Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
25372 | Journal of Biotechnology | 2007 | 6 Pages |
Xylitol dehydrogenase from Pichia stipitis (PsXDH) is one of the key enzymes for the bio-ethanol fermentation system from xylose. Previously, we constructed the C4 mutant (S96C/S99C/Y102C) with enhanced thermostability by introduction of structural zinc. In this study, for further improvement of PsXDH thermostability, we constructed the appropriate structural zinc-binding loop by comparison with other polyol dehydrogenase family members. A high thermostability of PsXDH was obtained by subsequent site-directed mutagenesis of the structural zinc-binding loop. The best mutant in this study (C4/F98R/E101F) showed a 10.8 °C higher thermal transition temperature (TCD) and 20.8 °C higher half denaturation temperature (T1/2) compared with wild-type.