A C-Terminal Proline-Rich Sequence Simultaneously Broadens the Optimal Temperature and pH Ranges and Improves the Catalytic Efficiency of Glycosyl Hydrolase Family 10 Ruminal Xylanases

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    Multiple sequence alignment of deduced XynA (AGC54685) with 12 xylanase homologs from the same sheep rumen (XynB, AGC54683), Bacteroides intestinalis CAG:564 (WP_021967563), Bacteroides cellulosilyticus CAG:158 (WP_022209201), Bacteroides eggerthii (WP_017140705), uncultured ruminal organisms (ADX05679, ADN44261, and ADX05692), Hymenobacter norwichensis (WP_022824491), Niastella koreensis GR20-10 (YP_005006534), Haliscomenobacter hydrossis DSM 1100 (YP_004446188), Spirosoma panaciterrae (WP_020601301), and Galbibacter sp. ck-I2-15 (WP_008991477) using the ClustalW program (Fig. S1); SDS–PAGE analysis of purified recombinant XynA, XynA-Tr, XynB, XynB-Fu, XynA-E151Q, and XynA-Tr-E151Q (Fig. S2); ability of XynA-E151Q to bind to soluble and insoluble polysaccharides on the native PAGE gel (Fig. S3); raw curves for binding of XynA and XynA-Tr to xylooligosaccharides, measured by ITC (Fig. S4); primers used in this study (Table S1).

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