Purification and general properties of L-fucose dehydrogenase from Pullularia pullalans
Arq. biol. tecnol
;
32(3): 575-87, ago. 1989. ilus, tab
Article
in English
| LILACS
| ID: lil-74261
RESUMEN
An inducible L-fucose dehydrogenase from the yeast-like fungus Pullularia pullulans was purified and studied. The enzyme catalyses the oxidation of L-fucose to L-fuconic acid possibly throught its unstable L-funcono-lactone. the enzyme was purified to hemogeneity by a sequence of protamine sulfate treatment, ammonium sulfate fractionation, gel filtration on Sephadex G-100, and DEAE-cellulose chromatography. This sequence resulted in an 87 fold purification. The apparent molecular weight determined by gel filtration and SDS polyacrilamide gel electrophoresis was 40 000. the dehydrogenase was NAD-especific and showed a high sugar substrate specificity. Of several D-and L-aldoses tested only L-fucose, L-galactose and-arabinose served as substrates. The maximum velocity for the reaction was at 33- and pH 10.5. Under these conditions, the Km values, and D-arabinose, respectively. The enzyme was strongly inhibited by thiol reagents, heavy metal ions, and was not particularly stable. At 4- it rapidly los activity, but remained active for two months when maintained at -20-. The enzyme has been used to quantativate L-fucose
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Index:
LILACS (Americas)
Main subject:
Carbohydrate Dehydrogenases
/
Fungi
Language:
English
Journal:
Arq. biol. tecnol
Journal subject:
Biology
/
Biotechnology
Year:
1989
Type:
Article
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