Hydrophobic interaction network analysis for thermostabilization of a mesophilic xylanase.
One widely known drawback of enzymes is their instability in diverse conditions. The thermostability of enzymes is particularly relevant for industrial applications because operation at high temperatures has the advantage of a faster reaction rate. Protein stability is mainly determined in this study by intra-molecular hydrophobic interactions that have a collective and 3-dimensional clustering effect. To interpret the thermostability of enzymes, network analysis was introduced into the protein structure, and a network parameter of structural hierarchy, k of k-clique, was used to discern more developed hydrophobic interaction clusters in the protein structure. The favorable clustering conformations of hydrophobic residues, which seemed to be important for protein thermostability, were discovered by the application of a network analysis to hydrophobic interactions of GH11 xylanases. Coordinating higher k-clique hydrophobic interaction clusters through the site-directed mutagenesis of the model enzyme, Bacillus circulans xylanase, stabilized the local structure and thus improved thermostability, such that the enzyme half-life and melting temperature increased by 78 fold and 8.8 °C, respectively. This study highlights the advantages of interpreting collective hydrophobic interaction patterns and their structural hierarchy and the possibility of applying network analysis to the thermostabilization of enzymes.
Interdisciplinary Program of Bioengineering, Seoul National University, Seoul 151-744, Republic of Korea.
Taeho
T
Kim
Jeong Chan
JC
Joo
Young Je
YJ
Yoo
D001407:Bacillus / Q000201:enzymology*; D001709:Biotechnology / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D043364:Endo-1,4-beta Xylanases / Q000235:genetics / Q000378:metabolism*; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D058503:Mesophyll Cells / Q000201:enzymology*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D055550:Protein Stability; D013696:Temperature
D017931:DNA Primers; D043364:Endo-1,4-beta Xylanases