Characterization of TEMPO-oxidized chitin nanofibers with various oxidation times and its application as an enzyme immobilization support

Rui Chen; Wen-Can Huang; Wei Wang; Xiangzhao Mao

doi:10.1007/s42995-020-00054-y

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Characterization of TEMPO-oxidized chitin nanofibers with various oxidation times and its application as an enzyme immobilization support

1 College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China
2 Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China

Corresponding author: Xiangzhao Mao, xzhmao@ouc.edu.cn
Received Date: 2019-12-13

Fund Project:
This work was supported by the National Key Research and Development Program of China (no. 2019YFD0901902), National Natural Science Foundation of China (31922072), China Agriculture Research System (CARS-48), Taishan Scholar Project of Shandong Province (tsqn201812020).

Abstract

Chitin nanofibers have recently received increased attention and are considered to be a promising material for a wide range of applications because of their excellent characteristics. In this study, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized chitin nanofibers (CNFs) with various oxidation times were prepared and characterized. CNFs with different oxidation times were then utilized for enzyme immobilization, using chymotrypsin as a model enzyme. The effects of oxidation time on enzyme immobilization were explored. Results showed characteristics of chitin nanofibers can be controlled by adjusting oxidation time. CNFs treated with TEMPO for 360 min showed the lowest crystallinity (79.13 ± 1.43%), the shortest length (241.70 ± 74.61 nm), the largest width (12.67 ± 3.43 nm), and the highest transmittance (73.01% at 800 nm). The activity of immobilized enzymes and enzyme loading showed good correlation to the carboxylate content of CNFs. The enzyme eiciency based on CNFs and the content of carboxylate groups peaked at the oxidization time of 60 min. When the additional amount of chymotrypsins (CTs) was 500 or 2000 mg/g carrier, the highest loading amount of CTs was 307.17 ± 4.08 or 726.82 ± 12.05 mg/g carrier, respectively.
- Chitin,
- Nanofiber,
- Chymotrypsin,
- Immobilization

References

Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

Deng Y, Deng C, Qi D, Liu C, Liu J, Zhang X, Zhao D (2009) Synthesis of core/shell colloidal magnetic zeolite microspheres for the immobilization of trypsin. Adv Mater 21:1377–1382

Fan Y, Saito T, Isogai A (2007) Chitin nanocrystals prepared by TEMPO-mediated oxidation of α-chitin. Biomacromol 9:192–198

Han H, Zhou Y, Li S, Wang Y, Kong XZ (2016) Immobilization of lipase from Pseudomonas luorescens on porous polyurea and its application in kinetic resolution of racemic 1-phenylethanol. ACS Appl Mater Interfaces 8:25714–25724

Hirano S (1999) Chitin and chitosan as novel biotechnological materials. Polym Int 48:732–734

Huang WC, Wang W, Xue C, Mao X (2018) Efective enzyme immobilization onto a magnetic chitin nanoiber composite. ACS Sustain Chem Eng 6:8118–8124

Ifuku S, Nogi M, Abe K, Yoshioka M, Morimoto M, Saimoto H, Yano H (2009) Preparation of chitin nanofibers with a uniform width as α-chitin from crab shells. Biomacromol 10:1584–1588

Ifuku S, Saimoto H (2012) Chitin nanofibers: preparations, modiications, and applications. Nanoscale 4:3308–3318

Iyer PV, Ananthanarayan L (2008) Enzyme stability and stabilization—aqueous and non-aqueous environment. Process Biochem 43:1019–1032

Jayakumar R, Prabaharan M, Nair SV, Tamura H (2010) Novel chitin and chitosan nanofibers in biomedical applications. Biotechnol Adv 28:142–150

Jia H, Zhu G, Vugrinovich B, Kataphinan W, Reneker DH, Wang P (2002) Enzyme-carrying polymeric nanofibers prepared via electrospinning for use as unique biocatalysts. Biotechnol Prog 18:1027–1032

Jiang J, Ye W, Yu J, Fan Y, Ono Y, Saito T, Isogai A (2018) Chitin nanocrystals prepared by oxidation of α-chitin using the O₂/laccase/TEMPO system. Carbohydr Polym 189:178–183

Kato Y, Kaminaga J, Matsuo R, Isogai A (2004) TEMPO-mediated oxidation of chitin, regenerated chitin and N-acetylated chitosan. Carbohydr Polym 58:421–426

Kawamura Y, Nakanishi K, Matsuno R, Kamikubo T (1981) Stability of immobilized α-chymotrypsin. Biotechnol Bioeng 23:1219–1236

Khoshnevisan K, Bordbar AK, Zare D, Davoodi D, Noruzi M, Barkhi M, Tabatabaei M (2011) Immobilization of cellulase enzyme on superparamagnetic nanoparticles and determination of its activity and stability. Chem Eng J 171:669–673

Kumar MNR (2000) A review of chitin and chitosan applications.React Funct Polym 46:1–27

Kurita K (2006) Chitin and chitosan: functional biopolymers from marine crustaceans. Mar Biotechnol 8:203

Mateo C, Palomo JM, Fernandez-Lorente G, Guisan JM, FernandezLafuente R (2007) Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme Microb Technol 40:1451–1463

