Department of Genomics and Genetic Engineering, Razi Vaccine and Serum Research Institute (RVSRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
Department of Genomics and Genetic Engineering, Razi Vaccine and Serum Research Institute (RVSRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.
Protein Expr Purif. 2020 Jun;170:105593. doi: 10.1016/j.pep.2020.105593. Epub 2020 Feb 4.
Cost-effectiveness is an important issue in biotechnological manufacturing industry and using alternative cheap materials with the same benefits has been noticed in most literatures. Isopropyl β-d-1-thiogalactopyranoside (IPTG), a well-known chemical element for induction of protein expression, has several disadvantages such as high expense and toxicity. In this study, we aimed to introduce skimmed milk as an alternative material for protein expression by induction of lac operon. In this way, Escherichia coli BL21 (DE3) bacteria were induced using 1 mM IPTG or 1.0% (w/v) skimmed milk. Protein purification was performed using Ni-NTA (nickel-nitrilotriacetic acid) for His-tagged recombinant proteins and protein purity was evaluated by SDS-PAGE. Results showed high level of recombinant protein expression using skimmed milk, and interestingly, the growth rate of bacteria improved. Our findings suggested that skimmed milk can be a suitable alternative for induction of recombinant protein expression, which has advantages such as more availability and affordability, in comparison to IPTG supplementation.
成本效益是生物技术制造业的一个重要问题，在大多数文献中都注意到使用具有相同效益的替代廉价材料。异丙基 β-D-1-硫代半乳糖吡喃糖苷（IPTG）是一种众所周知的诱导蛋白表达的化学元素，但存在费用高和毒性等缺点。在这项研究中，我们旨在通过诱导乳糖操纵子，将脱脂乳作为诱导蛋白表达的替代材料。通过使用 1mM IPTG 或 1.0%（w/v）脱脂乳，诱导大肠杆菌 BL21（DE3）细菌。使用 Ni-NTA（镍-亚氨二乙酸）对 His 标记的重组蛋白进行蛋白纯化，并通过 SDS-PAGE 评估蛋白纯度。结果表明，使用脱脂乳可实现高水平的重组蛋白表达，而且有趣的是，细菌的生长速度也有所提高。我们的研究结果表明，与 IPTG 补充剂相比，脱脂乳作为诱导重组蛋白表达的替代物具有更多的可用性和可负担性等优势。
