Abstract: Carbonic anhydrases (CAs) has been focused as
biological catalysis for CO2 sequestration process because it can
catalyze the conversion of CO2 to bicarbonate. Here, codon-optimized
sequence of α type-CA cloned from Duneliala species. (DsCAopt) was
constructed, expressed, and characterized. The expression level in E.
coli BL21(DE3) was better for codon-optimized DsCAopt than intact
sequence of DsCAopt. DsCAopt enzyme shows high-stability at pH
7.6/10.0. In final, we demonstrated that in the Ca2+ solution, DsCAopt
enzyme can catalyze well the conversion of CO2 to CaCO3, as the
calcite form.
Abstract: Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.