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- Time-resolved analysis of Bacillus subtilis DB104 Spo0A-mutant transcriptome profile and enhancement of recombinant protein release
- Ji-Su Jun, Soo Ji Kang, Kwang-Won Hong
- J. Microbiol. 2025;63(5):e2411032. Published online May 27, 2025
- DOI: https://doi.org/10.71150/jm.2411032
- 186 View
- 6 Download
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Abstract
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Supplementary Material -
Spo0A, the master regulator of sporulation initiation in Bacillus subtilis, controls over 500 genes directly or indirectly in early sporulation stages. Although the effects of Spo0A disruption on sporulation have been extensively studied, a comprehensive understanding of the genomic response throughout growth phases remain elusive. Here, we examined the transcriptomic changes in Spo0A mutant strain, R211E, and wild-type across a time-course RNA-seq to identify impacted biological processes and pathways. The R211E strain, which exhibits sporulation deficiency, was constructed using the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated protein (Cas)9 system, highlighting the critical role of proper Cas9 dosing in gene editing. Functional analysis of 3,010 differentially expressed genes (DEGs) showed significant alterations in sporulation, quorum sensing, metabolism, and biofilm formation. The R211E disrupted the Spo0A-AbrB regulatory pathway, reducing biofilm formation and enhancing flagellar gene expression. Up-regulated metabolic pathways, including glycolysis, histidine, and purine biosynthesis, increased cell numbers during vegetative growth. Further, the mutant displayed elevated vegetative autolysin expression, resulting in reduced cell viability in the stationary phase. We also introduce the novel potential of R211E in a recombinant protein expression system that facilitated protein release into the supernatant, providing valuable insight for future research in metabolic engineering and efficient production systems in B. subtilis.
- Gut microbiota metabolic characteristics in coronary artery disease patients with hyperhomocysteine
- Ran Tian , Hong-Hong Liu , Si-Qin Feng , Yi-Fei Wang , Yi-Yang Wang , Yu-Xiong Chen , Hui Wang , Shu-Yang Zhang
- J. Microbiol. 2022;60(4):419-428. Published online March 4, 2022
- DOI: https://doi.org/10.1007/s12275-022-1451-2
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- 8 Web of Science
- 7 Crossref
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Abstract
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Hyperhomocysteine (HHcy) is known as a risk factor for coronary
artery disease (CAD). Despite the knowledge that gut
microbiota related metabolism pathway shares metabolites
with that of Hcy, little has been shown concerning the association
between HHcy and gut microbiota. To explore their
relationship in the context of CAD, 105 patients and 14 healthy
controls were recruited from one single medical center located
in Beijing, China. Their serum and fecal samples were
collected, with multi-omics analyses performed via LC/MS/
MS and 16S rRNA gene V3-V4 region sequencing, respectively.
Participants from the prospective cohort were divided
into CAD, CAD & HHcy and healthy controls (HC) groups
based on the diagnosis and serum Hcy concentration. The
results
revealed significant different metabolic signatures between CAD and CAD & HHcy groups. CAD patients with HHcy suffered a heavier atherosclerotic burden compared to CAD patients, and the difference was closely associated to betaine-homocysteine S-methyltransferase (BHMT)-related metabolites and trimethylamine N-oxide (TMAO)-related metabolites. Dimethylglycine (DMG) exhibited a strong positive correlation with serum total Hcy (tHcy), and TMAO and trimethylysine (TML) were associated with heavier atherosclerotic burden. Multiple other metabolites were also identified to be related to distinct cardiovascular risk factors. Additionally, Clostridium cluster IV and Butyricimonas were enriched in CAD patients with elevated tHcy. Our study suggested that CAD patients with elevated tHcy were correlated with higher atherosclerotic burden, and the impaired Hcy metabolism and cardiovascular risk were closely associated with BHMT-related metabolites, TMAO-related metabolites and impaired gut microbiota homeostasis. -
Citations
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Electronic Journal of General Medicine.2024; 21(1): em556. CrossRef - Association of serum homocysteine levels with intestinal flora and cognitive function in schizophrenia
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