9JLT

GH57 family amylopullulanase D352N mutant from Aquifex aeolicus complex with alpha-cyclodextrin

Summary for 9JLT

• Entry DOI: 10.2210/pdb9jlt/pdb
• Related PRD ID: PRD_900015
• Descriptor: Glycoside hydrolase family 57 N-terminal domain-containing protein, Cyclohexakis-(1-4)-(alpha-D-glucopyranose), GLYCEROL, ... (4 entities in total)
• Functional Keywords: gh57 family, amylopullulanase, aquifex aeolicus, cyclodextrin, hydrolase
• Biological source: Aquifex aeolicus VF5
• Total number of polymer chains: 2
• Total formula weight: 117222.97

Authors

Zhu, Z.M.,Wang, W.W.,Li, M.J.,Xu, Q.,Zhou, H.,Huang, L.Q.,Wang, Q.S.,Yu, F. (deposition date: 2024-09-19, release date: 2025-06-04)

Primary citation

Zhu, Z.,Li, M.,Xu, Q.,Huang, L.,Zhou, H.,Wang, W.,Wang, Q.,Yu, F.
Mechanistic insights into cyclodextrins as substrates and inhibitors of GH57 family amylopullulanase from Aquifex aeolicus.
J.Struct.Biol., 217:108199-108199, 2025

PubMed Abstract: Maltooligosaccharides (MOs) have gained significant attention in the food and pharmaceutical industries owing to their valuable functional properties, including controlled sweetness, digestibility, and enhanced bioavailability. However, conventional MOs is production involves complex processing steps and significant production costs. A potential high-efficiency synthesis of specific MOs can be achieved through the ring-opening reaction of cyclodextrins (CDs) catalyzed by amylolytic enzymes. In this study, we analyze the catalytic conversion of α-, β-, and γ-CDs by a GH57 family amylopullulanase from Aquifex aeolicus (AaApu) using thin-layer chromatography (TLC). Our findings demonstrate that AaApu has a substrate specificity for γ-CD, while all three CDs exert competitive inhibition on pullulan hydrolysis. To elucidate the molecular mechanism of CDs as inhibitor and substrate of amylopullulanase, we determined high-resolution crystal structures of AaApu (wild-type and D352N) in complex with α-, β-, and γ-CD through co-crystallization. These findings establish a structure-function framework for understanding the bifunctional nature of CDs as both substrates and inhibitors in GH57 amylopullulanases.

PubMed: 40120836
DOI: 10.1016/j.jsb.2025.108199

Experimental method

X-RAY DIFFRACTION (1.56 Å)