NSF/EPA TSE: Improving Resistance to Enzyme Alkylation During Enzyme-Catalyzed Production of Acrylamide

利用丙烯腈的酶促水合生产丙烯酰胺提供了许多经济和环境优点，并且作为用于生产广泛使用的商品化学品的“绿色”方法的第一个实例。 基于酶的方法提供了优于传统化学方法的显著的成本和环境优势，但是受到丙烯腈底物对腈水合酶催化剂的温度依赖性失活的阻碍。 本项目中待检验的假设是，使用新的嗜热菌分离株BR 444对丙烯腈灭活腈水合酶进行分子表征，随后使用定点突变进行酶改变，将提供更具丙烯腈抗性的酶。 该项目预计将提供有关酶耐烷基化的基本信息，以及可能改进的丙烯酰胺生物生产催化剂。 该奖项是由NSF作为NSF/EPA可持续环境技术联合计划的一部分。

Production of acrylamide utilizing enzymatic hydration of acrylonitrile offers a number of economic and environmental advantages, and serves as a first example of a "green" process for production of an extensively-utilized commodity chemical. The enzyme-based process offers significant cost and environmental advantages over the traditional chemical process, but is hindered by temperature-dependent inactivation of the nitrile hydratase catalyst by the acrylonitrile substrate. The hypothesis to be tested in this project is that molecular characterization of the nitrile hydratase inactivation by acrylonitrile with a new thermophile isolate BR444 and subsequent enzyme alteration using site-directed mutation will provide a more acrylonitrile-resistant enzyme. The project is expected to provide basic information regarding enzyme resistance to alkylation in addition to possible improved catalysts for acrylamide bioproduction. This award is madeby NSF as part of the joint NSF/EPA Program on Technologies for aSustainable Environment.