8AIP

Crystal Structure of Two-domain bacterial laccase from the actinobacterium Streptomyces carpinensis VKM Ac-1300

8AIP の概要

• エントリーDOI: 10.2210/pdb8aip/pdb
• 分子名称: Two-Domain Laccase, COPPER (II) ION, OXYGEN MOLECULE, ... (4 entities in total)
• 機能のキーワード: two-domain laccase, laccase, streptomyces carpinensis, oxidoreductase
• 由来する生物種: Streptomyces carpinensis
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 32448.67

構造登録者

Gabdulkhakov, A.G.,Tishchenko, T.V.,Trubitsina, L.,Trubitsin, I.,Leontievsky, A.,Lisov, A. (登録日: 2022-07-27, 公開日: 2023-08-16, 最終更新日: 2024-02-28)

主引用文献

Trubitsina, L.I.,Trubitsin, I.V.,Lisov, A.V.,Gabdulkhakov, A.G.,Zavarzina, A.G.,Belova, O.V.,Larionova, A.P.,Tishchenko, S.V.,Leontievsky, A.A.
A Novel Two-Domain Laccase with Middle Redox Potential: Physicochemical and Structural Properties.
Biochemistry Mosc., 88:1658-1667, 2023

PubMed Abstract: The gene for a previously unexplored two-domain laccase was identified in the genome of actinobacterium Streptomyces carpinensis VKM Ac-1300. The two-domain laccase, named ScaSL, was produced in a heterologous expression system (Escherichia coli strain M15 [pREP4]). The enzyme was purified to homogeneity using affinity chromatography. ScaSL laccase, like most two-domain laccases, exhibited activity in the homotrimer form. However, unlike the most two-domain laccases, it was also active in multimeric forms. The enzyme exhibited maximum activity at 80°C and was thermally stable. Half-inactivation time of ScaSL at 80°C was 40 min. The laccase was able to oxidize a non-phenolic organic compound ABTS at a maximum rate at pH 4.7, and to oxidized a phenolic compound 2,6-dimethoxyphenol at a maximum rate at pH 7.5. The laccase stability was observed in the pH range 9-11. At pH 7.5, laccase was slightly inhibited by sodium azide, sodium fluoride, and sodium chloride; at pH 4.5, the laccase was completely inhibited by 100 mM sodium azide. The determined K and k of the enzyme for ABTS were 0.1 mM and 20 s, respectively. The K and k for 2,6-dimethoxyphenol were 0.84 mM and 0.36 s, respectively. ScaSL catalyzed polymerization of humic acids and lignin. Redox potential of the laccase was 0.472 ± 0.007 V. Thus, the ScaSL laccase is the first characterized two-domain laccase with a middle redox potential. Crystal structure of ScaSL was determined with 2.35 Å resolution. Comparative analysis of the structures of ScaSL and other two-domain laccases suggested that the middle potential of ScaSL may be associated with conformational differences in the position of the side groups of amino acids at position 230 (in ScaSL numbering), which belong to the second coordination sphere of the copper atom of the T1 center.

PubMed: 38105031
DOI: 10.1134/S0006297923100188

実験手法

X-RAY DIFFRACTION (2.35 Å)