9LF7

A PAE-hydrolyse Poc14

Summary for 9LF7

• Entry DOI: 10.2210/pdb9lf7/pdb
• Descriptor: Poc14_A, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID (3 entities in total)
• Functional Keywords: esterase, pae-hydrolyzing, hydrolysis, alpha/beita hydrolyse, hydrolase
• Biological source: Erythrobacter cryptus DSM 12079
• Total number of polymer chains: 2
• Total formula weight: 71872.47

Authors

Rong, Z.,Wu, Y.H. (deposition date: 2025-01-08, release date: 2025-02-19, Last modification date: 2025-09-03)

Primary citation

Rong, Z.,Hong, L.G.,Huo, Y.Y.,Li, J.,Zheng, D.Q.,Ha, Y.,Fan, J.,Xu, X.W.,Wu, Y.H.
Molecular Insight Into the Hydrolysis of Phthalate Esters by a Family IV Esterase.
Environ.Microbiol., 27:e70134-e70134, 2025

PubMed Abstract: Phthalate esters (PAEs) are prevalent environmental contaminants, with their biodegradation efficiently driven by microorganisms through ester bond hydrolysis. This study investigates the mechanism of Poc14, a novel family IV esterase, using x-ray crystallography, bioinformatics, biochemistry and site-directed mutagenesis. Phylogenetic analysis classifies Poc14 as a family IV esterase with conserved catalytic motifs crucial for its activity. Poc14 retains over 80% activity at 50°C for 4 h and tolerates up to 5% methanol or DMF, though surfactants like Tweens inhibit its function. Poc14 activity is independent of metal ions, and the addition of EDTA further enhances its activity by approximately 130%. The 1.8 Å crystal structure reveals a CAP domain and two substrate channels. Enzyme assays show Poc14 hydrolyses short-chain diethyl phthalate (DEP) (K = 0.068 mM, V = 9975 μM/min/mg) but not long-chain di(2-ethylhexyl) phthalate (DEHP) due to steric hindrance. Molecular docking assessed Poc14's potential to hydrolyse DEP and DEHP after residue mutations, resulting in the Poc14-AAG variant. Poc14-AAG could hydrolyse one bond of DEHP and diester bonds of DEP. Our study positions Poc14 as a promising enzyme for environmental remediation, with potential for optimising DEHP degradation and exploring dimerisation effects.

PubMed: 40600832
DOI: 10.1111/1462-2920.70134

Experimental method

X-RAY DIFFRACTION (1.8 Å)