Evaluation of fluoropyruvate as nucleophile in reactions catalysed by N-acetyl neuraminic acid lyase variants: scope, limitations and stereoselectivity. Issue 1 (5th November 2015)

• Record Type: Journal Article
• Authors: Stockwell, Jennifer; Daniels, Adam D.; Windle, Claire L.; Harman, Thomas A.; Woodhall, Thomas; Lebl, Tomas; Trinh, Chi H.; Mulholland, Keith; Pearson, Arwen R.; Berry, Alan; Nelson, Adam
• Is Part Of: Organic & biomolecular chemistry. Volume 14:Issue 1(2016)
• Page Start: 105
• Page End: 112
• Publication Date: 2015-11-05
• Subjects: Chemistry, Organic -- Periodicals; Bioorganic chemistry -- Periodicals; Chemistry, Physical organic -- Periodicals; 547
• DOI: 10.1039/c5ob02037a
• Languages: English
• ISSNs: 1477-0520

Abstract

Abstract : The stereochemical course of aldolase-catalysed reaction between fluoropyruvate and aldehydes is described. Abstract : The catalysis of reactions involving fluoropyruvate as donor by N -acetyl neuraminic acid lyase (NAL) variants was investigated. Under kinetic control, the wild-type enzyme catalysed the reaction between fluoropyruvate and N -acetyl mannosamine to give a 90 : 10 ratio of the (3 R, 4 R )- and (3 S, 4 R )-configured products; after extended reaction times, equilibration occurred to give a 30 : 70 mixture of these products. The efficiency and stereoselectivity of reactions of a range of substrates catalysed by the E192N, E192N/T167V/S208V and E192N/T167G NAL variants were also studied. Using fluoropyruvate and (2 R, 3 S )- or (2 S, 3 R )-2, 3-dihydroxy-4-oxo- N, N -dipropylbutanamide as substrates, it was possible to obtain three of the four possible diastereomeric products; for each product, the ratio of anomeric and pyranose/furanose forms was determined. The crystal structure of S. aureus NAL in complex with fluoropyruvate was determined, assisting rationalisation of the stereochemical outcome of C–C bond formation.