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据报道,儿茶酚-O-甲基转移酶(COMT,EC 2.1.1.6)与人类多种疾病(包括帕金森病)有关,这些疾病的最佳治疗方案取决于COMT抑制剂的疗效。因此,有必要研发具有COMT抑制活性的药物,而这些药物的研发需要制备重组COMT。本研究在甲醇诱导毕赤酵母(Pichia pastoris)培养基中发酵制备具有生物学活性的六组氨酸标记的COMT,并利用固定化金属亲和色谱纯化目标酶。该酶具有良好的催化活性和选择性。在最
It has been reported that catechol-O-methyltransferase (COMT, EC 2.1.1.6) is associated with a number of human diseases, including Parkinson’s disease, and the optimal treatment for these conditions depends on the efficacy of the COMT inhibitor. Therefore, it is necessary to develop drugs with COMT inhibitory activity, and the development of these drugs requires the preparation of recombinant COMT. In this study, a biologically active hexahistidine-tagged COMT was fermented in methanol-induced Pichia pastoris medium and the target enzyme was purified by immobilized metal affinity chromatography. The enzyme has good catalytic activity and selectivity. At the most