Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

doi:10.1016/j.cjche.2022.11.002

Chinese Journal of Chemical Engineering ›› 2023, Vol. 58 ›› Issue (6): 40-52.DOI: 10.1016/j.cjche.2022.11.002

Previous Articles Next Articles

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

Xiaohong Zhou¹, Wenfeng Zhou², Wei Zhuang², Chenjie Zhu², Hanjie Ying², Hongman Zhang¹

1. School of Chemistry and Molecular Engineering, Nanjing Tech University, Nanjing 211816, China;
2. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, National Engineering Technique Research Center for Biotechnology, Nanjing Tech University, Nanjing 211816, China

Received:2022-08-14 Revised:2022-11-01 Online:2023-08-31 Published:2023-06-28
Contact: Wei Zhuang,E-mail:weizhuang@njtech.edu.cn;Hongman Zhang,E-mail:hongmanzhang@njtech.edu.cn
Supported by:
This work was supported by grants from the National Key Research and Development Program of China (2021YFC2102805, 2019YFD1101204), the National Natural Science Foundation of China (21878142, 21776132), Key Research and Development Plan of Jiangsu Province (BE2020712), Key Research and Development Plan of Jiangsu Province (BE2019001), Jiangsu Natural Science Fund for Distinguished Young Scholars (BK20190035), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

Xiaohong Zhou¹, Wenfeng Zhou², Wei Zhuang², Chenjie Zhu², Hanjie Ying², Hongman Zhang¹

1. School of Chemistry and Molecular Engineering, Nanjing Tech University, Nanjing 211816, China;
2. State Key Laboratory of Materials-Oriented Chemical Engineering, College of Biotechnology and Pharmaceutical Engineering, National Engineering Technique Research Center for Biotechnology, Nanjing Tech University, Nanjing 211816, China

通讯作者: Wei Zhuang,E-mail:weizhuang@njtech.edu.cn;Hongman Zhang,E-mail:hongmanzhang@njtech.edu.cn
基金资助:
This work was supported by grants from the National Key Research and Development Program of China (2021YFC2102805, 2019YFD1101204), the National Natural Science Foundation of China (21878142, 21776132), Key Research and Development Plan of Jiangsu Province (BE2020712), Key Research and Development Plan of Jiangsu Province (BE2019001), Jiangsu Natural Science Fund for Distinguished Young Scholars (BK20190035), and the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD).

Abstract

Abstract: Cytidine 5'-monophosphate (5'-CMP) is an essential nucleotide for additives. In this study, enhanced production of 5'-CMP was realized by the transformation of cytidine using co-immobilized di-enzymes, uridine-cytidine kinase (UCK) and acetate kinase (AcK). The immobilization yield of the enzyme had a clear correlation with the surface charges as zeta potential (ξ). Among them, ε-polylysine-functionalized sepharose (SA-EPL, ξ = 9.31 mV) showed high immobilization yield (78.8%), which was 4.9-fold than that of nitrilotriacetic acid functionalized sepharose (SA-NTA, ξ = -12.6 mV). The residual activity of affinity co-immobilized enzyme (EPL-Ni/EPL@AcK-UCK) was higher than 70.6% after recycled 10 times. Thus, this study provides an effective approach for the production of 5'-CMP with the advantages of low adenosine 5'-triphosphate (ATP) consumption, reduced side reactions, and improved reusability by co-immobilized UCK and AcK on the functionalized Sepharose.

Key words: Sepharose, ε-Polylysine, Dual-enzyme cascade, Cytidine 5'-monophosphate, Enzyme immobilization

摘要： Cytidine 5'-monophosphate (5'-CMP) is an essential nucleotide for additives. In this study, enhanced production of 5'-CMP was realized by the transformation of cytidine using co-immobilized di-enzymes, uridine-cytidine kinase (UCK) and acetate kinase (AcK). The immobilization yield of the enzyme had a clear correlation with the surface charges as zeta potential (ξ). Among them, ε-polylysine-functionalized sepharose (SA-EPL, ξ = 9.31 mV) showed high immobilization yield (78.8%), which was 4.9-fold than that of nitrilotriacetic acid functionalized sepharose (SA-NTA, ξ = -12.6 mV). The residual activity of affinity co-immobilized enzyme (EPL-Ni/EPL@AcK-UCK) was higher than 70.6% after recycled 10 times. Thus, this study provides an effective approach for the production of 5'-CMP with the advantages of low adenosine 5'-triphosphate (ATP) consumption, reduced side reactions, and improved reusability by co-immobilized UCK and AcK on the functionalized Sepharose.

