The objective of the proposed research is to develop an efficient microbial process for commercial production of adhesive proteins with properties similar to the adhesive protein of the mussel Mytilis edulis. Unlike most adhesives, this strong adhesive car set and adhere to surfaces in wet environments. It should also be nontoxic. Thus, it is likely to have application in dental restorations, as a surgical soft tissue adhesive, and in a wide variety of other medical and specialized industrial applications. The natural mussel adhesive protein (MW 130,000) has been purified and characterized, and short analogs have been chemically synthesized. However, the most cost-effective method to produce a high molecular weight protein with the properties of the natural protein adhesive will undoubtedly be microbial production using a cloned gene. In Phase I, Genex isolated and sequenced cDNA clones encoding various portions of the mussel adhesive protein using Mytilis edulis phenol gland mRNA. In Phase II, several of the cDNA clones will be expressed in a Saccharomyces cerevisiae production strain; proprietary methods for fermentation' purification. hydroxylation and curing of adhesive protein analogs will be applied to the cDNA encoded proteins, further refined and scaled- up; and the potential for developing dental products based on these microbially produced proteins will be assessed.
