【作者】 肖安风；

【导师】 雷爱祖； 童张法；

【作者基本信息】 广西大学 ， 化学工艺， 2001， 硕士

【摘要】 利用酵母酶系催化一磷酸腺苷（AMP）磷酸化生成三磷酸腺苷（ATP），是最有工业前景的ATP生产方法之一。稳定细胞质量、掌握反应机理、优化生产控制和提高赡效益是目前ATP工业化生产急待解决的问题。本文针对存在的问题，从基础研究出发，选用一株具有较高ATP生产活力的酵母菌株，对影响酵母培养的环境因素、酵母的通透性与ATP生产活性的相关性进行了较深入的探讨和试验研究，系统地探索了酵母细胞酶系合成ATP的反应机理，同时，对应用活性炭吸附和离子交换树脂分离作了基础性研究。 在现有酵母培养工作中，对培养基中的酵母膏、（NH₄）₂SO₄、K₂HPO₄、葡萄糖四种因素不同浓度水平进行正交实验，研究其对培养酵母菌浓及 AMP转化活性的影响，确定了用于ATP生产的酵母细胞培养工艺的最佳配方，其结果为：m（葡萄糖）=4％，m（酵母膏）=0．5％，m（K₂HPO₄）=0．2％，m（（NH4）₂SO₄）=0．15％。 考察了丙酮处理、日光曝晒以及自然干燥三种酵母处理方法的AMP转化率，并且与新鲜酵母的AMP转化活性进行了比较，结果发现酵母需经过通透化处理后才具有AMP转化活性。 研究了利用酵母酶系生产ATP的反应机理，考察了多种因素对AMP转化情况的影响，结果表明：酵母酶系催化AMP转化为ATP的反应受到不同因素的影响，AMP转化反应首先是葡萄糖的酵解和含能磷酸化合物的积累；糖的磷酸化反应优先于AMP的磷酸化反应；Mg²⁺为有关酶的激活剂，高雌的Mg²⁺对反应转化率的影响不大，其它因素的最佳条件为：n(葡萄糖卜=150mmo l/L、n（磷酸缓冲液）=200mmol/L、p（酵母量）＝50g/L、搅拌速率为75r/min、温度35℃、PH=7．0。 查阅有关文献，对ATP的分离与纯化提出了探索性步骤，并对某些影响ATP提取效果的因素进行了初步研究，得出一系列有用的实验数据。更多还原

【Abstract】 Using yeast enzyme to yield adenosine triphosphate （ATP） from adenosinemonophosphate （AMP） is one of the most interesting techniques in ATP industryAt present, stabilizing yeast quality mastering reaction mechanism, optimizingsynthesis technique and increasing production scalc are some problems eagerly tosolve. At the beginning of basic research, a strain of yeast that is of high ATPproducing activity was selected to study the relation between AMP conversionactivity and yeast cultivation condition. Then the effect of permeability of yeastcells on AMP conversion activity was investigated. Finally, some separation andpurification methods of ATP was put forward.By orthogonal experiments, the cffects of four factors （glucose, ycast extract,（NH₄）₂SO₄, K₂HPO₄） at three levels on the AMP convcrsion activity wasinvcstigatcd and then the optimal ratio of components in the fcrment culturemedium was decided. The results are: m（glucose）=4%, m（yeast extract）=0.5%,m（K₂HPO₄）=0.2%, m（（NH₄）₂SO₄）=0.15%.The AMP conversion activity was mcasured through yeast treatcd withacetone washing, sunshine insolating and natural drying permeably Comparingwith that of fresh yeast, it was found that only the trcated yeast is of AMPconversion activityThere are somc factors may affect AMP conversion activity The expcrimentresuIts show that the concentration of Mg’+ affects ATP conversion rate Iittle.The optimal synthctic condition is as fOllows: c（glucosc）=l50mmoUl, c（pbosphatebuffer）=200mmollL, P （yeast amount）=50g/L, reaction temperature is 35C, andpH==7.0. On the basis of thc expcrimellt results, the mechanism of synthcsiringATP was investigated by use of yeast enZyme. The first process of synthesisingATP is glycolysis and phosphorylation of glucosc, and the second process is theaccumulation of ATP Md+ is the active factor of Embden-Meyerhof Passway（EMP）.By means of rcferences a proccss of separation and purification of ATP was putfOrward and got some useful data.更多还原