Journal Articles
- Antimicrobial Efficacy of Allium cepa and Zingiber officinale Against the Milk‑Borne Pathogen Listeria monocytogenes
-
Abirami Arasu , Nagaram Prabha , Durga Devi , Praveen Kumar Issac , Khaloud Mohammed Alarjani , Dunia A. Al Farraj , Reem A. Aljeidi , Dina S. Hussein , Magesh Mohan , Jehad Zuhair Tayyeb , Ajay Guru , Jesu Arockiaraj
-
J. Microbiol. 2023;61(11):993-1011. Published online December 4, 2023
-
DOI: https://doi.org/10.1007/s12275-023-00086-w
-
-
Abstract
- Listeria monocytogenes is an important food-borne pathogen that causes listeriosis and has a high case fatality rate despite
its low incidence. Medicinal plants and their secondary metabolites have been identified as potential antibacterial substances,
serving as replacements for synthetic chemical compounds. The present studies emphasize two significant medicinal plants,
Allium cepa and Zingiber officinale, and their efficacy against L. monocytogenes. Firstly, a bacterial isolate was obtained
from milk and identified through morphology and biochemical reactions. The species of the isolate were further confirmed
through 16S rRNA analysis. Furthermore, polar solvents such as methanol and ethanol were used for the extraction of secondary
metabolites from A. cepa and Z. officinale. Crude phytochemical components were identified using phytochemical
tests, FTIR, and GC–MS. Moreover, the antibacterial activity of the crude extract and its various concentrations were tested
against L. monocytogenes. Among all, A. cepa in methanolic extracts showed significant inhibitory activity. Since, the A.
cepa for methanolic crude extract was used to perform autography to assess its bactericidal activity. Subsequently, molecular
docking was performed to determine the specific compound inhibition. The docking results revealed that four compounds
displayed strong binding affinity with the virulence factor Listeriolysin-O of L. monocytogenes. Based on the above results,
it can be concluded that the medicinal plant A. cepa has potential antibacterial effects against L. monocytogenes, particularly
targeting its virulence.
- Vaginal Microbiome Dysbiosis is Associated with the Different Cervical Disease Status
-
Yingying Ma , Yanpeng Li , Yanmei Liu , Le Cao , Xiao Han , Shujun Gao , Chiyu Zhang
-
J. Microbiol. 2023;61(4):423-432. Published online April 3, 2023
-
DOI: https://doi.org/10.1007/s12275-023-00039-3
-
-
24
View
-
0
Download
-
3
Citations
-
Abstract
- Vaginal microbiome composition was demonstrated to be associated with cervical disease. The colonization characteristics
of vaginal microbes and their association with the different cervical disease status, especially cervical cancer (CC), are
rarely investigated. In this cross-sectional study, we characterized the vaginal microbiome of women with different status of
cervical diseases, including 22 NV + (normal tissue with HPV infection), low-grade squamous intraepithelial lesion (LSIL,
n = 45), high-grade squamous intraepithelial lesion (HSIL, n = 36) and CC (n = 27) using bacterial 16S DNA sequencing.
Thirty HPV-negative women with normal tissue were used as the control group. We found that higher diversity of microbiome
with gradual depletion of Lactobacillus, especially L. crispatus, was associated with the severity of cervical disease.
High-risk HPV16 infection was associated with higher microbiome diversity and depletion of Lactobacillus in high-grade
cervical diseases (i.e. HSIL and CC). The CC group was characterized by higher levels of Fannyhessea vaginae, Prevotella,
Bacteroides, Finegoldia, Vibrio, Veillonella, Peptostreptococcus, and Dialister. Co-occurrence network analyses showed that
negative correlations were exclusively observed between Lactobacillus and other bacteria, and almost all non-Lactobacillus
bacteria were positively correlated with each other. In particular, the most diverse and complex co-occurrence network of
vaginal bacteria, as well as a complete loss of L. crispatus, was observed in women with CC. Logistic regression model
identified HPV16 and Lactobacillus as significant risk and protective factors for CC, respectively. These results suggest that
specific Lactobacillus species (e.g. L. crispatus and L. iners) can be used as important markers to target prevention measures
prioritizing HPV16-infected women and other hrHPV-infected women for test, vaccination and treat initiatives.
