SBIR Phase I: Highly Active and Selective Hydroxylation Catalysts for Combinatorial Biocatalysis

这个小企业创新研究（SBIR）第一阶段项目将展示使用“定向进化”来创造高活性、选择性羟化酶的可行性，用于潜在药物的组合生物催化。这种蛋白质催化剂将能够直接羟基化非活性炭中心，而不会氧化其他敏感功能；这是一种用传统化学几乎不可能实现的转变。以高活性细菌酶为起点，随机产生突变体，并通过使用高通量筛选方法选择能够靶向羟基化的克隆。现有的细菌P450酶不具备所需的选择性。哺乳动物P450系统在体内有效工作，但难以表达且催化缓慢。这项技术在组合文库创建方面的潜力将通过药物支架来证明。该技术的商业应用最初将是为开发新的候选药物提供研究氧化催化剂。将准备用于测试的新型含氧候选者。随后，催化剂可用于新药的实际合成。

This Small Business Innovation Research (SBIR) Phase I project will demonstrate the feasibility of using "directed evolution" to create highly active, selective hydroxylation enzymes for use in combinatorial biocatalysis of potential drugs. Such protein catalysts will be capable of direct hydroxylation of non-activated carbon centers without oxidation of other sensitive functionality; a transformation that is virtually impossible using traditional chemistry. Using highly active bacterial enzymes as a starting point, random mutants will be generated and clones capable of targeted hydroxylations will be selected by using by using high-through-put screening methods. Existing bacterial P450 enzymes do not have the required selectivity. Mammalian P450 systems that work effectively in-vivo, are difficult to express and are slow in catalysis. The potential of this technology for combinatorial library creation will be demonstrated using drug scaffolds.The commercial application of the proposed technology will initially be to provide research oxidation catalysts for the development of new drug candidates. New classes of oxygenated candidates for testing will be prepared. Subsequently, the catalysts may be used in the actual synthesis of the new drugs.