Plastic pollution exacerbated by the excessive use of synthetic
plastics and its recalcitrance has been recognized among the
most pressing global threats. Microbial degradation of plastics
has gained attention as a possible eco-friendly countermeasure,
as several studies have shown microbial metabolic
capabilities as potential degraders of various synthetic plastics.
However, still defined biochemical mechanisms of biodegradation
for the most plastics remain elusive, because the
widely used culture-dependent approach can access only a
very limited amount of the metabolic potential in each microbiome.
A culture-independent approach, including metagenomics,
is becoming increasingly important in the mining
of novel plastic-degrading enzymes, considering its more expanded
coverage on the microbial metabolism in microbiomes.
Here, we described the advantages and drawbacks associated
with four different metagenomics approaches (microbial community
analysis, functional metagenomics, targeted gene sequencing,
and whole metagenome sequencing) for the mining
of plastic-degrading microorganisms and enzymes from
the plastisphere. Among these approaches, whole metagenome
sequencing has been recognized among the most powerful
tools that allow researchers access to the entire metabolic potential
of a microbiome. Accordingly, we suggest strategies
that will help to identify plastisphere-enriched sequences as
de novo plastic-degrading enzymes using the whole metagenome
sequencing approach. We anticipate that new strategies
for metagenomics approaches will continue to be developed
and facilitate to identify novel plastic-degrading microorganisms
and enzymes from microbiomes.
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Two strictly aerobic, Gram-staining-positive, non-spore-forming,
regular rod-shaped (approximately 0.7 × 1.9 mm)
bacteria (HY170T and HY001) were isolated from bat feces
collected from Chongzuo city, Guangxi province (22°2054N,
106°4920E, July 2011) and Chuxiong Yi Autonomous Prefecture,
Yunnan province (25°0910N, 102°0439E, October
2013) of South China, respectively. Optimal growth is obtained
at 25–28°C (range, 4–32°C) on BHI-5% sheep blood
plate with pH 7.5 (range, 5.0–10.0) in the presence of 0.5–
1.0% NaCl (w/v) (range, 0–15% NaCl [w/v]). The phylogenetic
and phylogenomic trees based respectively on the 16S
rRNA gene and 845 core gene sequences revealed that the
two strains formed a distinct lineage within the genus Brevibacterium,
most closely related to B. aurantiacum NCDO
739T (16S rRNA similarity, both 98.5%; dDDH, 46.7–46.8%;
ANI, 91.9–92.1%). Strain HY170T contained MK-8(H2), diphosphatidylglycerol
(DPG) and phosphatidylglycerol (PG),
galactose and ribose as the predominant menaquinone, major
polar lipids, and main sugars in the cell wall teichoic acids,
respectively. The meso-diaminopimelic acid (meso-DAP)
was the diagnostic diamino acid of the peptidoglycan found
in strain HY170T. Anteiso-C15:0 and anteiso-C17:0 were the
major fatty acids (> 10%) of strains HY170T and HY001, with
anteiso-C17:1A predominant in strain HY170T but absent in
strain HY001. Mining the genomes revealed the presence
of secondary metabolite biosynthesis gene clusters encoding
for non-alpha poly-amino acids (NAPAA), ectoine, siderophore,
and terpene. Based on results from the phylogenetic,
chemotaxonomic and phenotypic analyses, the two strains
could be classified as a novel species of the genus Brevibacterium,
for which the name Brevibacterium zhoupengii sp.
nov. is proposed (type strain HY170T = CGMCC 1.18600T
= JCM 34230T).
