7VQM

GH2 beta-galacturonate AqGalA in complex with galacturonide

Summary for 7VQM

• Entry DOI: 10.2210/pdb7vqm/pdb
• Descriptor: GH2 beta-galacturonate AqGalA, CHLORIDE ION, beta-D-galactopyranuronic acid, ... (4 entities in total)
• Functional Keywords: beta-galacturonate, complex, hydrolase
• Biological source: Aquimarina sp.
• Total number of polymer chains: 4
• Total formula weight: 285643.40

Authors

Yang, J. (deposition date: 2021-10-20, release date: 2021-11-24, Last modification date: 2023-11-29)

Primary citation

Yang, J.,Li, S.,Liu, Y.,Li, R.,Long, L.
Structural and Biochemical Basis of a Marine Bacterial Glycoside Hydrolase Family 2 beta-Glycosidase with Broad Substrate Specificity
Appl.Environ.Microbiol., 88:-, 2022

PubMed Abstract: Uronic acids are commonly found in marine polysaccharides and increase structural complexity and intrinsic recalcitrance to enzymatic attack. Glycoside hydrolase family 2 (GH2) includes proteins that target sugar conjugates with hexuronates and are involved in the catabolism and cycling of marine polysaccharides. Here, we report a novel GH2, GalA from a marine alga-associated organism with broad substrate specificity. Biochemical analyses revealed that GalA exhibits hydrolyzing activities against β-galacturonide, β-glucuronide, and β-galactopyranoside via retaining mechanisms. We solved the GalA crystal structure in complex with galacturonic acid (GalA) and determined (via mutagenesis) that charge characteristics at uronate-binding subsites controlled substrate selectivity for uronide hydrolysis. Additionally, conformational flexibility of the GalA active-site pocket was proposed as a key component for broad substrate enzyme selectivity. Our GalA structural and functional data augment the current understanding of substrate recognition of GH2 enzymes and provide key insights into the bacterial use of uronic acid-containing polysaccharides. The decomposition of algal glycans driven by marine bacterial communities represents one of the largest heterotrophic transformations of organic matter fueling marine food webs and global carbon cycling. However, our knowledge on carbohydrate cycling is limited due to structural complexity of marine polysaccharides and the complicated enzymatic machinery of marine microbes. To degrade algal glycan, marine bacteria such as members of the produce a complex repertoire of carbohydrate-active enzymes (CAZymes) matching the structural specificities of the different carbohydrates. In this study, we investigated an extracellular GH2 β-glycosidase, GalA from a marine organism, to identify the key components responsible for glycuronide recognition and hydrolysis. The broad substrate specificity of GalA against glycosides with diverse stereochemical substitutions indicates its potential in processing complex marine polysaccharides. Our findings promote a better understanding of microbially driven mechanisms of marine carbohydrate cycling.

PubMed: 34818100
DOI: 10.1128/AEM.02226-21

Experimental method

X-RAY DIFFRACTION (2.4 Å)