Functional expression of RubisCO by a high-speed carbon dioxide fixing microorganism

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A thermophilic hydrogen bacterium Pseudomonas hydrogenothermophila strain TH-1 possesses cbbQ gene under RubisCO gene (cbbLS). CbbQ protein was thought to participate in the post-translational activation/stabilization of RubisCO.We showed the presence of cbbO gene under cbbQ gene and clarified the gene sequence. In the next step, plasmids harboring cbbQ,cbbO,cbbQ-cbbO gene were prepared. By using one of the above plasmids and a plasmid harboring cbbLS,transformation of E.coli was performed. Then, difference in activity or expression of RubisCO was analyzed by measuring RubisCO activity or by electrophoreis, respectively. When only the plasmid harboring cbbLS was used, RubisCO activity was low and the RubisCO protein was detected as a smear band on polyacrylamide-gel electrophoresis. Contrary to this, in the presence of cbbQ or cbbO gene, the activity of RubisCO increased 2 to 3 fold, also the RubisCO protein was detected as a clear band on polyacrylamide-gel electrophoresis.In a separate experiments, three kinds of protein (CbbLS,CbbQ,and CbbO) were prepared for the experiment of in vitro RubisCO activation. However, successful results have not been obtained.During our research, CbbO protein itself was proved to possess CO_2 incorporation activity, which will be a source of thinking for future research.