A family 8 glycoside hydrolase from Bacillus halodurans C-125 (BH2105) is a reducing end xylose-releasing exo-oligoxylanase.
The gene encoding family 8 glycoside hydrolases from Bacillus halodurans C-125 (BH2105), an alkalophilic bacterium with a known genomic sequence, was expressed in Escherichia coli. The protein was expressed with the intact N-terminal sequence, suggesting that it did not possess a signal peptide and that it was an intracellular enzyme. The recombinant enzyme showed no hydrolytic activity on xylan, whereas it had been annotated as xylanase Y. It hydrolyzed xylooligosaccharide whose degree of polymerization is greater than or equal to 3 in an exo-splitting manner with anomeric inversion, releasing the xylose unit at the reducing end. Judging from its substrate specificity and reaction mechanism, we named the enzyme reducing end xylose-releasing exo-oligoxylanase (Rex). Rex was found to utilize only the beta-anomer of the substrate to form beta-xylose and alpha-xylooligosaccharide. The optimum pH of the enzymatic reaction (6.2-7.3) was found in the neutral range, a range beneficial for intracellular enzymes. The genomic sequence suggests that B. halodurans secretes two endoxylanases and possesses two alpha-arabinofuranosidases, one alpha-glucuronidase, and three beta-xylosidases intracellularly in addition to Rex. The extracellular enzymes supposedly hydrolyze xylan into arabino/glucurono-xylooligosaccharides that are then transported into the cells. Rex may play a role as a key enzyme in intracellular xylan metabolism in B. halodurans by cleaving xylooligosaccharides that were produced by the action of other intracellular enzymes from the arabino/glucurono-xylooligosaccharides.
Honda
Yuji
Y
National Food Research Institute, 2-1-12 Kannondai, Tsukuba, Ibaraki 305-8642, Japan.
Kitaoka
Motomitsu
M
D001407:Bacillus / Q000201:enzymology*; D002240:Carbohydrate Sequence; D002855:Chromatography, Thin Layer; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D043364:Endo-1,4-beta Xylanases / Q000737:chemistry / Q000502:physiology*; D004926:Escherichia coli / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000737:chemistry* / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008954:Models, Biological; D008956:Models, Chemical; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011108:Polymers / Q000737:chemistry; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000737:chemistry; D013696:Temperature; D014312:Trisaccharides / Q000737:chemistry; D014990:Xylans / Q000737:chemistry; D014994:Xylose / Q000737:chemistry; D014995:Xylosidases / Q000737:chemistry
D011108:Polymers; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D014312:Trisaccharides; D014990:Xylans; D004247:DNA; D014994:Xylose; D006026:Glycoside Hydrolases; D014995:Xylosidases; C096734:alpha-glucuronidase; C026579:exo-1,4-beta-D-xylosidase; D043364:Endo-1,4-beta Xylanases