混合模式扩张床吸附从枯草杆菌发酵液中分离纳豆激酶研究.pdfVIP

L027 混合模式扩张床吸附从枯草杆菌发酵液中分离纳豆激酶 贠 * 吕淼华 武元春 林东强 军贤 姚善泾 （浙江大学化学工程与生物工程系，浙江杭州 310027 ） 摘 要 用混合模式扩张床介质Fastline Pro ，通过静态吸附和固定床吸附实验，考察了pH 值和 离子强度对纳豆激酶吸附性能和洗脱特性的影响。结果表明，该介质对纳豆激酶的吸附能力较高， 特别是在高离子强度时。离子强度和pH 值均对吸附过程有影响，改变离子强度不能实现有效洗 脱，改变pH 值是洗脱的最佳途径。在此基础上，进行了扩张床吸附试验，从高离子强度的枯草 杆菌发酵液中直接分离得到了纳豆激酶，纯化因子高达12.3，该分离过程充分体现了扩张床吸附 的集成化优势。 关键词 扩张床吸附 混合模式介质 纳豆激酶 分离 SEPARATION OF NATTOKINASE FROM BACILLUS SUBTILIS FERMENTATION BROTH BY EXPANDED BED ADSORPTION WITH MIXED-MODE ADSORBENT LU Miaohua WU Yuanchun LIN Dongqiang YUN Junxian and YAO Shanjing (Department of Chemical and Biochemical Engineering, Zhejiang University, Hangzhou 310027,China) Abstract The influences of pH and ion concentration on the adsorption performance of nattokinase on a mixed-mode adsorbent Fastline Pro and the elution behaviors were investigated by batch and packed bed adsorption experiments, respectively. The results showed that the adsorption capacity of nattokinase with Fastline Pro was high, especially under high ion concentration. The adsorption processes were affected by either pH or ion concentration. A low elution efficacy was obtained by increasing NaCl concentration of the buffer, while the effective desorption of nattokinase was achieved by changing pH. Based on these results, nattokinase was directly recovered from Bacillus subtilis fermentation broth by expanded bed adsorption with the mixed-mode Fastline Pro. The purification factor reached 12.3, which demonstrated the integration advantage of expanded bed adsorption in enzyme separation. Keywords expanded bed adsorption, mixed-mode adsorbent, nattokinase, separation 引 言 扩张床吸附(Expanded Bed Adsorption，EBA)是一种新型的生化分离技术，能直接从发 酵液、细胞培养液或匀浆液中捕获目标产物，集固液分离、浓缩和初步纯化于一个操作单元 之中，因此可以减少操作步骤，提高产品收率，降低纯化费用，已有不少从细菌、酵母或动 物细胞中提取目标