豆油在红霉素发酵中作用及作用机制探究.docVIP

豆油在红霉素发酵中作用及作用机制探究 　 作者：沈兆兵 陈国豪 陈长华 毕业论文 【摘要】; 红色糖多孢菌D在15L发酵罐的基础培养基和发酵过程中补加豆油，当基础培养基中豆油量加倍时，效价提高43.1%；发酵过程补加豆油罐的红霉素效价、生产强度和Yp/s比不补豆油的分别提高了69.7%、69.6%和63.8%。通过实验测定发现，发酵过程补加豆油后，异柠檬酸脱氢酶、甲基丙二酰CoA异构酶的比活以及α?酮戊二酸的量均高于不补加豆油时的值，而琥珀酸的量低于不补加豆油时的值。推测豆油经过红色糖多孢菌代谢后进入红霉素合成的一条可能途径：豆油经过代谢后进入TCA循环，并强化了TCA循环的通量，产生了大量α?酮戊二酸，再生成琥珀酰CoA，琥珀酰CoA在甲基丙二酰CoA异构酶的作用下生成甲基丙二酰CoA作为红霉素合成的前体。 毕业论文 【关键词】; 豆油； 红色糖多孢菌； 红霉素发酵； α?酮戊二酸； 甲基丙二酰CoA； 琥珀酸 毕业论文 　　Study on effect of soybean oil on fermentation of　erythromycin and its mechanism 毕业论文 　　ABSTRACT; The final production of erythromycin by Saccharopolyspora erythraea was increased by 43.1% in 15L bioreactor with double quantily of soybean oil into culture medium. The final production, production capacity and Yp/s of erythromycin were improved 69.7%, 69.6% and 63.8%, respectively, in 15L bioreactor with added soybean oil during fermentation. Specific activity of NADP+?isocitrate dehydrogenase, methylmalonyl ?coenzyme A mutase and the concentration of α?ketoglutaric acid were higher with added soybean oil than that not added, and the concentration of succinic acid was lower than that without added soybean oil. Therefore, the metabolic pathway from soybean oil to erythromycin was presumed as follows: soybean oil is metabolized into TCA pathway to generate much α?ketoglutaric acid and then generate succinyl?CoA by strengthening TCA pathway, and the methylmalonyl?CoA is formed from succinyl?CoA by isomerization. Finally, methylmalonyl?CoA can be used as the precursor for the biosynthesis of erythromycin. 毕业论文 ; 　　KEY WORDS; Soybean oil Saccharopolyspora erythraea Erythromycin fermentation α?ketog?lutaric acid Methylmalonyl?CoA Succinic acid 毕业论文 　　红霉素的抗菌谱和青霉素相似，是治疗耐药性金葡菌和溶血性链球菌感染所引起疾病的首选药物，临床上可用于对青霉素过敏的患者［1］。红霉素的生物合成是由1个丙酰辅酶A开始的，依次接上6个2?甲基丙二酰辅酶A而形成的［2］。丙酰辅酶A的合成有2条途径：①丙酰辅酶A合成酶途径；②丙酸激酶与酰基磷酸辅酶A转移酶偶联也能够催化丙酸生成丙酰辅酶A。红色糖多孢菌合成红霉素所必须的另一个前体是甲基丙二酰辅酶A，合成甲基丙二酰辅酶A有3条途径［3］：①以三羧酸循环的琥珀酰辅酶A为底物的甲基丙二酰辅酶A变位酶和甲基丙二酰辅酶A消旋酶途径［4］；②以丙酰辅酶A为底物的甲基丙二酰辅酶A转羧基酶途径［5］