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. 2021 Mar 8;12:1513. doi: 10.1038/s41467-021-21632-3

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Influence of 1,3-propanediol on cell metabolism at the transcriptional level in H. bluephagenesis TDΔdddA. Volcano plot and fold change of gene expression of differentially expressed gene (DEG) distribution. A corrected P-value of 0.05 and log2 (fold change) of 1 were set as the thresholds for significantly differential expression. Red, upregulated genes; blue, downregulated genes. b 3HP production by H. bluephagenesis TDΔdddA overexpressing different biosynthetic genes using 1,3-propanediol as a carbon source. Plasmid pKS and p30 were used as the negative and positive controls, respectively. Gene dhaT_Pp encodes P. putida KT2440 alcohol dehydrogenase, aldD_Pp encodes P. putida KT2440 aldehyde dehydrogenase, aldD_Hb encodes H. bluephagenesis aldehyde dehydrogenase, GME_RS01345 encodes NAD(P)-dependent alcohol dehydrogenase; GME_05160 encodes zinc-binding alcohol dehydrogenase, GME_RS01585 encodes zinc-binding dehydrogenase, GME_RS00365 encodes alcohol dehydrogenase AdhP. c 3HP production and d cell growth (DCM) and PHB content under optimal expression levels of aldehyde dehydrogenase. A stronger RBS was cloned instead of the original construct p58 to form plasmid p59. A stronger promoter Pporin replaced the original construct p58 to generate plasmid p60 (Supplementary Table 5). Cells were grown in the defined minimal medium supplemented with 30 g L⁻¹ glucose, 10 g L⁻¹ 1,3-propanediol, and 3 g L⁻¹ acetic acid. All titers were obtained after 48 h cultivation at 200 r.p.m. and 37 °C. Initial pH of all shake-flask studies was 9. Two-tailed Student’s t-tests were performed to determine the statistical significance for two-group comparisons. e The enzyme activity of AldD_Hb was assayed using the recombinant H. bluephagenesis TDΔdddA harboring p58-p60 deleted with adhP gene cultivated in 60-LB medium for 24 h. The comparison of activities was conducted using a crude extract. All data represent the mean of n = 3 biologically independent samples and error bars show s.d.