Cheese-whey permeate improves the fitness of Escherichia coli cells during recombinant protein production.

Marcella de Divitiis, Diletta Ami, Alex Pessina, Alessandro Palmioli, Barbara Sciandrone, Cristina Airoldi, Maria Elena Regonesi, Luca Brambilla, Marina Lotti, Antonino Natalello, Stefania Brocca, Marco Mangiagalli
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引用次数: 4

Abstract

Background: Escherichia coli cells are the most frequently used hosts in recombinant protein production processes and mainly require molecules such as IPTG or pure lactose as inducers of heterologous expression. A possible way to reduce the production costs is to replace traditional inducers with waste materials such as cheese whey permeate (CWP). CWP is a secondary by-product generated from the production of the valuable whey proteins, which are obtained from ultrafiltration of cheese whey, a main by-product of the dairy industry, which is rich in lactose.

Results: The effects of CWP collected from an Italian plant were compared with those of traditional inducers on the production of two model proteins (i.e., green fluorescent protein and the toxic Q55 variant of ataxin-3), in E. coli BL21 (DE3) cells. It was found that the high lactose content of CWP (165 g/L) and the antioxidant properties of its micronutrients (vitamins, cofactors and osmolytes) sustain production yields similar to those obtained with traditional inducers, accompanied by the improvement of cell fitness.

Conclusions: CWP has proven to be an effective and low-cost alternative inducer to produce recombinant proteins. Its use thus combines the advantage of exploiting a waste product with that of reducing the production costs of recombinant proteins.

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在重组蛋白生产过程中,奶酪乳清渗透提高了大肠杆菌细胞的适应性。
背景:大肠杆菌细胞是重组蛋白生产过程中最常用的宿主,主要需要IPTG或纯乳糖等分子作为异源表达的诱导剂。降低生产成本的一种可能方法是用废弃材料如奶酪乳清渗透物(CWP)代替传统的诱导剂。CWP是生产有价值的乳清蛋白时产生的二级副产品,乳清蛋白是通过对奶酪乳清的超滤获得的,奶酪乳清是乳制品行业的主要副产品,富含乳糖。结果:比较了从意大利植物中收集的CWP与传统诱导剂对大肠杆菌BL21(DE3)细胞中产生两种模型蛋白(即绿色荧光蛋白和ataxin-3的毒性Q55变体)的影响。研究发现,CWP的高乳糖含量(165 g/L)及其微量营养素(维生素、辅因子和渗透液)的抗氧化特性维持了与传统诱导剂类似的产量,同时提高了细胞适应性。结论:CWP已被证明是一种有效且低成本的重组蛋白替代诱导剂。因此,它的使用结合了利用废物的优点和降低重组蛋白生产成本的优点。
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