Hong, Eun Hye;Lee, Sun Hee;Vendan, Regupathy Thamizh;Rhee, Young Ha
Korean Journal of Microbiology
/
v.48
no.4
/
pp.246-253
/
2012
The purpose of this study was to investigate the molecular diversity of rhizobacteria associated with ginseng of varying age levels and their plant benefiting attributes. A total of 143 different isolates belonging to 15 different bacterial genera were recovered. Although variation was found in the rhizobacterial community due to age of the plant, majority of bacteria belong to Firmicutes (58%). In which, Bacillus was found to be the predominant genus irrespective of age of the ginseng. To assess the plant benefiting attributes, 30 representative isolates were selected. The results indicated that some of the isolates could exhibit multiple plant growth promoting traits like secretion of cell wall degrading enzymes, production of indole-3-acetic acid, synthesis of siderophores, solubilization of phosphates and soil pathogens inhibition. It can be suggested that strains of B. subtilis, B. amyloliquefaciens, B. velezensis, and B. licheniformis were positive for all the above traits, which have potential to be used as plant growth promoting inoculants to improve ginseng crop in the future.
To investigate cellular regulation of phosphate metabolism between catabolically repressed and derepressed states in Saccharomyces uvarum, the activities of polyphosphatases, the analysis of polyphosphate and cytochemical observation of volutin granules were examined according to the culture phase and under various phosphate concentrations. As the results, tripolyphosphatase activity was increased more than six-fold during catabolic repression as compared with those of catabolic derepression and the polyphosphatase activity increased at the time of maximal accumulation of acid insoluble polyphosphate 'B'. Of the low molecular weight polyphosphates, tripolyphosphate was mainly detected by thin layer chromatography. When the synthesis of volutin granules in derepressed cells was observed cytochemically, acid insoluble polyphosphate localizing at the cell wall was primarily synthesized and then transferred into the cytoplasm, nucleus and/or vacuole.
Kim, Hye Rim;Kim, Hangeun;Jung, Bong Jun;You, Ga Eun;Jang, Soojin;Chung, Dae Kyun
Molecules and Cells
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v.38
no.2
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pp.163-170
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2015
Lipoteichoic acid (LTA) is a major component of the cell wall of Gram-positive bacteria. Its effects on living organisms are different from those of lipopolysaccharide (LPS) found in Gram-negative bacteria. LTA contributes to immune regulatory effects including anti-aging. In this study, we showed that LTA isolated from Lactobacillus plantarum (pLTA) inhibited melanogenesis in B16F10 mouse melanoma cells. pLTA reduced the cellular activity of tyrosinase and the expression of tyrosinase family members in a dose-dependent manner. The expression of microphthalmia- associated transcription factor (MITF), a key factor in the synthesis of melanin, was also decreased by pLTA. Further, we showed that pLTA activated melanogenesis signaling, such as extracellular signal-regulated kinase (ERK) and phosphatidylinositol 3-kinse (PI3K)/AKT. In addition, the expression of heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) and HuR, which are important RNA-binding proteins (RBPs), was reduced. pLTA likely degrades MITF via regulation of melanogenic signaling and RNA stability of melanogenic proteins, resulting in the reduction of melanin. Thus, our data suggest that pLTA has therapeutic potential for treating hyperpigmentation disorders and can also be used as a cosmetic whitening agent.
