FLUORESCENT ACTIVATED CELL SORTING FOR ENZYME SCREENING

用于酶筛选的荧光激活细胞分选

• 批准号: 2540800
• 负责人: MARTIN KELLER
• 金额: $ 9.91万
• 依托单位: DIVERSA CORPORATION
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-11-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2540800
• 关键词: bioassay; biological products; biotechnology; catalyst; endopeptidases; enzyme activity; esterase; flow cytometry; genetic library; method development; microorganism; recombinant proteins; unspecific monooxygenase

There has been increasing demand in the research reagent, diagnostic reagent and chemical process industries for protein-based catalysts possessing novel capabilities. At present, this need is largely addressed using enzymes purified from a variety of cultivated microorganisms, generally members of the domains Bacteria and Archaea. However, because less than 1% of naturally occurring microorganisms have been grown in culture, alternative techniques must be developed to exploit the full breadth of diversity for potentially valuable new products. We propose to develop fluorescent activated cell sorting for an extreme high throughput screening for recombinant expression clones from uncultivated organisms to discover a large number of unique, robust, specific biocatalysts for the cost effective synthesis of chiral synthons for use in production of pharmaceuticals, biological intermediates and natural products. PROPOSED COMMERCIAL APPLICATION: and chemical process industries has already begun to grow. Growth in these fields is limited by a lack of enzyme activities useful under the processing conditions required. RBI currently provides a number of enzyme libraries for various enzyme classes (Glycosidases; Esterases/Lipases; Aminotransferases; Phosphatases) suitable for screening. This project will serve to increase the number of enzymes available from RBI with which researchers and process engineers can design new methods.

对研究试剂、诊断试剂、 蛋白质基催化剂的试剂和化学加工工业 拥有新的能力。目前，这一需求在很大程度上得到了满足 使用从多种培养的微生物中纯化的酶， 通常属于细菌和细菌属。但由于 只有不到1%的天然微生物生长在 文化，必须开发替代技术来充分利用 具有潜在价值的新产品的多样性。我们建议 开发荧光激活细胞分选，以实现极高的通量 从未培养的生物体中筛选重组表达克隆， 发现大量独特的，强大的，特定的生物催化剂， 用于制备手性二氢吡喃酮的成本有效的合成 医药、生物中间体和天然产物。 拟定商业应用： 化学工业已经开始发展。增长在这些 领域是有限的，缺乏酶的活动下有用的， 加工条件要求。RBI目前提供多种酶 各种酶类别的文库（糖苷酶;酯酶/脂肪酶; 氨基转移酶;磷酸酶）。该项目将 用于增加可从RBI获得的酶的数量， 研究人员和工艺工程师可以设计新的方法。