【作者】 谢小瑜；

【导师】 雷爱祖； 童张法；

【作者基本信息】 广西大学 ， 化学工艺， 2006， 硕士

【摘要】 利用固定化酵母催化一磷酸腺苷（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。该法的中心内容包括：稳定的细胞质量、适宜的固定化形式、适宜的反应操作条件等。本文对酵母的培养条件、开孔明胶固定化细胞的制备条件以及ATP生产的反应条件进行了较深入的探讨和试验研究。此外还建立了ATP分离与提纯的实验方法，并通过实验考察了发酵液的ATP回收率、成品ATP的纯度以及ATP的实际产率。 首先选用了一株转化率较好但稳定性差的酵母菌株，对扩培培养基中的蔗糖蜜、蛋白胨、（NH₄）₂SO₄、KH₂PO₄四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ATP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较佳配方，其结果为：m（蔗糖蜜）=12％，m（蛋白胨）=0.70％，m（KH₂PO₄）=0.15％，m（（NH₄）₂SO₄）=0.25％。实验还确定了该啤酒酵母适宜的扩大培养时间是14～20小时。 研究了开孔明胶固定化啤酒酵母细胞的制备条件，结果表明15％的明胶和2％的海藻酸钠形成的复配胶包埋10％的啤酒酵母细胞制备成的固定化颗粒经2％戊二醛交联1小时具有较高的磷酸化活性。 利用该固定化颗粒研究了酵母酶系生产ATP的反应机理，考察了多种因素对ATP转化情况的影响，结果表明用开孔明胶包埋啤酒酵母制成的固定化细胞，在含有15g／LAMP，葡萄糖150mmol／L，硫酸镁16mmol／L，pH7.0的磷酸缓冲液250mmol／L中35℃振荡反应2小时，ATP转化率可高达95％更多还原

【Abstract】 Using immobilized yeast cells to yield adenosine tripHospHate （ATP） from adenosine monopHospHate （AMP） is one of the most interesting techniques in ATP industry. The key techniques in the ATP production are the steady of the cell quality, the appropriate method of immobilization and operational conditions. In the dissertation, the cultivation of yeast, the preparation of immobilized cells in open pore gelatin batrix and the operational conditions of ATP fermentation were investigated. Besides, the separation and purification technique of ATP production was built, the ATP recovery, the purity of ATP product and virtual productivity were also discussed with the experiment.First of all, a strain of beer yeast which has high conversion rate but low stability was selected. According to orthogonal experimental design, the effects of four factors （contents of cane sugar honey, peptone, （NH₄）SO₄ and KH₂PO₄） at three levels on the cells concentration and the ATP conversion activity were investigated and then the optimal ratio of components in the expanding culture medium were determined. The results are: m（cane sugar honey）=12%, m（peptone）=0.70%, m （（NH₄）₂SO₄）=0.25% and m（KH₂PO₄）=0.15%. Then, it was found that the appropriate expanding culture time was 14～20 hours.Secondly, it was selected to study the preparation of immobilized cells in更多还原