This review explores current advancements in microbiome functional analysis enabled by next-generation sequencing technologies, which have transformed our understanding of microbial communities from mere taxonomic composition to their functional potential. We examine approaches that move beyond species identification to characterize microbial activities, interactions, and their roles in host health and disease. Genome-scale metabolic models allow for in-depth simulations of metabolic networks, enabling researchers to predict microbial metabolism, growth, and interspecies interactions in diverse environments. Additionally, computational methods for predicting metabolite profiles offer indirect insights into microbial metabolic outputs, which is crucial for identifying biomarkers and potential therapeutic targets. Functional pathway analysis tools further reveal microbial contributions to metabolic pathways, highlighting alterations in response to environmental changes and disease states. Together, these methods offer a powerful framework for understanding the complex metabolic interactions within microbial communities and their impact on host physiology. While significant progress has been made, challenges remain in the accuracy of predictive models and the completeness of reference databases, which limit the applicability of these methods in under-characterized ecosystems. The integration of these computational tools with multi-omic data holds promise for personalized approaches in precision medicine, allowing for targeted interventions that modulate the microbiome to improve health outcomes. This review highlights recent advances in microbiome functional analysis, providing a roadmap for future research and translational applications in human health and environmental microbiology.
Systemic sclerosis (SSc) is a chronic autoimmune disorder characterised by skin fibrosis and internal organ involvement. Disruptions in the microbial communities on the skin may contribute to the onset of autoimmune diseases that affect the skin. However, current research on the skin microbiome in SSc is lacking. This study aimed to investigate skin microbiome associated with disease severity in SSc. Skin swabs were collected from the upper limbs of 46 healthy controls (HCs) and 36 patients with SSc. Metagenomic analysis based on the 16S rRNA gene was conducted and stratified by cutaneous subtype and modified Rodnan skin score (mRSS) severity. Significant differences in skin bacterial communities were observed between the HCs and patients with SSc, with further significant variations based on subtype and mRSS severity. The identified biomarkers were Bacteroides and Faecalibacterium for patients with diffuse cutaneous SSc with high mRSS (≥ 10) and Mycobacterium and Parabacteroides for those with low mRSS (< 10). Gardnerella, Abies, Lactobacillus, and Roseburia were the biomarkers in patients with limited cutaneous SSc (lcSS) and high mRSS, whereas Coprococcus predominated in patients with lcSS and low mRSS. Cutaneous subtype analysis identified Pediococcus as a biomarker in the HCs, whereas mRSS analysis revealed the presence of Pseudomonas in conjunction with Pediococcus. In conclusion, patients with SSc exhibit distinct skin microbiota compared with healthy controls. Bacterial composition varies by systemic sclerosis cutaneous subtype and skin thickness.
We isolated three novel strains, S1T, S2T, and S5T, from human oral cavities and identified them as distinct novel species. All these strains are facultatively anaerobic, Gram-stain-positive, and non-flagellated bacteria. Their optimal growth conditions for these strains were observed in Columbia broth (CB) at 37 °C, pH 7.0, and in the absence of NaCl. Phylogenetic analyses, employing the 16S rRNA gene and whole-genome sequencing, confirmed that all three strains belong to the genus Streptococcus. The 16S rRNA gene sequences of strains S1T, S2T, and S5T showed the highest similarities to Streptococcus parasanguinis, 98.57%, 99.05%, and 99.05%, respectively, and the orthologous average nucleotide identity (OrthoANI) values between the three strains and S. parasanguinis were 93.82%, 93.67%, and 94.04%, respectively. The pairwise OrthoANI values between the novel strains were 94.37% (S1T-S2T), 95.03% (S2T-S5T), and 94.71% (S1T-S5T). All strains had C20:1 ω9c and summed feature 8 (C18:1 ω7c and/or C18:1 ω6c) as major cellular fatty acids. Additionally, diphosphatidylglycerol (DPG) and hydroxyphosphatidylethanolamine (OH-PE) were identified as major polar lipids. Menaquinone was undetected in all strains. The results from the phylogenetic, phenotypic, chemotaxonomic, and genotypic analyses collectively indicated that strains S1T, S2T, and S5T represent three distinct novel species within the genus Streptococcus, and we propose the names Streptococcus dentalis sp. nov. for strain S1T (= KCTC 21234T = JCM 36526T), Streptococcus gingivalis sp. nov. for strain S2T (= KCTC 21235T = JCM 36527T), and Streptococcus lingualis sp. nov. for strain S5T (= KCTC 21236T = JCM 36528T).
