Fig. 3: Optimization of the SCTPK further enhanced AcP production.
From: Using a synthetic machinery to improve carbon yield with acetylphosphate as the core
a Genes that may shunt the SCTPK and will be repressed using antisense RNA (asRNA) interference. b Inhibition efficiency of asRNA with a paired termini was affected by the length and transcription level of the asRNA. The numbers below promoters represent the relative strengths of corresponding promoters, which were determined using GFP as a reporter and shown in Supplementary Fig. 5. c The intracellular AcP concentration of E. coli BW25113, Q3964 harboring the SCTPK, and three strains derived from Q3964 in which transcription of gapA, gapB, and pfkA was repressed strongly using a 100-nt asRNA transcribed from a promoter Pj23119, respectively (n = 3 biological independent samples). d The final cell density and specific growth rate of E. coli strains (n = 3 biological independent samples). e Relative mRNA level of gapA, gapB, and pfkA in strains with and without corresponding asRNA determined by qRT-PCR (n = 3 biological independent samples with two technical repeats). f Effect of combinational and hierarchical repression of gapA, gapB, and pfkA on the intracellular level of AcP (n = 3 biological independent samples). g The final cell density and specific growth rate of E. coli strains (n = 3 biological independent samples). h CO2 emission rate of E. coli BW25113 and Q4531 strain carrying optimized SCTPK (n = 2 biological independent samples). i Relative CO2 emission of BW25113 and Q4531 (n = 2 biological independent samples). j The intracellular pyruvate concentration of E. coli BW25113, Q3964, and Q4531 (n = 6 biological independent samples). k Acetate accumulation in cultures of E. coli BW25113 and Q4531 (n = 3 biological independent samples). Error bar, mean ± SD. Two-tailed Student’s t tests were performed to determine the statistical significance. Source data are provided as a Source Data file.
