期刊
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
卷 167, 期 -, 页码 326-334出版社
ELSEVIER
DOI: 10.1016/j.ijbiomac.2020.11.189
关键词
Mesorhizobium loti; Carbonic anhydrase; Chaperone GroELS; Inclusion bodies; Isothermal titration calorimetry; Biomineralization
资金
- Ministry of Science and Technology [MOST 108-2621-M-006-015, MOST 108-2221-E-006-004-MY3]
The study found that co-expressing MlCA with protein partners effectively increased the solubility, activity, and turnover rate, while also affecting its stability in temperature, pH range, and presence of metal ions.
Mesorhizobiumloti carbonic anhydrase (MlCA), an intrinsically high catalytic enzyme, has been employed for carbon dioxide capture and sequestration. However, recombinant expression of MlCA in Escherichia coli often forms inclusion bodies. Hence, protein partners such as fusion-tags and molecular chaperones are involved in regarding reduce the harshness of protein folding. TrxA-tag and GroELS have been chosen to co-express with MlCA in E. coli under an inducible T7 promoter or a constitutive J23100 promoter to compare productivity and activity. The results possessed that coupling protein partners effectively increased soluble MlCA up to 2.9-folds under T7 promoter, thus enhancing the CA activity by 120% and achieving a 5.2-folds turnover rate. Besides, it has also shifted the optimum temperature from 40 degrees C to 50 degrees C, promoted stability in the broad pH range (4.5 to 9.5) and the presence of various metal ions. Based on the in vitro assay and isothermal titration calorimetry (ITC) analysis, GroELS enhancing CA activity was due to change the intrinsic thermodynamic properties of the enzyme from endothermic to exothermic reaction (i.e.,Delta H = 89.8 to -121.8 kJ/mol). Therefore, the collaboration of TrxA-MlCA with GroELS successfully augmented CO2 biomineralization. (C) 2020 Elsevier B.V. All rights reserved.
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