PDBsum entry 4zrs

Hydrolase

PDB id

4zrs

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Contents

			Protein chains

					292 a.a.

			Ligands

					GOL ×2

			Waters ×615

PDB id:

4zrs

Links

Name:

Hydrolase

Title:

Crystal structure of a cloned feruloyl esterase from a soil metagenomic library

Structure:

Esterase. Chain: a, b. Synonym: feruloyl esterase. Engineered: yes. Mutation: yes

Source:

Uncultured bacterium. Organism_taxid: 77133. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

Resolution:

2.00Å

R-factor:

0.152

R-free:

0.199

Authors:

W.Xie,R.Chen,L.Cao,Y.Liu

Key ref:

L.C.Cao et al. (2015). Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis. J Agric Food Chem, 63, 8225-8233. PubMed id: 26329893 DOI: 10.1021/acs.jafc.5b03424

Date:

12-May-15

Release date:

03-Feb-16

PROCHECK

	Headers

	References

Protein chains

E7DJY5 (E7DJY5_9BACT) - Esterase from uncultured bacterium

Seq: Struc:					290 a.a. 292 a.a.*

Key:

PfamA domain

Secondary structure

* PDB and UniProt seqs differ at 7 residue positions (black crosses)

DOI no: 10.1021/acs.jafc.5b03424	J Agric Food Chem 63:8225-8233 (2015)
PubMed id: 26329893

Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis.
L.C.Cao, R.Chen, W.Xie, Y.H.Liu.

ABSTRACT

To improve the thermostability of EstF27, two rounds of random mutagenesis were performed. A thermostable mutant, M6, with six amino acid substitutions was obtained. The half-life of M6 at 55 °C is 1680 h, while that of EstF27 is 0.5 h. The Kcat/Km value of M6 is 1.9-fold higher than that of EstF27. The concentrations of ferulic acid released from destarched wheat bran by EstF27 and M6 at their respective optimal temperatures were 223.2 ± 6.8 and 464.8 ± 11.9 μM, respectively. To further understand the structural basis of the enhanced thermostability, the crystal structure of M6 is determined at 2.0 Å. Structural analysis shows that a new disulfide bond and hydrophobic interactions formed by the mutations may play an important role in stabilizing the protein. This study not only provides us with a robust catalyst, but also enriches our knowledge about the structure-function relationship of feruloyl esterase.