PRODUCTION OF NON-NATURALLY OCCURRING AMINO ACIDS

非天然氨基酸的生产

• 批准号: 6374135
• 负责人: JAMES DAVID ROZZELL
• 金额: $ 29.74万
• 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: 2000-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6374135
• 关键词: amines; aminoacid; chemical synthesis; decarboxylases; gene expression; immobilized enzymes; molecular cloning; oxaloacetates; peptide chemical synthesis; stereoisomer; technology /technique development; transaminases; transamination

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

非天然存在的氨基酸被用作组合合成的药物中间体和前体。最近大量基于非天然存在的氨基酸的候选药物对这些关键药物构建模块产生了强劲的需求。拟议的研究重点是开发用于生产非天然氨基酸的转氨过程。该过程是手性合成，而不是拆分，从而有效利用碳并减少浪费，并且该技术可以广泛应用于生产各种氨基酸靶标。高的化学产率可以从简单、廉价的前体获得。该方法已被证明与四个目标非天然存在的氨基酸具有直接的商业应用。为了提供足够的酶用于规模化反应并建立商业生产的方法，将克隆和表达编码来自不同类别的关键转氨酶（包括D-转氨酶）的基因。D-和L-氨基酸的生产将以100 g规模进行证明。将优化固定化方法。将通过将脱羧酶并入生物催化剂中来实施用于驱动反应完成的改进方法。最后，立体选择性生产的8个不同的15 N-标记的氨基酸将被证明。拟议的商业应用：生产用于药物中间体的非天然氨基酸;用于肽类似物和组合合成的结构单元