To elucidate the function of proteorhodopsin in Candidatus Puniceispirillum marinum strain IMCC1322, a cultivated representative of SAR116, we produced RNA-seq data under laboratory conditions. We examined the transcriptomes of six different cultures, including sets of expression changes under constant dark (DD), constant light (LL), and diel-cycled (LD; 14 h light: 10 h dark) conditions at the exponential and stationary/death phases. Prepared mRNA extracted from the six samples was analyzed on the Solexa Genome Analyzer with 36 cycles.
Differentially expressed genes on the IMCC1322 genome were distinguished as four clusters by K-mean clustering and each CDS (n = 2546) was annotated based on the KEGG BRITE hierarchy. Cluster 0 (n = 1573) covered most constitutive genes including proteorhodopsin, retinoids, and glycolysis/TCA cycle. Cluster 1 genes (n = 754) were upregulated in stationary/death phase under constant dark conditions and included genes associated with bacterial defense, membrane transporters, nitrogen metabolism, and senescence signaling. Cluster 2 genes (n = 197) demonstrated upregulation in exponential phase cultures and included genes involved in genes for oxidative phosphorylation, translation factors, and transcription machinery. Cluster 3 (n = 22) contained light-stimulated upregulated genes expressed under stationary/phases. Stringent response genes belonged to cluster 2, but affected genes spanned various cellular processes such as amino acids, nucleotides, translation, transcription, glycolysis, fatty acids, and cell wall components. The coordinated expression of antagonistic stringent genes, including mazG, ppx/gppA, and spoT/relA may provide insight into the controlled cultural response observed between constant light and constant dark conditions in IMCC1322 cultures, regardless of cell numbers and biomass.
Lipolanthine is a subclass of lanthipeptide that has the modification of lipid moiety at the N-terminus. A cryptic biosynthetic
gene cluster comprising four genes (sinA, sinKC, sinD, and sinE) involved in the biosynthesis of lipolanthine was identified in
the genome of an actinobacterium Sinosporangium siamense. Heterologous coexpression of a precursor peptide coding gene
sinA and lanthipeptide synthetase coding gene sinKC in the host Escherichia coli strain BL21(DE3) resulted in the synthesis
of a new lanthipeptide, sinosporapeptin. It contained unusual amino acids, including one labionin and two dehydrobutyrine
residues, as determined using NMR and MS analyses. Another coexpression experiment with two additional genes of decarboxylase
(sinD) and N-acetyl transferase (sinE) resulted in the production of a lipolanthine-like modified sinosporapeptin.
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Three novel bacterial strains, 321T,
335T,
and 353T,
were isolated from the intestines of Aegosoma sinicum larvae collected
from Paju-Si, South Korea. The strains were Gram-negative, obligate aerobe and had rod-shaped cells with a single flagellum.
The three strains belonged to the genus Luteibacter in the family Rhodanobacteraceae and shared < 99.2% similarity
in their 16S rRNA gene sequence and < 83.56% similarity in thier whole genome sequence. Strains 321T,
335T,
and 353T
formed a monophyletic clade with Luteibacter yeojuensis KACC 11405T,
L. anthropi KACC 17855T,
and L. rhizovicinus
KACC 12830T,
with sequence similarities of 98.77–98.91%, 98.44–98.58%, and 97.88–98.02%, respectively. Further
genomic analyses, including the construction of the Up-to-date Bacterial Core Gene (UBCG) tree and assessment of other
genome-related indices, indicated that these strains were novel species belonging to the genus Luteibacter. All three strains
contained ubiquinone Q8 as their major isoprenoid quinone and iso-C15:0 and summed feature 9 (
C16:0 10-methyl and/or
iso-C17:1 ω9c) as their major cellular fatty acids. Phosphatidylethanolamine and diphosphatidylglycerol were the major polar
lipids in all the strains. The genomic DNA G + C contents of strains 321T,
335T,
and 353T
were 66.0, 64.5, and 64.5 mol%,
respectively. Based on multiphasic classification, strains 321T,
335T,
and 353T
were classified into the genus Luteibacter
as the type strains of novel species, for which the names Luteibacter aegosomatis sp. nov., Luteibacter aegosomaticola sp.
nov., and Luteibacter aegosomatissinici sp. nov. are proposed, respectively.
