Year-end Sale % OFF 
Abstract
BackgroundCellulose is the most important component of lignocellulose, and its degradation requires three different types of enzymes to act synergistically. There have been reports of single gene duality, but no gene has been described to have more than two functions. Cloning and expression of fusion cellulases containing more than two kinds of catalytic domains has not been reported thus far.ResultsWe synthesized three different cellulase genes and linked the three catalytic domains with a (G4S)3 flexible linker. The trifunctional cellulase gene (BCE) containing three types of cellulase functions was constructed and expressed in S. cerevisiae successfully. The β-glucosidase, the exoglucanase and the endoglucanase activity of the trifunctional cellulase BCE reached 16.80 IU/mg, 2.26 IU/mg and 20.67 IU/mg, which was 46.27, 6.73 and 46.20% higher than the activities of the β-glucosidase BG, the endoglucanase CBH and the endoglucanase EG. The filter paper enzyme activity of BCE was higher than those of BG, CBH and EG, reached 2.04 IU/mg.ConclusionsThe trifunctional cellulase BCE was designed based on β-glucosidase BG, endoglucanase EG and exoglucanase CBH, and it possessed β-glucosidase activity, endoglucanase activity and exoglucanase activity simultaneously. The BCE has better filter paper activity, it means the potential practical application.
Highlights
Cellulose is the most important component of lignocellulose, and its degradation requires three different types of enzymes to act synergistically
Cellulases are mainly composed of various hydrolytic enzymes acting synergistically on cellulosic material which is the main ingredient in lignocellulose [1, 2]
Design of the trifunctional single gene In order to construct a single protein with three functional domains of cellulase enzymes, three different cellulase genes without cellulose binding domains were selected from different species
Summary
Cellulose is the most important component of lignocellulose, and its degradation requires three different types of enzymes to act synergistically. Cellulases are mainly composed of various hydrolytic enzymes acting synergistically on cellulosic material which is the main ingredient in lignocellulose [1, 2]. Considering the demand for a synergistic action of multiple enzymes, the expression of a variety of enzymes in a single cell is likely to improve the efficiency of cellulose degradation [10]. Wang et al constructed a co-expression vector containing two independent expression cassettes based on six restriction enzyme sites and achieved the co-expression of egv (GenBank: HG313887.1) and cbh (GenBank: HG313872.1) genes each controlled by a separate promoter in Trichoderma reesei [11]. Gong et al achieved the co-expression of eg (GenBank: M19373) and bgl (GenBank: TRU09580) in
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
