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DOI: 10.1055/sos-SD-214-00162

Wever, R.; Babich, L.; Hartog, A. F.Science of Synthesis: Biocatalysis in Organic Synthesis, (20151235.

Acid phosphatases catalyze the regioselective phosphorylation of the ribose group in inosine.[‌16‌,‌70‌] Later it was shown that many simple carbohydrates can also be phosphorylated using acid phosphatases from Shigella flexneri (PhoN-Sf) and Salmonella enterica (PhoN-Se)[‌70‌,‌77‌‌79‌] using pyrophosphate as the phosphate donor. In particular, d-glucose is phosphorylated in a very efficient manner, as has also been observed using the enzyme from Raoultella planticola.[‌80‌] The efficient conversion of glucose into d-glucose 6-phosphate (4) has been used in a three-enzyme cascade system containing glucose 6-phosphate dehydrogenase (EC to regenerate nicotinamide adenine dinucleotide phosphate (NADPH), which was used to produce chiral alcohols from ketones.[‌78‌] d-Glucose 6-phosphate (4) has also been synthesized in a preparative manner by this enzymatic method (Scheme 10).[‌79‌] The yield is 76% based upon pyrophosphate; however, on the basis of the input of glucose, the yield is considerably less and amounts to only 7.6%. Much better conversions (19%) are obtained in fed-batch reactions with acid phosphatase from Shigella flexneri (PhoN-Sf) immobilized on Immobeads.[‌77‌] Interestingly, unlike other primary phosphorylated carbohydrates, d-glucose 6-phosphate (4) once formed is hardly dephosphorylated by PhoN-Sf and the incubation time is not very critical. The productivity of the synthesis of d-glucose 6-phosphate using a continuous-flow reactor has also been tested and high yields were obtained.[‌77‌] Using this reactor, 2-acetamido-2-deoxy-d-glucose can also be phosphorylated, providing product 7 (R1 = NHAc) in 53% yield (Scheme 11).[‌77‌] Considering the ease of preparation, these continuous-flow reactors with immobilized acid phosphatase have significant advantages compared to classical synthetic chemistry methods. The success of this continuous-flow system is mainly due to very high stability of the immobilized acid phosphatase. During turnover the system is stable for at least 2 weeks and the column can be repetitively used for the preparation of other phosphorylated carbohydrates and alcohols.

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