c glutamicum atcc 13032  (ATCC)

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Improving the tolerance to formaldehyde via ALE. (A) Growth of FM-1 in minimal medium with 10 g/L glucose and different formaldehyde concentrations. 0 mM (square), 0.5 mM (triangle), 0.8 mM (circle), and 1 mM (inverted triangle). (B) ALE procedure of culture-1 in CGXII minimal medium supplemented with different formaldehyde concentrations and 10 g/L glucose. (C) Growth curve of the evolved mutants in CGXII minimal medium supplemented with 10 g/L glucose and 0.8 mM formaldehyde. (D) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose and 1.6 mM formaldehyde. (E) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose. (F) Formaldehyde degradation during cell growth of wild-type C. glutamicum ATCC 13032, FM-1 and FM-3. Values and error bars reflect the mean ± s.d. of three biological replicates (N = 3).

Article Snippet: The mutant cgl1590 fragment was amplified from the genomic DNA of C. glutamicum ATCC 13032 using the primer pairs cgl1590 750insG -F1/ cgl1590 750insG - R1, cgl1590 750insG -F2/ cgl1590 750insG -R2, and then ligated with the Bam HI-linearized pK18 mobsacB using the ClonExpress MultiS One Step Cloning Kit (Vazyme Biotech, China).

Techniques: Mutagenesis

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Improving the tolerance to formaldehyde via ALE. (A) Growth of FM-1 in minimal medium with 10 g/L glucose and different formaldehyde concentrations. 0 mM (square), 0.5 mM (triangle), 0.8 mM (circle), and 1 mM (inverted triangle). (B) ALE procedure of culture-1 in CGXII minimal medium supplemented with different formaldehyde concentrations and 10 g/L glucose. (C) Growth curve of the evolved mutants in CGXII minimal medium supplemented with 10 g/L glucose and 0.8 mM formaldehyde. (D) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose and 1.6 mM formaldehyde. (E) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose. (F) Formaldehyde degradation during cell growth of wild-type C. glutamicum ATCC 13032, FM-1 and FM-3. Values and error bars reflect the mean ± s.d. of three biological replicates (N = 3).

Article Snippet: Each evolved strain possesses more than 100 mutations including single nucleotide polymorphisms (SNPs), insertions, and deletions, which were aligned against the C. glutamicum ATCC 13032 reference genome (GenBank accession number GCA_000011325.1).

Techniques: Mutagenesis

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Improving the tolerance to formaldehyde via ALE. (A) Growth of FM-1 in minimal medium with 10 g/L glucose and different formaldehyde concentrations. 0 mM (square), 0.5 mM (triangle), 0.8 mM (circle), and 1 mM (inverted triangle). (B) ALE procedure of culture-1 in CGXII minimal medium supplemented with different formaldehyde concentrations and 10 g/L glucose. (C) Growth curve of the evolved mutants in CGXII minimal medium supplemented with 10 g/L glucose and 0.8 mM formaldehyde. (D) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose and 1.6 mM formaldehyde. (E) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose. (F) Formaldehyde degradation during cell growth of wild-type C. glutamicum ATCC 13032, FM-1 and FM-3. Values and error bars reflect the mean ± s.d. of three biological replicates (N = 3).

Article Snippet: To assess the toxicity of formaldehyde in C. glutamicum ATCC 13032 lacking the formaldehyde dissimilation pathway, we cultivated a previously developed C. glutamicum FM-1 strain ( C. glutamicum ATCC 13032 Δ adhE Δ ald ), which was referred to as MX-1 in our previous study [ ], in CGXII medium supplemented with various concentrations of formaldehyde ( A).

Techniques: Mutagenesis

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Effects of single-site mutations on formaldehyde tolerance. (A) Frequency of mutations of four bases in evolved strain FM-3. (B) Growth of strain FM-1 and its derivatives harboring single-site mutations on CGXII minimal agar medium supplemented with 10 g/L glucose and 1 mM formaldehyde. (C) Growth curves of strain FM-1 and its derivatives in CGXII medium supplemented with 10 g/L glucose and 0.8 mM formaldehyde. Values and error bars reflect the mean ± s.d. of three biological replicates (N = 3). Statistical significance at 31 h between FM-1- cgl1199 1015−1032del and FM-1 was determined by unpaired two-tailed Student's t -test: ∗∗∗P < 0.001.

Article Snippet: To assess the toxicity of formaldehyde in C. glutamicum ATCC 13032 lacking the formaldehyde dissimilation pathway, we cultivated a previously developed C. glutamicum FM-1 strain ( C. glutamicum ATCC 13032 Δ adhE Δ ald ), which was referred to as MX-1 in our previous study [ ], in CGXII medium supplemented with various concentrations of formaldehyde ( A).

Techniques: Two Tailed Test

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Transcriptome analysis of FM-3 and FM-1 cultivated with or without formaldehyde stress. (A) Volcano plots of differential transcription levels in 1F vs. 1N, 3F vs. 3N, 3N vs. 1N, and 3F vs. 1F. (B) Changes in mRNA levels of genes involved in central metabolism and the respiratory chain between FM-3 and FM-1. Only significant changes (log 2 (fold change) ≥1 or ≤ −1, FDR≤0.05) are shown. Upregulated and downregulated genes are indicated with red and blue, respectively. 1F, FM-1 cultivated with formaldehyde stress. 1N, FM-1 cultivated without formaldehyde stress. 3F, FM-3 cultivated with formaldehyde stress. 3N, FM-3 cultivated without formaldehyde stress.

