The stable human haemoglobin variant was obtained through the design of novel high-throughput screening strategies and methods. Furthermore, the supply level of heme in the production host was enhanced. Ultimately, an robust human haemoglobin was achieved with stronger stability, lower heme loss rate, lower autoxidation rate, more efficient oxygen transport capacity and higher activity.
ABSTRACT
Recombinant human haemoglobin (rHb) is a tetramer protein with heme as cofactors, which have extensive applications in the fields of biomaterials and biomedical therapeutics. However, due to the poor structural stability, the dissociation of heme, weak oxygen transport efficiency, and lower activity, the utilisation of rHb is severely limited in artificial oxygen carriers. Herein, based on the novel developed high-throughput screening strategies and semi-rational design, the engineered rHb mutant with strong stability and heme-binding ability was obtained. In addition, through the homology alignment and rational design, the oxygen transport capacity of rHb was significantly enhanced. Furthermore, the bottlenecks of heme supply were overcome by applying the fine-tuned heme synthesis in Escherichia coli. Finally, the robust and high-activity rHb mutant was synthesised and can be used as a new generation of artificial oxygen carriers.
Microbial Biotechnology, Volume 18, Issue 3, March 2025. Read More
