In vitro fecal fermentation is an assay that uses fecal microbes
to ferment foods, the results of which can be used to
evaluate the potential of prebiotic candidates. To date, there
have been various protocols used for in vitro fecal fermentation-
based assessments of food substances. In this study,
we investigated how personal gut microbiota differences and
external factors affect the results of in vitro fecal fermentation
assays. We used Cheonggukjang (CGJ), a Korean traditional
fermented soybean soup that is acknowledged as
healthy functional diet. CGJ was digested in vitro using acids
and enzymes, and then fermented with human feces anaerobically.
After fecal fermentation, the microbiota was analyzed
using MiSeq, and the amount of short chain fatty acids
(SCFAs) were measured using GC-MS. Our results suggest
that CGJ was effectively metabolized by fecal bacteria to produce
SCFAs, and this process resulted in an increase in the
abundance of Coprococcus, Ruminococcus, and Bifidobacterium
and a reduction in the growth of Sutterella, an opportunistic
pathogen. The metabolic activities predicted from the
microbiota shifts indicated enhanced metabolism linked to
methionine biosynthesis and depleted chondroitin sulfate
degradation. Moreover, the amount of SCFAs and microbiota
shifts varied depending on personal microbiota differences.
Our findings also suggest that in vitro fecal fermentation of
CGJ for longer durations may partially affect certain fecal
microbes. Overall, the study discusses the usability of in vitro
gastrointestinal digestion and fecal fermentation (GIDFF)
to imitate the effects of diet-induced microbiome modulation
and its impact on the host.
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Protein lysine acetylation influences many physiological functions,
such as gene regulation, metabolism, and disease in
eukaryotes. Although little is known about the role of lysine
acetylation in bacteria, several reports have proposed its importance
in various cellular processes. Here, we discussed the
function of the protein lysine acetylation and the post-translational
modifications (PTMs) of histone-like proteins in bacteria
focusing on Salmonella pathogenicity. The protein lysine
residue in Salmonella is acetylated by the Pat-mediated enzymatic
pathway or by the acetyl phosphate-mediated non-enzymatic
pathway. In Salmonella, the acetylation of lysine 102
and lysine 201 on PhoP inhibits its protein activity and DNAbinding,
respectively. Lysine acetylation of the transcriptional
regulator, HilD, also inhibits pathogenic gene expression.
Moreover, it has been reported that the protein acetylation
patterns significantly differ in the drug-resistant and
-sensitive Salmonella strains. In addition, nucleoid-associated
proteins such as histone-like nucleoid structuring protein
(H-NS) are critical for the gene silencing in bacteria, and
PTMs in H-NS also affect the gene expression. In this review,
we suggest that protein lysine acetylation and the post-translational
modifications of H-NS are important factors in understanding
the regulation of gene expression responsible
for pathogenicity in Salmonella.
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The DNA damage checkpoint signaling pathway is a highly
conserved surveillance mechanism that ensures genome integrity
by sequential activation of protein kinase cascades.
In mammals, the main pathway is orchestrated by two central
sensor kinases, ATM and ATR, that are activated in response
to DNA damage and DNA replication stress. Patients
lacking functional ATM or ATR suffer from ataxia-telangiectasia
(A-T) or Seckel syndrome, respectively, with pleiotropic
degenerative phenotypes. In addition to DNA strand
breaks, ATM and ATR also respond to oxidative DNA damage
and reactive oxygen species (ROS), suggesting an unconventional
function as regulators of intracellular redox status.
Here, we summarize the multiple roles of ATM and ATR, and
of their orthologs in Saccharomyces cerevisiae, Tel1 and Mec1,
in DNA damage checkpoint signaling and the oxidative stress
response, and discuss emerging ideas regarding the possible
mechanisms underlying the elaborate crosstalk between those
pathways. This review may provide new insights into the integrated
cellular strategies responsible for maintaining genome
stability in eukaryotes with a focus on the yeast model
organism.
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Pif1 DNA helicase is the prototypical member of a 5' to 3' helicase superfamily conserved from bacteria to humans. In Saccharomyces cerevisiae, Pif1 and its homologue Rrm3, localize in both mitochondria and nucleus playing multiple roles in the maintenance of genomic homeostasis. They display relatively weak processivities in vitro, but have largely non-overlapping functions on common genomic loci such as mitochondrial DNA, telomeric ends, and many replication forks especially at hard-to-replicate regions including ribosomal DNA and G-quadruplex structures. Recently, emerging evidence shows that Pif1, but not Rrm3, has a significant new role in repair-associated DNA synthesis with Polδ during homologous recombination stimulating D-loop migration for conservative DNA replication. Comparative genetic and biochemical studies on the structure and function of Pif1 family helicases across different biological systems are further needed to elucidate both diversity and specificity of their mechanisms of action that contribute to genome stability.
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DNA-unwinding activity of Saccharomyces cerevisiae Pif1 is modulated by thermal stability, folding conformation, and loop lengths of G-quadruplex DNA Lei Wang, Qing-Man Wang, Yi-Ran Wang, Xu-Guang Xi, Xi-Miao Hou Journal of Biological Chemistry.2018; 293(48): 18504. CrossRef
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Yeast Pif1 Accelerates Annealing of Complementary DNA Strands Ramanagouda Ramanagoudr-Bhojappa, Alicia K. Byrd, Christopher Dahl, Kevin D. Raney Biochemistry.2014; 53(48): 7659. CrossRef