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Purification and Characterization of 1,3-β-<scp>D</scp>-Glucan Phosphorylase fromOchromonas danica
- Source :
- Bioscience, Biotechnology, and Biochemistry. 77:1949-1954
- Publication Year :
- 2013
- Publisher :
- Informa UK Limited, 2013.
-
Abstract
- 1,3-β-D-glucan phosphorylase (BGP) is an enzyme that catalyzes the reversible phosphorolysis of 1,3-β-glucosidic linkages to form α-D-glucose 1-phosphate (G1P). Here we report on the purification and characterization of BGP from Ochromonas danica (OdBGP). The purified enzyme preparation showed three bands (113, 118, and 124 kDa) on SDS-polyacrylamide gel electrophoresis. The optimum pH and temperature were 5.5 and 25 °C-30 °C. OdBGP phosphorolysed laminaritriose, larger laminarioligosaccharides, and laminarin, but not laminaribiose. In the synthesis reaction, laminarin and laminarioligosaccharides served as good acceptors, but OdBGP did not act on glucose. Kinetic analysis indicated that the phosphorolysis reaction of OdBGP follows a sequential Bi Bi mechanism. The equilibrium of the enzymatic reaction indicated that OdBGP favors the reaction in the synthetic direction. Overnight incubation of OdBGP with laminaribiose and G1P resulted in the formation of precipitates, which were probably 1,3-β-glucans.
- Subjects :
- beta-Glucans
Phosphorylases
Kinetics
Biology
Disaccharides
Ochromonas
Applied Microbiology and Biotechnology
Biochemistry
Chemical synthesis
Substrate Specificity
Analytical Chemistry
Laminarin
chemistry.chemical_compound
Glycogen phosphorylase
Molecular Biology
Laminaribiose
Phosphorolysis
chemistry.chemical_classification
Gel electrophoresis
Organic Chemistry
Glucosephosphates
Temperature
General Medicine
Hydrogen-Ion Concentration
Enzyme
chemistry
Proteoglycans
Biotechnology
Subjects
Details
- ISSN :
- 13476947 and 09168451
- Volume :
- 77
- Database :
- OpenAIRE
- Journal :
- Bioscience, Biotechnology, and Biochemistry
- Accession number :
- edsair.doi.dedup.....843b27e24db5f3addba2c3f98e09f0ce