Hyper-expression of functional human basic fibroblast growth factor 2 in Escherichia coli

Area of Science:

• Biotechnology
• Cell Biology
• Biochemistry

Background:

• Human basic fibroblast growth factor 2 (hbFGF2) is vital for cell functions but expensive, hindering cultivated meat (CM) production.
• High cost of commercial hbFGF2 significantly impacts the economic viability and scalability of CM media.

Purpose of the Study:

• To develop a cost-effective method for high-yield production of active hbFGF2.
• To optimize expression and purification of hbFGF2 for potential use in CM production.

Main Methods:

• A codon-optimized hbFGF2 variant (hbFGF2-G3) was expressed in Escherichia coli T7 Express using the pET-28a(+) plasmid.
• Cultivation parameters were optimized in shake flasks and batch bioreactors, followed by His-trap affinity and size exclusion chromatography for purification.
• Functional activity was confirmed via FGF/FGFR activation assays and cell proliferation studies.

Main Results:

• Optimized conditions achieved a yield of 1.3 g/L in shake flasks and 2.95 g/L in a batch bioreactor.
• Purified hbFGF2-G3 demonstrated functional activity in cell-based assays.
• The study presents a scalable and cost-effective production strategy for active hbFGF2.

Conclusions:

• A high-titer, cost-effective method for producing active hbFGF2-G3 from E. coli was established.
• This advancement significantly addresses the economic challenges in scaling up cultivated meat production.
• The developed process offers a viable solution for reducing media costs in the cultivated meat industry.