Coupled Enzyme Process for Tryptamine Synthesis

色胺合成的偶联酶工艺

• 批准号: 6622212
• 负责人: JAMES L KILGORE
• 金额: $ 35.84万
• 依托单位: BIOCATALYTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-07-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6622212
• 关键词: Escherichia coli; aromatic L aminoacid decarboxylase; chemical synthesis; directed evolution; enzyme activity; enzyme mechanism; enzyme substrate; immobilized enzymes; indoles; melatonin; method development; sumatriptan; thin layer chromatography; tryptamines; tryptophan synthase

DESCRIPTION (provided by applicant): Neuroactive tryptamine derivatives can be prepared by a new coupled-enzyme process developed in Phase 1 by combining tryptophan-synthetic and -decarboxylating enzymes. This comprises the first reported single-pot aminoethylation process. The tryptamine products can thereby be conveniently produced from readily-available indole precursors. Expression systems which provide higher enzyme yields will be utilized in order to permit scaleup to preparative reaction systems. The gene encoding Camptothectin accuminata tryptophan decarboxylase 1 (TDC 1) will be redesigned to optimize expression in E. coli. Stepwise binding of tryptophan synthase subunits and mutation of specific sites on the enzyme surface in order to improve enzyme activity following immobilization. Directed evolution will be explored in an effort to broaden the substrate ranges of two enzymes. New substrates will be tested he process will be utilized in the preparation of the migraine drugs rizatriptan, and sumatriptan, as well as melatonin and a series of melatonin receptor active compounds. PROPOSED COMMERCIAL APPLICATION: The coupled aminoethylation process will be used to make intermediates for neuropharmaceuticals, including both currently-approved and developmental neuroactive drugs.

描述（由申请人提供）：神经活性色胺衍生物可以是 通过在阶段1中开发的新偶联酶工艺制备， 色氨酸合成和脱羧酶。这包括第一个 报道了一锅氨乙基化工艺。色胺产品可以 从而方便地由容易获得的吲哚前体制备。 将使用提供更高酶产量的表达系统， 以允许放大到制备反应系统。编码基因 喜树碱色氨酸脱羧酶1（TDC 1）将被重新设计 优化E.杆菌色氨酸合酶的逐步结合 亚基和酶表面上特定位点的突变， 固定化后提高酶活性。定向进化将是 探索在努力扩大底物范围的两种酶。新 将对基质进行测试，该过程将用于制备 偏头痛药物利扎曲普坦和舒马曲普坦，以及褪黑素和一系列 褪黑激素受体活性化合物。 拟定商业应用： 偶联的氨乙基化过程将用于制备中间体， 神经药物，包括目前批准的和开发的 神经活性药物