Process Biochemistry
Process Biochemistry
5 years ago

Recombinant expression insulin-like growth factor 1 in Bacillus subtilis using a low-cost heat-purification technology

Recombinant expression insulin-like growth factor 1 in Bacillus subtilis using a low-cost heat-purification technology
Insulin-like growth factor 1 (IGF-1) has many positive effects on neonates’ intestinal health. However, the production of IGF-1 involves complex and costly chromatography-based purification devices that greatly increase the cost of IGF-1. In this study, we applied the DAMP4 fusion technology to express IGF-1 in Bacillus subtilis cell factories without using these purification devices. After optimizing the induction conditions in a shaker culture procedure, Western blotting results indicated that the DAMP4-IGF-1 fusion protein was successfully secreted into the culture medium. Approximately 17mg/L DAMP4-IGF-1 was obtained by a simple heat-purification method. The subsequent cleavage of purified fusion protein by enterokinase produced approximately 2.5mg/L of tag-free recombinant IGF-1 with immunogenic activity. Additionally, the recombinant IGF-1 had the biological activity to promote the proliferation of Caco-2 cells. This approach provides a safe, versatile and cost-effective protocol to produce milkborne growth factors in B. subtilis as well as other functional low molecular weight peptide products.

Publisher URL: www.sciencedirect.com/science

DOI: S1359511317308589

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