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Toxin-antitoxin (TA) systems are widespread in bacteria and archaea plasmids and genomes to regulate DNA replication,
gene transcr!ption, or protein translation. Higher eukaryotic and prokaryotic nucleotide-binding (HEPN) and minimal
nucleotidyltransferase (MNT) domains are prevalent in prokaryotic genomes and constitute TA pairs. However, three gene
pairs (MTH304/305, 408/409, and 463/464) of Methanothermobacter thermautotropicus ΔH HEPN-MNT family have not
been studied as TA systems. Among these candidates, our study characterizes the MTH463/MTH464 TA system. MTH463
expression inhibited Escherichia coli growth, whereas MTH464 did not and blocked MTH463 instead. Using site-directed
MTH463 mutagenesis, we determined that amino acids R99G, H104A, and Y106A from the R[ɸX]4-6H motif are involved
with MTH463 cell toxicity. Furthermore, we established that purified MTH463 could degrade MS2 phage RNA, whereas
purified MTH464 neutralized MTH463 activity in vitro. Our results indicate that the endonuclease toxin MTH463 (encoding
a HEPN domain) and its cognate antitoxin MTH464 (encoding the MNT domain) may act as a type II TA system in
M. thermautotropicus ΔH. This study provides initial and essential information studying TA system functions, primarily
archaea HEPN-MNT family.
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Mycobacterium tuberculosis is the causative agent of tuberculosis. M. tuberculosis can survive in a dormant state within
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to antibiotics and stress conditions, and thus the transition of M. tuberculosis to the nonreplicating dormant state acts as
an obstacle to tuberculosis treatment. M. tuberculosis in the granuloma encounters hostile environments such as hypoxia,
nitric oxide, reactive oxygen species, low pH, and nutrient deprivation, etc., which are expected to inhibit respiration of M.
tuberculosis. To adapt to and survive in respiration-inhibitory conditions, it is required for M. tuberculosis to reprogram its
metabolism and physiology. In order to get clues to the mechanism underlying the entry of M. tuberculosis to the dormant
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across different growth stages was identified through transcriptome data analysis. The candidate promoters were functionally
characterized in A. oryzae by transcriptional control of β-glucuronidase (GUS) as a reporter. The results showed that
the glyceraldehyde triphosphate dehydrogenase promoter (PgpdA1) of A. oryzae with a unique structure displayed the most
robust strength in constitutively controlling the expression compared to the PgpdA2 and other putative promoters tested. In
addition, the ubiquitin promoter (Pubi) of A. oryzae exhibited a moderate expression strength. The deletion analysis revealed
that the 5' untranslated regions of gpdA1 and ubi with the length of 1028 and 811 nucleotides, counted from the putative
translation start site (ATG), respectively, could efficiently drive the GUS expression. Interestingly, both promoters could
function on various carbon sources for cell growth. Glucose was the best fermentable carbon source for allocating high constitutive
expressions during cell growth, and the high concentrations (6–8% glucose, w/v) did not repress their functions. It
was also demonstrated that the secondary metabolite gene coding for indigoidine could express under the control of PgpdA1
or Pubi promoter. These strong and moderate promoters of A. oryzae provided beneficial options in tuning the transcriptional
expression for leveraging the metabolic control towards the targeted products.
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of coronaviruses encodes several enzymes, including RNAdependent
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systems have been developed to identify potential inhibitors.
In this review, we describe the natural compounds that
have been shown to exert inhibitory effects on coronavirus enzymes.
Although computer-aided molecular structural studies
have predicted several antiviral compound candidates, the current
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Oligopeptides with functional activities are of current interest
in the nutraceutical and medical sectors. The development of
the biosynthetic process of oligopeptides through a nonribosomal
peptide synthetase (NRPS) system has become more
challenging. To develop a production platform for nonribosomal
peptides (NRPs), reprogramming of transcriptional
regulation of the acv gene encoded ACV synthetase (ACVS)
was implemented in Aspergillus oryzae using the CRISPRCas9
system. Awakening silent acv expression was successfully
achieved by promoter substitution. Among the three exchanged
promoters, AoPgpdA, AoPtef1, and PtPtoxA, the
replacement of the native promoter with AoPgpdA led to the
highest ACV production in A. oryzae. However, the ACV production
of the AoPGpdA strain was also dependent on the
medium composition, in which urea was the best nitrogen
source, and a C:N ratio of 20:1 was optimal for tripeptide production.
In addition to cell growth, magnesium ions are an
essential element for ACV production and might participate
in ACVS activity. It was also found that ACV was the growthassociated
product of the engineered strain that might be a result of constitutive transcriptional control by the AoPgpdA
promoter. This study offers a potential strategy for nonribosomal
ACV production using the fungal system, which is applicable
for redesigning bioactive oligopeptides with industrial
relevance.
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Yvh1 is a dual-specificity phosphatase (DUSP) that is evolutionarily
conserved in eukaryotes, including yeasts and humans.
Yvh1 is involved in the vegetative growth, differentiation,
and virulence of animal and plant fungal pathogens.
