Integrated Fermentation Engineering Enables High‐Level Leghemoglobin Production in Kluyveromyces marxianus via Metabolic Rewiring of Central Carbon Metabolism and Amino Acid Utilization

) rose over 100%. Notably, the optimized approach extended the cellular high productivity phase and enhanced the product yield from 0.011 g/g glucose to 0.026 g/g glucose. Furthermore, targeted metabolomics analysis of central carbon and amino acid metabolism provided mechanistic insights into the improved production performance. The optimized strategy reshaped the cellular metabolic network, enhanced carbon conversion efficiency toward LBA synthesis, and thereby improved the production performance of the microbial cell factory. These results highlight the potential of coordinated control of nutritional and physical parameters to enhance LBA biosynthesis in K. marxianus, providing a promising strategy for industrial production.