The importance of the complex interplay between the microbiome and mucosal immunity, particularly within the respiratory tract, has gained significant attention due to its potential implications for the severity and progression of lung diseases. Therefore, this review summarizes the specific interactions through which the respiratory tract-specific microbiome influences mucosal immunity and ultimately impacts respiratory health. Furthermore, we discuss how the microbiome affects mucosal immunity, considering tissue-specific variations, and its capacity in respiratory diseases containing asthma, chronic obstructive pulmonary disease, and lung cancer. Additionally, we investigate the external factors which affect the relationship between respiratory microbiome and mucosal immune responses. By exploring these intricate interactions, this review provides valuable insights into the potential for microbiome-based interventions to modulate mucosal immunity and alleviate the severity of respiratory diseases.
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Pseudomonas stutzeri strain AJR13 was isolated for growth on the related compounds biphenyl (BPH) and diphenylmethane
(DPM). The BPH and DPM degradative pathway genes are present on an integrative and conjugative element (ICE) in the
chromosome. Examination of the genome sequence of AJR13 revealed a gene encoding a salicylate 1-monooxygenase (salA)
associated with the ICE even though AJR13 did not grow on salicylate. Transfer of the ICE to the well-studied Pseudomonas
putida KT2440 resulted in a KT2440 strain that could grow on salicylate. Knockout mutagenesis of the salA gene on the
ICE in KT2440 eliminated the ability to grow on salicylate. Complementation of the knockout with the cloned salA gene
restored growth on salicylate. Transfer of the cloned salA gene under control of the lac promoter to KT2440 resulted in a
strain that could grow on salicylate. Heterologous expression of the salA gene in E. coli BL21 DE3 resulted in the production
of catechol from salicylate, confirming that it is indeed a salicylate 1-monooxygenase. Interestingly, transfer of the cloned
salA gene under control of the lac promoter to AJR13 resulted in a strain that could now grow on salicylate, suggesting that
gene expression for the downstream catechol pathway is intact.
Potyvirids, members of the family Potyviridae, produce the P3N-PIPO protein, which is crucial for the cell-to-cell transport
of viral genomic RNAs. The production of P3N-PIPO requires an adenine (A) insertion caused by RNA polymerase slippage
at a conserved GAA AAA A (
GA6) sequence preceding the PIPO open reading frame. Presently, the slippage rate of
RNA polymerase has been estimated in only a few potyvirids, ranging from 0.8 to 2.1%. In this study, we analyzed publicly
available plant RNA-seq data and identified 19 genome contigs from 13 distinct potyvirids. We further investigated the RNA
polymerase slippage rates at the GA6
motif. Our analysis revealed that the frequency of the A insertion variant ranges from
0.53 to 4.07% in 11 potyviruses (genus Potyvirus). For the two macluraviruses (genus Macluravirus), the frequency of the
A insertion variant was found to be 0.72% and 10.96% respectively. Notably, the estimated RNA polymerase slippage rates
for 12 out of the 13 investigated potyvirids were reported for the first time in this study. Our findings underscore the value of
plant RNA-seq data for quantitative analysis of potyvirid genome variants, specifically at the GA6
slippage site, and contribute
to a more comprehensive understanding of the RNA polymerase slippage phenomenon in potyvirids.
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Streptococcus suis serotype 2 (SS2) is an important zoonotic pathogen that can infect humans in contact with infected pigs
or their byproducts. It can employ different types of genes to defend against oxidative stress and ensure its survival. The
thioredoxin (Trx) system is a key antioxidant system that contributes adversity adaptation and pathogenicity. SS2 has been
shown to encode putative thioredoxin genes, but the biological roles, coding sequence, and underlying mechanisms remains
uncharacterized. Here, we demonstrated that SSU05_0237-ORF, from a clinical SS2 strain, ZJ081101, encodes a protein
of 104 amino acids with a canonical CGPC active motif and an identity 70–85% similar to the thioredoxin A (TrxA) in
other microorganisms. Recombinant TrxA efficiently catalyzed the thiol-disulfide oxidoreduction of insulin. The deletion
of TrxA led to a significantly slow growth and markedly compromised tolerance of the pathogen to temperature stress,
as well as impaired adhesion ability to pig intestinal epithelial cells (IPEC-J2). However, it was not involved in H2O2
and
paraquat-induced oxidative stress. Compared with the wild-type strain, the ΔTrxA strain was more susceptible to killing by
macrophages through increasing NO production. Treatment with TrxA mutant strain also significantly attenuated cytotoxic
effects on RAW 264.7 cells by inhibiting inflammatory response and apoptosis. Knockdown of pentraxin 3 in RAW 264.7
cells was more vulnerable to phagocytic activity, and TrxA promoted SS2 survival in phagocytic cells depending on pentraxin
3 activity compared with the wild-type strain. Moreover, a co-inoculation experiment in mice revealed that TrxA mutant
strain is far more easily cleared from the body than the wild type strain in the period from 8–24 h, and exhibits significantly
attenuated oxidative stress and liver injury. In summary, we reveal the important role of TrxA in the pathogenesis of SS2.
