Production of bacteria and yeast of hemoglobin and analogues thereof.

摘要

Alpha subunits of hemoglobin are provided as a novel recombinant di-alpha globin polypeptide comprising the two alpha subunits connected directly via peptide bond or indirectly by a flexible amino-acid or peptide linker. Di-alpha globin may be combined in vivo or in vitro with beta globin and heme to form hemoglobin. Tetrameric human hemoglobin and di-alpha/beta2 hemoglobin are produced in S. cerevisiae by three types of expression vectors: 1) two separate plasmids containing respectively alpha and beta globin genes expressed in diploid strains. 2) a single plasmid comprising alpha and beta globin genes expressed in either haploid or diploid strains. 3) a single plasmid containing di-alpha and beta globin genes expressed in haploid strains. Tetrameric form or separate subunits can be recovered from the soluble fraction. So, 3 types of hemoglobin-like molecules can be produced: di-alpha/two beta, di-beta/two alpha or di-alpha/di-beta, with a long half life.