A Collaborative Proposal: Engineering Disulfide Formation Kinetics to Enhance Heterologous Secretions in Saccharomyces Cerevisiae

合作提案：改造二硫键形成动力学以增强酿酒酵母的异源分泌

• 批准号: 0120690
• 负责人: Ronald Raines
• 金额: $ 25万
• 依托单位: University of Wisconsin-Madison
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-05-01 至 2005-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0120690&HistoricalAwards=false
• 关键词: Collaborative; Proposal; Engineering; Disulfide; Formation

The secretion of heterologous proteins that contain disulfide bonds is not at its full potential. Evidence suggests that the regulation of the redox state of the endoplasmic reticulum may be a bottleneck. A multifaceted approach is proposed to probe and manage the redox state in yeast in order to investigate the bottleneck hypothesis further and to ultimately increase secretion efficiency. The four elements of the proposed work are: (1) amplify the products of the individual genes now thought to be involved redox management to assess the effect, (2) use display methods to uncover possibly missing pathway elements, (3) add redox altering compounds to the growth medium to alter redox state, and (4) develop a tachyplesin I substrate-based diagnostic system.

含有二硫键的异源蛋白的分泌并不具有全部潜力。 有证据表明，内质网的氧化还原状态可能是瓶颈。 提出了一种多方面的方法来探测和管理酵母中的氧化还原状态，以进一步研究瓶颈假说并最终提高分泌效率。 拟议工作的四个要素是：（1）扩大了现在认为涉及氧化还原管理以评估效果的单个基因的产物，（2）使用显示方法来发现可能缺少的途径元素，（3）在生长培养基中添加氧化还原化合物以改变氧化还原状态，并改变氧化还原状态，（（4）开发基于tachypals i subtrate i subtrate Indrotrate诊断系统。