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Cover Picture: Enzymatic Thioxyloside Synthesis: Characterization of Thioglycoligase Variants Identified from A Site‐Saturation Mutagenesis Library of Bacillus Circulans Xylanase (ChemBioChem 4/2010)
Author(s) -
Armstrong Zachary,
Reitinger Stephan,
Kantner Terrence,
Withers Stephen G.
Publication year - 2010
Publication title -
chembiochem
Language(s) - English
Resource type - Reports
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201090010
Subject(s) - saturated mutagenesis , bacillus circulans , mutagenesis , chemistry , active site , protein engineering , biochemistry , enzyme , site directed mutagenesis , stereochemistry , mutant , gene
The cover picture shows the reaction intermediate formed at the active site of a mutant of the Bacillus circulans xylanase that has been optimized for the synthesis of thioglycoside products. Saturation mutagenesis of the codon corresponding to its catalytic acid/base residue generated a library of transformants. Screening of crude cell extracts in situ for activity by TLC analysis and for protein concentration by active‐site titration identified the best mutants without the need for purification. The thioglycoside products are resistant to enzymatic degradation, thus they serve as inhibitors of the wild‐type enzyme for structural studies. For more information, see the paper by S. G. Withers et al. on p. 533 ff.