| Products/Services Used | Details | Operation |
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| Gene Synthesis | All the DNA work was performed using E. coli TOP10 (Invitrogen, USA). E. coli BL21 (DE3) (Novagen, USA) was used as the host for heterologous protein expression. Cells were routinely grown in Luria-Bertani (LB) medium (Usb Corp., USA) containing appropriate antibiotics (50 μg/mL ampicillin or 10 μg/mL streptomycin) at 37 °C, 220 rpm. The protein sequence information was obtained from the National Center for Biotechnology Information (NCBI). The genes encoding pmCA and taCA were chemically synthesized with codon optimization for E. coli(Genscript, USA). The putative signal sequences indicated in Fig. 1 were not included in the synthetic genes. The synthetic CA gene sequences were designed to contain a NdeI restriction site that half overlaps the 5′ start codon and a XhoI site at their 3′ termini without a stop codon. The genes were excised from the pUC57-based plasmids by digestion with NdeI and XhoI and were subcloned into the expression vector pET-22b(+) (Novagen) treated with the same restriction enzymes, resulting in pET-pmCA and pET-taCA. The recombinant CA genes have hexahistidine (His6)-tag sequences at their 3′ termini. | Get A Quote |
Carbonic anhydrase (CA) is an enzyme that rapidly catalyzes the reversible hydration of carbon dioxide (CO2) in aqueous solutions. Recent progresses in CA research suggest that this biocatalyst can be used as an alternative or an additive for chemical CO2 capture, which is considered an essential green route for the reduction of anthropogenic CO2 emissions. However, the facilities in which CA will most likely be used usually operate under high temperature conditions, requiring a highly thermostable CA. Here, we attempted to find and characterize α-CAs originated from Persephonella marina and Thermovibrio ammonificans, thermophilic Gram-negative bacteria in deep-sea hydrothermal vents. The recombinant α-C... More
