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过程工程学报 ›› 2019, Vol. 19 ›› Issue (4): 728-734.DOI: 10.12034/j.issn.1009-606X.218311

• 反应与分离 • 上一篇    下一篇

大孔树脂吸附法提取羟基钴胺素

杨 军1,2， 王自强2， 王云山2*， 张万忠1， 苏志国2   

  1. 1. 沈阳化工大学制药与生物工程学院，辽宁 沈阳 110142 2. 中国科学院过程工程研究所生化工程国家重点实验室，北京 100190
  • 收稿日期:2018-11-06 修回日期:2018-12-25 出版日期:2019-08-22 发布日期:2019-08-15
  • 通讯作者: 王云山 yswang@ipe.ac.cn
  • 基金资助:
    国家自然科学基金资助项目 (U1302274)

Extraction of hydroxycobalamin with the method of macroporous resin adsorption

Jun YANG1,2, Ziqiang WANG2, Yunshan WANG2*, Wanzhong ZHANG1, Zhiguo SU2   

  1. 1. College of Pharmaceutial and Biological Engineering, Shenyang University of Chemical Technology, Shenyang, Liaoning 110142, China 2. National Key Laboratory Biochemistry Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China
  • Received:2018-11-06 Revised:2018-12-25 Online:2019-08-22 Published:2019-08-15
  • Supported by:
    Projects (U1302274) supported by the National Science Foundation of China

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摘要： 比较了8种大孔树脂对费氏丙酸杆菌发酵提取液光解转化后形成的羟基钴胺素的吸附性能，考察了上柱料液pH值对吸附量的影响，分析了不同浓度和pH下乙醇溶液作为洗脱剂的解吸情况，研究了上柱速率对吸附量和解吸率的影响。结果表明，静态实验中，LX-50B型大孔树脂对羟基钴胺素的吸附性能最好，当上柱溶液pH为6时，LX-50B树脂饱和吸附量最高，为203.87 mg/g。用pH为2的60%乙醇溶液作洗脱剂时解吸效果最好，羟基钴胺素提取液以2 BV(床柱体积)/h速度上柱吸附饱和，用60%乙醇溶液以0.5 BV/h速度洗脱后，羟基钴胺素浓度提高至1863.32 mg/L，为原提取液浓度的35倍，纯度由0.38%提升至15.73%，提取纯化效果良好。

关键词: 脱氧腺苷钴胺素, 羟基钴胺素, 大孔树脂, 提取

Abstract: Propionibacterium freudenreichii is regarded as a GRAS (generally recognized as safe) strain, and has been successfully applied to industrial anaerobic production of deoxyadenosylcobalamin, known as the natural structure of vitamin B12. Generally, due to the unstable of deoxyadenosylcobalamin, it needs to be converted to cyanocobalamin by cyanidation during the subsequent extraction and purification steps. Hydroxycobalamin is another form of vitamin B12 and widely used in the treatment of neurological diseases and cyanide poisoning. For the industrial production of hydroxycobalamin, cyanocobalamin is usually used as the raw material and then deacylated to form hydroxycobalamin. According to this principle, the production of hydroxycobalamin involves multiple chemical reactions, including cyanidation and deacylation, along with the high cost, environmental pollution, and hazardous conditions to the operators. To avoid these disadvantages, the photolysis production of hydroxycobalamin directly from deoxyadenosylcobalamin was proposed. In this work, the adsorption and extraction of hydroxycobalamin in the deoxyadenosylcobalamin photolysis solution were studied. The adsorption properties of 8 kinds of different macroporous resins on hydroxycobalamin were compared, and the effects of pH and ethanol concentration on hydroxycobalamin adsorption and desorption were discussed. The loading and elution velocity were optimized to improve the hydroxycobalamin yield. The results showed that LX-50B macroporous resin had the best adsorption performance for hydroxycobalamin in the static adsorption process. The adsorption capacity of LX-50B macroporous resin reached 203.87 mg/g at pH 6.0. For hydroxycobalamin eluent, 60% ethanol solution with pH 2.0 was the best choice. The optimal loading velocity and elution velocity were 2 and 0.5 BV(bed volume)/h, respectively. Under the above conditions, the concentration of hydroxycobalamin in the elution was 1863.32 mg/L, improved by 35 times compared with the original photolysis solution, while the purity of hydroxycobalamin increased from 0.38% to 15.73%, with an acceptable extraction yield of 92.46%, which achieved to a good extraction and preliminary purification result.

Key words: deoxyadenosylcobalamin, hydroxycobalamin, resin, extraction