Systematic engineering of Saccharomyces cerevisiae for efficient synthesis of hemoglobins and myoglobins

[Display omitted] •Various hemoglobins and myoglobins were synthetized in S. cerevisiae.•Subcellular localization of all enzymes involved in heme synthesis were confirmed.•The spatial barrier was removed by reconstructing heme synthetic pathway.•Enhancing precursor supply and assembling key enzymes to increase heme supply.•The biochemical properties of synthesized hemoglobins and myoglobins were detected. Hemoglobin (Hb) and myoglobin (Mb) are kinds of heme-binding proteins that play crucial physiological roles in different organisms. With rapid application development in food processing and biocatalysis, the requirement of biosynthetic Hb and Mb is increasing. However, the production of Hb and Mb is limited by the lower expressional level of globins and insufficient or improper heme supply. After selecting an inducible strategy for the expression of globins, removing the spatial barrier during heme synthesis, increasing the synthesis of 5-aminolevulinate and moderately enhancing heme synthetic rate-limiting steps, the microbial synthesis of bovine and porcine Hb was firstly achieved. Furthermore, an engineered Saccharomyces cerevisiae obtained a higher titer of soybean (108.2 ± 3.5 mg/L) and clover (13.7 ± 0.5 mg/L) Hb and bovine (68.9 ± 1.6 mg/L) and porcine (85.9 ± 5.0 mg/L) Mb. Therefore, this systematic engineering strategy will be useful to produce other hemoproteins or hemoenzymes with high activities.