Article Snippet: To assess the toxicity of formaldehyde in C. glutamicum ATCC 13032 lacking the formaldehyde dissimilation pathway, we cultivated a previously developed C. glutamicum FM-1 strain ( C. glutamicum ATCC 13032 Δ adhE Δ ald ), which was referred to as MX-1 in our previous study [ ], in CGXII medium supplemented with various concentrations of formaldehyde ( A).

Techniques:

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Comparison of proteomes between FM-3 and FM-1 cultivated with formaldehyde or without formaldehyde stress. Functional classification of transcriptome differences based on KEGG_small_class annotation in 3N vs. 1N (A), 3F vs. 1F (B). Proteins with differentially expression in 1F vs. 1N (C) and 3F vs. 3N (D). Only significant changes (log 2 (fold change) ≥1 or ≤ −1, FDR≤0.05) are shown. Proteins exhibiting increased or decreased abundance are highlighted in red and blue, respectively. 1F, FM-1 cultivated with formaldehyde stress. 1N, FM-1 cultivated without formaldehyde stress. 3F, FM-3 cultivated with formaldehyde stress. 3N, FM-3 cultivated without formaldehyde stress.

Article Snippet: To assess the toxicity of formaldehyde in C. glutamicum ATCC 13032 lacking the formaldehyde dissimilation pathway, we cultivated a previously developed C. glutamicum FM-1 strain ( C. glutamicum ATCC 13032 Δ adhE Δ ald ), which was referred to as MX-1 in our previous study [ ], in CGXII medium supplemented with various concentrations of formaldehyde ( A).

Techniques: Comparison, Functional Assay, Expressing

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Effects of Cgl1590 mutations on formaldehyde tolerance. (A) Amino acid sequence alignment between Cgl1590 and its derivatives. (B) Effects of cgl1590 truncation on formaldehyde tolerance. (C) Effects of cgl1590 knock-out on formaldehyde tolerance. (D) Effects of cgl1590 overexpression on formaldehyde tolerance. Cells were treated with 0.8 mM formaldehyde as stress condition. Values and error bars reflect the mean ± s.d. of three biological replicates (N = 3). Analysis of cell length and morphology of FM-1 without formaldehyde stress (E), FM-1 containing cgl1590 750insG mutation without formaldehyde stress (F), FM-1 with formaldehyde stress (G), and FM-1 strain containing cgl1590 750insG mutation with formaldehyde stress (H). All strains were grown in CGXII minimal medium supplemented with 10 g/L glucose, with or without 0.8 mM formaldehyde, and examined by SEM. Cell length was determined by measuring 70 cells of each strain and analyzed using ImageJ software. Statistical significance at 36 h between FM-1-△ cgl1590 and FM-1 was determined by two-tailed Student's t -test: ∗∗∗P < 0.001.

Article Snippet: To assess the toxicity of formaldehyde in C. glutamicum ATCC 13032 lacking the formaldehyde dissimilation pathway, we cultivated a previously developed C. glutamicum FM-1 strain ( C. glutamicum ATCC 13032 Δ adhE Δ ald ), which was referred to as MX-1 in our previous study [ ], in CGXII medium supplemented with various concentrations of formaldehyde ( A).

Techniques: Sequencing, Knock-Out, Over Expression, Mutagenesis, Software, Two Tailed Test

Journal: Synthetic and Systems Biotechnology

Article Title: Multi-omics analyses of evolved Corynebacterium glutamicum mutants reveal the molecular responses to formaldehyde stress

doi: 10.1016/j.synbio.2026.01.020

Figure Lengend Snippet: Improving the tolerance to formaldehyde via ALE. (A) Growth of FM-1 in minimal medium with 10 g/L glucose and different formaldehyde concentrations. 0 mM (square), 0.5 mM (triangle), 0.8 mM (circle), and 1 mM (inverted triangle). (B) ALE procedure of culture-1 in CGXII minimal medium supplemented with different formaldehyde concentrations and 10 g/L glucose. (C) Growth curve of the evolved mutants in CGXII minimal medium supplemented with 10 g/L glucose and 0.8 mM formaldehyde. (D) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose and 1.6 mM formaldehyde. (E) Growth curve of evolved mutant in CGXII minimal medium supplemented with 10 g/L glucose. (F) Formaldehyde degradation during cell growth of wild-type C. glutamicum ATCC 13032, FM-1 and FM-3. Values and error bars reflect the mean ± s.d. of three biological replicates (N = 3).

Article Snippet: The pCas9gRNA- cgl0752 and the ssDNA containing cgl0752 G761T mutation were co-transformed into C. glutamicum ATCC 13032 (pRecT) via electroporation. cgl0752 edited mutants were screened following the procedure described previously [ ]. cgl0942 G443T and cgl1199 1015−1032del nucleotide mutations were introduced following a similar procedure.

Techniques: Mutagenesis