 |
PDBsum entry 4zrs
|
|
|
|
References listed in PDB file
|
 |
|
Key reference
|
|
|
Title
|
|
Enhancing the thermostability of feruloyl esterase estf27 by directed evolution and the underlying structural basis.
|
|
|
Authors
|
|
L.C.Cao,
R.Chen,
W.Xie,
Y.H.Liu.
|
|
|
Ref.
|
|
J Agric Food Chem, 2015,
63,
8225-8233.
[DOI no: ]
|
|
|
PubMed id
|
|
|
|
|
|
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.
|
|
|
|
|
|
|
