导　　师： 张积仁

学科专业： 100201

授予学位： 硕士

作　　者： ;

机构地区： 中国人民解放军第一军医大学

摘　　要： 研究背景和研究目的： 生命现象的实质是能量的不断流动，生物的生命维持、细胞生长和更新，都需能量才能进行。还原型烟酰胺腺嘌呤二核苷酸/(NADH/)广泛存在于一切活细胞中，是细胞能量代谢所必需的辅酶。1分子NADH还原可产生3分子ATP，36千卡能量。它在维持细胞生长、分化和能量代谢中起十分重要的作用。 NADH不仅是酶促反应中一种重要的生化辅因子，同时也是一种有效的细胞保护剂。科学研究证明，包括心血管疾病、癌症以及许多老年病/(如Alzheimer's病、Parkinson's症和其它神经退行性变/)在内的八十多种疾病与自由基损伤有关，NADH是体内最有效的抗氧化剂，可使机体免受自由基损伤。作为一种新型细胞保护剂，它在修复细胞损伤；提高细胞应激反应能力；降低化疗药物、放射线及化学毒剂对正常组织和细胞的毒性损伤作用等方面，有直接而有效的作用，作为一种独特的天然物质，具有非常广泛的应用前景。因此，研究一种能经济、大规模、高效地制备、提取纯化NADH的新方法，具有重大的社会效益和经济效益。 我们希望通过本实验的研究，可以寻找出更为经济、高效、 简便的提取、分离和纯化辅酶NADH的工艺条件并进行优化，为 下一步中试放大和工业化生产积累经验。进而研制出以NADH为 主体的新型细胞保护剂，为今后临床疾病的细胞保护治疗及军事 医学领域细胞损伤的防护提供更加有效的措施和手段。 方法和结果： 1．NADH粗提方法 使用化学渗透法处理啤酒酵母细胞hoe切romyces cerev。s。e， 采用Lud正交设计，比较酵母细胞浓度、乙醚用量、提取时间 对NADH得率的影响，得出最佳提取条件。实验结果表� Background and Objective: Life is a constant flow of energy,with which ensure the maintenance,gowth and reproduction of the organisms. Nicotinamide adenine dinucleotide reduced /(NADH/) is present in every living cell and is essential to cellular energy production. One mole of NADH produces three moles of ATP and 36 kilocalories. It plays a very important role in cell growth,cell differentiation and energy metabolism. NADH serves as not only a important biochemical cofactor in enzymatic catalysis,but also a potent cytoprotective agent. Medical science proved that free radicals are involved in more than 81 diseases,including heart /& cardiovascular disease,cancer and the diseases we associate with aging /(like Alzheimer's,Parkinson's and other neuro-degenerative disorders/). On the basis of its biochemical properties,NADH is the most biological potent antioxidant found in the body and has the ability to protect the body from the damages of free radicals. Science is learning that NADH may have a direct positive involvement in enhancing the level of energy metabolism,repairing cellular damage,improving cellular stress ability,decreasing cytotoxicity caused by chemotherapy drugs,chemical toxic agents and radiation. It has a prospect of wide application as a remarkable natural substance,so there have significant social effects and economic returns to find a new technique of extraction,isolation and purification NADH in a inexpens ive,large-scaled and high-yielding way. We want to find and optimize a technique which is simple,inexpensive and high-yielding,and it should be able to be scaled up to deal with large volumes in industrial operation. An available way can be provided by producing cytoprotective drugs contained NADH in cytoprotection treatment for clinical and military medical cytotoxicity. Methods and Results:l.The method to crude extraction of NADH Saccharomyces cerevisiae cells were treated by chemical permeabilization to release

关 键 词： 还原型烟酰胺腺嘌呤二核苷酸 下游加工工程 化学渗透法 分离 纯化 亲和层析 膜分离技术 亲和超滤

领　　域： [化学工程]