Laccase is a multinuclear copper-containing oxidoreductase that performs one-electron oxidation of environmentally relevant pollutants and is known as a green catalyst/green tool. With its advanced protein engineering platform established over the years, Creative BioMart has successfully developed engineered laccases with specificity to target alien organisms and tolerance to conditions using biocatalysts. Our scientists are committed to providing fully customized engineered laccases production solutions for environment to customers around the world.

Introduction of Laccases Production

The current environmental pollution is increasing over time, and laccase is an oxidoreductase that plays an important role in remediation. Produced in fungi, plants and bacteria, laccase catalyzes the oxidation of a wide variety of phenolic and non-phenolic compounds, using oxygen as an electron acceptor and producing water as the only by-product. It is currently investigated for various industrial applications, including dye decolorization, detoxification of environmental pollutants, and reuse of waste and wastewater. Laccases offer exciting prospects for the green industry, but the potential of laccases remains untapped. For example, some laccases lack biocatalysts with the required selectivity, compatibility with stringent process conditions, etc. In recent years, scientists have increasingly focused on improving the performance of laccases based on protein engineering. Researchers have extensively employed molecular engineering approaches to design efficient laccases through rational and random mutagenesis and enhance laccase tolerance under stressful conditions.

Fig 1. Target regions for laccase mutations. (Stanzione I, et al., 2020)

Solutions 

Laccases are promising tools in pollutant detoxification and bioremediation of phenolic compounds, which play an important role in improving environmental quality. Creative BioMart has successfully utilized a variety of protein engineering techniques to produce engineered laccases to improve the properties of laccases, including exogenous functional expression, activity and thermostability, tolerance to non-natural media (organic solvents, ionic liquids, physiological liquids) and resistance to different types of inhibitors. Our customers have widely used improved laccases for biofuel production, bioremediation, organic synthesis, pulp biobleaching, textiles, beverage and food industries, biosensors and biofuel cell development. In order to provide customized high-quality laccases to global customers, we take the following solutions:

• Integrating targeted mutations into laccase scaffolds to study the effects of hotspot amino acid residues on laccase properties, such as redox potential, catalytic efficiency, or thermal stability.
• Improving the functional expression and activity of laccase through directed evolution combined with hybridization strategy, and adapt to non-natural environment.
• Designing laccase chimeras with combined traits.
• Implementing a suitable heterologous expression system for high laccase expression levels (ie, low metabolic drainage and/or high resistance to laccase toxicity).

Applications of Protein Engineering in Laccases Production

To meet the environmental demands for high catalytic activity and stability in the treatment of pollutants, while reducing production costs, our protein engineers are revolutionizing the properties of bacterial and fungal laccases through protein engineering. Rational, semi-rational and directed evolution approaches have been used to finally convert laccases into high value-added biocatalysts.

• Rational approaches
  Site-directed mutagenesis to select specific residues based on previous structural information is a common strategy to study the structure-function relationship of laccases. Our protein engineers use computational analysis to reliably predict mutation sites in the laccase region, enabling the design of efficient, tailored biocatalysts for specific applications.
• Semi-rational approaches
  Saturation mutagenesis is often used to explore the properties of laccases on hotspot residues identified by rational analysis or directed evolution. Our protein engineers have successfully performed saturation mutagenesis of laccases to improve their stability and catalytic efficiency.
• Directed evolution and hybrid approaches
  Directed molecular evolution has become a powerful tool for developing laccases with improved characteristics or new functions. Our protein engineers use directed evolution combined with semi-rational methods to improve or create properties of laccases, including: functional heterologous expression and activity, performance in non-natural environments such as organic solvents.

Creative BioMart has in-depth knowledge and experience of the tools and processes involved in engineered laccases development, and provides professional laccases production solutions for the environment. Whether your laccases are in the discovery and screening stages, or are planning engineered laccases production, please contact us to discuss further details to ensure your next success.

References

1. Stanzione I, Pezzella C, Giardina P, et al.. (2020) Beyond natural laccases: extension of their potential applications by protein engineering[J]. Applied Microbiology and Biotechnology. 104(3): 915-924.
2. Mate D M, Alcalde M. (2015) Laccase engineering: from rational design to directed evolution[J]. Biotechnology advances. 33(1): 25-40.

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