Various mutations by using yeast gene for protein-engineering.
For construct an efficient-ethanol fermentation system from xylose by using yeast, several protein-engineering of xylitol dehydrogenase (XDH) were carried out. At first, we created a NADP+-dependent XDH by multiple site-directed mutagenesis. Furthermore, we succeeded to improve the thermostability by introduction of the structural zinc atom into XDH which does not possess a second zinc atom natively. Finally, the introduction of a structural zinc atom into the NADP+-dependent XDH mutant increased the thermostability of this mutant and improved further the catalytic efficiency with NADP+.
Watanabe
Seiya
S
Institute of Advanced Energy, Kyoto University, Uji 611-0011, Japan.
Kodaki
Tsutomu
T
Makino
Keisuke
K
D003067:Coenzymes / Q000378:metabolism; D050542:D-Xylulose Reductase / Q000378:metabolism; D004795:Enzyme Stability; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D050505:Mutant Proteins / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D009249:NADP / Q000378:metabolism; D010843:Pichia / Q000201:enzymology / Q000235:genetics*; D015202:Protein Engineering; D013379:Substrate Specificity; D044366:Transition Temperature; D015032:Zinc / Q000378:metabolism
D003067:Coenzymes; D050505:Mutant Proteins; D009249:NADP; D050542:D-Xylulose Reductase; D015032:Zinc