All Yvh1 orthologs have a conserved DUSP catalytic domain
at the N-terminus and a zinc-binding (ZB) domain with two
zinc fingers (ZFs) at the C-terminus. Although the DUSP domain
is implicated in the regulation of MAPK signaling in
humans, only the ZB domain is essential for most cellular
functions of Yvh1 in fungi. This study aimed to analyze the
functions of the DUSP and ZB domains of Yvh1 in the human
fungal pathogen Cryptococcus neoformans, whose Yvh1
(CnYvh1) contains a DUSP domain at the C-terminus and
a ZB domain at the N-terminus. Notably, CnYvh1 has an extended
internal domain between the two ZF motifs in the ZB
domain. To elucidate the function of each domain, we constructed
individual domain deletions and swapping strains
by complementing the yvh1Δ mutant with wild-type (WT)
or mutated YVH1 alleles and examined their Yvh1-dependent
phenotypes, including growth under varying stress conditions,
mating, and virulence factor production. Here, we found
that the complementation of the yvh1Δ mutant with the mutated
YVH1 alleles having two ZFs of the ZB domain, but not
the DUSP and extended internal domains, restored the WT
phenotypic traits in the yvh1Δ mutant. In conclusion, the
ZB domain, but not the N-terminal DUSP domain, plays a
pivotal role in the pathobiological functions of cryptococcal
Yvh1.
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several elaborate strategies to colonize a wide range of natural
or clinical niches and to overcome the neighboring bacterial
competitors in polymicrobial communities. However,
the relationship and interaction mechanism of P. aeruginosa
with other bacterial pathogens remains largely unexplored.
Here we explore the interaction dynamics of P. aeruginosa and
Escherichia coli, which frequently coinfect the lungs of immunocompromised
hosts, by using a series of on-plate proximity
assays and RNA-sequencing. We show that the extracellular
products of P. aeruginosa can inhibit the growth of
neighboring E. coli and induce a large-scale of transcriptional
reprogramming of E. coli, especially in terms of cellular respiration-
related primary metabolisms and membrane components.
In contrast, the presence of E. coli has no significant
effect on the growth of P. aeruginosa in short-term culture,
but causes a dysregulated expression of genes positively controlled
by the quorum-sensing (QS) system of P. aeruginosa
during subsequent pairwise culture. We further demonstrate
that the divergent QS-regulation of P. aeruginosa may be related
to the function of the transcriptional regulator PqsR,
which can be enhanced by E. coli culture supernatant to increase
the pyocyanin production by P. aeruginosa in the absence
of the central las-QS system. Moreover, the extracellular
products of E. coli promote the proliferation and lethality
of P. aeruginosa in infecting the Caenorhabditis elegans
model. The current study provides a general characterization
of the extracellular products-mediated interactions between
P. aeruginosa and E. coli, and may facilitate the understanding
of polymicrobial infections.
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Histone acetylation/deacetylation represent a general and
efficient epigenetic mechanism through which fungal cells control
gene expression. Here we report developmental requirement
of MoHOS2-mediated histone deacetylation (HDAC)
for the rice blast fungus, Magnaporthe oryzae. Structural similarity
and nuclear localization indicated that MoHOS2 is an
ortholog of Saccharomyces cerevisiae Hos2, which is a member
of class I histone deacetylases and subunit of Set3 complex.
Deletion of MoHOS2 led to 25% reduction in HDAC
activity, compared to the wild-type, confirming that it is a
bona-fide HDAC. Lack of MoHOS2 caused decrease in radial
growth and impinged dramatically on asexual sporulation.
Such reduction in HDAC activity and phenotypic defects of
ΔMohos2 were recapitulated by a single amino acid change
in conserved motif that is known to be important for HDAC
activity. Expression analysis revealed up-regulation of MoHOS2
and concomitant down-regulation of some of the key genes
involved in asexual reproduction under sporulation-promoting
condition. In addition, the deletion mutant exhibited defect
in appressorium formation from both germ tube tip and
hyphae. As a result, ΔMohos2 was not able to cause disease
symptoms. Wound-inoculation showed that the mutant is
compromised in its ability to grow inside host plants as well.
We found that some of ROS detoxifying genes and known
effector genes are de-regulated in the mutant. Taken together,
our data suggest that MoHOS2-dependent histone deacetylation
is pivotal for proper timing and induction of transcription
of the genes that coordinate developmental changes
and host infection in M. oryzae.
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Primary infections with the varicella-zoster virus (VZV) result
in varicella, while latent reactivation leads to herpes zoster.
Both varicella and zoster can be prevented by live attenuated
vaccines. There have been reports suggesting that both clinical
VZV strains and those in vaccine preparations are genetically
polymorphic, containing mixtures of both wild-type
and vaccine-type sequences at certain vaccine-specific sites.
