A novel bacterial GH30 xylobiohydrolase from Hungateiclostridium clariflavum

Typical bacterial GH30 xylanases are glucuronoxylanases requiring 4-O-methylglucuronic acid (MeGlcA) substitution of a xylan main chain for their action. They do not exhibit a significant activity on neutral xylooligosaccharides, arabinoxylan (AraX), or rhodymenan (Rho). In this work, the biochemical characterization of the bacterial Clocl_1795 xylanase from Hungateiclostridium (Clostridium) clariflavum DSM 19732 (HcXyn30A) is presented. Amino acid sequence analysis of HcXyn30A revealed that the enzyme does not contain amino acids known to be responsible for MeGlcA coordination in the -2b subsite of glucuronoxylanases. This suggested that the catalytic properties of HcXyn30A may differ from those of glucuronoxylanases. HcXyn30A shows similar specific activity on glucuronoxylan (GX) and Rho, while the specific activity on AraX is about 1000 times lower. HcXyn30A releases Xyl(2) as the main product from the non-reducing end of different polymeric and oligomeric substrates. Catalytic properties of HcXyn30A resemble the properties of the fungal GH30 xylobiohydrolase from Acremonium alcalophilum, AaXyn30A. HcXyn30A is the first representative of a prokaryotic xylobiohydrolase. Its unique specificity broadens the catalytic diversity of bacterial GH30 xylanases.

Automated summary

The study presents the biochemical characterization of the bacterial Clocl_1795 xylanase from Hungateiclostridium clariflavum DSM 19732 (HcXyn30A), revealing its unique catalytic properties and specificity compared to glucuronoxylanases. HcXyn30A releases Xyl(2) as the main product, expanding the catalytic diversity of bacterial GH30 xylanases. This enzyme is the first representative of a prokaryotic xylobiohydrolase, showing similarities with the fungal GH30 xylobiohydrolase Acremonium alcalophilum, AaXyn30A.