Fig. 4: Characterisation of limonene-producing strains with limonene synthetic genes on a 2μ plasmid (LIM141R and LIM141R2) or integrated at amplified RPL25 locus (LIM141M and LIM141MH).
From: An in vivo gene amplification system for high level expression in Saccharomyces cerevisiae
LIM141R2 is one of LIM141R biological replicates. a Schematic map of genetic vectors used to introduce limonene synthetic genes into yeast. b–f Strain characterisation in two-phase flask cultivation with 20 g l−1 glucose and dodecane overlay. Synthetic auxin 1-Naphthaleneacetic acid (NAA) was added to 1 mM at the late exponential growth phase (OD > 4). Y-FAST fluorescence was measured after 4-hydroxy-3-methylbenzylidene rhodanine (HMBR) with final concentration 20 μM was added to the yeast samples before flow cytometry assay and is expressed as fold-change of exponential-phase auto-fluorescence of the reference strain GH430. Limonene and geraniol production at 96 h was shown. Mean values ± standard deviations are shown (N = 3 independent biological replicates for LIM141R, LIM141M and three independent cultures for LIM141R2 in b–f. N = 4 independent biological replicates in b–e and three independent biological replicates in f for LIM141MH). Source data are provided as a Source Data file.
