EC Number |
Protein Variants |
Reference |
---|
1.1.1.271 | C109A |
lower Km than wild-type enzyme |
389518 |
1.1.1.271 | C109S |
active site mutant, produces GDP-6-deoxy-D-altrose as its major product (75%) besides GDP-L-fucose (25%) out of GDP-4-keto-6-deoxy-D-mannose |
698467 |
1.1.1.271 | C109S |
produces GDP-6-deoxy-D-altrose as its major product, thus C-3'' epimerization occurs first and premature reduction of the GDP-4-keto-6-deoxy-D-altrose intermediate becomes competitive with GDP-L-fucose production. Results in the appearance of a kinetic isotope effect when [3''-2H]-GDP-6-deoxy-4-ketomannose is used as a substrate. The mutant catalyzes a rapid wash-in of solvent derived deuterium into the C-5'' position of GDP-fucose in the presence of NADP+ |
698467 |
1.1.1.271 | H179N |
lower Km than wild-type enzyme |
389518 |
1.1.1.271 | H179Q |
is inactive, readily catalyzes the wash-out of deuterium from the C-3'' position of [3''-2H]-GDP-6-deoxy-4-keto-mannose |
698467 |
1.1.1.271 | H179Q |
the mutant catalyzes isotope exchange into starting material indicates that its active site has not been dramatically perturbed and it is still able to bind substrate and catalyze deprotonation events |
698467 |
1.1.1.271 | K140R |
higher Km than wild-type enzyme |
389518 |
1.1.1.271 | K140S |
higher Km than wild-type enzyme |
389518 |
1.1.1.271 | more |
effective GDP-L-fucose production in recombinant Escherichia coli by expression of GDP-D-mannose-4,6-dehydratase and GDP-4-keto-6-deoxymannose 3,5-epimerase 4-reductase, method optimization at 25°C and 0.1 mM isopropyl-beta-D-thioglucopyranoside, overview |
677753 |
1.1.1.271 | R187A |
lower Km than wild-type enzyme |
389518 |