Enhanced Production of a Thermostable Carbonic Anhydrase in Escherichia coli by Using a Modified NEXT Tagopen access
- Authors
- Hwang, In Seong; Kim, Joo Hyeon; Jo, Byung Hoon
- Issue Date
- Oct-2021
- Publisher
- MDPI
- Keywords
- recombinant protein; solubility enhancer; NEXT tag; carbonic anhydrase; Thermovibrio ammonificans; CO2 capture
- Citation
- MOLECULES, v.26, no.19
- Indexed
- SCIE
SCOPUS
- Journal Title
- MOLECULES
- Volume
- 26
- Number
- 19
- URI
- https://scholarworks.gnu.ac.kr/handle/sw.gnu/3203
- DOI
- 10.3390/molecules26195830
- ISSN
- 1420-3049
1420-3049
- Abstract
- Carbonic anhydrase (CA) is an ultrafast enzyme that catalyzes the reversible conversion of carbon dioxide (CO2) to bicarbonate. CA is considered to be a green catalyst for enzyme-based CO2 capture and utilization. In particular, the CA of Thermovibrio ammonificans (taCA) has attracted increasing attention as a highly stable enzyme. However, the poor solubility and the low expression level in Escherichia coli have hampered further utilization of taCA. In a recent study, these limitations were partly resolved by using a small solubility-enhancing fusion tag named NEXT, which originates from the N-terminal extension of Hydrogenovibrio marinus CA. In this study, the NEXT tag was engineered by adding small peptides to the N terminus to further increase the production yield of NEXT-tagged taCA. The addition of ng3 peptide (His-Gly-Asn) originating from the N-terminal sequence of Neisseria gonorrhoeae CA improved the expression of NEXT-taCA, while the previously developed translation-enhancing element (TEE) and Ser-Lys-Ile-Lys (SKIK) tag were not effective. The expression test with all 16 codon combinations for the ng3 sequence revealed that the change in translation initiation rate brought about by the change in nucleotide sequence was not the primary determinant for the change in expression level. The modified ng3-NEXT tag may be applied to increase the production yields of various recombinant proteins.</p>
- Files in This Item
- There are no files associated with this item.
- Appears in
Collections - 자연과학대학 > Division of Life Sciences > Journal Articles

Items in ScholarWorks are protected by copyright, with all rights reserved, unless otherwise indicated.