MOESM1 of Coupling gene regulatory patterns to bioprocess conditions to optimize synthetic metabolic modules for improved sesquiterpene production in yeast
2017-02-21T05:00:00Z (GMT) by
Additional file 1: Table S1. Primers and PCR fragments amplified/used in this work. PXXX, promoter of gene XXX; T XXX , terminator of gene XXX; Y-GDNA, CEN.PK113-7D genomic DNA; sequence annealing to template in primers is shown in red and italics; over-lap sequence for over-lap extension PCR and Gibson Assembly is underlined; restriction sites used in cloning are shown in bold. Table S2. Molecular construction of plasmids used in this work. Figure S1. Plasmid rearrangement in the strain NC1D. Plasmid pPMVAd36 and [TRP1] from NC1D were digested by restriction enzymes NotI, SalI, SphI, BamHI and SbfI and gel figure was shown as the right-bottom figure. Figure S2. The growth profile of strain GH4 [CEN.PK113-5D derivative; ura3(1, 704 )::KlURA3] (1) on sucrose. The cells were pre-cultured on 40 g L-1 glucose. Mean values from duplicate experiments are shown. Figure S3. Logic charts for fed-batch feeding scripts: (a) carbon-source-restricted/DO-triggered fed-batch cultivation; (b) carbon-source-overflowed/carbon-source-pulsing fed-batch cultivation. Fs, feeding flow storage value; DOt, dissolved oxygen on-line value at time t; DOL, lowest dissolved oxygen storage value; T1, storage time; t, on-line time; Îź, specific rate of feeding flow increasement; N, agitatation speed; Nmax, the maxium agitation speed; FVs, feeding volume storage value; FVt, feeding volume on-line value; Vt, culture volume on-line value. Figure S4. Growth (OD600) and process values (Dissovled oxygen, DO; oxygen transfer rate, OTR; carbon transfer rate; CTR; respiration quotient, RQ) in fed-batch cultivation for strain N391DA, with feeding logics in Fig. S1a employed. (a&b), 600 g L-1 glucose feeding; (c&d), 600 g L-1 sucrose feeding; (e&f), 400 g L-1 glucose and 158 g L-1 ethanol feeding. Figure S5. Growth (OD600) and process values (Dissovled oxygen, DO; oxygen transfer rate, OTR; carbon transfer rate; CTR; respiration quotient, RQ) in fed-batch cultivation for strain N391DA, with feeding logics in Fig. S1b employed. (a&b), 600 g L-1 glucose feeding with 10 g L-1 glucose pulse; (c&d), 600 g L-1 glucose feeding with 20 g L-1 glucose pulse; (e&f), 600 g L-1 sucrose feeding with 20 g L-1 sucrose pulse. Figure S6. The influence of nerolidol on yeast growth. Synthetic minimal medium was used, which contained 6.7 g L-1 yeast nitrogen base (Sigma-Aldrich #Y0626; pH 6.0) and 20 g L-1 glucose. Isomer-mixed nerolidol (Sigma-Aldrich #H59605) was used. Tween 80 was added to homogenize nerolidol into liquid medium. Mean values Âą standard deviations are shown (N = 3).