- Gamma-glutamyltransferase of Helicobacter pylori alters the proliferation, migration, and pluripotency of mesenchymal stem cells by affecting metabolism and methylation status
-
Zeyu Wang , Weijun Wang , Huiying Shi , Lingjun Meng , Xin Jiang , Suya Pang , Mengke Fan , Rong Lin
-
J. Microbiol. 2022;60(6):627-639. Published online April 18, 2022
-
DOI: https://doi.org/10.1007/s12275-022-1575-4
-
-
18
View
-
0
Download
-
6
Citations
-
Abstract
- Virulence factor gamma-glutamyltransferase (GGT) of H.
pylori consumes glutamine (Gln) in the stomach to decrease
the tricarboxylic acid metabolite alpha-ketoglutarate (α-kg)
and alter the downstream regulation of α-kg as well as cellular
biological characteristics. Our previous research indicated
that under H. pylori infection, mesenchymal stem cells
(MSCs) migrated to the stomach and participated in gastric
cancer (GC) development either by differentiating into epithelial
cells or promoting angiogenesis. However, how MSCs
themselves participate in H. pylori-indicated GC remains
unclear. Therefore, a GGT knockout H. pylori strain (Hp-
KS-1) was constructed, and downstream histone H3K9 and
H3K27 methylation and the PI3K/AKT signaling pathway
of α-kg were detected using Western blotting. The biological
characteristics of MSCs were also examined. An additive α-kg
supplement was also added to H. pylori-treated MSCs to investigate
alterations in these aspects. Compared to the control
and Hp-KS-1 groups, H. pylori-treated MSCs reduced Gln
and α-kg, increased H3K9me3 and H3K27me3, activated the
PI3K-AKT signaling pathway, and promoted the proliferation,
migration, self-renewal, and pluripotency of MSCs. The
addition of α-kg rescued the H. pylori-induced alterations.
Injection of MSCs to nude mice resulted in the largest tumors
in the H. pylori group and significantly reduced tumor sizes
in the Hp-KS-1 and α-kg groups. In summary, GGT of H.
pylori affected MSCs by interfering with the metabolite α-kg
to increase trimethylation of histone H3K9 and H3K27, activating
the PI3K/AKT signaling pathway, and promoting
proliferation, migration, self-renewal, and pluripotency in tumorigenesis,
elucidating the mechanisms of MSCs in GC
development.
- Alcohol dehydrogenase 1 and NAD(H)-linked methylglyoxal oxidoreductase reciprocally regulate glutathione-dependent enzyme activities in Candida albicans
-
Sa-Ouk Kang , Min-Kyu Kwak
-
J. Microbiol. 2021;59(1):76-91. Published online December 23, 2020
-
DOI: https://doi.org/10.1007/s12275-021-0552-7
-
-
16
View
-
0
Download
-
2
Citations
-
Abstract
- Glutathione reductase (Glr1) activity controls cellular glutathione
and reactive oxygen species (ROS). We previously
demonstrated two predominant methylglyoxal scavengers–
NAD(H)-linked methylglyoxal oxidoreductase (Mgd1) and
alcohol dehydrogenase 1 (Adh1)–in glutathione-depleted γ-
glutamyl cysteinyl synthetase-disrupted Candida albicans.
However, experimental evidence for Candida pathophysiology
lacking the enzyme activities of Mgd1 and Adh1 on glutathione-
dependent redox regulation remains unclear. Herein,
we have aimed to demonstrate that glutathione-dependent
enzyme activities coupled with cellular ROS changes is regulated
by methylglyoxal accumulation in Δmgd1/Δadh1 double
disruptants. Δmgd1/Δadh1 showed severe growth defects
and G1-phase cell cycle arrest. The observed complementary
and reciprocal methylglyoxal-oxidizing and methylglyoxalreducing
activities between Δmgd1 and Δadh1 were not always
exhibited in Δmgd1/Δadh1. Although intracellular accumulation
of methylglyoxal and pyruvate was shown in all
disruptants, to a greater or lesser degree, methylglyoxal was
particularly accumulated in the Δmgd1/Δadh1 double disruptant.
