肺炎链球菌磷酸甘油氧化酶的表征及在稀有糖合成中的应用

In this paper, the expression of glycerophosphate oxidase (GPO) from Streptococcus pneumoniae and its application for rare sugar synthesis were investigated. The gene glpo encoding GPO was amplified and cloned into the expression plasmid pET-28a. The recombinant plasmid was transformed into Escherichia coli Rosetta (DE3) to express the protein after induction with IPTG. After purified by Ni2+ chelating affinity column chromatography, GPO pure enzyme was characterized and employed in the rare sugar synthesis using one-pot four-enzyme strategy. The results demonstrated that the molecular weight of GPO is about 65 kD and the coenzyme FAD is easily dissociable from GPO. In addition, the optimum pH is 8.0 and the optimum temperature is 30℃. Compared with GPO from Streptococcus thermophilus, this GPO has similar or even higher activity, as the production yields were about 50% or higher. To the best of our knowledge, this is the first time to characterize GPO from Streptococcus pneumoniae and it's of great significance for further study.