Modifications of laccase activities of copper efflux oxidase, CueO by synergistic mutations in the first and second coordination spheres of the type I copper center

Kataoka, Keisuke; Kogi, Hiroki; Tsujimura, Seiya; Sakurai, Takeshi

doi:10.1016/j.bbrc.2013.01.040

Cited by 23 publications

(12 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Compared with the wild-type laccase, the mutants displayed significant changes in the activity and E 0' T1, which were in correlation with the characteristics of their circular dichroism, absorption and electron paramagnetic resonance spectra. More recently, CueO variants with single, double and triple mutations in the first and second coordination spheres of the T1 Cu have been studied (Kataoka et al, 2013). Specifically, four single mutants (D439A, P444A, M510L and M510Q), five double mutants (D439A/P444A, D439A/M510L, D439A/M510Q, P444A/M510L and P444A/M510Q) and two triple variants (D439A/P444A/M510L and D439A/P444A/M510Q) were evaluated.…”

Section: Rational Approachesmentioning

confidence: 99%

Laccase engineering: From rational design to directed evolution

Maté

Alcalde

2015

Biotechnology Advances

271

207

View full text Add to dashboard Cite

Laccases are multicopper oxidoreductases considered by many in the biotechonology field as the ultimate "green catalysts". This is mainly due to their broad substrate specificity and relative autonomy (they use molecular oxygen from air as an electron acceptor and they only produce water as by-product), making them suitable for a wide array of applications: biofuel production, bioremediation, organic synthesis, pulp biobleaching, textiles, the beverage and food industries, biosensor and biofuel cell development. Since the beginning of the 21st century, specific features of bacterial and fungal laccases have been exhaustively adapted in order to reach the industrial demands for high catalytic activity and stability in conjunction with reduced production cost. Among the goals established for laccase engineering, heterologous functional expression, improved activity and thermostability, tolerance to non-natural media (organic solvents, ionic liquids, physiological fluids) and resistance to different types of inhibitors are all challenges that have been met, while obtaining a more comprehensive understanding of laccase structure-function relationships. In this review we examine the most significant advances in this exciting research area in which rational, semi-rational and directed evolution approaches have been employed to ultimately convert laccases into high value-added biocatalysts.

show abstract

Section: Rational Approachesmentioning

confidence: 99%

Laccase engineering: From rational design to directed evolution

Maté

Alcalde

2015

Biotechnology Advances

271

207

View full text Add to dashboard Cite

show abstract

“…17 The electrochemical studies realized thereafter on CueO did not take into consideration or even mentioned the Met-rich helix. [18][19][20][21] Even less studies reported the behavior of Tt Lac on electrochemical interfaces. The first study was realized in 2011 with Tt Lac drop casted from an agarose solution on glassy carbon electrodes.…”

Section: Introductionmentioning

confidence: 99%

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

et al. 2019

View full text Add to dashboard Cite

Multicopper oxidases (MCOs) catalyze the oxidation of a variety of substrates while reducing oxygen into water through four copper atoms. As an additional feature, some MCOs display an enhanced activity in solution in the presence of Cu 2+. This is the case of the hyperthermophilic laccase HB27 from Thermus thermophilus, the physiologic role of which is unknown. As a particular feature, this enzyme presents a methionine rich domain proposed to be involved in copper interaction. In this work, laccase from T. thermophilus was produced in E. coli, and the effect of Cu 2+ on its electroactivity at carbon nanotube modified electrodes was investigated. Direct O2 electroreduction is strongly dictated by carbon nanotube surface chemistry in accordance with the enzyme dipole moment. In the presence of Cu 2+ , an additional low potential cathodic wave occurs, which was never described earlier. Analysis of this wave as a function 2 of Cu 2+ availability allows us to attribute this wave to a cuprous oxidase activity displayed by the laccase and induced by copper binding close to the Cu T1 center. A mutant lacking the methionine-rich hairpin domain characteristic of this laccase conserves its copper activity suggesting a different site of copper binding. This study provides new insight into the copper effect in methionine rich MCOs, and highlights the utility of the electrochemical method to investigate cuprous oxidase activity and to understand the physiological role of these MCOs.

show abstract

“…The precise binding site(s) of syringaldazine and other organic substrates in CueO is unknown, although mechanistic considerations imply proximity to the sCu/T1 coppers. Mutations in the vicinity of the CueO sCu/T1 binding sites can impact Cu binding and enzyme redox potentials both positively and negatively (24, 40, 45). The reduced T1 copper occupancy observed in CueO-PM2 likely accounts for its reduced k cat , with this phenotype contingent on either all or a subset of the Phe/Trp/Leu residues in the PM2 insert.…”

Section: Discussionmentioning

confidence: 99%

Development and structural characterization of an engineered multi-copper oxidase reporter of protein–protein interactions

Sana

Chee

Wongsantichon

et al. 2019

Journal of Biological Chemistry

View full text Add to dashboard Cite

Protein–protein interactions (PPIs) are ubiquitous in almost all biological processes and are often corrupted in diseased states. A detailed understanding of PPIs is therefore key to understanding cellular physiology and can yield attractive therapeutic targets. Here, we describe the development and structural characterization of novel Escherichia coli CueO multi-copper oxidase variants engineered to recapitulate protein–protein interactions with commensurate modulation of their enzymatic activities. The fully integrated single-protein sensors were developed through modular grafting of ligand-specific peptides into a highly compliant and flexible methionine-rich loop of CueO. Sensitive detection of diverse ligand classes exemplified by antibodies, an E3 ligase, MDM2 proto-oncogene (MDM2), and protease (SplB from Staphylococcus aureus ) was achieved in a simple mix and measure homogeneous format with visually observable colorimetric readouts. Therapeutic antagonism of MDM2 by small molecules and peptides in clinical development for treatment of cancer patients was assayed using the MDM2-binding CueO enzyme. Structural characterization of the free and MDM2-bound CueO variant provided functional insight into signal-transducing mechanisms of the engineered enzymes and highlighted the robustness of CueO as a stable and compliant scaffold for multiple applications.

show abstract

Modifications of laccase activities of copper efflux oxidase, CueO by synergistic mutations in the first and second coordination spheres of the type I copper center

Cited by 23 publications

References 25 publications

Laccase engineering: From rational design to directed evolution

Laccase engineering: From rational design to directed evolution

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction

Development and structural characterization of an engineered multi-copper oxidase reporter of protein–protein interactions

Contact Info

Product

Resources

About

Modifications of laccase activities of copper efflux oxidase, CueO by synergistic mutations in the first and second coordination spheres of the type I copper center

Cited by 23 publications

References 25 publications

Laccase engineering: From rational design to directed evolution

Laccase engineering: From rational design to directed evolution

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O2 Reduction

Development and structural characterization of an engineered multi-copper oxidase reporter of protein–protein interactions

Contact Info

Product

Resources

About

Interplay between Orientation at Electrodes and Copper Activation of Thermus thermophilus Laccase for O₂ Reduction