关键词: Sepharose, ε-Polylysine, Dual-enzyme cascade, Cytidine 5'-monophosphate, Enzyme immobilization

Xiaohong Zhou, Wenfeng Zhou, Wei Zhuang, Chenjie Zhu, Hanjie Ying, Hongman Zhang. Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose[J]. Chinese Journal of Chemical Engineering, 2023, 58(6): 40-52.

References

[1] A.J. Deoda, R.S. Singhal, 5'-Phosphodiesterase (5'-PDE) from germinated barley for hydrolysis of RNA to produce flavour nucleotides, Bioresour. Technol. 88 (3) (2003) 245-250.
[2] M. Yoshikawa, T. Kato, T. Takenishi, Studies of phosphorylation. III. selective phosphorylation of unprotected nucleosides, Bull. Chem. Soc. Jpn. 42 (12) (1969) 3505-3508.
[3] A.R. van Rompay, A. Norda, K. Lindén, M. Johansson, A. Karlsson, Phosphorylation of uridine and cytidine nucleoside analogs by two human uridine-cytidine kinases, Mol. Pharmacol. 59 (5) (2001) 1181-1186.
[4] C. Ingram-Smith, S.R. Martin, K.S. Smith, Acetate kinase: Not just a bacterial enzyme, Trends Microbiol. 14 (6) (2006) 249-253.
[5] U. Hanefeld, L. Gardossi, E. Magner, Understanding enzyme immobilisation, Chem. Soc. Rev. 38 (2) (2009) 453-468.
[6] W.P. Jin, Z.F. Wang, D.F. Peng, W.Y. Shen, Z.Z. Zhu, S.Y. Cheng, B. Li, Q.R. Huang, Effect of linear charge density of polysaccharides on interactions with α-amylase: Self-Assembling behavior and application in enzyme immobilization, Food Chem. 331 (2020) 127320.
[7] J.D. Cui, S.R. Jia, Optimization protocols and improved strategies of cross-linked enzyme aggregates technology: Current development and future challenges, Crit. Rev. Biotechnol. 35 (1) (2015) 15-28.
[8] R.C. Rodrigues, C. Ortiz, Á. Berenguer-Murcia, R. Torres, R. Fernández-Lafuente, Modifying enzyme activity and selectivity by immobilization, Chem. Soc. Rev. 42 (15) (2013) 6290-6307.
[9] J.H. Yi, M.Y. Qiu, Z.B. Zhu, X.L. Dong, E. Andrew Decker, D.J. McClements, Robust and recyclable magnetic nanobiocatalysts for extraction of anthocyanin from black rice, Food Chem. 364 (2021) 130447.
[10] R.A. Sheldon, S. van Pelt, Enzyme immobilisation in biocatalysis: Why, what and how, Chem. Soc. Rev. 42 (15) (2013) 6223-6235.
[11] S. Arana-Peña, D. Carballares, R. Morellon-Sterlling, Á. Berenguer-Murcia, A.R. Alcántara, R.C. Rodrigues, R. Fernandez-Lafuente, Enzyme co-immobilization: Always the biocatalyst designers' choice…or not? Biotechnol. Adv. 51 (2021) 107584.
[12] C. Garcia-Galan, Á. Berenguer-Murcia, R. Fernandez-Lafuente, R.C. Rodrigues, Potential of different enzyme immobilization strategies to improve enzyme performance, Adv. Synth. Catal. 353 (16) (2011) 2885-2904.
[13] E. Hwang, S. Lee, Multienzymatic cascade reactions via enzyme complex by immobilization, ACS Catal, (2019)
[14] S. Datta, L.R. Christena, Y.R. Rajaram, Enzyme immobilization: An overview on techniques and support materials, 3 Biotech 3 (1) (2013) 1-9.
[15] A.I. Benítez-Mateos, M.L. Contente, Agarose vs. methacrylate as material supports for enzyme immobilization and continuous processing, Catalysts 11 (7) (2021) 814.
[16] C. Engelmann, N. Ekambaram, J. Johannsen, O. Fellechner, T. Waluga, G. Fieg, A. Liese, P. Bubenheim, Enzyme immobilization on synthesized nanoporous silica particles and their application in a Bi-enzymatic reaction, ChemCatChem 12 (8) (2020) 2245-2252.
[17] N.C. Ricardi, L.T. Arenas, E.V. Benvenutti, R. Hinrichs, E.E.E. Flores, P.F. Hertz, T.M.H. Costa, High performance biocatalyst based on β-d-galactosidase immobilized on mesoporous silica/titania/chitosan material, Food Chem 359 (2021) 129890.
[18] J.D. Cui, Y.X. Feng, S.R. Jia, Silica encapsulated catalase@metal-organic framework composite: A highly stable and recyclable biocatalyst, Chem. Eng. J. 351 (2018) 506-514.
[19] Y. Chen, Y. Xin, H.L. Yang, L. Zhang, Y.R. Zhang, X.L. Xia, Y.J. Tong, W. Wang, Immobilization and stabilization of cholesterol oxidase on modified sepharose particles, Int. J. Biol. Macromol. 56 (2013) 6-13.
[20] P. Zucca, R. Fernandez-Lafuente, E. Sanjust, Agarose and its derivatives as supports for enzyme immobilization, Molecules 21 (11) (2016) 1577.
[21] Y.A. Ramírez Tapias, C.W. Rivero, F.L. Gallego, J.M. Guisán, J.A. Trelles, Stabilization by multipoint covalent attachment of a biocatalyst with polygalacturonase activity used for juice clarification, Food Chem. 208 (2016) 252-257.