While cellular ROS significantly increased in Δmgd1
and Δadh1 as compared to the wild-type, Δmgd1/Δadh1 underwent
a decrease in ROS in contrast to Δadh1. Despite the
experimental findings underlining the importance of the
undergoing unbalanced redox state of Δmgd1/Δadh1, glutathione-
independent antioxidative enzyme activities did not
change during proliferation and filamentation. Contrary to
the significantly lowered glutathione content and Glr1 enzyme
activity, the activity staining-based glutathione peroxidase
activities concomitantly increased in this mutant. Additionally,
the enhanced GLR1 transcript supported our results in
Δmgd1/Δadh1, indicating that deficiencies of both Adh1 and
Mgd1 activities stimulate specific glutathione-dependent enzyme
activities. This suggests that glutathione-dependent redox
regulation is evidently linked to C. albicans pathogenicity
under the control of methylglyoxal-scavenging activities.
Retracted Publication
- Cryptic prophages in a blaNDM-1-bearing plasmid increase bacterial survival against high NaCl concentration, high and low temperatures, and oxidative and immunological stressors
-
So Yeon Kim , Kwan Soo Ko
-
J. Microbiol. 2020;58(6):483-488. Published online March 28, 2020
-
DOI: https://doi.org/10.1007/s12275-020-9605-6
-
-
19
View
-
0
Download
-
4
Citations
-
Abstract
- In this study, we investigated the effect of cryptic prophage
regions in a blaNDM-1-bearing plasmid, which was identified in
a patient from South Korea, on the survival of bacteria against
adverse environmental conditions. First, we conjugated the
intact plasmid and plasmids with deleted cryptic prophages
into Escherichia coli DH5α. The E. coli transconjugants carrying
the plasmid with intact cryptic prophages showed increased
survival during treatment with a high concentration
of NaCl, high and low temperatures, an oxidative stressor
(H2O2), and an immunological stressor (human serum). By
contrast, the transconjugants carrying the plasmid with a
single-cryptic prophage knockout did not show any change
in survival rates. mRNA expression analyses revealed that the
genes encoding sigma factor proteins were highly upregulated
by the tested stressors and affected the expression of
various proteins (antioxidant, cell osmosis-related, heat shock,
cold shock, and universal stress proteins) associated with the
specific defense against each stress. These findings indicate
that a bacterial strain carrying a plasmid with intact carbapenemase
gene and cryptic prophage regions exhibited an increased
resistance against simulated environmental stresses,
and cryptic prophages in the plasmid might contribute to this
enhanced stress resistance. Our study indicated that the coselection
of antibiotic resistance and resistance to other stresses
may help bacteria to increase survival rates against adverse
environments and disseminate.
Journal Articles
- Methyltransferase of a cell culture-adapted hepatitis E inhibits the MDA5 receptor signaling pathway
-
Jinjong Myoung , Jeong Yoon Lee , Kang Sang Min
-
J. Microbiol. 2019;57(12):1126-1131. Published online November 22, 2019
-
DOI: https://doi.org/10.1007/s12275-019-9478-8
-
-
17
View
-
0
Download
-
7
Citations
-
Abstract
- Hepatitis E virus (HEV) is a causative agent of acute hepatitis
and jaundice. The number of human infections is approximated
to be over 20 million cases per year. The transmission
is mainly via the fecal-oral route and contaminated water and
food are considered to be a major source of infection. As a
mouse model is not available, a recent development of a cell
culture-adapted HEV strain (47832c) is considered as a very
important tools for molecular analysis of HEV pathogenesis
in cells. Previously, we demonstrated that HEV-encoded methyltransferase
(MeT) encoded by the 47832c strain inhibits
MDA5- and RIG-I-mediated activation of interferon β (IFN-β)
promoter. Here, we report that MeT impairs the phosphorylation
and activation of interferon regulatory factor 3 and the
p65 subunit of NF-κB in a dose-dependent manner. In addition,
the MeT encoded by the 47832c, but not that of HEV
clinical or field isolates (SAR-55, Mex-14, KC-1, and ZJ-1),
displays the inhibitory effect. A deeper understanding of MeTmediated
suppression of IFN-β expression would provide
basis of the cell culture adaptation of HEV.
