Design and Characterization of Biosensors for the Screening of Modular Assembled Naringenin Biosynthetic Library in Saccharomyces cerevisiae

• DOI: 10.1021/acssynbio.9b00212
• Publication Date: 2019-08-21T19:57:06Z
• Authors: Rufeng Wang; Brady F. Cress; Zheng Yang; John C. Hordines; Shujuan Zhao; Gyoo Yeol Jung; Zhengtao Wang; Mattheos A. G. Ko...
• Keywords: Metabolic Engineering; Synthetic Biology

A common challenge in the assembly and optimization of plant natural product biosynthetic pathways recombinant hosts is identification gene orthologues that will result best production titers. Here, we describe modular a naringenin pathway Saccharomyces cerevisiae was facilitated by optimized naringenin-inducible prokaryotic transcription activators used as biosensors. The biosensors were designed developed S. multiparametric engineering strategy, which further applied for vivo, high-throughput screening established yeast library. workflow assembling involved Golden gate-directed combinatorial genes promoters, resulting strain library ideally covering 972 combinations cerevisiae. For improving performance our biosensor, series fundamental components optimized, affecting efficiency biosensor such nuclear localization signal (NLS), detector module effector module. One (pTDH3_NLS_FdeR-N_tPGK1-pGPM1-fdeO_mcherry_tTDH1-MV2) showed better performance, defined dynamic range sensitivity than others this study well other previously reported Using able to identify most efficient candidate from This approach can be exploited metabolites derived flavonoid more importantly employed characterization putative genes.