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Acid phosphatase project started in 1998 Acid phosphatases were originally studied because the binding pocket for vanadate in vanadium CPO was shown by us to be the same as that in a group of acid phosphatases. Two recombinant acid phosphatases were successfully expressed in E.coli en we have isolated these enzymes in good yield. We study the phosphorylation of a variety of alcohols and carbohydrates using cheap pyrophosphate. We have also shown that these enzymes are able to phosphorylate dihydroxyacetone to dihydroxyacetone phosphate using pyrophosphate. Dihydroxyacetone phosphate is a substrate for aldolase and we have developed a one-pot two-enzyme procedure to synthesize a variety of compounds from aldehydes that up to now could only be synthesized via a number of steps. We have also studied the enantioselective dephosphorylation of racemic DL threonine and DL serine. By a directed evolution approach we have obtained mutants that show an increases enantioselectivity. We expect that these acid phosphatases will be of growing importance for synthetic applications.

References:
N. Tanaka, V . Dumay, Q. Liao, A.J. Lange and R. Wever.
Bromoperoxidase activity of vanadate-substituted acid phosphatases from Shigella flexneri and Salmonella enterica ser. typhimurium.
Eur. J. Biochem. 269 (2002) 2162-2167

N. Tanaka, Z. Hasan, A. F. Hartog, T.van Herk, R.-J. Sanders and R. Wever.
Phosphorylation and dephosphorylation of polyhydroxy compounds by class A bacterial phosphatases.
Org. Biomol. Chem. 1 (2003) 2833-2839

T. van Herk , A. F. Hartog, A.M. Van der Burg and R. Wever.
Regio-selective phosphorylation of carbohydrates and various alcohols by bacterial acid phosphatases; probing the substrate specificity of the enzym from Shigella flexneri.
Adv. Syn. Catal. 347 (2005) 1155-1162

T. van Herk, A. F. Hartog, H. E. Schoemaker and R. Wever.
A simple enzymatic synthesis of dihydroxyacetone phosphate and its use in a cascade reaction for the synthesis of carbohydrates.
J. Org. Chem. 71 (2006) 6244-6247

T. van Herk , A.F. Hartog and R. Wever.
Enzymatic method for producing dihydroxyacetone phosphate.
International patent application. WO 2007/003574 A1

Teunie van Herk, Aloysius F. Hartog, Harald J.Ruijssenaars, Richard Kerkman, Hans E. Schoemaker and Ron Wever.
Optimization of the kinetic resolution of the DL-phosphomonoesters of threonine and serine by random mutagenesis of the acid phosphatase from Salmonella enterica.
Advanced Synthesis and Catalysis (2007), 349, 1349-1352

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