- Biofilm characterization of Fusarium solani keratitis isolate: increased resistance to antifungals and UV light
-
Itzel Margarita Córdova-Alcántara , Diana Laura Venegas-Cortés , María Ángeles Martínez-Rivera , Néstor Octavio Pérez , Aida Verónica Rodriguez-Tovar
-
J. Microbiol. 2019;57(6):485-497. Published online May 27, 2019
-
DOI: https://doi.org/10.1007/s12275-019-8637-2
-
-
10
View
-
0
Download
-
42
Citations
-
Abstract
- Fusarium solani has drawn phytopathogenic, biotechnological,
and medical interest. In humans, it is associated with
localized infections, such as onychomycosis and keratomycosis,
as well as invasive infections in immunocompromised
patients. One pathogenicity factor of filamentous fungi is biofilm
formation. There is still only scarce information about
the in vitro mechanism of the formation and composition of
F. solani biofilm. In this work, we describe the biofilm formed
by a clinical keratomycosis isolate in terms of its development,
composition and susceptibility to different antifungals and
ultraviolet light (UV) at different biofilm formation stages.
We found five biofilm formation stages using scanning electron
microscopy: adherence, germination, hyphal development,
maturation, and cell detachment. Using epifluorescence
microscopy with specific fluorochromes, it was elucidated
that the extracellular matrix consists of carbohydrates, proteins,
and extracellular DNA. Specific inhibitors for these
molecules showed significant biofilm reductions. The antifungal
susceptibility against natamycin, voriconazole, caspofungin,
and amphotericin B was evaluated by metabolic activity
and crystal violet assay, with the F. solani biofilm preformation
to 24 h increased in resistance to natamycin, voriconazole,
and caspofungin, while the biofilm preformation
to 48 h increased in resistance to amphotericin B. The preformed
biofilm at 24 h protected and reduced UV light
mortality. F. solani isolate could produce a highly structured
extra biofilm; its cellular matrix consists of carbohydrate polymers,
proteins, and eDNA. Biofilm confers antifungal resistance
and decreases its susceptibility to UV light. The fungal
biofilm functions as a survival strategy against antifungals
and environmental factors.
- Gentic overexpression increases production of hypocrellin A in Shiraia bambusicola S4201
-
Dan Li , Ning Zhao , Bing-Jing Guo , Xi Lin , Shuang-Lin Chen , Shu-Zhen Yan
-
J. Microbiol. 2019;57(2):154-162. Published online January 31, 2019
-
DOI: https://doi.org/10.1007/s12275-019-8259-8
-
-
14
View
-
0
Download
-
15
Citations
-
Abstract
- Hypocrellin A (HA) is a perylenequinone (PQ) isolated from
Shiraia bambusicola that shows antiviral and antitumor activities,
but its application is limited by the low production
from wild fruiting body. A gene overexpressing method was
expected to augment the production rate of HA in S. bambusicola.
However, the application of this molecular biology
technology in S. bambusicola was impeded by a low genetic
transformation efficiency and little genomic information. To
enhance the plasmid transformant ratio, the Polyethylene
Glycol-mediated transformation system was established and
optimized. The following green fluorescent protein (GFP)
analysis showed that the gene fusion expression system we
constructed with a GAPDH promoter Pgpd1 and a rapid 2A
peptide was successfully expressed in the S. bambusicola S4201
strain. We successfully obtained the HA high-producing strains
by overexpressing O-methyltransferase/FAD-dependent monooxygenase
gene (mono) and the hydroxylase gene (hyd),
which were the essential genes involved in our putative HA
biosynthetic pathway. The overexpression of these two genes
increased the production of HA by about 200% and 100%,
respectively. In general, this study will provide a basis to identify
the genes involved in the hypocrellin A biosynthesis. This
improved transformation method can also be used in genetic
transformation studies of other fungi.
