ABSTRACT Enhancing the nutritional and sensory qualities of microbial single‐cell proteins (SCPs) requires strategies to increase heme content in edible microorganisms. We adapted the Growth‐Acceleration Targeting Evolution (GATE) platform, initially developed in Corynebacterium glutamicum , for use in Saccharomyces cerevisiae . By engineering a plasmid that connects the heme‐responsive CYC1 promoter to the growth‐promoting PTH1 gene, we established a feedback loop that links intracellular heme levels to accelerated cell proliferation. After 100 h of continuous culture under growth‐selective pressure, we cured out the plasmid to isolate an Evol‐GATE strain. Compared to the parental type, Evol‐GATE displayed a five‐fold increase in intracellular heme, a slight reduction in biomass, and coordinated upregulation of the heme biosynthetic pathway. Transcriptome analysis confirmed increased expression of heme biosynthesis and associated respiratory genes in Evol‐GATE. Whole‐genome sequencing revealed only a small number of dispersed variants, and no residual plasmid sequences, supporting its classification as a non‐GMO mutant. Our results demonstrate that GATE can effectively select yeast mutants with significantly improved heme productivity, providing a promising approach to develop non‐GMO SCPs enriched in heme for next‐generation meat analogues.
Bang et al. (Thu,) studied this question.
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