Expression, characterization, and application of human-like recombinant gelatin

Area of Science:

• Biomaterials Science
• Biotechnology
• Tissue Engineering

Background:

• Gelatin, derived from collagen, is a vital biomaterial in biomedical science and tissue engineering.
• Existing gelatin sources have limitations, driving the need for recombinant alternatives.

Purpose of the Study:

• To engineer and express recombinant human-like gelatin using Pichia pastoris.
• To characterize the recombinant gelatin and evaluate its suitability for cell culture.

Main Methods:

• Designed a hexameric amino acid sequence based on human collagen type III.
• Inserted the corresponding nucleotide sequence into Pichia pastoris for expression.
• Purified recombinant gelatin using affinity chromatography.
• Analyzed composition, structure (SEM), and cell viability (confocal microscopy).

Main Results:

• Achieved a yield of 0.057 g/L with up to 95% purity.
• Confirmed identical amino acid composition and molecular weight to the designed sequence.
• Demonstrated a porous structure in freeze-dried hydrogels.
• Observed viable, 3D cell growth within gelatin microspheres.

Conclusions:

• Successfully produced recombinant gelatin with high purity and desired characteristics.
• Recombinant gelatin supports cell viability and three-dimensional growth, showing promise for biomedical applications.
• This engineered gelatin offers a potential alternative for tissue engineering and regenerative medicine.