摘要
为了有效提取基因工程菌DHPYAAS-T7发酵液中的莽草酸,文章采用了离子交换色谱法对发酵液中莽草酸进行分离纯化。包括发酵液的预处理、离子交换树脂的选择、动态吸附、洗脱、脱色、脱盐以及纯度检测等过程。研究结果表明,所选的4种树脂中,717型强碱性阴离子交换树脂的吸附容量最大,发酵液的浓缩上样浓度为6.46 g/L。当调节发酵液pH值为6.0时,莽草酸成完全解离状态,能被树脂完全吸附。以1.0 mol/L的NH4Cl溶液洗脱,莽草酸回收率为91.7%。以HZ-816大孔吸附树脂进行脱色,莽草酸回收率为98.8%。以732型强酸性阳离子交换树脂进行脱盐,回收率为98.3%。将得到的莽草酸溶液进行结晶,HPLC检测纯度,晶体纯度达到98.6%。该工艺能有效的从发酵液中分离纯化高纯度的莽草酸,为莽草酸生产工业化奠定了基础。
In order to extract Shikimic acid（SA） in fermentation liquid of Escherichia coli DHPYAAS-T7,in the present study, ion exchange chromatography was selected, including fermentation liquid pretreatment, selecting ion exchange chromatography, dynamic adsorption,elution, decoloration, desalination and purity detection processes. The data showed that 717 anion exchange resin was found to be suitable for isolation and purification of SA in the four selected resins. Using 717 anion exchange resins, the concentration of fermentation liquid was 6.46g/L. When the pH of fermentation liquid was 6.0 ,SA existed as the dianion for completely adsorbed by resin. When eluted with 1.0 mol/L NH4CI, decolorated with HZ-816 microporous adsorption resin and desalinated with 732 cation exchange resin,the recovery of SA was 91.7% ,98.3% and 98.3% respectively. The SA eluate was then crystallized and the purity of the SA crystal was 98.6% by HPLC. High purity SA was obtained from fermentation liquid through this process, which laid the foun- dation for large-scale production of SA.
出处
《药物生物技术》
CAS
2013年第6期528-531,共4页
Pharmaceutical Biotechnology
基金
国家自然科学基金项目(No.30780049)
国家973计划项目(No.2011CBA00801)
关键词
莽草酸
发酵液
离子交换
分离
纯化
工艺优化
Shikimic acid, Ion exchange chromatography, Isolation, Purification, Fermentation liquid, Process optimization
