Journal
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 105, Issue 3, Pages 1133-1145Publisher
SPRINGER
DOI: 10.1007/s00253-020-11077-x
Keywords
Endoglucanase; Cellulose; Nanocellulose; Fiber; Molecular evolution
Categories
Funding
- Bio Based Industries Joint Undertaking under the European Union's Horizon 2020 research and innovation program, project SWEETWOODS [792061]
- H2020 Societal Challenges Programme [792061] Funding Source: H2020 Societal Challenges Programme
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The study successfully addressed the issue of endoglucanase being diverted by other substrates through enzyme engineering and evolution, leading to improved substrate specificity and enhanced activity towards high molecular weight substrates. These findings hold potential applications in the bioeconomy sector, such as second-generation biofuels and biomaterials from lignocellulosic biomass.
Endoglucanases are key elements in several industrial applications, such as cellulosic biomass hydrolysis, cellulose fiber modification for the production paper and composite materials, and in nanocellulose production. In all of these applications, the desired function of the endoglucanase is to create nicks in the amorphous regions of the cellulose. However, endoglucanase can be diverted from its activity on the fibers by other substrates-soluble oligosaccharides. This issue was addressed in the current study using enzyme engineering and an enzyme evolution approach. To this end, a hypothetical endoglucanase from a thermostable bacterium Spirochaeta thermophila was for the first time cloned and characterized. The wild-type enzyme was used as a starting point for mutagenesis and molecular evolution toward a preference for the higher molecular weight substrates. The best of the evolved enzymes was more active than the wild-type enzyme toward high molecular weight substrate at temperatures below 45 degrees C (3-fold more active at 30 degrees C) and showed little or no activity with low molecular weight substrates. These findings can be instrumental in bioeconomy sectors, such as second-generation biofuels and biomaterials from lignocellulosic biomass.
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