Low temperature promotes the production and efflux of terpenoids in yeast

Qin, Lei; Ma, Dongshi; Lin, Guangyuan; Sun, Wentao; Li, Chun

doi:10.1016/j.biortech.2024.130376

Cited by 3 publications

(1 citation statement)

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“…Decreasing cultivation temperature has been used in production of recombinant proteins for improved protein solubility and stability, reduced protein aggregation and protease activity, and enhanced cell viability ( 7–12 ). Decreasing cultivation temperature has also been employed to optimize production of valuable secondary metabolites, including terpenoids ( 13 ), and especially in cases where pathway enzymes show higher activity at lower temperature ( 12 , 14–17 ). In these cases, low-temperature-triggered genetic circuits can be applied, with additional advantages of avoiding inducers and modification of the fermenters to fit specific conditions for induction.…”

Section: Introductionmentioning

confidence: 99%

LowTempGAL: a highly responsive low temperature-inducible GAL system in Saccharomyces cerevisiae

Lu,

Shen,

Bandari

et al. 2024

Nucleic Acids Research

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Temperature is an important control factor for biologics biomanufacturing in precision fermentation. Here, we explored a highly responsive low temperature-inducible genetic system (LowTempGAL) in the model yeast Saccharomyces cerevisiae. Two temperature biosensors, a heat-inducible degron and a heat-inducible protein aggregation domain, were used to regulate the GAL activator Gal4p, rendering the leaky LowTempGAL systems. Boolean-type induction was achieved by implementing a second-layer control through low-temperature-mediated repression on GAL repressor gene GAL80, but suffered delayed response to low-temperature triggers and a weak response at 30°C. Application potentials were validated for protein and small molecule production. Proteomics analysis suggested that residual Gal80p and Gal4p insufficiency caused suboptimal induction. ‘Turbo’ mechanisms were engineered through incorporating a basal Gal4p expression and a galactose-independent Gal80p-supressing Gal3p mutant (Gal3Cp). Varying Gal3Cp configurations, we deployed the LowTempGAL systems capable for a rapid stringent high-level induction upon the shift from a high temperature (37–33°C) to a low temperature (≤30°C). Overall, we present a synthetic biology procedure that leverages ‘leaky’ biosensors to deploy highly responsive Boolean-type genetic circuits. The key lies in optimisation of the intricate layout of the multi-factor system. The LowTempGAL systems may be applicable in non-conventional yeast platforms for precision biomanufacturing.

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