“…In an effort to evolve the cyclization specificity of the Thermoanerobacterium thermosulfurigenes strain EM1 (Tabium) CGTase into R-amylase like hydrolytic activity on starch, we separately applied error-prone PCR (epPCR) and saturated mutagenesis, also evaluating the effectiveness of directed evolution techniques for interconversion of reaction specificities. Directed evolution has emerged as a powerful method for the creation of proteins with altered properties, e.g., tighter binding, resistance to denaturing conditions, and modified and/or improved reaction specificities (8)(9)(10)(11)(12)(13). Several directed evolution methods are commonly used nowadays, epPCR, DNA shuffling, site-saturation mutagenesis, etc., in the generation of genetic diversity, as reviewed (14,15).…”