Structure of hydroxynitrile lyase from Manihot esculenta in complex with substrates acetone and chloroacetone: implications for the mechanism of cyanogenesis.
The crystal structures of hydroxynitrile lyase from Manihot esculenta (MeHNL) complexed with the native substrate acetone and substrate analogue chloroacetone have been determined and refined at 2.2 A resolution. The substrates are positioned in the active site by hydrogen-bond interactions of the carbonyl O atom with Thr11 OG, Ser80 OG and, to a lesser extent, Cys81 SG. These studies support a mechanism for cyanogenesis as well as for the stereospecific MeHNL-catalyzed formation of (S)-cyanohydrins, which closely resembles the base-catalyzed chemical reaction of HCN with carbonyl compounds.
Lauble
H
H
Universität Stuttgart, Institut für Organische Chemie, Pfaffenwaldring 55, D-70569 Stuttgart, Germany. peterlauble@t-online.de
Förster
S
S
Miehlich
B
B
Wajant
H
H
Effenberger
F
F
D000096:Acetone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D000446:Aldehyde-Lyases / Q000737:chemistry* / Q000378:metabolism*; D003460:Crystallization; D018360:Crystallography, X-Ray; D008958:Models, Molecular; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D019685:Rosales / Q000201:enzymology*; D013379:Substrate Specificity
D011994:Recombinant Proteins; D000096:Acetone; C006973:chloroacetone; D000446:Aldehyde-Lyases; C079359:hydroxymandelonitrile lyase