Characterization of the bga1‐encoded glycoside hydrolase family 35 β‐galactosidase of Hypocrea jecorina with galacto‐β‐d‐galactanase activity

• Published in: The FEBS journal 2007-04, Vol.274 (7), p.1691-1700
• Main Authors: Gamauf, Christian, Marchetti, Martina, Kallio, Jarno, Puranen, Terhi, Vehmaanperä, Jari, Allmaier, Günter, Kubicek, Christian P, Seiboth, Bernhard
• Format: Article
• Language: English
• Subjects: beta-Galactosidase - chemistry; beta-Galactosidase - isolation & purification; beta-Galactosidase - metabolism; Catalysis; Disaccharides - chemistry; Disaccharides - metabolism; Enzyme Stability; Fungal Proteins - chemistry; Fungal Proteins - isolation & purification; Fungal Proteins - metabolism; galactanase; Galactans - chemistry; Galactans - metabolism; Galactose - analogs & derivatives; Galactose - chemistry; Galactose - metabolism; Glycoproteins - chemistry; Glycoproteins - metabolism; Glycoside Hydrolases - chemistry; Glycoside Hydrolases - metabolism; Glycosylation; Hydrogen-Ion Concentration; Hypocrea - chemistry; Hypocrea - enzymology; Hypocrea jecorina; Kinetics; Lactose - chemistry; Lactose - metabolism; Lactulose - chemistry; Lactulose - metabolism; Molecular Weight; Nitrophenylgalactosides - chemistry; Nitrophenylgalactosides - metabolism; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Substrate Specificity; Temperature; transglycosylation; β‐galactosidase
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/j.1742-4658.2007.05714.x

The extracellular bga1‐encoded β‐galactosidase of Hypocrea jecorina (Trichoderma reesei) was overexpressed under the pyruvat kinase (pki1) promoter region and purified to apparent homogeneity. The monomeric enzyme is a glycoprotein with a molecular mass of 118.8 ± 0.5 kDa (MALDI‐MS) and an isoelectric point of 6.6. Bga1 is active with several disaccharides, e.g. lactose, lactulose and galactobiose, as well as with aryl‐ and alkyl‐β‐d‐galactosides. Based on the catalytic efficiencies, lactitol and lactobionic acid are the poorest substrates and o‐nitrophenyl‐β‐d‐galactoside and lactulose are the best. The pH optimum for the hydrolysis of galactosides is ∼ 5.0, and the optimum temperature was found to be 60 °C. Bga1 is also capable of releasing d‐galactose from β‐galactans and is thus actually a galacto‐β‐d‐galactanase. β‐Galactosidase is inhibited by its reaction product d‐galactose and the enzyme also shows a significant transferase activity which results in the formation of galacto‐oligosaccharides.