Two new xylanases with different substrate specificities from the human gut bacterium Bacteroides intestinalis DSM 17393.
Xylan is an abundant plant cell wall polysaccharide and is a dominant component of dietary fiber. Bacteria in the distal human gastrointestinal tract produce xylanase enzymes to initiate the degradation of this complex heteropolymer. These xylanases typically derive from glycoside hydrolase (GH) families 10 and 11; however, analysis of the genome sequence of the xylan-degrading human gut bacterium Bacteroides intestinalis DSM 17393 revealed the presence of two putative GH8 xylanases. In the current study, we demonstrate that the two genes encode enzymes that differ in activity. The xyn8A gene encodes an endoxylanase (Xyn8A), and rex8A encodes a reducing-end xylose-releasing exo-oligoxylanase (Rex8A). Xyn8A hydrolyzed both xylopentaose (X5) and xylohexaose (X6) to a mixture of xylobiose (X2) and xylotriose (X3), while Rex8A hydrolyzed X3 through X6 to a mixture of xylose (X1) and X2. Moreover, rex8A is located downstream of a GH3 gene (xyl3A) that was demonstrated to exhibit β-xylosidase activity and would be able to further hydrolyze X2 to X1. Mutational analyses of putative active site residues of both Xyn8A and Rex8A confirm their importance in catalysis by these enzymes. Recent genome sequences of gut bacteria reveal an increase in GH8 Rex enzymes, especially among the Bacteroidetes, indicating that these genes contribute to xylan utilization in the human gut.
Department of Animal Sciences, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA.
Pei-Ying
PY
Hong
Michael
M
Iakiviak
Dylan
D
Dodd
Meiling
M
Zhang
Roderick I
RI
Mackie
Isaac
I
Cann
D019943:Amino Acid Substitution; D001439:Bacteroides / Q000201:enzymology* / Q000302:isolation & purification; D020134:Catalytic Domain; D004252:DNA Mutational Analysis; D041981:Gastrointestinal Tract / Q000382:microbiology*; D006801:Humans; D013379:Substrate Specificity; D014990:Xylans / Q000378:metabolism*; D014995:Xylosidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism*
D014990:Xylans; D014995:Xylosidases