7NN3

A carbohydrate esterase family 15 (CE15) glucuronoyl esterase from Caldicellulosiruptor kristjansonii

Summary for 7NN3

• Entry DOI: 10.2210/pdb7nn3/pdb
• Descriptor: Beta-xylanase, TRIETHYLENE GLYCOL, 1,2-ETHANEDIOL, ... (6 entities in total)
• Functional Keywords: hydrolase
• Biological source: Caldicellulosiruptor kristjanssonii (strain ATCC 700853 / DSM 12137 / I77R1B)
• Total number of polymer chains: 4
• Total formula weight: 180900.89

Authors

Krska, D.,Mazurkewich, S.,Navarro Poulsen, J.,Larsbrink, J.,Lo Leggio, L. (deposition date: 2021-02-24, release date: 2021-07-07, Last modification date: 2024-01-31)

Primary citation

Krska, D.,Mazurkewich, S.,Brown, H.A.,Theibich, Y.,Poulsen, J.N.,Morris, A.L.,Koropatkin, N.M.,Lo Leggio, L.,Larsbrink, J.
Structural and Functional Analysis of a Multimodular Hyperthermostable Xylanase-Glucuronoyl Esterase from Caldicellulosiruptor kristjansonii .
Biochemistry, 60:2206-2220, 2021

PubMed Abstract: The hyperthermophilic bacterium encodes an unusual enzyme, Xyn10C-GE15A, which incorporates two catalytic domains, a xylanase and a glucuronoyl esterase, and five carbohydrate-binding modules (CBMs) from families 9 and 22. The xylanase and glucuronoyl esterase catalytic domains were recently biochemically characterized, as was the ability of the individual CBMs to bind insoluble polysaccharides. Here, we further probed the abilities of the different CBMs from Xyn10C-GE15A to bind to soluble poly- and oligosaccharides using affinity gel electrophoresis, isothermal titration calorimetry, and differential scanning fluorimetry. The results revealed additional binding properties of the proteins compared to the former studies on insoluble polysaccharides. Collectively, the results show that all five CBMs have their own distinct binding preferences and appear to complement each other and the catalytic domains in targeting complex cell wall polysaccharides. Additionally, through renewed efforts, we have achieved partial structural characterization of this complex multidomain protein. We have determined the structures of the third CBM9 domain (CBM9.3) and the glucuronoyl esterase (GE15A) by X-ray crystallography. CBM9.3 is the second CBM9 structure determined to date and was shown to bind oligosaccharide ligands at the same site but in a different binding mode compared to that of the previously determined CBM9 structure from . GE15A represents a unique intermediate between reported fungal and bacterial glucuronoyl esterase structures as it lacks two inserted loop regions typical of bacterial enzymes and a third loop has an atypical structure. We also report small-angle X-ray scattering measurements of the N-terminal CBM22.1-CBM22.2-Xyn10C construct, indicating a compact arrangement at room temperature.

PubMed: 34180241
DOI: 10.1021/acs.biochem.1c00305

Experimental method

X-RAY DIFFRACTION (1.88948348333 Å)