High-throughput screening method for glycolipids based on substrate modification and their efficient biomanufacturing.

The development of a high-throughput screening (HTS) method is crucial for boosting microbial synthesis. However, in non-model strains, genetic editing-based HTS is often not feasible. Here, a substrate-modified HTS strategy using Starmerella bombicola PL0120, a sophorolipids (SLs)-producing strain, was designed. First, the medium was optimized to increase the SLs yield to 248 g/L. Then, 2-benzylstearic acid was synthesized, which PL0120 can use to make SLs analogs. Subsequently, by selecting dark-phenotype strains and using 254 nm absorption, a 51.1 % high-yielding SLs strain screening rate was achieved. Among these strains, strain A6 yielded a concentration of 324 g/L. Moreover, this method has been extended to rhamnolipids (RLs). By engineering a microbubble reactor, the yield of RLs was increased to 70.3 g/L. This HTS methodology is instrumental in augmenting the production output of microbial fermentation products, which in turn, facilitates cost-effective biomanufacturing.