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Microbial communities in the rhizosphere play a crucial role
in determining plant growth and crop yield. A few studies
have been performed to evaluate the diversity and co-occurrence
patterns of rhizosphere microbiomes in soybean (Glycine
max) at a regional scale. Here, we used a culture-independent method to compare the bacterial communities of the
soybean rhizosphere between Nebraska (NE), a high-yield
state, and Oklahoma (OK), a low-yield state. It is well known
that the rhizosphere microbiome is a subset of microbes that
ultimately get colonized by microbial communities from the
surrounding bulk soil. Therefore, we hypothesized that differences
in the soybean yield are attributed to the variations in
the rhizosphere microbes at taxonomic, functional, and community
levels. In addition, soil physicochemical properties
were also evaluated from each sampling site for comparative
study. Our result showed that distinct clusters were formed
between NE and OK in terms of their soil physicochemical
property. Among 3 primary nutrients (i.e., nitrogen, phosphorus,
and potassium), potassium is more positively correlated
with the high-yield state NE samples. We also attempted
to identify keystone communities that significantly affected the
soybean yield using co-occurrence network patterns. Network
analysis revealed that communities formed distinct clusters
in which members of modules having significantly positive
correlations with the soybean yield were more abundant in
NE than OK. In addition, we identified the most influential
bacteria for the soybean yield in the identified modules. For
instance, included are class Anaerolineae, family Micromonosporaceae,
genus Plantomyces, and genus Nitrospira in the
most complex module (ME9) and genus Rhizobium in ME23.
This research would help to further identify a way to increase
soybean yield in low-yield states in the U.S. as well as worldwide
by reconstructing the microbial communities in the
rhizosphere.
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Aspergillus section Nigri is a fungus used industrially because
of its ability to produce enzymes such as cellulolytic, amylolytic
and proteolytic enzymes. In this study, we obtained twentyeight
strains of Aspergillus section Nigri from the traditional
Korean fermentation starter, nuruk, which is known as a mixed
culture of enzymatic filamentous fungi and yeasts. All strains
were identified as Aspergillus section Nigri through combined
phylogenetic analysis using partial β-tubulin and calmodulin
gene sequences. The cellulase, amylase and protease activities
of Korean strains were measured and compared with ten reference
strains of Aspergillus niger. Most Korean strains showed
higher cellulolytic activity than reference strains, and Aspergillus
neoniger KCN5 showed the highest β-glucosidase activity.
Two-thirds of the Korean strains showed similar levels
of α- and glucoamylase activity as the reference strains. The
protease activity of Aspergillus section Nigri strains was the
highest at pH 3.0, and A. niger KSJ2 showed the highest acidic
protease activity. By comparing ten reference strains and
twenty-eight Korean strains, our results suggested useful Aspergillus
section Nigri strains from nuruk with high enzyme
activity, such as KCN5 and KSJ2, and their potential for industrial
applications as enzyme producers.
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A novel haloarchaeal species designated as MBLA0099T was
isolated from seawater near Yeongheung Island. Cells were
Gram-negative, non-motile, red-pigmented, and rod-shaped.
They grew at 10–45°C, within pH 5.5–9.0, and between 7.5%
and 30% NaCl concentrations. Cells were able to grow without
Mg2+ and were lysed in distilled water. The size of the
whole-genome and G + C content of DNA was 3.02 Mb and
68.9 mol%, respectively. Phylogenetic analysis shows that
the strain MBLA0099T belongs to the genus Halorubrum.
The average nucleotide and amino acid identity, and in silico
DNA-DNA hybridization values were below the species delineation
threshold. Pan-genomic analysis revealed that 3.2%
of all genes present in strain MBLA0099T were unique to the
strain. The red carotenoid produced by strain MBLA0099T
was subjected to spectrometric and chromatographic analyses
and confirmed to be bacterioruberin as C50 carotenoid.
Mevalonic acid, terpenoid backbone, and carotenoid biosynthesis
pathway were annotated for strain MBLA0099T. The
C50 carotenoid production by strain MBLA0099T was also enhanced
under various stress conditions including relatively
netural pH, high oxidative and salinity conditions. Additionally,
the strain MBLA0099T-derived bacterioruberin showed
the antioxidant activity with EC50 value of 12.29 μg/ml, based
on the evaluation of DPPH free radical scavenging activity.
The present study would be the first report on the identification
of C50 carotenoid from the strain MBLA0099T representing
a novel species of the genus Halorubrum, for which
the name Halorubrum ruber sp. nov. is proposed. The typestrain
used was MBLA0099T (= KCTC 4296T = JCM 34701T).
