Acquisition of Protein Purification Instrumentation for Isoflavonoid Research

获取用于异黄酮研究的蛋白质纯化仪器

• 批准号: 0421379
• 负责人: Nancy Paiva
• 金额: $ 6.75万
• 依托单位: Southeastern Oklahoma State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0421379&HistoricalAwards=false
• 关键词: Acquisition; Protein; Purification; Instrumentation; Isoflavonoid

This award supports the acquisition of a protein chromatography system and associated protein purification and analysis equipment to be used in research activities and undergraduate education at Southeastern Oklahoma State University. The instrumentation will be used to study isoflavonoids and isoflavonoid conjugates which are natural products produced in many legume plant species. Isoflavonoids have been proposed to play several roles in plant survival, including defense against fungal pathogens and recruitment of beneficial nitrogen-fixing bacteria. Specific enzymatic hydrolysis of isoflavonoid conjugates may be essential during these processes. Isoflavonoids are also potentially important constituents of human foods, in that diets rich in isoflavonoids are thought to decrease rates of certain cancers, osteoporosis, and other diseases. Hydrolysis of isoflavonoid conjugates before or during digestion may greatly affect the uptake, bioavailability and stability of isoflavonoids. Despite their potential importance, little has been published on beta-glucosidases which cleave isoflavonoid beta-glucosides. The instrumentation will facilitate the biochemical analysis of plant beta-glucosidases with high specificity towards isoflavonoid conjugates, for which clones were recently isolated from Medicago truncatula. Conditions will be investigated for expression and purification of large amounts of recombinant enzymes for use in animal diet studies designed to determine the relative importance of isoflavonoid conjugate hydrolysis in nutrition and disease prevention. The instrumentation will also allow the purification and eventual cloning of the enzyme 7,2'-dihydroxy-4'-methoxy-isoflavanol dehydratase (DMID). This enzyme catalyzes the final step in the biosynthesis of the antifungal isoflavonoid medicarpin, and may be involved in the biosynthesis of other important isoflavonoid derivatives such as coumestans. Acquisition of this instrumentation will also facilitate the training of students in current biochemical and molecular biology techniques, and will directly involve undergraduates in functional genomics research. The instrumentation will be extremely beneficial to revisions of the existing Biochemistry laboratory courses and the initiation of a new Advanced Protein Techniques half-course, for biology, chemistry and biotechnology majors. Southeastern Oklahoma State University serves a diverse student body, including Native American students, students from rural areas, and first generation college students. The instrumentation will aid in preparing these students for biochemistry, biotechnology and interdisciplinary research careers.

该奖项支持在东南俄克拉荷马州州立大学的研究活动和本科教育中使用的蛋白质色谱系统和相关蛋白质纯化和分析设备的收购。 该仪器将用于研究在许多豆科植物物种中产生的天然产物-。 已经提出异黄酮在植物存活中发挥几种作用，包括防御真菌病原体和招募有益的固氮细菌。 在这些过程中，特异性的酶促水解可能是必不可少的。 异黄酮也是人类食物中潜在的重要成分，因为富含异黄酮的饮食被认为可以降低某些癌症，骨质疏松症和其他疾病的发病率。 在消化前或消化过程中，β-内酰胺酶结合物的水解可能极大地影响β-内酰胺酶的吸收、生物利用度和稳定性。 尽管它们具有潜在的重要性，但很少有关于切割β-葡糖苷的β-葡糖苷酶的报道。 该仪器将促进植物β-葡萄糖苷酶的生化分析，具有高特异性对葡糖苷酶缀合物，其克隆最近从蒺藜苜蓿中分离出来。 将研究用于动物饮食研究的大量重组酶的表达和纯化的条件，所述动物饮食研究旨在确定重组酶缀合物水解在营养和疾病预防中的相对重要性。 该仪器还将允许纯化和最终克隆酶7，2 '-二羟基-4'-甲氧基-藜芦醇脱氢酶（DMID）。 这种酶催化抗真菌的阿托伐他汀的生物合成的最后一步，并可能参与其他重要的阿托伐他汀衍生物如香豆烷的生物合成。 这种仪器的收购也将促进当前的生物化学和分子生物学技术的学生的培训，并将直接参与功能基因组学研究的本科生。 仪器将是非常有益的现有的生物化学实验室课程的修订和启动一个新的高级蛋白质技术半课程，生物，化学和生物技术专业。 东南俄克拉荷马州州立大学为多样化的学生群体提供服务，包括美国原住民学生，农村地区的学生和第一代大学生。 仪器将帮助这些学生为生物化学，生物技术和跨学科研究职业做好准备。