<p>Leghemoglobin (LegH) is a monomer heme-binding protein in legume rhizomes, which possesses essential functions in cellular oxygen transport and provision of bioavailable iron. Meanwhile, it has great potential application in food additive for the promising artificial meat, endowing the products realistic color and taste. Compared with the traditional extraction method, the production of LegH by microbial biosynthesis has advantages in short production period, low cost and high yield. In this review, we summarize the recent strategies for the optimization of LegH expression in different microorganisms, the metabolic regulation of heme synthetic pathway, and the progress in the improvement of fermentation process. Compared with the previous reviews, this review covers numerous prokaryotic and eukaryotic microorganisms used for LegH production, and also takes into account the improvement of LegH secretory expressional level. Additionally, this review summarizes the key technological breakthroughs and existing challenges in the field of current LegH microbial synthesis, providing a reference for the subsequent industrial production and application of LegH and other high-value-added hemoproteins.</p>