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of feeding intolerance (FI) remains unclear. Understanding
the role of the gut microbiota could provide a new avenue for
microbiota-targeted therapeutics. This study aimed to explore
the associations between aberrant gut microbiota and FI in
very low or extremely low birth weight (VLBW/ELBW) preterm
infants. In this observational case-control study, VLBW/
ELBW infants were divided into two groups: FI group and
feeding tolerance (FT) group. 16S rRNA gene sequencing was
performed to analyze the gut microbial diversity and composition
of the infants. The differences in the gut microbiota of
the two groups were compared. In total, 165 stool samples
were obtained from 44 infants, among which, 31 developed
FI and 13 served as controls. Alpha diversity was the highest
in the meconium samples of the two groups. LEfSe analysis
revealed that the abundances of Peptostreptococcaceae, Clostridiales
and Clostridia in the FT group were significantly higher
than in the FI group. At the phylum level, the FI group was dominated
by Proteobacteria, and the FT group was dominated
by Firmicutes. The meconium samples of the FI group had
higher proportions of γ-proteobacteria and Escherichia-Shigella
and a lower proportion of Bacteroides compared with the FT
group. Kyoto Encyclopedia of Genes and Genomes (KEGG)
analysis demonstrated that aberrant gut bacteria in the FI group
were strongly associated with dysregulation of C5-Brancheddibasic-
acid-metabolism, protein kinases, and sporulation.
These findings reveal candidate microbial markers to prevent
FI. Increased relative abundances of γ-proteobacteria
and Escherichia-Shigella and decreased abundance of Bacteroides
in meconium were associated with an increased risk
of FI, while Peptostreptococcaceae, Clostridiales and Clostridia
reduced the risk of FI in VLBW/ELBW infants.
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Glycoprotein (G protein)-based DNA vaccines are effective
in protecting aquaculture fish from rhabdoviruses but the degree
of immune response they elicit depends on plasmid concentration
and antigen cassette. Here, we developed a DNA
vaccine using the viral hemorrhagic septicemia virus G (VG)
gene and chemokine (C-C motif) ligand 19 (CCL19)a.2 regulated
by the CMV promoter as the molecular adjuvant. After
transfection of the prepared plasmid (pVG + CCL19) into epithelioma
papulosum cyprini cells, mRNA expression was confirmed
through quantitative real-time polymerase chain reaction.
The vaccine was intramuscularly injected into zebrafish
(Danio rerio), and 28 days after immunization, viral hemorrhagic
septicemia virus (105 TCID50/10 μl/fish) was intraperitoneally
injected. A survival rate of 68% was observed in the
pVG + CCL19 group but this was not significantly different
from the survival rate of fish treated with pVG alone, that is,
without the adjuvant. However, the expression of interferonand
cytokine-related genes in the spleen and kidney tissues
of zebrafish was significantly increased (p < 0.05) on days 1,
3, 7, and 14 after immunization. Thus, CCL19a.2 induced an
initial immune response as a molecular adjuvant, which may
provide initial protection against virus infection before vaccination-
induced antibody formation. This study provides insights
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Hepatitis B virus (HBV) infection highly increases the risk
for liver cirrhosis and hepatocellular carcinoma (HCC). The
clinical manifestation of HBV infection is determined by the
mutual interplay of the viral genotype, host genetic factors,
mode of transmission, adaptive mutations, and environmental
factors. Core promoter activation plays a critical role in the
pre-genomic RNA transcription of HBV for HBV replication.
The mutations of core promoter have been implicated in HCC
development. We had obtained HBV genes from Myanmar
HBV infectants and identified gene variations at the core promoter
region. For measuring the relative transactivation activity
on core promoter, we prepared the core-promoter reporter
construct. Both of A1762T and G1764A mutation were
consistently found in the HBV genes with hepatocellular carcinoma.
The A1762T/G1764A mutation was corresponding
to K130M/V131I of HBx protein. We prepared the core promoter-
luciferase reporter construct containing the double
A1762T/G1764A mutation and the K130M/V131I HBx protein
expression construct. The A1762T/G1764A mutation
highly was responsive to core promoter transactivation by
HBx, regardless of HBx mutation. The A1762T/G1764A mutation
newly created hepatocyte nuclear factor 1 (HNF1) responsive
element. Ectopic expression of HNF1 largely increased
the HBV core promoter containing A1762T/G1764A
mutation. In addition, hepatic rich fatty acid, palmitic acid
and oleic acid, increased K130M/V131I HBx level by core
promoter activation. These results provide biological properties
and clinical significance of specific HBV core promoter
mutants related with HCC development.
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