6EEJ
Streptomyces bingchenggensis Aldolase-Dehydratase in covalent complex with dienone product.
Summary for 6EEJ
| Entry DOI | 10.2210/pdb6eej/pdb |
| Descriptor | Acetoacetate decarboxylase, (3E,5E)-6-(4-nitrophenyl)-2-oxohexa-3,5-dienoic acid, FORMIC ACID, ... (5 entities in total) |
| Functional Keywords | secondary metabolism, aldolase, dehydratase, lyase |
| Biological source | Streptomyces bingchenggensis (strain BCW-1) |
| Total number of polymer chains | 4 |
| Total formula weight | 113998.25 |
| Authors | Mydy, L.S.,Silvaggi, N.R. (deposition date: 2018-08-14, release date: 2019-08-14, Last modification date: 2024-10-09) |
| Primary citation | Mydy, L.S.,Hoppe, R.W.,Hagemann, T.M.,Schwabacher, A.W.,Silvaggi, N.R. Mechanistic Studies of theStreptomyces bingchenggensisAldolase-Dehydratase: Implications for Substrate and Reaction Specificity in the Acetoacetate Decarboxylase-like Superfamily. Biochemistry, 58:4136-4147, 2019 Cited by PubMed Abstract: The acetoacetate decarboxylase-like superfamily (ADCSF) is a little-explored group of enzymes that may contain new biocatalysts. The low level of sequence identity (∼20%) between many ADCSF enzymes and the confirmed acetoacetate decarboxylases led us to investigate the degree of diversity in the reaction and substrate specificity of ADCSF enzymes. We have previously reported on Sbi00515, which belongs to Family V of the ADCSF and functions as an aldolase-dehydratase. Here, we more thoroughly characterize the substrate specificity of Sbi00515 and find that aromatic, unsaturated aldehydes yield lower and higher values compared to those of other small electrophilic substrates in the condensation reaction. The roles of several active site residues were explored by site-directed mutagenesis and steady state kinetics. The lysine-glutamate catalytic dyad, conserved throughout the ADCSF, is required for catalysis. Tyrosine 252, which is unique to Sbi00515, is hypothesized to orient the incoming aldehyde in the condensation reaction. Transient state kinetics and an intermediate-bound crystal structure aid in completing a proposed mechanism for Sbi00515. PubMed: 31524380DOI: 10.1021/acs.biochem.9b00652 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.892 Å) |
Structure validation
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