Laccases are multicopper-containing enzymes which catalyze the oxidation of phenolic and nonphenolic compounds with the concomitant reduction of molecular oxygen. They often occur as isoenzymes, either constitutive or inducible, that oligomerize to multilateral complexes, what allow for penetration to the woody cell wall structure. White rot basidiomycete fungi may produce a number of laccase isoenzymes, some constitutively and others after induction. Fungal laccase is commonly induced by many ions, such as $Cu^{2+}$, $Cd^{2+}$$Ca^{2+}$, $Li^+$, $Mn^{2+}$, $Ag^+$, $Hg^{2+}$, Mn and $Fe^{3+}$, phenolic compounds, some organic compounds, such as ethanol, isopropanol, cAMP, caffeine, p-anisidine, viscosinamide and paraquat, and nitrogens and even heat shock. A combination of Cu and pHB (p-hydroxybenzoic acid) made it possible to extend the inducible laccase activities over 30-fold. But the most effective inducer of laccase in the basidiomycete and other higher fungi is 2,5-xylidine, over 160-fold stimulation of laccase activity. The laccases are frequently encoded by gene families, as e.g. in Pycnoporus cinnabarinus, from which the lcc3-1 or the allelic form lac1 and lac3-2 have been cloned and sequenced. In the case of inducible forms the post-inductional laccase formation depends upon the synthesis of mRNA and the induction is due to the synthesis of a new protein.
Lignin is a complex phenylpropanoid polymer abundant in the cell walls of vascular plants. It is mainly presented in conducting and supporting tissues, assisting in water transport and mechanical strength. Lignification is also utilized as a defense mechanism against pathogen infection or wounding to protect plant tissues. The monolignol precursors of lignin are synthesized by cinnamyl alcohol dehydrogenase (CAD). CAD catalyzes cinnamaldehydes to cinnamyl alcohols, such as p-coumaryl, coniferyl, and sinapyl alcohols. CAD exists as a multigenic family in angiosperms, and CAD isoforms with different functions have been identified in different plant species. Multiple isoforms of CAD genes are differentially expressed during development and upon environmental cues. CAD enzymes having different functions have been found so far, showing that one of its isoforms may be involved in developmental lignification, whereas others may affect the composition of defensive lignins and other wall-bound phenolics. Substrate specificity appears differently depending on the CAD isoform, which contributes to revealing the biochemical properties of CAD proteins that regulate lignin synthesis. In this review, details regarding the expression and regulation of the CAD family in lignin biosynthesis are discussed. The isoforms of the CAD multigenic family have complex genetic regulation, and the signaling pathway and stress responses of plant development are closely linked. The synthesis of monolignol by CAD genes is likely to be regulated by development and environmental cues as well.
The vancomycin, one of the family of glycopeptide antibiotics, inhibits the synthesis of bacterial cell wall peptidoglycan and has been widely used against gram-positive bacterial infections, especially for a treatment of methicillin resistant S. aureus infection. However, clinical isolate which was intermediately resistant to vancomycin (Mu50: MIC 8 $\mu\textrm{g}$/ml) was isolated in recent years. In this study we performed vancomycin susceptibility test with the increment method and population analysis with clinical isolates S. aureus. Also we did several kinds of tests with three selected isolates (s129: MIC 7 $\mu\textrm{g}$/ml, s134: MIC 7 $\mu\textrm{g}$/ml, s135: MIC 8 $\mu\textrm{g}$/ml) to find out possible mechanism of vancomycin resistance. As a result, the prevalence of vancomycin resistant S. aureus isolates among S. aureus strains resistant to methicillin was 23.3% (25/107). The vancomycin resistances of isolated strains of S. aureus were between those of Mu5O and Mu3 strains. By PCR analysis, none of the isolates with decreased vancomycin susceptibility contained known vancomycin resistant genes such as vanA, vanB, vanC1, vanC2, and vanH. Major bands of 81 kDa, 58 kDa, 33 kDa, 28 kDa were demonstrable in whole cell lysates by SDS-PAGE from all three isolates as well as reference strains. And especially,45 kDa protein was overproduced in Mu50 strains. Among them increased production of NAD$^{+}$-linked-$_{D}$-lactate dehydrogenase (dnLDH) were detected from one clinical strain (s135) and Mu5O strain. From these data, we suggest that the mechanism of vancomycin resistance in these isolates are distinct from that in enterococci.