- Crystal structure of the inactive state of the receiver domain of Spo0A from Paenisporosarcina sp. TG-14, a psychrophilic bacterium isolated from an Antarctic glacier
-
Chang Woo Lee , Sun-Ha Park , Sung Gu Lee , Seung Chul Shin , Se Jong Han , Han-Woo Kim , Hyun Ho Park , Sunghwan Kim , Hak Jun Kim , Hyun Park , HaJeung Park , Jun Hyuck Lee
-
J. Microbiol. 2017;55(6):464-474. Published online March 9, 2017
-
DOI: https://doi.org/10.1007/s12275-017-6599-9
-
-
14
View
-
0
Download
-
2
Citations
-
Abstract
- The two-component phosphorelay system is the most pre-valent mechanism for sensing and transducing environ-mental signals in bacteria. Spore formation, which relies on the two-component phosphorelay system, enables the long- term survival of the glacial bacterium Paenisporosarcina sp. TG-14 in the extreme cold environment. Spo0A is a key re-sponse regulator of the phosphorelay system in the early stage of spore formation. The protein is composed of a regu-latory N-terminal phospho-receiver domain and a DNA- binding C-terminal activator domain. We solved the three- dimensional structure of the unphosphorylated (inactive) form of the receiver domain of Spo0A (PaSpo0A-R) from Paenisporosarcina sp. TG-14. A structural comparison with phosphorylated (active form) Spo0A from Bacillus stearo-thermophilus (BsSpo0A) showed minor notable differences. A molecular dynamics study of a model of the active form and the crystal structures revealed significant differences in the α4 helix and the preceding loop region where phosphorylation occurs. Although an oligomerization study of PaSpo0A-R by analytical ultracentrifugation (AUC) has shown that the protein is in a monomeric state in solution, both crosslinking and crystal-packing analyses indicate the possibility of weak dimer formation by a previously undocumented mechanism. Collectively, these observations provide insight into the me-chanism of phosphorylation-dependent activation unique to Spo0A.
- Heterologous expression and enzymatic characterization of γ-glutamyltranspeptidase from Bacillus amyloliquefaciens
-
Jung-Min Lee , Jaejung Lee , Gyeong-Hwa Nam , Byung-Sam Son , Myoung-Uoon Jang , So-Won Lee , Byung-Serk Hurh , Tae-Jip Kim
-
J. Microbiol. 2017;55(2):147-152. Published online January 26, 2017
-
DOI: https://doi.org/10.1007/s12275-017-6638-6
-
-
12
View
-
0
Download
-
16
Citations
-
Abstract
- γ-Glutamyltranspeptidase (GGT) catalyzes the cleavage of γ-
glutamyl compounds and the transfer of γ-glutamyl moiety
to water or to amino acid/peptide acceptors. GGT can be utilized
for the generation of γ-glutamyl peptides or glutamic
acid, which are used as food taste enhancers. In the present
study, Bacillus amyloliquefaciens SMB469 with high GGT activity
was isolated from Doenjang, a traditional fermented soy
food of Korea. The gene encoding GGT from B. amyloliquefaciens
SMB469 (BaGGT469) was cloned from the isolate, and
heterologously expressed in E. coli and B. subtilis. For comparison,
three additional GGT genes were cloned from B.
subtilis 168, B. licheniformis DSM 13, and B. amyloliquefaciens
FZB42. The BaGGT469 protein was composed of 591
amino acids. The final protein comprises two separate polypeptide
chains of 45.7 and 19.7 kDa, generated via autocatalytic
cleavage. The specific activity of BaGGT469 was determined
to be 17.8 U/mg with γ-L-glutamyl-p-nitroanilide
as the substrate and diglycine as the acceptor. GGTs from B.
amyloliquefaciens showed 1.4- and 1.7-fold higher transpeptidase
activities than those from B. subtilis and B. licheniformis,
respectively. Especially, recombinant B. subtilis expressing
BaGGT469 demonstrated 11- and 23-fold higher GGT
activity than recombinant E. coli and the native B. amyloliquefaciens,
respectively, did. These results suggest that BaGGT469
can be utilized for the enzymatic production of various γ-
glutamyl compounds.
