Patent ID: 11859223
Assignee: THE TRUSTEES OF PRINCETON UNIVERSITY
Field: Biotechnology (Chemistry)
Classification: CPC C  Y | IPC C

Claim 0:
1. A yeast cell comprising:
a plurality of genes capable of being controlled bi-directionally with at least one wavelength of light, the plurality of genes capable of expressing enzymes for a metabolic pathway and expressing enzymes for an engineered biosynthetic pathway for overproduction of a chemical of interest, the plurality of genes comprising:
a first gene configured such that the first gene is expressed when exposed to the at least one wavelength of light, and the first gene is not expressed when not exposed or when exposed to a different wavelength of light; and
a second gene configured such that the second gene is not expressed when exposed to the at least one wavelength of light, and the second gene is expressed when not exposed or when exposed to a different wavelength of light,

wherein the plurality of genes includes:
a first sequence comprising a nucleotide sequence that encodes a light-activated transcription factor that binds to a first promoter and initiates transcription under certain wavelengths;
a second sequence comprising a second promoter which can be activated by the light-activated transcription factor encoded by the first sequence, the second sequence further comprising a nucleotide sequence that encodes for a first metabolic enzyme;
a third sequence comprising a third promoter which can be activated by the light-activated transcription factor encoded by the first sequence, and further comprising a nucleotide sequence that encodes a repressor; and
a fourth sequence comprising a fourth promoter which can be repressed by the repressor encoded by the third sequence, and further comprising a nucleotide sequence that encodes a second metabolic enzyme,

wherein the yeast cell is an S. cerevisiae cell, and
wherein the first promoter is a constitutive promoter, the light-activated transcription factor is derived from a light-oxygen voltage (LOV) sensing domain, CRY2, CM, or the phytochrome B (PhyB) and PIF3 binding domain, the first metabolic enzyme is required for cell growth, the repressor is GAL80, and the fourth promoter is a galactose-inducible promoter via GAL4, and the second metabolic enzyme is an enzyme that drives the desired metabolic pathway to completion.