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Title: PROPERTIES OF THE MACROPHOMINA PHASEOLINA ENDOGLUCANASE (EGL 1) GENE PRODUCT IN BACTERIAL AND YEAST EXPRESSION SYSTEMS

Author
item WANG, HAIYIN - UNIV. CALIF.-BERKELEY
item Jones, Richard

Submitted to: Applied Biochemistry and Biotechnology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/27/1999
Publication Date: N/A
Citation: N/A

Interpretive Summary: Cellulases are a group of enzyme proteins which facilitate the breakdown of plant material which is found in foods and fibers. By breaking apart plant material, the enzymes help to produce paper in an environmentally friendly way. These enzymes also help to improve the consistency and flavor of fruit juices. A new use for the enzymes is found in laundry detergents where they aid in maintaining color quality in plant derived cotton fabrics. Large-scale production of these important cellulase enzymes requires a thorough knowledge of the enzyme and adaptation of current production methods. This report furthers our ability to produce the enzymes in a commercially feasible manner. We used cloned DNA to compare production of a cellulase in bacteria and yeast. Important features of cellulase production were identified which include temperature and pH tolerances. These features are important when using cellulase in laundry detergents where they are subject to high temperatures and high pH during clothes washing. We concluded that the bacterial production system is best suited for large-scale commercial production of cellulase enzymes. This will be useful to scientists and industries that produce cellulase enzymes.

Technical Abstract: Functional expression of a Beta-D-1,4 glucanase-encoding gene (egl1) from a filamentous fungus was achieved in both Escherichia coli and Saccharomyces cerevisiae using a modified version of pRS413. Optimal activity of the E. coli expressed enzyme was found at incubation temperatures of 60C, while the enzyme activity was optimal at 40C when expressed by S. cerevisiae. Enzyme activity at different pH levels was similar for both bacterial and yeast, being highest at 5.0. Yeast expression resulted in a highly glycosylated protein of approximately 60 kDa, compared to bacterial expression which resulted in a protein of 30 kDa. The hyperglycosylated protein had reduced enzyme activity, indicating that E. coli is a preferred vehicle for production scale-up.