- Alteration in the ultrastructural morphology of mycelial hyphae and the dynamics of transcriptional activity of lytic enzyme genes during basidiomycete morphogenesis
-
Elena Vetchinkina , Maria Kupryashina , Vladimir Gorshkov , Marina Ageeva , Yuri Gogolev , Valentina Nikitina
-
J. Microbiol. 2017;55(4):280-288. Published online January 26, 2017
-
DOI: https://doi.org/10.1007/s12275-017-6320-z
-
-
12
View
-
0
Download
-
11
Citations
-
Abstract
- The morphogenesis of macromycetes is a complex multilevel process resulting in a set of molecular-genetic, physiological- biochemical, and morphological-ultrastructural changes in the cells. When the xylotrophic basidiomycetes Lentinus edodes, Grifola frondosa, and Ganoderma lucidum were grown on wood waste as the substrate, the ultrastructural morphology of the mycelial hyphal cell walls differed considerably between mycelium and morphostructures. As the macromycetes passed from vegetative to generative development, the expression of the tyr1, tyr2, chi1, chi2, exg1, exg2, and exg3 genes was acti-vated. These genes encode enzymes such as tyrosinase, chi-tinase, and glucanase, which play essential roles in cell wall growth and morphogenesis.
Review
- MINIREVIEW] High-resolution imaging of the microbial cell surface
-
Ki Woo Kim
-
J. Microbiol. 2016;54(11):703-708. Published online October 29, 2016
-
DOI: https://doi.org/10.1007/s12275-016-6348-5
-
-
15
View
-
0
Download
-
11
Citations
-
Abstract
- Microorganisms, or microbes, can function as threatening
pathogens that cause disease in humans, animals, and plants;
however, they also act as litter decomposers in natural ecosystems.
As the outermost barrier and interface with the environment,
the microbial cell surface is crucial for cell-to-cell
communication and is a potential target of chemotherapeutic
agents. Surface ultrastructures of microbial cells have typically
been observed using scanning electron microscopy (SEM)
and atomic force microscopy (AFM). Owing to its characteristics
of low-temperature specimen preparation and superb
resolution (down to 1 nm), cryo-field emission SEM has
revealed paired rodlets, referred to as hydrophobins, on the
cell walls of bacteria and fungi. Recent technological advances
in AFM have enabled high-speed live cell imaging in liquid
at the nanoscale level, leading to clear visualization of celldrug
interactions. Platinum-carbon replicas from freeze-fractured
fungal spores have been observed using transmission
electron microscopy, revealing hydrophobins with varying
dimensions. In addition, AFM has been used to resolve bacteriophages
in their free state and during infection of bacterial
cells. Various microscopy techniques with enhanced spatial
resolution, imaging speed, and versatile specimen preparation
are being used to document cellular structures and
events, thus addressing unanswered biological questions.
Journal Article
- Identification of D-amino acid dehydrogenase as an upstream regulator of the autoinduction of a putative acyltransferase in Corynebacterium glutamicum
-
Jung-Hoon Lee , Yong-Jae Kim , Hee-Sung Shin , Heung-Shick Lee , Shouguang Jin , Un-Hwan Ha
-
J. Microbiol. 2016;54(6):432-439. Published online May 27, 2016
-
DOI: https://doi.org/10.1007/s12275-016-6046-3
-
-
13
View
-
0
Download
-
1
Citations
-
Abstract
- Expression of a putative acyltransferase encoded by NCgl-
0350 of Corynebacterium glutamicum is induced by cell-free
culture fluids obtained from stationary-phase growth of both
C. glutamicum and Pseudomonas aeruginosa, providing evidence
for interspecies communication. Here, we further confirmed
that such communication occurs by showing that acyltransferase
expression is induced by culture fluid obtained
from diverse Gram-negative and -positive bacterial strains,
including Escherichia coli, Salmonella Typhimurium, Bacillus
subtilis, Staphylococcus aureus, Mycobacterium sp. strain JC1,
and Mycobacterium smegmatis. A homologous acyltransferase
encoded by PA5238 of P. aeruginosa was also induced by
fluids obtained from P. aeruginosa as well as other bacterial
strains, as observed for NCgl0350 of C. glutamicum. Because
C. glutamicum is difficult to study using molecular approaches,
the homologous gene PA5238 of P. aeruginosa was used to
identify PA5309 as an upstream regulator of expression. A
homologous D-amino acid dehydrogenase encoded by NCgl-
2909 of C. glutamicum was cloned based on amino acid similarity
to PA5309, and its role in the regulation of NCgl0350
expression was confirmed. Moreover, NCgl2909 played positive
roles in growth of C. glutamicum. Thus, we identified a
D-amino acid dehydrogenase as an upstream regulator of the
autoinduction of a putative acyltransferase in C. glutamicum.
