绿色分离纯化生产高纯度维生素K_(2)  

Green separation and purification of high purity vitamin K_(2)

作　　者：马国良 郑之明 王鹏 王丽 赵根海 王晗 MA Guoliang;ZHENG Zhiming;WANG Peng;WANG Li;ZHAO Genhai;WANG Han(Institute of Intelligent Machinery,Hefei Institutes of Physical Science,Chinese Academy of Sciences,Hefei 230031,China;Science Island Branch of Graduate School,University of Science and Technology of China,Hefei 230026,China)

机构地区：[1]中国科学院合肥物质科学研究院智能机械研究所,合肥230031 [2]中国科学技术大学研究生院科学岛分院,合肥230026

出　　处：《生物学杂志》2023年第3期107-112,共6页Journal of Biology

基　　金：安徽省重点研究与开发计划:“针对骨质疏松症的维生素K 2制剂开发和应用研究”(No.202004b11020014);国家重点研发计划合成生物学专项:“智能工业菌株高效合成技术体系”(No.2019YFA0904300,No.2019YFA0904304);安徽省科技重大专项(No.202103a06020003);安徽省重点研发计划项目(No.1804b06020342);安徽省自然科学基金项目(No.1908085MB48,No.1908085MB43)。

摘　　要：为克服维生素K_(2)(Vitamin K_(2),VK_(2))提取工艺有毒溶剂用量大、有毒物质残留等缺点,建立一种高纯度VK_(2)绿色分离纯化工艺方法。使用天然油剂与甲醇、乙醇等有机试剂分别萃取纳豆芽孢杆菌干菌体内的VK_(2),结果表明,中链甘油三酯的萃取效果最好,最大提取量为2.7 mg/g。通过使用7种不同型号的大孔树脂分离中链甘油三酯溶剂相中的VK_(2),以及利用静态吸附与解吸附实验,筛选出最优型大孔树脂HC-200S。进一步优化大孔树脂的动态吸附和解吸附工艺,并且绘制大孔树脂的动态解吸附曲线,结果表明:上样液吸附流速为2.0 mL/min、解吸剂为无水乙醇/乙酸丁酯(1∶2,体积比)的混合溶液、解吸剂流速为1.0 mL/min时,HC-200S型大孔树脂对VK 2的吸附和解吸附效果最佳;VK_(2)纯度较纯化前提升4倍,达到87.8%,回收率达到93.1%。将树脂分离纯化得到的VK_(2)解吸液经过冷冻结晶和溶剂挥发结晶,分别得到纯度为96.7%和97.1%的VK_(2)晶体。研究将为VK 2的工业生产和绿色分离纯化提供方法依据,同时也为其他脂溶性维生素的分离纯化提供借鉴。In order to overcome the shortcomings of high purity Vitamin K_(2) extraction process,such as large amount of toxic solvent,serious pollution and toxic substance residues,a green separation and purification process of high purity Vitamin K_(2) was established.Natural oil,methanol,ethanol and other organic reagents were used to extract VK_(2) from Bacillus natto.The results showed that medium chain triglyceride had the best extraction effect,and the maximum extraction amount was 2.7 mg/g.Seven types of macroporous resins were used to separate VK_(2) from medium chain triglyceride solvent phase,and the optimal macroporous resin HC-200S was selected by static adsorption and desorption experiments.The dynamic adsorption and desorption process of macroporous resin was optimized,and the dynamic desorption curve of macroporous resin was drawn.The results showed that when the adsorption flow rate of sample solution was 2.0 mL/min,the desorption agent was anhydrous ethanol/butyl acetate mixture solution(1∶2,volume ratio)and the flow rate of desorption agent was 1.0 mL/min,the adsorption and desorption effect of HC-200S macroporous resin on VK_(2) was the best.At this time,the purity of vitamin K_(2) was 4 times higher than that of the before purification,reaching 87.8%,and the recovery rate reached 93.1%.VK 2 desorption solutions obtained by resin separation and purification were frozen crystallization and solvent volatilization crystallization,respectively,to obtain VK_(2) crystals with purity of 96.7% and 97.1%.This study provided a method basis for the industrial production and green separation and purification of VK_(2).

关 键 词：维生素K_(2) 大孔树脂 分离纯化 绿色提取 MK-7晶体 

分 类 号：Q568[生物学—生物化学]