基因工程菌表达维生素b,2条件优化研究-生物化学与分子生物学专业论文.docxVIP

摘 摘 要 利用基因重组技术构建的基因工程菌虽然在实验室中很容易做到，但如何 使它走出实验室，真正应用于生产实践，则需要对其利用什么手段，采用什么 技术，以及培养条件和控制方法进行深入细致的研究。 本文主要是对通过基冈重组手段构建的基因工程菌Y24／PM×45，利用发 酵手段来表达核黄素的条件进行优化，使之能够充分发挥基因工程菌的能力。 本文分别就基因工程菌的抗性、稳定性进行了验证，证实了基因工程菌Y24／PM ×45分别对红霉素和链霉素具有抗性；通过摇瓶试验考察了对基因工程菌的保 存条件、碳源、氮源，确定了基因工程菌Y24／PM×45的保藏条件和最佳碳源 为蔗糖，最佳氮源为酵母粉和硫酸铵并确定了原料的最佳配比；通过1L发酵 罐分别对原料的产地、通风量和pH范围等条件进行了优化，并将优化的条件 放大在30L的发酵罐表达核黄素，培养65小时，可以得到9～10∥l的核黄素， 为进一步扩大到工业化生产创立了模式和奠定了基础。 关键词：核黄素发酵基因工程菌 The The study of optimum conditions for the riboflavin expression with genetically engineered strain Abstracts Although it is casy obtain genetically engineered strain with genie recombination technology in laboratory，however,there is long way make it walk of laboratory and put it into production massively．That is say,we must study varies of conditions and means deeply,including methods of technology,culture and controlling In this paper， we have optimized varies of conditions for the riboflavin expression with genetically engineered strain Y24／PM×45 obtained by genic recombination technology by taking fermentation method，only making full of the effects of it． Also，we have verified the resistance and stability of genetically engineered strain and confirmed that the strain has resistance to erythromyein and streptomycin． What’S more，we have researched the reserving conditions，carbonic nitrogenous by shaking flask test，the results showed th．at the optimum carbonic is sucrose，the optimum nitrogenous is yeast powder and (NH4)2S04，and the optimum ratios of the carbonic and the nitrogenous With 1 liter fermentor，we have optimized the aerial flux，the ranges of pH and the materials of molasses and established the fermentation technology．When we put it into production by scaling-up these conditions，the yield is up to 9～109／l riboflavin in 30 1iter fermentor after 65h of culture． Key words：riboflavin；fermentation；genetically engineered strain 南开大学坝士研究生学位论文 南开大学坝士研究生学位论文 基因工程菌表达维生素B：条件优化研究 一、前 言 1．概述 维生素B：，俗称核黄素，是生物生命活动不可缺少的维生素之一。分子结 构中含有7，8一二甲基异咯嗪和