In Situ Magnetic Separation for Extracellular Protein Production
In Situ Magnetic Separation for Extracellular Protein Production
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DOI：
10.1002/bit.22064
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发表时间：
2009-02-01
影响因子：
3.8
通讯作者：
Posten, Clemens
中科院分区：
文献类型：
--
作者：
Kaeppler, Tobias;Cerff, Martin;Posten, Clemens
提出了一种利用高梯度磁分离技术从生物反应器中原位去除产物的新方法。该分离过程用于吸附去除地衣芽孢杆菌分泌的蛋白酶。在发酵过程中直接在发酵液中使用小的无孔杆菌肽连接的磁性吸附剂，然后进行原位磁性分离。首先在摇瓶培养中证明了概念的证明，然后在3.7 L生物反应器中的补料分批培养期间放大并应用。说明B.地衣状菌不受原位产物去除步骤的影响。分离步骤后蛋白酶产量也保持不变。此外，降解的蛋白酶，遵循一级动力学，减少使用该方法。通过理论建模，我们可以证明，通过使用原位磁分离，蛋白酶的总产量得到了提高。本文所述的工艺是一种有前途的技术，可提高NO生产工艺的总产率，该工艺通常因下游操作薄弱而受到限制。可以克服生物工艺期间遇到的潜在限制，例如产物抑制或降解。讨论了在工业生产中实现原位磁分离需要研究的关键问题。
A new approach for in situ product removal from bioreactors is presented in which high-gradient magnetic separation is used. This separation process was used for the adsorptive removal of proteases secreted by Bacillus licheniformis. Small, non-porous bacitracin linked magnetic adsorbents were employed directly in the broth during the fermentation, followed by in situ magnetic separation, Proof of the concept was first demonstrated in shake flask culture, then scaled up and applied during a fed batch cultivation ill a 3.7 L bioreactor. It could be demonstrated that growth of B. licheniformis was not influenced by the in situ product removal step. Protease production also remained the same after the separation step. Furthermore, degradation of the protease, which followed first order kinetics, was reduced by using the method. Using a theoretical modeling approach, we Could show that protease yield in total was enhanced by using in situ magnetic separation. The process described here is a promising technique to improve overall yield in No production processes which are often limited due to weak downstream operations, Potential limitations encountered during a bioprocess can be overcome such as product inhibition or degradation. We also discuss the key points where research is needed to implement in situ magnetic separation in industrial production.