Abstract
Non-digestible oligosaccharides (NDOs) are likely prebiotic candidates that have been related to the prevention of intestinal infections and other disorders for both humans and animals. Lignocellulosic biomass is the largest carbon source in the biosphere, therefore cello-oligosacharides (COS), especially cellobiose, are potentially the most widely available choice of NDOs. Production of COS and cellobiose with enzymes offers numerous benefits over acid-catalyzed processes, as it is milder, environmentally friendly and produces fewer by-products. Cellobiohydrolases (CBHs) and a class of endoglucanases (EGs), namely processive EGs, are key enzymes for the production of COS, as they have higher preference toward glycosidic bonds near the end of cellulose chains and are able to release soluble products. In this work, we describe the heterologous expression and characterization of two CBHs from the filamentous fungus Thermothelomyces thermophila, as well as their synergism with proccessive EGs for cellobiose release from organosolv pretreated spruce and birch. The properties, inhibition kinetics and substrate specific activities for each enzyme are described in detail. The results show that a combination of EGs belonging to Glycosyl hydrolase families 5, 6, and 9, with a CBHI and CBHII in appropriate proportions, can enhance the production of COS from forest materials, underpinning the potential of these biocatalysts in the production of NDOs.
Highlights
In food and nutraceutical industry, the development of compounds that have the potential to reduce disease risk and thereby enhance human health has attracted much interest
The efficient cellobiose production from lignocellulosic materials requires enzymes that act on the reducing and non-reducing cellulose chains
TtCBH6 and TtCBH7 from T. thermophila are two CBHs that were heterologously produced in P. pastoris and fully characterized with the aim to be tested in optimized enzyme mixtures for cellobiose production from organosolv pretreated spruce and birch
Summary
In food and nutraceutical industry, the development of compounds that have the potential to reduce disease risk and thereby enhance human health has attracted much interest. Diet-related modulations of this microflora offers promise for reducing pathogens/mediated gut disorders linked to a variety of chronic diseases, including obesity, type 2 diabetes, and cardiovascular diseases. This has resulted in the development of a prebiotic concept. Prebiotics are dietary ingredients that are not digested and selectively promote the growth and the activity of the bacteria in the colon (Gibson and Roberfroid, 1995). The level of active prebiotics that can be obtained through the diet is too low to have a significant impact; a sustainable production of prebiotics with a defined action is to be preferred
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