Purification, characterization and specificity of a new GH family 35 galactosidase from Aspergillus awamori.
IR@CSIR-CFTRI
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Relation |
http://ir.cftri.com/14662/
https://doi.org/10.1016/j.ijbiomac.2020.04.013 0141-8130/ |
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Title |
Purification, characterization and specificity of a new GH family 35 galactosidase from Aspergillus awamori. |
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Creator |
Vidya, C. H.
Gnanesh Kumar, B. S. Chinmayee, Cirium V. Sridevi Annapurna, Singh |
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Subject |
04 Fermentation Technology
05 Enzymes |
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Description |
Galactosidases, ubiquitous in nature, are complex carbohydrate-active enzymes and find extensive applications in food, pharma, and biotechnology industries. The present study deals with the production of galactosidases from fungi by solid-state fermentation. Fifteen fungi were screened and Aspergillus awamori (MTCC 548), exhibited the highest α and β-galactosidase activities of 75.11±0.29 U/g and 155.34± 1.26 U/g, respectively. 30 g of wheat bran substituted with 6% defatted soy flour, at 28°C, pH 5.0 for 120 h, was established as the optimum production conditions by one-factor approach. The enzyme was purified to homogeneity with an apparent mass of 118 ± 2 kDa by ammonium sulfate precipitation (50–80%), ion exchange and hydrophobic interaction chromatography. Specific activities for α and β-galactosidase were 22 and 74 U/mg, respectively. Optimum temperature and pH ranges for enzyme activities were 55–60 °C, 5.0–5.5, respectively. The thermal inactivation mid-point was 65 °C. The purified enzyme not only exhibited α and β-galactosidase activities, but also exhibited β-xylosidase and β-glucosidase activities, indicating the enzyme has broad substrate specificity. Sequence analysis by in-gel digestion and tandem mass spectrometry (MS/MS) revealed that the enzyme was a probable β-galactosidase A, belonging to glycoside hydrolase 35 family, and is being reported for the first time. |
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Date |
2020
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Type |
Article
PeerReviewed |
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Format |
pdf
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Language |
en
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Identifier |
http://ir.cftri.com/14662/1/International%20Journal%20of%20Biological%20Macromolecules%20156%20%282020%29%20885%E2%80%93895.pdf
Vidya, C. H. and Gnanesh Kumar, B. S. and Chinmayee, Cirium V. and Sridevi Annapurna, Singh (2020) Purification, characterization and specificity of a new GH family 35 galactosidase from Aspergillus awamori. International Journal of Biological Macromolecules, 156. pp. 885-895. ISSN 0141-8130 |
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