SALT-ACTIVATED ENZYMES FOR ORGANIC SYNTHESIS

酶能在非水介质中发挥作用的发现为酶在合成化学中的应用提供了巨大的潜力。然而，酶在有机介质中的活性几乎总是比在水溶液中低得多。由于酶催化转化的许多理想目标不溶于水，但可溶于有机溶剂，因此在有机介质中的低催化活性限制了酶在合成化学中的商业应用。最近的研究已经表明，在盐基质中冻干丝氨酸蛋白酶以>1000的因子活化酶以用于在有机溶剂中的催化。第一阶段的研究目标是完善制备盐活化酶的方法，通过测试不同反应类别的酶（包括脂肪酶、金属蛋白酶、酰胺酶、糖苷酶、氨基转移酶和糖苷酶）来展示范围的广度，并测量各种盐活化酶在有机溶剂中的稳定性。如果成功，拟议的研究将导致在非水溶剂中使用酶的广泛适用的方法。作为具有商业重要性的一个具体例子，将在两种关键抗生素前体的酰化中评价盐活化青霉素酰胺酶。该反应将确立用于半合成β-内酰胺抗生素的酶促合成的改进方法的可行性。拟议的商业应用：生产医药中间体和精细化学品;开发改进的生物催化剂

The discovery that enzymes can function in non-aqueous media offers the potential for expanding dramatically the usefulness of enzymes in synthetic chemistry. However, the activity of enzymes in organic media is almost always much lower than that in aqueous solutions. Since many desirable targets for enzyme-catalyzed transformations are insoluble in water, but soluble in organic solvents, low catalytic activity in organic media has thus far limited commercial applications of enzymes in synthetic chemistry. Recent research has shown that lyophilization of serine proteases in a salt matrix activates the enzymes by factors of >1000 for catalysis in organic solvents. The Phase 1 research goals are to refine the methods for preparing salt-activated enzymes, demonstrate the breadth of scope by testing enzymes from different reaction classes, including lipases, metallo-proteases, amidases, dehydrogenases, aminotransferases, and glycosidases, and measure the stability of various salt-activated enzymes in organic solvents. The proposed research would, if successful, result in a broadly applicable method for using enzymes in non-aqueous solvents. As a specific example of commercial importance, salt-activated penicillin amidase will be evaluated in the acylation of two key antibiotic precursors. This reaction will establish the feasibility of an improved method for the enzymatic synthesis of semi-synthetic beta-lactam antibiotics. PROPOSED COMMERCIAL APPLICATIONS: Production of pharmaceutical intermediates and fine chemicals; development of improved biocatalysts