Induction of Gene Expression Using a High Concentration Sugar Mixture

Abstract

Described herein is a composition useful for inducing expression of genes whose expression is under control of an inducible promoter sequence and methods for the compositions preparation and use.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority to U.S. Provisional Application No. 60 / 409,466, filed Sep. 10, 2002 (Attorney Docket No. GC774P), which is herein incorporated in its entirety by reference.STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT[0002]Portions of this work were funded by Subcontract No. ZCO-0-30017-01 with the National Renewable Energy Laboratory under Prime Contract No. DE-AC36-99GO10337 with the U.S. Department of Energy. Accordingly, the United States Government may have certain rights in this invention.FIELD OF THE INVENTION[0003]This invention relates to methods for improved production of proteins from a cell culture. The inventors have discovered culture components and conditions that dramatically increase the amount of protein produced from genes under the control of cellulase gene promoter sequences. The improved methods can be used for the production of proteins encoded by natur...

AI Technical Summary

Benefits of technology

[0015]It has now been discovered that when a whole cellulase preparation is added to a concentrated glucose solution, and the composition is incubated for at least two days at 50° C. to about 65° C., a sugar mixture containing appreciable quantities of an inducer of cellulase gene expression is produced. Surprisingly, the resulting complex mixture is sufficient to induce cellulase production as is without further purification. This discovery is surprising since glucose acts as a repressor of cellulase genes in Trichoderma reesei. This discovery provides an inducer of cellulase gene expression that is an inexpensive alternative to lactose or purified sophorose and a less cumbersome alternative to solid cellulose for the production of cellulases in Trichoderma reesei.

Problems solved by technology

The most problematic and expensive aspect of industrial cellulase production is providing the appropriate inducer to Trichoderma.

Unfortunately on an industrial scale, both methods of induction have drawbacks which result in high costs being associated with cellulase production.

Although cellulose is an effective and inexpensive inducer, controlling the glucose concentration when Trichoderma is grown on solid cellulose can be problematic.

At low concentrations of cellulose, glucose production may be too slow to meet the metabolic needs of active cell growth and function.

Thus, expensive process control schemes are required to provide slow substrate addition and monitoring of glucose concentration (Ju and Afolabi, Biotechnol. Prog., 91-97, 1999).

Moreover, the slow continuous delivery of substrate can be difficult to achieve as a result of the solid nature of the cellulosic materials.

Purification of the β-glucosidase is both time-consuming and expensive.

Sophorose is a more potent inducer than cellulose, but sophorose is expensive and difficult to manufacture.

Thus, while it is easier to handle and control than solid cellulose, sophorose can nonetheless make the cost of producing cellulases prohibitively expensive and, thus, is impractical for commercial cellulase production.

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