Defining a process operating window for the synthesis of 5-methyluridine by transglycosylation of guanosine and thymine.
This paper describes a high yielding coupled enzymatic reaction using Bacillus halodurans purine nucleoside phosphorylase (PNP) and E. coli uridine phosphorylase (UP) for synthesis of 5-methyluridine (5-MU) by transglycosylation. Key parameters such as reaction temperature, pH, reactant loading, reactor configuration and enzyme loading were investigated. A guanosine conversion of 95% and a 5-MU yield of 85% were achieved at 1l scale, with a productivity of 10 g l⁻¹ h⁻¹.
CSIR Biosciences, Private Bag X2, Modderfontein, Johannesburg 1645, South Africa.
Gregory E R
GE
Gordon
Daniel F
DF
Visser
Dean
D
Brady
Nelly
N
Raseroka
Moira L
ML
Bode
D001407:Bacillus / Q000201:enzymology / Q000378:metabolism; D019149:Bioreactors / Q000382:microbiology*; D006031:Glycosylation; D006151:Guanosine / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D011683:Purine-Nucleoside Phosphorylase / Q000378:metabolism; D013696:Temperature; D013941:Thymine / Q000737:chemistry / Q000378:metabolism*; D014529:Uridine / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D014543:Uridine Phosphorylase / Q000378:metabolism
D006151:Guanosine; C009182:ribothymidine; D011683:Purine-Nucleoside Phosphorylase; D014543:Uridine Phosphorylase; D013941:Thymine; D014529:Uridine