Purification, characterization and specificity of a new GH family 35 galactosidase from Aspergillus awamori.

Purification, characterization and specificity of a new GH family 35 galactosidase from Aspergillus awamori.

Int J Biol Macromol. 2020 Apr 08;:

Authors: Vidya CH, Gnanesh Kumar BS, Chinmayee CV, Singh SA

Abstract
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.

PMID: 32277984 [PubMed – as supplied by publisher]

Source: Industry