A proposed feeding strategy for the overproduction of recombinant proteins in Escherichia coli
A proposed feeding strategy for the overproduction of recombinant proteins in Escherichia coli
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DOI：
10.1042/ba20070089
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发表时间：
2008-02-01
影响因子：
2.8
通讯作者：
Tabandeh, Fatemeh
中科院分区：
文献类型：
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作者：
Babaeipour, Valiollah;Shojaosadati, Seyed Abbas;Tabandeh, Fatemeh
研究了在重组大肠杆菌 BL21(DE3)（质粒 pET3a-ifn gamma）中使用异丙基 β-D-硫代半乳糖苷诱导表达系统生产人干扰素 γ 的不同饲喂策略。设计了四种补料分批模式，以比较其在大肠杆菌高细胞密度培养物的化学诱导前后对重组蛋白生产、底物消耗、副产物形成和质粒稳定性的影响（比生长速率）。结果发现，干扰素γ的产物/底物产率Y-p/s在整个过程中受其显着影响，但在预诱导阶段产物/生物质产率(Y-p/x)受mu影响。通过应用有效的补料策略，将重组蛋白维持在最大可达到的水平，重组蛋白的积累量达到细胞总蛋白的 60%，并且其生产率显着提高。在这种情况下，生物量和重组蛋白的总体生产率分别为6.36和2.1，而指数饲喂期间的1.91和0.16 g center dot h(-1) center dot litre(-1)，在整个过程中保持恒定。
Different feeding strategies for the production of human interferon-gamma using an isopropyl beta-D-thio-galactoside-inducible expression system in recombinant Escherichia coli BL21(DE3) (plasmid pET3a-ifn gamma) were studied. Four fed-batch modes were designed to compare the effect of it (specific growth rate) on recombinant-protein production, substrate consumption, by-product formation and plasmid stability during pre- and post-chemical induction in high-cell-density cultures of E. coli. It was found that Y-p/s, the product/substrate yield of interferon-gamma was significantly affected by it throughout the process, but product/biomass yield (Y-p/x) was influenced by mu at the pre-induction stage. By, applying an efficient feeding strategy, in which the it was maintained at the maximum attainable level, recombinant protein was accumulated up to a level of 60% of the total cell protein and its productivity was increased significantly. In this case, the overall productivities of biomass and recombinant protein were 6.36 and 2.1 respectively, in comparison with 1.91 and 0.16 g center dot h(-1) center dot litre(-1) during exponential feeding, in which It was kept constant throughout the entire process.