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

利用丙烯腈的酶促水合作用生产丙烯酰胺具有许多经济和环境优势，是生产一种广泛利用的商品化学品的“绿色”工艺的第一个例子。与传统的化学工艺相比，基于酶的工艺具有显著的成本和环境优势，但由于丙烯腈底物对腈水合酶催化剂的温度依赖性失活而受到阻碍。在这个项目中要测试的假设是，用新的嗜热性分离物BR444对丙烯腈使丁腈水合酶失活进行分子表征，随后使用位点定向突变对酶进行改变，将提供更具丙烯腈抗性的酶。该项目预计将提供有关酶抗烷基化的基本信息，以及可能改进的丙烯酰胺生物生产催化剂。该奖项由美国国家科学基金会颁发，作为美国国家科学基金会/美国环保署可持续环境技术联合项目的一部分。

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.