STTR Phase I: Microbial Production of a Multifunctional Natural Product 5-Hydroxytryptophan

STTR 第一阶段：多功能天然产物 5-羟基色氨酸的微生物生产

• 批准号: 1520927
• 负责人: Yuheng Lin
• 金额: $ 22.5万
• 依托单位: BiotecEra Inc.
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1520927&HistoricalAwards=false
• 关键词: STTR; Phase; Microbial; Production; Multifunctional

The broader impact/commercial potential of this Small Business Technology Transfer (STTR) project will be to develop a microbe to produce 5-hydroxytryptophan (5-HTP) for use as a dietary supplement, and as an animal feed supplement. 5-HTP is a multifunctional natural product that has been used as antidepressant, sleep aid, and appetite suppressant worldwide. In addition, 5-HTP can be used to prevent milk fever, a common disease occurring among farm animals, which causes substantial economic losses. Currently, the only commercial production of 5-HTP is through the extraction from the seeds of Griffonia simplicifolia, a woody climbing shrub grown in west and central Africa. However, the insufficient and unstable supply of the raw material has resulted in a high and volatile price of 5-HTP, which has greatly constrained the development of its market. The technology to be developed is expected to lower the production cost by up to 90% compared with the current botanical extraction method, secure the stable production of 5-HTP, and satisfy the market demands.This STTR Phase I project proposes to create an optimal 5-HTP-producing strain that can be readily used for large scale production. The strain is expected to have the following desired features: 1) Eliminates the use of antibiotics; 2) eliminates the use of the expensive inducer IPTG; and 3) provides an optimized co-factor generation and recycling system. The company has achieved the microbial production of 5-HTP at over 1 g/L from tryptophan in shake flasks, which demonstrates great commercialization potential. However, this approach still uses antibiotics and the expensive inducer IPTG, which can increase the cost of large scale production by about 44%. In addition, extensive use of antibiotics in large scale production may raise environmental concerns. To address these issues and achieve the goal of this STTR Phase I project, the following three research objectives will be pursued: 1) Construction of an antibiotic-free plasmid selection system; 2) development of an inducer-free expression system with incremental strengths; and 3) optimization of the generation and recycling of cofactor MH4.

该小企业技术转让 (STTR) 项目更广泛的影响/商业潜力将是开发一种微生物来生产 5-羟基色氨酸 (5-HTP)，用作膳食补充剂和动物饲料补充剂。 5-HTP 是一种多功能天然产物，在世界范围内被用作抗抑郁剂、助眠剂和食欲抑制剂。此外，5-HTP还可用于预防奶热病，这是一种农场动物常见的疾病，造成巨大的经济损失。目前，5-HTP 的唯一商业化生产是通过从加纳单叶植物 (Griffonia simplicifolia) 的种子中提取，加纳植物是一种生长在西非和中非的木本攀缘灌木。但由于原料供应不足且不稳定，导致5-HTP价格居高不下且波动较大，极大地制约了其市场的发展。与目前的植物提取方法相比，拟开发的技术预计可降低生产成本高达90%，确保5-HTP的稳定生产，满足市场需求。该STTR一期项目旨在创建一个最佳的5-HTP生产工艺。 5-HTP产生菌株，易于大规模生产。该菌株预计具有以下所需特征：1）消除抗生素的使用； 2) 消除了昂贵的诱导剂IPTG的使用； 3) 提供优化的辅因子生成和回收系统。公司已实现摇瓶中色氨酸微生物生产5-HTP超过1g/L，具有巨大的商业化潜力。然而，这种方法仍然使用抗生素和昂贵的诱导剂IPTG，这会使大规模生产的成本增加约44%。此外，大规模生产中抗生素的大量使用可能会引起环境问题。为了解决这些问题并实现STTR一期项目的目标，将追求以下三个研究目标：1）构建无抗生素质粒选择系统； 2) 开发具有增量强度的无诱导物表达系统； 3) 辅因子 MH4 的生成和回收优化。