PRODUCTION OF NON-NATURALLY OCCURRING AMINO ACIDS

非天然存在的氨基酸用作组合合成的药物中间体和前体。最近大量基于非天然氨基酸的候选药物对这些关键的药物构建块产生了强烈的需求。提出的研究重点是开发一种用于生产非天然存在的氨基酸的转氨过程。该过程是一种手性合成，而不是一种分解，导致碳的有效利用和减少浪费，该技术可以广泛应用于生产各种氨基酸目标。从简单、廉价的前体可以获得高的化学产量。该方法已被证明与四种目标非天然存在的氨基酸具有直接的商业应用。为了为制备规模的反应提供足够的酶并建立商业化生产的手段，将克隆和表达来自不同类别的编码关键转氨酶的基因，包括d -转氨酶。D-氨基酸和l -氨基酸的生产将以100克的规模进行演示。优化固定方法。通过在生物催化剂中加入脱羧酶，将实现一种驱动反应完成的改进方法。最后，将展示8种不同的15n标记氨基酸的立体选择性生产。建议的商业应用：用于制药中间体的非天然氨基酸的生产；构建块肽类似物和组合合成

Non-naturally occurring amino acids are used as pharmaceutical intermediates and precursors for combinatorial synthesis. The large number of recent drug candidates based on non-naturally occurring amino acids has created strong demand for these key pharmaceutical building blocks. The proposed research focuses on the development of a transamination process for the production of non-naturally occurring amino acids. The process is a chiral synthesis, not a resolution, leading to efficient use of carbon and reduced waste, and the technology may be applied broadly to produce a wide range of amino acid targets. High chemical yields can be achieved from simple, inexpensive precursors. The method has been demonstrated with four target non-naturally occurring amino acids having immediate commercial applications. To provide sufficient enzyme for preparative-scale reactions and establish the means for commercial production, genes encoding key transaminases from different classes and including D-transaminases will be cloned and expressed. The production of both D- and L-amino acids will be demonstrated on the 100 gram scale. Immobilization methods will be optimized. An improved method for driving the reaction to completion will be implemented by incorporating a decarboxylase into the biocatalyst. Finally, the stereoselective production of 8 different 15N-labeled amino acids will be demonstrated. PROPOSED COMMERCIAL APPLICATIONS: Production of non-naturally occurring amino acids for pharmaceutical intermediates; building blocks for peptide analogs and for combinatorial synthesis