FIGURE 2.

Work flow of combining genetically encoded erythritol biosensor with high-throughput screening to improve erythritol production in Y. lipolytica. The erythritol biosensor was developed in Escherichia coli BL21 (DE3). Transcriptional repressor EryD and cognate DNA-binding site were used to control the expression of enhanced green fluorescence protein (GEFP). After mutagenesis, mutant library of erythritol producing Y. lipolytica was combined with biosensor and submitted for microplate-based high-throughput screening. High performers can be screened out according to the fluorescent signals. T7, T7 promoter; DBS, EryD DNA-binding site; RFI, relative fluorescence intensity.