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Two Gram-stain-positive, motile, endospore-forming, facultatively anaerobic strains, designated MEBiC13591T
and
MEBiC13594T,
were isolated from tidal flat sediment of the Incheon City on the west coast of Korea. Growth of both
novel strains was observed at pH 5–9 (optimum, pH 7–7.5), and in 0–8% NaCl (optimum, 2% for MEBiC13591T
and
3% for MEBiC13594T).
Strains MEBiC13591T
and MEBiC13594T
grew optimally at 50 °C, (37.5–56.1 °C) and 44 °C
(20.7–50.7 °C), respectively. The main cellular fatty acids of strain MEBiC13591T
were iso-C15: 0, anteiso-C15: 0, iso-C16: 0,
iso-C17: 0 and anteiso-C17: 0, while those for strain MEBiC13594T
were C14:
0, iso-C14: 0, iso-C15: 0, anteiso-C15: 0 and C16:
0. In
both taxa, the major isoprenoid was MK-7. The genomic DNA G + C contents were 34.1 and 37.0 mol% for MEBiC13591T
and MEBiC13594T,
respectively. A 16S rRNA gene sequence analysis revealed that the novel strains showed high similarity
with members of the genera Aeribacillus (95.0%) and Caldibacillus (93.5–94.5%); however, showed lower than 90%
with Caldibacillus debilis TfT.
Phylogenetic and Phylogenomic analysis revealed that two novel strains comprised distinct
phyletic line with members formerly assigned to Caldibacillus. Based on genomic indices, such as AAI and ANI, members
formerly affiliated with Caldibacillus and Bacillus as well as the novel strains should be classified into five independent
genera. Based on the phenotypic, genomic and biochemical data, strains MEBiC13591T
and MEBiC13594T
represent two
novel species in the novel genus, for which the names Fervidibacillus albus gen. nov., sp. nov. (
MEBiC13591T [= KCCM
43317T
= KCTC 43181T
= JCM 33662T
= MCCC 1K04565T]),
and Fervidibacillus halotolerans sp. nov. (
MEBiC13594T
[= KCTC 43182T
= JCM 34001T])
are proposed. Three additional genera Caldifermentibacillus, Palidibacillus, and Perspicuibacillus
are also proposed by reclassification of the several species with valid names that formerly affiliated with the
genera Caldibacillus.
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Microbiome research has been on the rise recently for a more
in-depth understanding of gout. Meanwhile, there is a need to
understand the gut microbiome related to uric acid-lowering
drug resistance. In this study, 16S rRNA gene-based microbiota
analysis was performed for a total of 65 stool samples
from 17 healthy controls and 48 febuxostat-treated gout patients
(including 28 controlled subjects with decreased uric
acid levels and 20 uncontrolled subjects with non-reduced
uric acid levels). Alpha diversity of bacterial community decreased
in the healthy control, controlled, and uncontrolled
groups. In the case of beta diversity, the bacterial community
was significantly different among groups (healthy control, controlled,
and uncontrolled groups). Taxonomic biomarker analysis
revealed the increased population of g-Bifidobacterium
in healthy controls and g-Prevotella in uncontrolled patients.
PCR further confirmed this result at the species level. Additionally,
functional metagenomics predictions led to the exploration
of various functional biomarkers, including purine
metabolism. The results of this study can serve as a basis
for developing potential new strategies for diagnosing and
treating gout from microbiome prospects.
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Terpenes have many applications and are widely found in
nature, but recent progress in synthetic biology has enabled
the use of microorganisms as chassis cells for the synthesis
of these compounds. Candida glycerinogenes (C. glycerinogenes)
is an industrial strain that may be developed as a chassis
for the synthesis of terpenes since it has a tolerance to hyperosmolality
and high sugar, and has a complete mevalonate
(MVA) pathway. However, monoterpenes such as pinene are
highly toxic, and the tolerance of C. glycerinogenes to pinene
was investigated. We also measured the content of mevalonate
and squalene to evaluate the strength of the MVA pathway.
To determine terpene synthesis capacity, a pathway for the synthesis
of pinene was constructed in C. glycerinogenes. Pinene
production was improved by overexpression, gene knockdown
and antisense RNA inhibition. Pinene production was mainly
enhanced by strengthening the upstream MVA pathway and
inhibiting the production of by-products from the downstream
pathway. With these strategies, yield could be increased
by almost 16 times, to 6.0 mg/L. Overall, we successfully constructed
a pinene synthesis pathway in C. glycerinogenes and
enhanced pinene production through metabolic modification.
Citations
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