<p>Manipulation of enzyme behaviour
represents a sustainable technology that can be harnessed to enhance the
production of valuable metabolites and chemical precursors. b-glucose 1,6-bisphosphate (bG16BP) is a native reaction
intermediate in the catalytic cycle of b-phosphoglucomutase
(bPGM) that has been proposed as a treatment
for human congenital disorder of glycosylation involving phosphomannomutase 2
(PMM2). Studies of both bPGM
and PMM2 could benefit from a green and high-yielding method for bG16BP production. Three strategies
have been reported previously for the synthesis of bG16BP; however, each of these
methods either delivers low yields or uses chemicals and procedures with significant
environmental impacts. Herein, through combined use of NMR spectroscopy, kinetic assays and site-directed mutagenesis, we report the efficient enzymatic synthesis of anomer-specific
bG16BP using a variant of bPGM. Further purification,
employing a simple environmentally considerate precipitation procedure
requiring only a standard biochemical toolset, results in a product with high purity
and yield. Moreover, this synthesis strategy illustrates how manipulation of
the catalytic magnesium coordination of an enzyme can be utilised to generate large
quantities of a valuable metabolite.</p>
Enzymatic Production of β-Glucose 1,6-Bisphosphate Through Manipulation of Catalytic Magnesium Coordination
