Site-directed incorporation of non-natural amino acid into Streptomyces xylanase.
A key residue, W26G existing around the substrate-binding cleft of beta-1,4-xylanase from Streptomyces olivaceoviridis E-86 (SoXyn10A), was ascertained in this study. Several kinds of W266 mutants substituted by Trp analogs were synthesized by a site-directed incorporation method to elucidate the functional role of this aromatic residue. Significant decreases of hydrolytic activities were observed in all mutants, even though the substitution of the aromatic side chain was not so drastic, suggesting that the W266 residue had a fatal role in the hydrolytic activity of SoXyn10A.
Ichikawa
Takayuki
T
School of Life and Environmental Science, Tsukuba University, 1-1-1 Tennoudai, Tsukuba 305-8577,Japan.
Kuno
Atsushi
A
Taki
Masumi
M
Hohsaka
Takahiro
T
Sisido
Masahiko
M
Kaneko
Satoshi
S
Taira
Kazunari
K
Kobayashi
Hideyuki
H
Hasegawa
Tsunemi
T
D000596:Amino Acids / Q000378:metabolism*; D004187:Disaccharides / Q000378:metabolism; D043364:Endo-1,4-beta Xylanases / Q000378:metabolism*; D006868:Hydrolysis; D013302:Streptomyces / Q000201:enzymology*; D013379:Substrate Specificity; D014364:Tryptophan / Q000031:analogs & derivatives / Q000737:chemistry
C414004:4-methylumbelliferyl-xylobioside; D000596:Amino Acids; D004187:Disaccharides; D014364:Tryptophan; D043364:Endo-1,4-beta Xylanases