Abstract
A type II arabinogalactan-degrading enzyme, termed Exo-1,3-Gal, was purified to homogeneity from the culture filtrate of Sphingomonas sp. 24T. It has an apparent molecular mass of 48 kDa by SDS–PAGE. Exo-1,3-Gal was stable from pH 3 to 10 and at temperatures up to 40 °C. The optimum pH and temperature for enzyme activity were pH 6 to 7 and 50 °C, respectively. Galactose was released from β-1,3-d-galactan and β-1,3-d-galactooligosaccharides by the action of Exo-1,3-Gal, indicating that the enzyme was an exo-β-1,3-d-galactanase. Analysis of the reaction products of β-1,3-galactotriose by high-performance anion-exchange chromatography revealed that the enzyme hydrolyzed the substrate in a non-processive mode. Exo-1,3-Gal bypassed the branching points of β-1,3-galactan backbones in larch wood arabinogalactan (LWAG) to produce mainly galactose, β-1,6-galactobiose, and unidentified oligosaccharides 1 and 2 with the molar ratios of 7:19:62:12. Oligosaccharides 1 and 2 were enzymatically determined to be β-1,6-galactotriose and β-1,6-galactotriose substituted with a single arabinofuranose residue, respectively. The ratio of side chains enzymatically released from LWAG was in good agreement with the postulated structure of the polysaccharide previously determined by chemical methods.
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The authors thank Mr. Y. Taniguchi, Osaka Prefecture University, for assistance in screening experiments.
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Sakamoto, T., Tanaka, H., Nishimura, Y. et al. Characterization of an exo-β-1,3-d-galactanase from Sphingomonas sp. 24T and its application to structural analysis of larch wood arabinogalactan. Appl Microbiol Biotechnol 90, 1701–1710 (2011). https://doi.org/10.1007/s00253-011-3219-1
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DOI: https://doi.org/10.1007/s00253-011-3219-1