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

doi:10.1016/j.cjche.2022.11.002

中国化学工程学报 ›› 2023, Vol. 58 ›› Issue (6): 40-52.DOI: 10.1016/j.cjche.2022.11.002

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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

收稿日期:2022-08-14 修回日期:2022-11-01 出版日期:2023-06-28 发布日期:2023-08-31
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).

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:2023-06-28 Published:2023-08-31
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).

摘要/Abstract

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

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

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]. 中国化学工程学报, 2023, 58(6): 40-52.

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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

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