Characterization and structural study of a novel β‐ N ‐acetylgalactosaminidase from Niabella aurantiaca - Archive ouverte HAL Access content directly
Journal Articles FEBS Journal Year : 2024

Characterization and structural study of a novel β‐ N ‐acetylgalactosaminidase from Niabella aurantiaca

Eduardo Sebastian Moreno Prieto
Sune Fjermedal
  • Function : Author
Suzana Siebenhaar
  • Function : Author
Marlène Vuillemin
  • Function : Author
Jesper Holck
  • Function : Author
Garry Gippert
  • Function : Author
Casper Wilkens
  • Function : Author
Jens Preben Morth
  • Function : Author
Bernard Henrissat
  • Function : Author

Abstract

We report here the identification, characterization and three‐dimensional (3D) structure determination of Na Nga, a newly identified β‐ N ‐acetylgalactosaminidase from the Gram‐negative soil bacterium Niabella aurantiaca DSM 17617. When recombinantly expressed in Escherichia coli , the enzyme selectively cleaved 4‐nitrophenyl‐ N ‐acetyl‐β‐ d ‐galactosamine ( p NP‐β‐ d ‐Gal p NAc). The X‐ray crystal structure of the protein was refined to 2.5 Å and consists of an N‐terminal β‐sandwich domain and a (β/α) 8 barrel catalytic domain. Despite a mere 22% sequence identity, the 3D structure of Na Nga is similar to those previously determined for family GH123 members, suggesting it also employs the same substrate‐assisted catalytic mechanism. Inhibition by N ‐acetyl‐galactosamine thiazoline (GalNAc‐thiazoline) supports the suggested mechanism. A phylogenetic analysis of its proximal sequence space shows significant clustering of unknown sequences around Na Nga with sufficient divergence with previously identified GH123 members to subdivide this family into distinct subfamilies. Although the actual biological substrate of our enzyme remains unknown, examination of the active site pocket suggests that it may be a β‐ N ‐acetylgalactosaminide substituted by a monosaccharide at O‐3. Analysis of the genomic context suggests, in turn, that this substituted β‐ N ‐acetylgalactosaminide may be appended to a d ‐arabinan from an environmental Actinomycete .
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hal-04382377 , version 1 (09-01-2024)

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Eduardo Sebastian Moreno Prieto, Sune Fjermedal, Suzana Siebenhaar, Marlène Vuillemin, Jesper Holck, et al.. Characterization and structural study of a novel β‐ N ‐acetylgalactosaminidase from Niabella aurantiaca. FEBS Journal, 2024, ⟨10.1111/febs.17042⟩. ⟨hal-04382377⟩
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