QC, Dong XY, Tang H, Huang FH (2018) A pH-responsive gel macrosphere based on sodium alginate and cellulose nanofiber for potential intestinal delivery of probiotics. ACS Sustain Chem Eng 6:13924–13931. https://doi.org/10.1021/acssuschemeng.8b02237
Zhou Q, Bao YP, Zhang H, Luan Q, Tang H, Li XT (2019) Regenerated cellulose-based composite membranes as adsorbent for protein adsorption. Cellulose 27:335–345. https://doi.org/10.1007/s10570-019-02761-x
Acknowledgments
This work was supported by the Open Project from Beijing Advanced Innovation Center for Food Nutrition and Human Health, Beijing Technology and Business University (BTBU) (No.20182013), National Natural Science Foundation of China (31701560).
Author information
Authors and Affiliations
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval
This article does not contain any studies with human participants or animals performed by any of the authors.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Luan, Q., Zhang, H., Lei, Y. et al. Microporous regenerated cellulose-based macrogels for covalent immobilization of enzymes. Cellulose 28, 5735–5744 (2021). https://doi.org/10.1007/s10570-021-03887-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10570-021-03887-7