Transcription Factor-Based Biosensor for Dynamic Control in Yeast for Natural Product Synthesis

Abstract

The synthesis of natural products in yeast has gained remarkable achievements with intensive metabolic engineering efforts. In particular, transcription factor (TF)-based biosensors for dynamic control of gene circuits could facilitate strain evaluation, high-throughput screening (HTS), and adaptive laboratory evolution (ALE) for natural product synthesis. In this review, we summarized recent developments of several TF-based biosensors for core intermediates in natural product synthesis through three important pathways, i.e., fatty acid synthesis pathway, shikimate pathway, and methylerythritol-4-phosphate (MEP)/mevalonate (MVA) pathway. Moreover, we have shown how these biosensors are implemented in synthetic circuits for dynamic control of natural product synthesis and also discussed the design/evaluation principles for improved biosensor performance.

Keywords: MEP/MVA pathway; biosensor; fatty acids; natural products; shikimate pathway; transcription factor; yeast.

FIGURE 1

Transcription factor (TF)-based biosensors involved in synthesis pathways of natural products in yeast. E4P, erythrose-4-phosphate; G3P, glyceraldehyde-3-phosphate; PEP, phosphoenolpyruvate; IPP, isopentenyl pyrophosphate; DMAPP, dimethylallyl pyrophosphate; DAHP, 3-deoxy-D-arabino-2-heptulosonic acid 7-phosphate; CHR, chorismic acid; MEP, methylerythritol-4-phosphate; MVA, mevalonate; Trp, tryptophan; Tyr, tyrosine; Phe, phenylalanine.

FIGURE 2

Dynamic regulation of gene circuits through TF-based biosensors. (A) Activated regulation of gene circuits. Transcription factor (TF) functions as a repressor to inhibit gene transcription (OFF), and ligand binding induces a conformational shift in TF and activates gene transcription (ON). (B) Repressive regulation of gene circuits. TF functions as an activator of gene transcription (ON), and ligand binding induces a conformational shift in TF and inhibit gene transcription (OFF). DBD, DNA binding domain; LBD, ligand binding domain; RNA Pol, RNA polymerase.