Team:British Columbia/Project/S-Layer/Laccases
Laccase
S-Layer Engineering
Abstract
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Key Achievements
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Introduction
Current pretreatment and hydrolysis approaches to biomass depolymerization involve thermochemical methods that apply alkaline chemicals, in addition to high heat and pressure (1). These methods enhance the fractionation of lignin and hemicellulose from the plant cell wall, thereby allowing enzymes access to the cellulosic fraction for downstream processing. Although these methods are effective, their high cost to industries and to the environment requires that alternative methods to remove the more recalcitrant components of lignocellulosic biomass, are erected. We intend to produce a system that minimally relies on external thermochemical treatments to make the cellulosic component of biomass available for valuable-chemical production. To achieve this, we attempted to engineer surface layer expression of a small laccase derived from Amycolatopsis sp. 75iv2, in Caulobacter crescentus.
Laccases belong to a superfamily of enzymes called multicopper oxidases (MCO), and are expressed by several rot-fungi and soil bacteria (2). As an MCO, they catalyze the one-electron oxidation of substrates through associated four-electron reductions of molecular oxygen to water, using four copper ions coordinated in designated copper centers (3, 4). Their broad phenolic and polyphenolic substrate specificity allows for direct transformation of lignin, or removal of toxic phenols that arise during lignocellulosic biomass pre-treatment (5).
Design
WE MUST ADD THIS SECTION :)
Methods
Results
Conclusion
References
- 1. Chundawat S, Beckham G, Himmel M, Dale B. 2011. Deconstruction of Lignocellulosic Biomass to Fuels and Chemicals. Annual Review of Chemical and Biomolecular Engineering 2:121-145.
- 2. Sirim D, Wagner F, Wang L, Schmid R, Pleiss J. 2011. The Laccase Engineering Database: a classification and analysis system for laccases and related multicopper oxidases. Database 2011:bar006.
- 3. Jones Solomon E. 2015. Electron transfer and reaction mechanism of laccases. Cell. Mol. Life Sci. 72:869-883.
- 4. Reiss R, Ihssen J, Richter M, Eichhorn E, Schilling B, Thöny-Meyer L. 2013. Laccase versus Laccase-Like Multi-Copper Oxidase: A Comparative Study of Similar Enzymes with Diverse Substrate Spectra. PLoS ONE 8:e65633.
- 5. Kunamneni A, Ballesteros A, Plou FJ, Alcalde M. 2007. Fungal laccase – a versatile enzyme for biotechnological applications, p 233-245. In Mendez-Vilas A (ed). Communicating current research and educational topics and trends in applied microbiology. FORMATEX, Badajoz.
- 6. Majumdar S, Lukk T, Solbiati J, Bauer S, Nair S, Cronan J, Gerlt J. 2014. Roles of Small Laccases from Streptomyces in Lignin Degradation. Biochemistry 53:4047-4058.
- 7. More S, Malini S. 2011. Isolation, Purification, and Characterization of Fungal Laccase from Pleurotus sp. Enzyme Research 2011:1-7.
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