[22] S. Hosseinkhani, R. Szittner, M. Nemat-Gorgani, E.A. Meighen, Adsorptive immobilization of bacterial luciferases on alkyl-substituted sepharose 4B, Enzyme Microb. Technol. 32 (1) (2003) 186-193.
[23] Y.F. Zhang, Q. Wang, H. Hess, Increasing enzyme cascade throughput by pH-engineering the microenvironment of individual enzymes, ACS Catal. 7 (3) (2017) 2047-2051.
[24] S. Hudson, J. Cooney, B.K. Hodnett, E. Magner, Chloroperoxidase on periodic mesoporous organosilanes: Immobilization and reuse, Chem. Mater. 19 (8) (2007) 2049-2055.
[25] P. Zucca, E. Sanjust, Inorganic materials as supports for covalent enzyme immobilization: Methods and mechanisms, Molecules 19 (9) (2014) 14139-14194.
[26] H. Zheng, S.J. Yang, Y.C. Zheng, Y. Cui, Z. Zhang, J.Y. Zhong, J. Zhou, Electrostatic effect of functional surfaces on the activity of adsorbed enzymes: Simulations and experiments, ACS Appl. Mater. Interfaces 12 (31) (2020) 35676-35687.
[27] Y. Wang, X.F. Zhang, N.Y. Han, Y.S. Wu, D.X. Wei, Oriented covalent immobilization of recombinant protein A on the glutaraldehyde activated agarose support, Int. J. Biol. Macromol. 120 (Pt A) (2018) 100-108.
[28] L.L. Yu, X.Y. Dong, Y. Sun, Ion-exchange resins facilitate like-charged protein refolding: Effects of porous solid phase properties, J. Chromatogr. A 1225 (2012) 168-173.
[29] C. Mateo, J.M. Bolivar, C.A. Godoy, J. Rocha-Martin, B.C. Pessela, J.A. Curiel, R. Muñoz, J.M. Guisan, G. Fernández-Lorente, Improvement of enzyme properties with a two-step immobilizaton process on novel heterofunctional supports, Biomacromolecules 11 (11) (2010) 3112-3117.
[30] R. Wu, F.X. Liu, Q.H. Dong, Y.Y. Huang, Y.B. Qiu, Y.Y. Sun, E.Z. Su, Combination of adsorption and cellulose derivative membrane coating for efficient immobilization of laccase, Appl. Biochem. Biotechnol. 193 (2) (2021) 446-462.
[31] L.T. Wu, S.S. Wu, Z. Xu, Y.B. Qiu, S. Li, H. Xu, Modified nanoporous titanium dioxide as a novel carrier for enzyme immobilization, Biosens. Bioelectron. 80 (2016) 59-66.
[32] D.H. Zhang, Y.Q. Li, L.J. Peng, N. Chen, Lipase immobilization on magnetic microspheres via spacer arms: Effect of steric hindrance on the activity, Biotechnol. Bioprocess Eng. 19 (5) (2014) 838-843.
[33] Y.T. Zhang, Y.K. Yang, W.F. Ma, J. Guo, Y. Lin, C.C. Wang, Uniform magnetic core/shell microspheres functionalized with Ni²⁺-iminodiacetic acid for one step purification and immobilization of his-tagged enzymes, ACS Appl. Mater. Interfaces 5 (7) (2013) 2626-2633.
[34] Q. Xiao, H.F. Weng, G. Chen, A.F. Xiao, Preparation and characterization of octenyl succinic anhydride modified agarose derivative, Food Chem. 279 (2019) 30-39.
[35] S.C. Liufu, H.N. Xiao, Y.P. Li, Thermal analysis and degradation mechanism of polyacrylate/ZnO nanocomposites, Polym. Degrad. Stab. 87 (1) (2005) 103-110.
[36] C. Chao, J.D. Liu, J.T. Wang, Y.W. Zhang, B. Zhang, Y.T. Zhang, X. Xiang, R.F. Chen, Surface modification of halloysite nanotubes with dopamine for enzyme immobilization, ACS Appl. Mater. Interfaces 5 (21) (2013) 10559-10564.
[37] H.S. Lee, J.S. Kim, K. Shim, J.W. Kim, K. Inouye, H. Oneda, Y.W. Kim, K.A. Cheong, H. Cha, E.J. Woo, J.H. Auh, S.J. Lee, J.W. Kim, K.H. Park, Dissociation/association properties of a dodecameric cyclomaltodextrinase. Effects of pH and salt concentration on the oligomeric state, FEBS J. 273 (1) (2006) 109-121.
[38] Y.F. Zhang, J. Ge, Z. Liu, Enhanced activity of immobilized or chemically modified enzymes, ACS Catal. 5 (8) (2015) 4503-4513.
[39] S. Zofair, A. Arsalan, M.A. Khan, F.A. Alhumaydhi, H. Younus, Immobilization of laccase on Sepharose-linked antibody support for decolourization of phenol red, Int. J. Biol. Macromol. 161 (2020) 78-87.
[40] R.M. da Silva, L. Gonçalves, S. Rodrigues, Different strategies to co-immobilize dextransucrase and dextranase onto agarose based supports: Operational stability study, Int. J. Biol. Macromol. 156 (2020) 411-419.
[41] J.Y. Liao, S.S. Han, X.L. Li, J. He, F. Secundo, H. Liang, Co-immobilization of two-component hydroxylase monooxygenase by functionalized magnetic nanoparticles for preserving high catalytic activity and enhancing enzyme stabilty, Int. J. Biol. Macromol. 164 (2020) 3163-3170.

