4UB9

Structural and catalytic characterization of molinate hydrolase

Summary for 4UB9

• Entry DOI: 10.2210/pdb4ub9/pdb
• Descriptor: Molinate hydrolase, ZINC ION (3 entities in total)
• Functional Keywords: amidohydrolase molinate thiocarbamate, hydrolase
• Biological source: Gulosibacter molinativorax
• Total number of polymer chains: 8
• Total formula weight: 437955.77

Authors

Leite, J.P.,Duarte, M.,Paiva, A.,Ferreira-da-Silva, F.,Matias, P.M.,Nunes, O.,Gales, L. (deposition date: 2014-08-12, release date: 2015-06-24, Last modification date: 2024-05-01)

Primary citation

Leite, J.P.,Duarte, M.,Paiva, A.M.,Ferreira-da-Silva, F.,Matias, P.M.,Nunes, O.C.,Gales, L.
Structure-guided engineering of molinate hydrolase for the degradation of thiocarbamate pesticides.
Plos One, 10:e0123430-e0123430, 2015

PubMed Abstract: Molinate is a recalcitrant thiocarbamate used to control grass weeds in rice fields. The recently described molinate hydrolase, from Gulosibacter molinativorax ON4T, plays a key role in the only known molinate degradation pathway ending in the formation of innocuous compounds. Here we report the crystal structure of recombinant molinate hydrolase at 2.27 Å. The structure reveals a homotetramer with a single mononuclear metal-dependent active site per monomer. The active site architecture shows similarities with other amidohydrolases and enables us to propose a general acid-base catalysis mechanism for molinate hydrolysis. Molinate hydrolase is unable to degrade bulkier thiocarbamate pesticides such as thiobencarb which is used mostly in rice crops. Using a structural-based approach, we were able to generate a mutant (Arg187Ala) that efficiently degrades thiobencarb. The engineered enzyme is suitable for the development of a broader thiocarbamate bioremediation system.

PubMed: 25905461
DOI: 10.1371/journal.pone.0123430

Experimental method

X-RAY DIFFRACTION (2.27 Å)