Research Support, Non-U.S. Gov'ts
- Role of bacterial γ-glutamyltranspeptidase as a novel virulence factor in bone-resorbing pathogenesis
-
Jinmoon Kim , Sungil Jang , Aeryun Kim , Hanfu Su , Niluka Gunawardhana , Yeong-Eui Jeon , Eun Jung Bak , Ji-Hye Kim , Jeong-Heon Cha
-
J. Microbiol. 2016;54(5):396-402. Published online April 20, 2016
-
DOI: https://doi.org/10.1007/s12275-016-6137-1
-
-
11
View
-
0
Download
-
5
Citations
-
Abstract
- Mammalian γ-glutamyltranspeptidase (GGT) has been identified
as a bone-resorbing factor. Since GGT of Bacillus subtilis
exhibits similarity in their primary structure and enzymatic
characteristics with mammalian GGTs, the bone-resorbing
activity of bacterial GGT was examined in this study. Osteoclastogenesis
was performed in a co-culture system of mouse
calvaria-derived osteoblasts and bone marrow cells. A conditioned
medium from GGT-overproducing B. subtilis culture
showed significantly higher activity of osteoclast formation
than a conditioned medium from wild-type B. subtilis culture.
Recombinant GGT (rGGT) of wild-type B. subtilis and an
enzymatic activity-defected rGGT of B. subtilis 2288 mutant
were expressed in Escherichia coli and purified using His tag.
Both purified rGGTs induced similar levels of osteoclastogenesis,
suggesting that B. subtilis GGT possesses virulent boneresorbing
activity and its activity is probably independent of
its enzymatic activity. Furthermore, a recombinant protein of
B. subtilis GGT heavy subunit (Bs rGGT/H) showed strong
activity of osteoclastogenesis while the light subunit failed to
show strong activity, suggesting that the bone-resorbing activity
is mainly located at the heavy subunit. More importantly,
the GGT enzymatic activity may not be required for this virulence
activity since the light subunit contains the catalytic
pocket. In addition, B. subtilis rGGT stimulated mRNA expressions
of receptor activator of nuclear factor kappa-B ligand
(RANKL) and cyclooxygenase-2 (COX-2), while an
osteoprotegerin inhibited the osteoclast formation induced by Bs rGGT/H. This is the first demonstration that bacterial
GGT itself is sufficient to act as a bone-resorbing virulence
factor via RANKL-dependent pathway. Therefore, it can be
hypothesized that GGT of periodontopathic bacteria may play
an important role as a virulence factor in bone destruction.
- Bacillus cheonanensis sp. nov. Isolated from Near Poultry Farm Soil
-
Hyun-Ju Kim , Cheol-Su Park , Siwon Lee , Tae-Young Ahn
-
J. Microbiol. 2014;52(7):554-558. Published online May 30, 2014
-
DOI: https://doi.org/10.1007/s12275-014-3458-9
-
-
11
View
-
0
Download
-
1
Citations
-
Abstract
- A novel bacterial strain, designated PFS-5T, was isolated from the soil environment with feces of a live poultry farm located in Cheonan, Republic of Korea. Strain PFS-5T was Gram-staining-positive, motile, strictly aerobic bacterium, rod-shaped, and endospore-forming. The strain contained meso-diaminopimelic acid in their peptidoglycan and MK-7 menaquinone. The major fatty acids were anteiso-C15:0 (44.2%), C16:0 (22.2%), and iso-C15:0 (16.7%). The DNA G+C content was 40.1 mol%. Comparative 16S rRNA gene sequence analysis identified strain PFS-5T in the genus Bacillus, exhibiting the highest level of sequence similarity with type strain of B. herbersteinensis D-1,5aT (96.9%), B. humi LMG 22167T (96.7%), B. alkalitelluris BA288T (96.1%), B. litoralis SW-211T (96.0%), and B. luteolus YIM93174T (95.5%). The major polar lipids of PFS-5T were diphosphatidylglycerol and phosphatidylglycerol. On the basis of result from poly-phasic data, strain PFS-5T represents a novel species, for which the name Bacillus cheonanensis sp. nov. is proposed (Type strain PFS-5T= KACC 17469T= JCM19333T).