6EEJ

Streptomyces bingchenggensis Aldolase-Dehydratase in covalent complex with dienone product.

6EEJ の概要

• エントリーDOI: 10.2210/pdb6eej/pdb
• 分子名称: Acetoacetate decarboxylase, (3E,5E)-6-(4-nitrophenyl)-2-oxohexa-3,5-dienoic acid, FORMIC ACID, ... (5 entities in total)
• 機能のキーワード: secondary metabolism, aldolase, dehydratase, lyase
• 由来する生物種: Streptomyces bingchenggensis (strain BCW-1)
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 113998.25

構造登録者

Mydy, L.S.,Silvaggi, N.R. (登録日: 2018-08-14, 公開日: 2019-08-14, 最終更新日: 2024-10-09)

主引用文献

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

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: 31524380
DOI: 10.1021/acs.biochem.9b00652

実験手法

X-RAY DIFFRACTION (1.892 Å)