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Characterization of novel α-galactosidase in glycohydrolase family 97 from Bacteroides thetaiotaomicron and its immobilization for industrial application
- Source :
- International Journal of Biological Macromolecules. 152:727-734
- Publication Year :
- 2020
- Publisher :
- Elsevier BV, 2020.
-
Abstract
- Bacteroides thetaiotaomicron (B. thetaiotaomicron), which resides in the human intestinal tract, has a number of carbohydrate enzymes, including glycoside hydrolase (GH) family 97. Only a few GH 97 enzymes have been characterized to date. In this study, a novel α-galactosidase (Bt_3294) was cloned from B. thetaiotaomicron, expressed in Escherichia coli, and purified using affinity chromatography. This novel enzyme showed optimal activity at 60 °C and pH 7.0. Enzyme activity was reduced by 94.4% and 95.7% in the presence of 5 mM Ca2+ and Fe2+, respectively. It is interesting that Bt_3294 specifically hydrolyzed shorter α-galactosyl oligosaccharides, such as melibiose and raffinose. The D-values of Bt_3294 at 40 °C and 50 °C were about 107 and 6 min, respectively. After immobilization of Bt_3294, the D-values at 40 °C and 50 °C were about 37.6 and 29.7 times higher than those of the free enzyme, respectively. As a practical application, the immobilized Bt_3294 was used to hydrolyze raffinose family oligosaccharides (RFOs) in soy milk, decreasing the RFOs by 98.9%.
- Subjects :
- Glycoside Hydrolases
Immobilized enzyme
Oligosaccharides
02 engineering and technology
Biochemistry
03 medical and health sciences
chemistry.chemical_compound
Raffinose
Affinity chromatography
Structural Biology
Glycoside hydrolase
Melibiose
Molecular Biology
030304 developmental biology
chemistry.chemical_classification
0303 health sciences
biology
General Medicine
Hydrogen-Ion Concentration
Enzymes, Immobilized
021001 nanoscience & nanotechnology
Enzyme assay
Soy Milk
Bacteroides thetaiotaomicron
Enzyme
chemistry
alpha-Galactosidase
biology.protein
0210 nano-technology
Subjects
Details
- ISSN :
- 01418130
- Volume :
- 152
- Database :
- OpenAIRE
- Journal :
- International Journal of Biological Macromolecules
- Accession number :
- edsair.doi.dedup.....c3a3bdb3b67efa735fd83ef428de768e