Acquisition of Protein Purification Instrumentation for Isoflavonoid Research

该奖项支持收购蛋白质色谱系统和相关的蛋白质纯化和分析设备，用于俄克拉荷马州东南部州立大学的研究活动和本科教育。该仪器将用于研究多种豆科植物中产生的天然产物异黄酮和异黄酮缀合物。异黄酮类化合物在植物的生存中起着多种作用，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，异黄酮缀合物的特殊酶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。异黄酮缀合物在消化前或消化过程中的水解会极大地影响异黄酮的吸收、生物利用度和稳定性。尽管它们具有潜在的重要性，但很少有关于裂解异黄酮-葡萄糖苷的-葡萄糖苷酶的报道。该仪器将有助于植物β -葡萄糖苷酶的生化分析，对异黄酮缀合物具有高特异性，这是最近从药用苜蓿中分离到的克隆。为了确定异黄酮缀合物水解在营养和疾病预防中的相对重要性，将研究用于动物饲料研究的大量重组酶的表达和纯化条件。该仪器还将允许纯化和最终克隆酶7,2'-二羟基-4'-甲氧基-异黄烷醇脱水酶（DMID）。该酶催化抗真菌异黄酮类化合物medicarpin生物合成的最后一步，并可能参与其他重要的异黄酮类化合物的生物合成，如coumestans。该仪器的获得还将促进学生在当前生化和分子生物学技术方面的培训，并将直接使本科生参与功能基因组学的研究。该仪器将非常有助于修订现有的生物化学实验课程，并为生物学、化学和生物技术专业开设新的高级蛋白质技术半课程。东南俄克拉荷马州立大学拥有多元化的学生群体，包括美国原住民学生、来自农村地区的学生和第一代大学生。这些仪器将帮助这些学生为生物化学、生物技术和跨学科研究事业做好准备。

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.