Mohapatra S, Pramanik N, Mukherjee S, Ghosh SK, Pramanik P (2007) A simple synthesis of amine-derivatised superparamagnetic iron oxide nanoparticles for bioapplications. J Mater Sci 42:7566–7574

Mushi NE, Utsel S, Berglund LA (2014) Nanostructured biocomposite ilms of high toughness based on native chitin nanofibers and chitosan. Front Chem 2:99

Muzzarelli RAA, Muzzarelli C, Cosani A, Terbojevich M (1999) 6-Oxychitins, novel hyaluronan-like regiospeciically carboxylated chitins. Carbohydr Polym 39:361–367

Pan C, Hu B, Li W, Sun Y, Ye H, Zeng X (2009) Novel and eicient method for immobilization and stabilization of β-D-galactosidase by covalent attachment onto magnetic Fe₃O₄-chitosan nanoparticles. J Mol Catal B Enzym 61:208–215

Rinaudo M (2006) Chitin and chitosan: properties and applications.Prog Polym Sci 31:603–632

Saito T, Isogai A (2004) TEMPO-mediated oxidation of native cellulose. The effect of oxidation conditions on chemical and crystal structures of the water-insoluble fractions. Biomacromol 5:1983–1989

Shahidi F, Arachchi JKV, Jeon YJ (1999) Food applications of chitin and chitosans. Trends Food Sci Technol 10:37–51

Sheldon RA (2007) Enzyme immobilization: the quest for optimum performance. Adv Synth Catal 349:1289–1307

Shigemasa Y, Matsuura H, Sashiwa H, Saimoto H (1996) Evaluation of different absorbance ratios from infrared spectroscopy for analyzing the degree of deacetylation in chitin. Int J Biol Macromol 18:237–242

Tamura H, Furuike T, Nair SV, Jayakumar R (2011) Biomedical applications of chitin hydrogel membranes and scafolds. Carbohydr Polym 84:820–824

Tamura H, Nagahama H, Tokura S (2006) Preparation of chitin hydrogel under mild conditions. Cellulose 13:357–364

Wang Q, Yan X, Chang Y, Ren L, Zhou J (2018a) Fabrication and characterization of chitin nanofibers through esteriication and ultrasound treatment. Carbohydr Polym 180:81–87

Wang W, Guo N, Huang WC, Zhang Z, Mao X (2018b) Immobilization of chitosanases onto magnetic nanoparticles to enhance enzyme performance. Catalysts 8:401

Wang Y, Caruso F (2005) Mesoporous silica spheres as supports for enzyme immobilization and encapsulation. Chem Mater 17:953–961

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

Xiao A, Xiao Q, Lin Y, Ni H, Zhu Y, Cai H (2017) Eicient immobilization of agarase using carboxyl-functionalized magnetic nanoparticles as support. Electron J Biotechnol 25:13–20

Zhang YT, Zhi TT, Zhang L, Huang H, Chen HL (2009) Immobilization of carbonic anhydrase by embedding and covalent coupling into nanocomposite hydrogel containing hydrotalcite. Polymer 50:5693–5700

Zhang Y, Xue C, Xue Y, Gao R, Zhang X (2005) Determination of the degree of deacetylation of chitin and chitosan by X-ray powder difraction. Carbohydr Res 340:1914–1917

Zheng H, Zhou J, Du Y, Zhang L (2002) Cellulose/chitin ilms blended in NaOH/urea aqueous solution. J Appl Polym Sci 86:1679–1683

Zhong C, Cooper A, Kapetanovic A, Fang Z, Zhang M, Rolandi M (2010) A facile bottom-up route to self-assembled biogenic chitin nanofibers. Soft Matter 6:5298–5301

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Characterization of TEMPO-oxidized chitin nanofibers with various oxidation times and its application as an enzyme immobilization support

Corresponding author: Xiangzhao Mao, xzhmao@ouc.edu.cn

1 College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China

2 Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao 266200, China

Received Date: 2019-12-13

Fund Project: This work was supported by the National Key Research and Development Program of China (no. 2019YFD0901902), National Natural Science Foundation of China (31922072), China Agriculture Research System (CARS-48), Taishan Scholar Project of Shandong Province (tsqn201812020).

Abstract: Chitin nanofibers have recently received increased attention and are considered to be a promising material for a wide range of applications because of their excellent characteristics. In this study, 2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPO)-oxidized chitin nanofibers (CNFs) with various oxidation times were prepared and characterized. CNFs with different oxidation times were then utilized for enzyme immobilization, using chymotrypsin as a model enzyme. The effects of oxidation time on enzyme immobilization were explored. Results showed characteristics of chitin nanofibers can be controlled by adjusting oxidation time. CNFs treated with TEMPO for 360 min showed the lowest crystallinity (79.13 ± 1.43%), the shortest length (241.70 ± 74.61 nm), the largest width (12.67 ± 3.43 nm), and the highest transmittance (73.01% at 800 nm). The activity of immobilized enzymes and enzyme loading showed good correlation to the carboxylate content of CNFs. The enzyme eiciency based on CNFs and the content of carboxylate groups peaked at the oxidization time of 60 min. When the additional amount of chymotrypsins (CTs) was 500 or 2000 mg/g carrier, the highest loading amount of CTs was 307.17 ± 4.08 or 726.82 ± 12.05 mg/g carrier, respectively.

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