8P9V

Crystal Structure of Two-Domain Laccase mutant M199G/R240H/D268N from Streptomyces griseoflavus

Summary for 8P9V

• Entry DOI: 10.2210/pdb8p9v/pdb
• Descriptor: Two-domain laccase, OXYGEN MOLECULE, COPPER (II) ION, ... (5 entities in total)
• Functional Keywords: two-domain laccase, enzyme engineering, sgfsl, tnc, proton transport, laccase trinuclear cluster, oxygen reduction, structural biology, oxidoreductase
• Biological source: Streptomyces griseoflavus
• Total number of polymer chains: 6
• Total formula weight: 182852.29

Authors

Kolyadenko, I.A.,Tishchenko, S.V.,Gabdulkhakov, A.G. (deposition date: 2023-06-06, release date: 2024-04-10)

Primary citation

Kolyadenko, I.,Tishchenko, S.,Gabdulkhakov, A.
Structural Insight into the Amino Acid Environment of the Two-Domain Laccase's Trinuclear Copper Cluster.
Int J Mol Sci, 24:-, 2023

PubMed Abstract: Laccases are industrially relevant enzymes. However, their range of applications is limited by their functioning and stability. Most of the currently known laccases function in acidic conditions at temperatures below 60 °C, but two-domain laccases (2D) oxidize some substrates in alkaline conditions and above 70 °C. In this study, we aim to establish the structural factors affecting the alkaline activity of the 2D laccase from (SgfSL). The range of methods used allowed us to show that the alkaline activity of SgfSL is influenced by the polar residues located close to the trinuclear center (TNC). Structural and functional studies of the SgfSL mutants Met199Ala/Asp268Asn and Met199Gly/Asp268Asn revealed that the substitution Asp268Asn (11 Å from the TNC) affects the orientation of the Asn261 (the second coordination sphere of the TNC), resulting in hydrogen-bond-network reorganization, which leads to a change in the SgfSL-activity pH profile. The combination of the Met199Gly/Arg240His and Asp268Asn substitutions increased the efficiency (k/K) of the 2,6-DMP oxidation by 34-fold compared with the SgfSL. Our results extend the knowledge about the structure and functioning of 2D laccases' TNC active sites and open up new possibilities for the directed engineering of laccases.

PubMed: 37569288
DOI: 10.3390/ijms241511909

Experimental method

X-RAY DIFFRACTION (2.2 Å)