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: 6555857
• 负责人: KIMBERLY M ANDERSON
• 金额: $ 2.22万
• 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/6555857
• 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 梭菌培养液中的新型糖苷内切酶（Endo-ABase） 产气荚膜杆菌。本研究计划的长期目标是分离和 EndoABase 的特征，一种医学上有用的糖苷酶，可能能够 将A型和B型红细胞转化为血型不敏感细胞 用于输血。为了实现这一目标，Endo-ABase 必须大规模生产 数量足以破坏红细胞的 A 型和 B 型血活性 输血前。该项目提出了四个具体目标。 (1) Endo-ABase将从梭菌培养上清液中纯化 通过一系列凝胶过滤、离子交换和亲和来去除产气荚膜菌 色谱法。 (2) Endo-ABase 的特性为 特点——稳定性、最佳pH值。底物特异性。 (3) 肽类 从 Endo-ABase 生成的基因将被测序，感兴趣的基因将被 克隆，重组酶将在大肠杆菌中大规模表达。 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。