枯草杆菌重组水蛭素的冷冻干燥工艺及热稳定性研究  

Studies on the Freeze Drying Technology and Thermal Stability of Recombinant Hirudin from Bacillus subtilis

作　　者：陈华友[1,2] 耿旭[3] 齐向辉[1,2] 徐庆刚[1] 

机构地区：[1]江苏大学,江苏镇江212013 [2]南京工业大学,江苏南京210009 [3]河南大学医学院,河南开封475004

出　　处：《安徽农业科学》2009年第35期17695-17697,共3页Journal of Anhui Agricultural Sciences

基　　金：国家"863"资助项目(2006AA03Z0453);江苏省高校自然科学研究项目资助(09KJB230001);国家"973"资助项目(2009CB724700);江苏大学校基金资助项目(08JDG009)

摘　　要：[目的]研究枯草杆菌重组水蛭素的冷冻干燥优化工艺及在酸碱中的热稳定性。[方法]测定不同填充剂、保护剂浓度组合下的共熔点,根据共熔点最高的原则确定其组合浓度,据此再设计水蛭素的冷冻干燥工艺。[结果]优化的冷冻干燥工艺为:填充剂浓度4%,保护剂浓度1.2%,共熔点为-30℃,装样量为3.0 ml,冷冻温度-40℃,冷冻时间2 h,升华干燥温度-32℃,升华时间24 h,再干燥温度25℃,时间5 h,活力回收为96.2%,含水量为1.48%。100℃时,酸性条件下,水蛭素非常稳定;加热结合碱性的条件下水蛭素才相当不稳定。[结论]可为进一步工业化冷冻干燥水蛭素研究提供借鉴。[ Objective ] The research aimed to study the optimized freeze drying technology and thermal stability of hirudin produced from Bacillus subtilis.[ Method ] Through measuring the euteetie melting point of different concentration of bulking agent and protective agent, and deciding their concentrations in principle of the highest euteetie melting point, the freeze drying technologies of hirudin were designed. [ Result ] The optimized freeze drying procedures were as follows： concentrations of bulking agent （mannitol）and protective agent （human serum albumin）were 4% and 1.2% respectively. And the corresponding eutectic melting point was 30 ℃. Freezing temperature was -40 ℃. Freezing time was 2 h. Sublimation drying temperature was - 32 ℃. Sublimation drying time was 24 h. Redrying temperature was 25℃. Redrying time was 5 h. The proper loading volume was 3.0 ml, and the corresponding recovery was 96.2% , and containing water was l. 48%. Hirudin was stable in 100℃ and acidity condition, but it was unstable under the condition of both heating and alkalinity. [ Conclusion] The research provided the reference for the further industrialization freeze drying of hiruin.

关 键 词：水蛭素 枯草杆菌 共熔点 冷冻干燥 

分 类 号：Q813.1[生物学—生物工程]