4ZRS

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

Summary for 4ZRS

• Entry DOI: 10.2210/pdb4zrs/pdb
• Descriptor: Esterase, GLYCEROL (3 entities in total)
• Functional Keywords: feruloyl esterase, metagenomic library, hydrolase
• Biological source: uncultured bacterium
• Total number of polymer chains: 2
• Total formula weight: 67091.69

Authors

Xie, W.,Chen, R.,Cao, L.,Liu, Y. (deposition date: 2015-05-12, release date: 2016-02-03, Last modification date: 2023-11-08)

Primary citation

Cao, L.C.,Chen, R.,Xie, W.,Liu, Y.H.
Enhancing the Thermostability of Feruloyl Esterase EstF27 by Directed Evolution and the Underlying Structural Basis
J.Agric.Food Chem., 63:8225-8233, 2015

PubMed 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.

PubMed: 26329893
DOI: 10.1021/acs.jafc.5b03424

Experimental method

X-RAY DIFFRACTION (2 Å)