Purification and Partial Characterization of a Novel β-1,3-Endoglucanase from Streptomyces rutgersensis

• Published in: Protein Journal Vol. 32; no. 5; pp. 411 - 417
• Main Authors: Javmen, Artur, Grigiškis, Saulius, Rudenkov, Mark, Mauricas, Mykolas
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.06.2013
• Subjects: Animal Anatomy; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Biochemistry; Bioorganic Chemistry; Cellulase - chemistry; Cellulase - isolation & purification; Cellulase - metabolism; Chemistry; Chemistry and Materials Science; Enzyme Stability; Histology; Hot Temperature; Hydrogen-Ion Concentration; Isoelectric Point; Molecular Weight; Morphology; Organic Chemistry; Saccharomyces cerevisiae; Streptomyces; Streptomyces - chemistry; Streptomyces - enzymology; Substrate Specificity; Endoglucanase; Purification; β-Glucan
• ISSN: 1572-3887; 1573-4943; 1875-8355
• DOI: 10.1007/s10930-013-9500-7

A β-1,3-endoglucanase produced by Streptomyces rutgersensis was purified to a homogeneity by the fractional precipitation with ammonium sulfate, ion exchange chromatography on Q-Sepharose and hydrophobic chromatography on Butyl Sepharose. A typical procedure provided 11.74-fold purification with 12.53 % yield. SDS-PAGE of the purified protein showed one protein band. The exact molecular mass of the enzyme obtained by mass spectrometry was 41.25 kDa; the isoelectric point was between pH 4.2–4.4. The optimal β-glucanase catalytic activity was at pH 7 and 50 °C. An enzyme was only active toward glucose polymers containing β-1,3 linkages and hydrolyzed Saccharomyces cerevisiae cell wall β-glucan in an endo-like way: reaction products were different molecular size β-glucans, which were larger than glucose.