Studies of a novel A and B blood group cleaving enzyme

新型A型和B型血型裂解酶的研究

• 批准号: 6647162
• 负责人: KIMBERLY M ANDERSON
• 金额: $ 2.4万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6647162
• 关键词: Clostridium; acidity /alkalinity; blood group antigens; enzyme activity; enzyme substrate; expression cloning; glycosidases; protein purification; protein sequence; thermodynamics

DESCRIPTION (provided by the applicant): Our laboratory has recently discovered a novel endo-glycosidase (Endo-ABase) in the culture fluid of Clostridium perfringens. The long-term goal of this research proposal is to isolate and characterize EndoABase, a medically useful glycosidase that may be capable of converting type A and type B red blood cells into blood type insensitive cells for transfusion. To achieve this goal, Endo-ABase must be produced in large enough quantity to destroy the blood group A and B activity of red blood cells before transfusion. Four specific aims have been proposed for this project. (1) Endo-ABase will be purified from the culture supenatant of Clostridium perfringens by a series of gel filtration, ion-exchange, and affinity chromatographies. (2) The properties of Endo-ABase will be characterized--stability, optimal pH. substrate specificity. etc. (3) Peptides generated from Endo-ABase will be sequenced, the gene of interest will be cloned, and the recombinant enzyme will be expressed on a large scale in E. coli or other suitable expression system. Finally, the recombinant enzyme will be used to treat type A and type B erythrocytes.

描述（由申请人提供）：我们的实验室最近发现了 梭状芽胞杆菌培养液中一种新型的内糖苷酶（内碱） 灌注。该研究建议的长期目标是隔离和 表征内核，一种医学上有用的糖苷酶，可能能够 将A型和B型红细胞转换为血型不敏感细胞 输血。为了实现这一目标，必须大量生产内部库 足够数量来破坏红细胞A和B活性 输血之前。该项目提出了四个具体目标。 （1） 内骨将从梭状芽孢杆菌的培养物中纯化 通过一系列凝胶过滤，离子 - 交换和亲和力进行渗透 色谱。 （2）内部库的特性将是 特征 - 稳定性，最佳pH。底物特异性。 （3）肽 将从胚胎库产生的测序，感兴趣的基因将是 克隆，重组酶将在E中大规模表达。 大肠杆菌或其他合适的表达系统。最后，重组酶将 用于治疗A型和B型红细胞。