通过筛选血红蛋白基因及分泌元件，在银耳拟酵母细胞中生产重组血红蛋白

Abstract

Hemoglobins have attracted increasing interest for their potential applications in alternative protein foods and food biotechnology, but the use of fungal yeast-like cells for recombinant hemoglobin production remains underexplored. In this study,Tremella fuciformisyeast-like cells (YLCs) were evaluated as a potential host system by screening candidate hemoglobins and secretion elements. Clover hemoglobin fromTrifolium subterraneum(CHb),Vitreoscillahemoglobin (VHb), andVigna unguiculataleghemoglobin 2 (Lb2) were heterologously expressed inT. fuciformisYLCs. The estimated yields of CHb, VHb, and Lb2 were 32.00, 19.03, and 23.51 mg/L, respectively, with CHb showing the highest observed yield among the three tested hemoglobins. CHb was then used to evaluate the CysR signal peptide, α-mating factor prepro leader (Amf), and glucoamylase signal peptide (Gla). All three elements mediated CHb secretion, with Amf producing the highest extracellular CHb yield of 106.43 mg/L and total CHb production of 133.02 mg/L, approximately 4.16 times that of CHb without a secretion element. CysR and Gla produced extracellular CHb yields of 72.54 and 49.50 mg/L, respectively. These results identify CHb and Amf as a promising expression combination inT. fuciformisYLCs, but direct characterization of heme incorporation and functional properties is required before food applications can be evaluated.