TY - JOUR
T1 - Enhanced recombinant Sulfurihydrogenibium yellowstonense carbonic anhydrase activity and thermostability by chaperone GroELS for carbon dioxide biomineralization
AU - Wahyu Effendi, Sefli Sri
AU - Tan, Shih I.
AU - Ting, Wan Wen
AU - Ng, I. Son
N1 - Funding Information:
The authors are grateful to the financial support for this study provided by the Ministry of Science and Technology ( MOST 108-2621-M-006-015 , MOST 108-2221-E-006-004-MY3 and MOST 109-2218-E-006-015 ) in Taiwan.
Funding Information:
The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: The authors are grateful for the financial support received from the Ministry of Science and Technology ( MOST 108-2221-E-006-004-MY3 , MOST 108-2621-M-006-015 and MOST 109-2218-E-006-015 ) in Taiwan.
Publisher Copyright:
© 2020 Elsevier Ltd
PY - 2021/5
Y1 - 2021/5
N2 - Biological carbon fixation is a feasible strategy to reduce atmospheric carbon dioxide levels (CO2). In this platform, carbonic anhydrase (CA) enzyme is employed to accelerate the sequestration of CO2. The present work explored the effect of chaperone GroELS and TrxA-tag on improving soluble expression of the recombinant Sulfurihydrogenibium yellowstonense CA which activity and biomineralization capability were taken into consideration. At first, the expression of GroELS using the inducible T7 promoter and constitutive J23100 promoter were investigated. The results indicated that 1.4 folds increment of soluble protein and 100% of CA activity enhancement were achieved with GroELS co-expression driven by J23100 promoter. Furthermore, the involvement of TrxA fusion tag displayed a significant enhancement of soluble protein production which was about 2.67 times higher than that of original SyCA. Besides, co-expression with GroELS intensified the thermostability of SyCA at 60 °C owing to changes in the structural conformation of the protein, which was proved by an in vitro assay. The SyCA was further entrapped and immobilized into polyacrylamide gel (i.e., PAGE-SyCA). The biomineralization capability of the PAGE-SyCA and whole-cell (WC) was compared in a two-column system. After 5 cycles of reuse, PAGE-SyCA maintained 29.8% activity and formed 774 mg of CaCO3 solids in the B::JG strain. This study presents the recombinant engineering strategies to improve SyCA productivity, activity, thermostability, and effective carbon dioxide conversion.
AB - Biological carbon fixation is a feasible strategy to reduce atmospheric carbon dioxide levels (CO2). In this platform, carbonic anhydrase (CA) enzyme is employed to accelerate the sequestration of CO2. The present work explored the effect of chaperone GroELS and TrxA-tag on improving soluble expression of the recombinant Sulfurihydrogenibium yellowstonense CA which activity and biomineralization capability were taken into consideration. At first, the expression of GroELS using the inducible T7 promoter and constitutive J23100 promoter were investigated. The results indicated that 1.4 folds increment of soluble protein and 100% of CA activity enhancement were achieved with GroELS co-expression driven by J23100 promoter. Furthermore, the involvement of TrxA fusion tag displayed a significant enhancement of soluble protein production which was about 2.67 times higher than that of original SyCA. Besides, co-expression with GroELS intensified the thermostability of SyCA at 60 °C owing to changes in the structural conformation of the protein, which was proved by an in vitro assay. The SyCA was further entrapped and immobilized into polyacrylamide gel (i.e., PAGE-SyCA). The biomineralization capability of the PAGE-SyCA and whole-cell (WC) was compared in a two-column system. After 5 cycles of reuse, PAGE-SyCA maintained 29.8% activity and formed 774 mg of CaCO3 solids in the B::JG strain. This study presents the recombinant engineering strategies to improve SyCA productivity, activity, thermostability, and effective carbon dioxide conversion.
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U2 - 10.1016/j.chemosphere.2020.128461
DO - 10.1016/j.chemosphere.2020.128461
M3 - Article
C2 - 33131750
AN - SCOPUS:85094599521
SN - 0045-6535
VL - 271
JO - Chemosphere
JF - Chemosphere
M1 - 128461
ER -