8YAG

Cryo-electron microscopic structure of an amide hydrolase from Pseudoxanthomonas wuyuanensis

Summary for 8YAG

• Entry DOI: 10.2210/pdb8yag/pdb
• EMDB information: 39098
• Descriptor: Imidazolonepropionase, ZINC ION (2 entities in total)
• Functional Keywords: amide hydrolase, iron-binding, ochratoxin a, zn, hydrolase
• Biological source: Pseudoxanthomonas wuyuanensis
• Total number of polymer chains: 8
• Total formula weight: 348396.61

Authors

Dai, L.H.,Xu, Y.H.,Hu, Y.M.,Niu, D.,Yang, X.C.,Shen, P.P.,Li, X.,Xie, Z.Z.,Li, H.,Guo, R.-T.,Chen, C.-C. (deposition date: 2024-02-09, release date: 2024-10-09)

Primary citation

Hu, Y.,Dai, L.,Xu, Y.,Niu, D.,Yang, X.,Xie, Z.,Shen, P.,Li, X.,Li, H.,Zhang, L.,Min, J.,Guo, R.T.,Chen, C.C.
Functional characterization and structural basis of an efficient ochratoxin A-degrading amidohydrolase.
Int.J.Biol.Macromol., 278:134831-134831, 2024

PubMed Abstract: Ochratoxin A (OTA) contamination in various agro-products poses a serious threat to the global food safety and human health, leading to enormous economic losses. Enzyme-mediated OTA degradation is an appealing strategy, and the search for more efficient enzymes is a prerequisite for achieving this goal. Here, a novel amidohydrolase, termed PwADH, was demonstrated to exhibit 7.3-fold higher activity than that of the most efficient OTA-degrading ADH3 previously reported. Cryo-electron microscopy structure analysis indicated that additional hydrogen-bond interactions among OTA and the adjacent residue H163, the more compact substrate-binding pocket, and the wider entry to the substrate-access cavity might account for the more efficient OTA-degrading activity of PwADH compared with that of ADH3. We conducted a structure-guided rational design of PwADH and obtained an upgraded variant, G88D, whose OTA-degrading activity was elevated by 1.2-fold. In addition, PwADH and the upgraded G88D were successfully expressed in the industrial yeast Pichia pastoris, and their catalytic activities were compared to those of their counterparts produced in E. coli, revealing the feasibility of producing PwADH and its variants in industrial yeast strains. These results illustrate the structural basis of a novel, efficient OTA-degrading amidohydrolase and will be beneficial for the development of high-efficiency OTA-degrading approaches.

PubMed: 39163957
DOI: 10.1016/j.ijbiomac.2024.134831

Experimental method

ELECTRON MICROSCOPY (2.33 Å)