2V3W
Crystal structure of the benzoylformate decarboxylase variant L461A from Pseudomonas putida
Summary for 2V3W
| Entry DOI | 10.2210/pdb2v3w/pdb |
| Related | 1BFD 1MCZ 1PI3 1PO7 1Q6Z 1YNO |
| Descriptor | BENZOYLFORMATE DECARBOXYLASE, MAGNESIUM ION, SULFATE ION, ... (5 entities in total) |
| Functional Keywords | lyase, calcium, magnesium, flavoprotein, thiamine pyrophosphate, rational protein design, aromatic hydrocarbons catabolism, thdp-dependent, mandelate pathway, metal-binding, decarboxylase, carboligation |
| Biological source | PSEUDOMONAS PUTIDA |
| Total number of polymer chains | 4 |
| Total formula weight | 227674.01 |
| Authors | Gocke, D.,Walter, L.,Gauchenova, K.,Kolter, G.,Knoll, M.,Berthold, C.L.,Schneider, G.,Pleiss, J.,Mueller, M.,Pohl, M. (deposition date: 2007-06-25, release date: 2008-01-22, Last modification date: 2023-12-13) |
| Primary citation | Gocke, D.,Walter, L.,Gauchenova, K.,Kolter, G.,Knoll, M.,Berthold, C.L.,Schneider, G.,Pleiss, J.,Muller, M.,Pohl, M. Rational Protein Design of Thdp-Dependent Enzymes-Engineering Stereoselectivity. Chembiochem, 9:406-, 2008 Cited by PubMed Abstract: Benzoylformate decarboxylase (BFD) from Pseudomonas putida is an exceptional thiamin diphosphate-dependent enzyme, as it catalyzes the formation of (S)-2-hydroxy-1-phenylpropan-1-one from benzaldehyde and acetaldehyde. This is the only currently known S-selective reaction (92 % ee) catalyzed by this otherwise R-selective class of enzymes. Here we describe the molecular basis of the introduction of S selectivity into ThDP-dependent decarboxylases. By shaping the active site of BFD through the use of rational protein design, structural analysis, and molecular modeling, optimal steric stabilization of the acceptor aldehyde in a structural element called the S pocket was identified as the predominant interaction for adjusting stereoselectivity. Our studies revealed Leu461 as a hot spot for stereoselectivity in BFD. Exchange to alanine and glycine resulted in variants that catalyze the S-stereoselective addition of larger acceptor aldehydes, such as propanal with benzaldehyde and its derivatives-a reaction not catalyzed by the wild-type enzyme. Crystal structure analysis of the variant BFDL461A supports the modeling studies. PubMed: 18224647DOI: 10.1002/CBIC.200700598 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.2 Å) |
Structure validation
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