Production of Heterologous Proteins by Recombinant Sccharomyces Cerevisiade

重组酿酒酵母生产异源蛋白

• 批准号: 8913497
• 负责人: Alkis Constantinides
• 金额: $ 9.9万
• 依托单位: Rutgers University New Brunswick
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-01 至 1992-02-29
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=8913497&HistoricalAwards=false
• 关键词: Production; Heterologous; Proteins; Recombinant; Sccharomyces

The object of this project is to determine and quantify the environmental factors that influence the productivity of a cloned protein by a recombinant yeast population. The system used as a model is Saccharomyces cerevisiae transformed with a 2u hybrid plasmid. This is a host- vector system with significant practical applicability due to the potential of 2u-based plasmids for good stability and relatively high copy numbers. The effects of the bioculturing environment on host cell physiology and recombinant productivity will be studied for the system S. cerevisiae - 2u hybrid plasmid under the control of constitutive and inducible promoters. The identity and concentration of medium components, the nature of nutrient limitation in the chemostat, and the bioreactor operation strategy will be examined systematically in order to assess quantitatively the influence of these parameters on plasmid stability, copy number, growth rate, and cloned protein expression rate. Saccharomyces cerevisiae (baker's yeast) is an excellent microorganism for the production of recombinant proteins since it is non-pathogenic to humans and has been grown at an industrial scale for centuries. Large-scale fermentation of recombinant yeast is currently used for the production of several valuable products, including therapeutic proteins. The proposed research program will provide a better understanding of the factors that influence the efficiency of recombinant protein production in yeast using a system of commercial significance, the 2u hybrid plasmid. This project will also suggest appropriate manipulations of the various fermentation parameters that can result in more economic production of biomedically important proteins through this biotechnological route. The generation of new knowledge in this area and the potential development of industrial operational strategies will increase the competitive edge of the United States in biotechnology.

本项目的目标是确定和量化 影响生产力的环境因素 通过重组酵母群体克隆蛋白质。 的 用作模型的系统是酿酒酵母 用2u杂交质粒转化。 这是一个主机- 矢量系统具有重要的实际适用性， 基于2u的质粒具有良好稳定性的潜力 和相对高的拷贝数。 的影响 生物培养环境对宿主细胞生理学的影响， 将研究系统S的重组生产率。 cerevisiae - 2u杂合质粒 组成型和诱导型启动子。 身份和 培养基成分浓度、营养物质性质 恒化器和生物反应器操作的限制 将系统地审查战略，以评估 定量分析了这些参数对质粒的影响 稳定性、拷贝数、生长速率和克隆蛋白 表达率 酿酒酵母（Saccharomyces cerevisiae）是一种优良的 用于生产重组蛋白的微生物 因为它对人类无致病性， 几个世纪以来的工业规模。 大规模 重组酵母的发酵目前用于 生产多种有价值的产品，包括 治疗性蛋白质 拟议的研究计划将 更好地了解影响 影响重组蛋白生产的效率 在使用具有商业意义的系统的酵母中， 杂交质粒 该项目还将建议适当的 各种发酵参数的操作， 可以带来更经济的生物医学生产， 重要的蛋白质。 这一领域新知识的产生以及 工业经营战略的潜在发展 将提高美国的竞争力， 生物技术.