방향적 진화로 개량된 Desulfovibrio vulgaris 유래 carbonic anhydrase의 NEXT 태그 융합에 의한 대장균 재조합 발현 향상

초록

The CO2-hydrating enzyme carbonic anhydrase (CA) has gained much attention as a green and economical catalyst for expediting CO2 capture. Stable CAs are desirable, as they can withstand the high temperatures of CO2-capturing facilities. One of the most stable CAs is the engineered β-CA from Desulfovibrio vulgaris (DvCA10). However, the low bacterial expression level of DvCA10 must be addressed for industrial applications. In this study, the NEXT tag, an intrinsically disordered peptide tag, was fused to the N terminus of DvCA10 to improve its soluble expression. Following the fusion of the NEXT tag, along with a flexible linker, the soluble expression of DvCA10 was dramatically improved (6-fold) compared to the untagged DvCA10, demonstrating the powerful performance of the NEXT tag. Although the activity of NEXT-DvCA10 was approximately 20% lower than that of DvCA10, their thermal stabilities assessed at 80℃ were essentially the same, exhibiting biphasic inactivation kinetics. This result shows that the NEXT tag can be used as a fusion tag to improve the production yield of DvCA10 without compromising its enzymatic properties. The NEXT-tagged DvCA10 is expected to be used as an economical and practical enzyme in future industrial CO2 capture applications.

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