Abstract
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 Xyl2 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.
Key points
• Bacterial GH30 xylobiohydrolase from H. clariflavum (HcXyn30A) has been characterized.
• HcXyn30A releases xylobiose from the non-reducing end of different substrates.
• HcXyn30A is the first representative of bacterial xylobiohydrolase.
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Acknowledgments
We are very grateful to Prof. Carlos M.G.A. Fontes (NZYTech, Lisboa, Portugal) for a gift of HcXyn30A. We thank the Slovak Research and Development Agency (the contract No. APVV-0602-12), and Scientific Grant Agency (the contract No. 2/0016/18) for the financial support.
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This work was supported by the Slovak Research and Development Agency under the contract No. APVV-0602-12, and by Scientific Grant Agency under the contract No. 2/0016/18.
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PB and KŠ conceived research, KŠ and VP performed experiments and analyzed data, and KŠ wrote the manuscript with inputs from all authors. All authors read and approved the manuscript.
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Šuchová, K., Puchart, V. & Biely, P. A novel bacterial GH30 xylobiohydrolase from Hungateiclostridium clariflavum. Appl Microbiol Biotechnol 105, 185–195 (2021). https://doi.org/10.1007/s00253-020-11023-x
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DOI: https://doi.org/10.1007/s00253-020-11023-x