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 4

Recommended Articles

Metrics

Comments

[1]	Jelena Bebić, Katarina Banjanac, Marija Ćorović, Ana Milivojević, Milica Simović, Aleksandar Marinković, Dejan Bezbradica. Immobilization of laccase from Myceliophthora thermophila on functionalized silica nanoparticles: Optimization and application in lindane degradation [J]. Chinese Journal of Chemical Engineering, 2020, 28(4): 1136-1144.
[2]	Zhenhua Wu, Yan Nan, Yang Zhao, Xueying Wang, Shouying Huang, Jiafu Shi. Immobilization of carbonic anhydrase for facilitated CO₂ capture and separation [J]. Chinese Journal of Chemical Engineering, 2020, 28(11): 2817-2831.
[3]	ZHOU Huacong, LI Wei, SHOU Qinghui, GAO Hongshuai, XU Peng, DENG Fuli, LIU Huizhou. Immobilization of Penicillin G Acylase on Magnetic Nanoparticles Modified by Ionic Liquids [J]. Chin.J.Chem.Eng., 2012, 20(1): 146-151.
[4]	WANG Anming a,b , ZHOU Cheng a , WANG Hua a , SHEN Shubao a , XUE Jianyue b , OUYANG Pingkai a . Covalent assembly of penicillin acylase in mesoporous silica based on macromolecular crowding theory [J]. , 2007, 15(6): 788-790.