Uridine phosphorylase from Escherichia coli was evolved by iterative saturation mutagenesis. The best mutant showed a temperature optimum of 60C and a half-life of 17.3 h at 60C. The mutant enzyme, as well as a purine nucleoside phosphorylase from Bacillus halodurans, were immobilised as SpherezymesTM. Immobilisation of the mutant enzyme provided a further increase in thermostability. When combined with the purine nucleoside phosphorylase from B. halodurans, productivity of 5-methyluridine, a pharmaceutical intermediate, was increased from 10 to 31 g.l-1.h-1.
Reference:
Visser, DF, Hennessy, F, Rashamuse, J, et al. 2011. Stabilization of Escherichia coli uridine phosphorylase by evolution and immobilization. Journal of Molecular Catalysis B: Enzymatic, Vol. 68(3/4), pp. 279-285
Visser, D. F., Hennessy F, Rashamuse J, Pletschke B, & Brady D (2010). Stabilization of Escherichia coli uridine phosphorylase by evolution and immobilization. http://hdl.handle.net/10204/5109
Visser, Daniel F, Hennessy F, Rashamuse J, Pletschke B, and Brady D "Stabilization of Escherichia coli uridine phosphorylase by evolution and immobilization." (2010) http://hdl.handle.net/10204/5109
Visser DF, Hennessy F, Rashamuse J, Pletschke B, Brady D. Stabilization of Escherichia coli uridine phosphorylase by evolution and immobilization. 2010; http://hdl.handle.net/10204/5109.
Copyright: 2010 Elsevier Publishers. This is a pre print version of the work. The definitive version is published in the Journal of Molecular Catalysis B: Enzymatic, Vol. 68(3/4), pp. 279-285
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