In this study, the genetic polymorphism of the VZV genome
was examined by analyzing the frequencies of minor alleles
at each nucleotide position. Next-generation sequencing of
the clinical VZV strain YC02 passaged in an in vitro cell culture
was used to identify genetically polymorphic sites (GPS),
where the minor allele frequency (MAF) exceeded 5%. The
number of GPS increased by 7.3-fold at high passages (p100)
when compared to low passages (p17), although the average
MAF remained similar. GPS were found in 6 open reading
frames (ORFs) in p17, 35, and 54 ORFs in p60 and p100, respectively.
GPS were found more frequently in the dispensable
gene group than the essential gene group, but the average MAF
was greater in the essential gene group. The most common
two major/minor base pairs were A/g and T/c. GPS were found
in all three passages at 16 positions, all located in the reiterated
(R) region. The population diversity as measured by Shannon
entropy increased in p60 and p100. However, the entropy
remained unchanged in the R regions.
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In the present study, a laccase gene (BaLc) from a lignin degrading
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and expressed in Escherichia coli. The optimal catalytic activity
of the protein was achieved at 5.5 pH and 35°C temperature,
measured by oxidation of ABTS. The Km and Vmax
values were determined as 1.42 mM and 4.16 μmole/min, respectively.
To achieve the enzyme recovery, the biocatalyst
(BaLc) was covalently attached onto the functionalized iron
magnetic-nanoparticles. The nanoparticles were characterized
by zeta-potential and FTIR analyses. The immobilized BaLc
enzyme was physico-kinetically characterized, exhibiting retention
of 60% of the residual activity after ten reaction cycles
of ABTS oxidation. The immobilized biocatalyst system was
tested for its biotechnological exploitability in plant juice
processing, achieving 41–58% of phenol reduction, 41–58%
decolorization, 50–59% turbidity reduction in the extracts of
banana pseudo-stem and sweet sorghum stalk, and apple fruit
juice. This is the first study to demonstrate the use of nanoparticle-
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Strain IMCC1322 was isolated from a surface water sample
from the East Sea of Korea. Based on 16S rRNA analysis,
IMCC1322 was found to belong to the OCS28 sub-clade of
SAR116. The cells appeared as short vibrioids in logarithmicphase
culture, and elongated spirals during incubation with
mitomycin or in aged culture. Growth characteristics of strain
IMCC1322 were further evaluated based on genomic information;
proteorhodopsin (PR), carbon monoxide dehydrogenase,
and dimethylsulfoniopropionate (DMSP)-utilizing
enzymes. IMCC1322 PR was characterized as a functional
retinylidene protein that acts as a light-driven proton pump
in the cytoplasmic membrane. However, the PR-dependent
phototrophic potential of strain IMCC1322 was only observed
under CO-inhibited and nutrient-limited culture conditions.
A DMSP-enhanced growth response was observed in addition
to cultures grown on C1 compounds like methanol, formate,
and methane sulfonate. Strain IMCC1322 cultivation
analysis revealed biogeochemical processes characteristic of
the SAR116 group, a dominant member of the microbial community
in euphotic regions of the ocean. The polyphasic taxonomy
of strain IMCC1322 is given as Candidatus Puniceispirillum
marinum, and was confirmed by chemotaxonomic
tests, in addition to 16S rRNA phylogeny and cultivation
analyses.
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Characteristics of naringinase nano-encapsulated forms on
different carrier materials (chitosan and alginate polymers)
were investigated in this study. Screening of twelve fungal isolates
for naringinase production indicated that Trichoderma
longibrachiatum was the most promising. Grapefruit rind was
used as a substrate containing naringin for naringinase production.
TEM micrographs showed that chitosan nano-capsules
were applied for the production of morphologically homogeneous
enzymatic nano-particles with high enzyme encapsulation
efficiency, small asymmetric sizes (from 15.09 to
27.07 nm with the mean of 21.8 nm) and rough surfaces compared
to nano-encapsulated naringinase in alginate which
showed nano-particle size (from 33.37 to 51.01 nm with the
mean of 43.03 nm). It was revealed that the highest naringinase
activity was found in case of chitosan nano-capsule naringinase
compared to alginate nano-capsule one. Thermogram
analysis (TGA) showed that the free enzyme loses about
92% of its weight at approximately 110°C, while the nanoencapsulated
ones show more stability at higher temperatures.
Conclusively, the nano-capsulation process improves the kinetics
and operational stability so could be useful as a debittering
agent for various thermal processing applications in
citrus juices industries which makes the fruit juice more acceptable
and cost-effective to the consumer.
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