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Enzymatic Production of β-Glucose 1,6-Bisphosphate Through Manipulation of Catalytic Magnesium Coordination

revised on 29.09.2020, 13:52 and posted on 30.09.2020, 09:50 by Henry P Wood, Nicola J Baxter, Clare R Trevitt, F Aaron Cruz-Navarrete, Andrea M Hounslow, Jonathan P Waltho

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.


BBSRC X/009906-20-26

BBSRC BB/M021637/1

BBSRC BB/S007965/1

BBSRC BB/P007066/1

CONACYT 472448


Email Address of Submitting Author


University of Manchester


United Kingdom

ORCID For Submitting Author


Declaration of Conflict of Interest

There are no conflicts to declare.

Version Notes

Version 2.0