LAPSE:2023.36134
Published Article
LAPSE:2023.36134
Effects of His-Tag Length on the Soluble Expression and Selective Immobilization of D-Amino Acid Oxidase from Trigonopsis variabilis: A Preliminary Study
July 4, 2023
His-tags are widely used for the purification of recombinant proteins. High-cost carriers functionalized with nickel ions are commonly required for the selective immobilization of His-tagged enzymes. In this study, His-tags of varying lengths were fused to the N-terminus of D-amino acid oxidase (DAO) from Trigonopsis variabilis. The attachment of a His6 tag significantly improved the solubility of the recombinant DAO expressed in Escherichia coli. By modulating the tag lengths, a better balance between cell growth and protein solubility was achieved, resulting in a higher volume activity (His3). Furthermore, the fusion of longer tags (His6 and His9) facilitated the rapid immobilization of DAOs onto a commercial epoxy carrier without metal bearing, resulting in more selective immobilization. In conclusion, the modulation of His-tag length was preliminarily demonstrated as a simple and cost-effective approach to achieve efficient expression, as well as fast and selective immobilization of DAO.
Record ID
Keywords
D-amino acid oxidase, His-tags, immobilization, recombinant expression
Subject
Suggested Citation
Yan Z, Zhu Q, Ma L, Li G, Su E, Zeng J, Chen Y, Meng E, Deng S. Effects of His-Tag Length on the Soluble Expression and Selective Immobilization of D-Amino Acid Oxidase from Trigonopsis variabilis: A Preliminary Study. (2023). LAPSE:2023.36134
Author Affiliations
Yan Z: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Zhu Q: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Ma L: National Engineering Research Center of Oiltea Camellia, Research Institute of Oiltea Camellia, Hunan Academy of Forestry, Shao Shan South Road, No. 658, Changsha 410004, China
Li G: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Su E: Enzyme and Fermentation Technology Laboratory, College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, China [ORCID]
Zeng J: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Chen Y: National Engineering Research Center of Oiltea Camellia, Research Institute of Oiltea Camellia, Hunan Academy of Forestry, Shao Shan South Road, No. 658, Changsha 410004, China
Meng E: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Deng S: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China; National Engineering Research Center of Oiltea Camellia, Researc
Zhu Q: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Ma L: National Engineering Research Center of Oiltea Camellia, Research Institute of Oiltea Camellia, Hunan Academy of Forestry, Shao Shan South Road, No. 658, Changsha 410004, China
Li G: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Su E: Enzyme and Fermentation Technology Laboratory, College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, China [ORCID]
Zeng J: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Chen Y: National Engineering Research Center of Oiltea Camellia, Research Institute of Oiltea Camellia, Hunan Academy of Forestry, Shao Shan South Road, No. 658, Changsha 410004, China
Meng E: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China
Deng S: Hunan Key Laboratory of Economic Crops Genetic Improvement and Integrated Utilization, School of Life and Health Sciences, Hunan University of Science and Technology, Xiangtan 411201, China; National Engineering Research Center of Oiltea Camellia, Researc
Journal Name
Processes
Volume
11
Issue
6
First Page
1588
Year
2023
Publication Date
2023-05-23
Published Version
ISSN
2227-9717
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Original Submission
Other Meta
PII: pr11061588, Publication Type: Journal Article
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Published Article
LAPSE:2023.36134
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External Link
doi:10.3390/pr11061588
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Version History
[v1] (Original Submission)
Jul 4, 2023
Verified by curator on
Jul 4, 2023
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v1
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https://psecommunity.org/LAPSE:2023.36134
Original Submitter
Calvin Tsay
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