Polysialic acid production using Escherichia coli K1 in a disposable bag reactor
Polysialic acid production using Escherichia coli K1 in a disposable bag reactor
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DOI：
10.1002/elsc.201600220
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发表时间：
2017-07-01
影响因子：
2.7
通讯作者：
Scheper, Thomas
中科院分区：
文献类型：
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作者：
de Vries, Ingo;Busse, Christoph;Scheper, Thomas
聚唾液酸（polySia），由α-（2，8）-连接的N-乙酰神经氨酸单体组成，在许多生物过程中起着至关重要的作用。本研究提出了一种新的方法，用于生产内源性polySia使用大肠杆菌K1在一次性袋式反应器与波诱导混合。一次性袋式反应器在GMP、灵活性等监管要求方面提供了简单快速的生产，并且不仅可以通过规模化而且可以通过并行化轻松调整到更大的生产能力。由于与搅拌釜反应器相比差的氧转移速率，在培养期间加入纯氧以避免氧限制。在指数生长期，生长速率为0.61 h（-1）。从细胞表面的应力相关的产品释放的调查表明，与波诱导混合的一次性袋式反应器和搅拌釜反应器之间没有显着差异。分批培养后，达到6.8 g L-1的细胞干重和245 mg L-1的polySia浓度。上清液中的总蛋白浓度为132 mg L-1。在高效和省时的下游加工后，最终产品的表征显示蛋白质含量低于0.04 mg（蛋白质）/g（polySia），最大链长接近90聚合度。
Polysialic acid (polySia), consisting of alpha-(2,8)-linked N-acetylneuraminic acid monomers plays a crucial role in many biological processes. This study presents a novel process for the production of endogenous polySia using Escherichia coli K1 in a disposable bag reactor with wave-induced mixing. Disposable bag reactors provide easy and fast production in terms of regulatory requirements as GMP, flexibility, and can easily be adjusted to larger production capacities not only by scale up but also by parallelization. Due to the poor oxygen transfer rate compared to a stirred tank reactor, pure oxygen was added during the cultivation to avoid oxygen limitation. During the exponential growth phase the growth rate was 0.61 h(-1). Investigation of stress-related product release from the cell surface showed no significant differences between the disposable bag reactor with wave-induced mixing and the stirred tank reactor. After batch cultivation a cell dry weight of 6.8 g L-1 and a polySia concentration of 245 mg L-1 were reached. The total protein concentration in the supernatant was 132 mg L-1. After efficient and time-saving downstream processing characterization of the final product showed a protein content of below 0.04 mg(protein)/g(polySia) and a maximal chain length of similar to 90 degree of polymerization.