Aromatic hydrocarbons which are not easily degraded by microorganisms can be accumulated in the conlaminated environment for a long lime, producing toxic effects on wild lives and humans. However, the sublethal concentrations of the chemicals induce the synthesis of stress-shock proteins in the cells and increase the adaptability of the organisms to the environmental stresses. In this study, therefore, the cells of Psezido~nonus sp. DJ- 12 treated with catechol at various concentrations were inveshgated for their survival, biodegtadability of catechol, production of stress-shock proteins, and cytological changes. The organisms were capable of degrading catechol at the range of 0.5 to 1.0 mM concentration wilhin 6 hours incubation, but they were killed by $10^2$-10$^3$ celllinl at 3 mM or higel- concentration without any catechol degradation. These cells treated with catechol begm lo produce DnaK and GroEL at 1 mM and 0.5 mM. respectively. Pseudumonas sp. DJ-12 treated with 10 mM catechol for I hour exhihiled some punctuated pores on the cell wall and contortion of the rod shape. The cells treated with he sublethal concentration of catechol showed the increased tolerance for suvival when exposed to the lethal concentration, and such tolerant effects were functioned crossly among benzoate, 4-chlorobenzoate, 'and catechol.
To know how the ribosomes involved in secretory protein synthesis were attached to the cytoplasmic membrane in Bacillus amyloliquefaciens, the cells were treated with puromycin combinated with magnesium at the logarithmic phase, and the variation of cell-bound and extracellular $\alpha$-amylase activity was assayed for determining the $\alpha$-amylase translocation blocking through the cytoplasmic membrane. In the abnormal $\alpha$-amylase producing mutant in which the C-terminal of the $\alpha$-amylase structure was deleted, B. umytotiquefaciens CH10-2, the $\alpha$-amylase was translocated normally through the cytoplasmic membranes, and the translocation blocking by puromycin was revealed to have a similar pattern as that in the wild type. This means that the C-terminal part of the enzyme structure may not have a signal for secretion. The cell death of the logarithmic phase cells in both strains was not affected much under 20$\mu\textrm{g}$/$m\ell$ of puromycin, however, the $\alpha$-amylase translocation was blocked markedly under less than 10$\mu\textrm{g}$/$m\ell$ of the puromycin concentration. The blocking of the enzyme secretion by puromycin may be due to the detachment of the ribosomes from cytoplasmic membranes by disturbing the nascent polypeptide synthesis. Further evidence for confirming this was that the detachment was increased in 50 mM of magnesium ion because the extracellular $\alpha$-amylase activity was decreased more under this condition. If the cells were treated with trypsin combinated with Iysozyme, the extracellular $\alpha$-amylase activity from the cultured medium was reduced markedly, however, the activity from the cells treated with trypsin only was not reduced. This means that the nascent polypeptides protruding from the cytoplasmic membrane were sensitive to the trypsin digestion, whereas the matured ones were not. Therefore, the protruding polypeptides from the cytoplasmic membranes may be truncated by trypsin before forming their final tertiary structures by folding in the cell wall layer.
Cephradine is a first generation cephalosporin and has broad spectrum antibacterial activity against gram-positive and gram-negative microorganisms, through inhibition of bacterial cell wall synthesis. Cephradine is useful for treatment of infections of the urinary and respiratory tract, skin and soft tissues. The purpose of the present study was to evaluate the bioequivalence of two cephradine capsules, Cefradine Yuhan (YuHan Corporation) and Broadcef (Ilsung Pharmaceuticals Co. Ltd.), according to the guidelines of Korea Food and Drug Administration (KFDA). The cephradine release from the two cephradine capsules in vitro was tested using KP VII Apparatus II method with various different kinds of dissolution media (pH 1.2, 4.0, 6.8 buffer solution and water). Twenty normal male volunteers, $23.10{\pm}2.90$ years in age and $67.69{\pm}8.04\;kg$ in body weight, were divided into two groups and a randomized $2{\times}2$ cross-over study was employed. After one capsule containing 500 mg as cephradine was orally administered, blood was taken at predetermined time intervals and the concentrations of cephradine in serum were determined using HPLC method with UV detector. The dissolution profiles of two cephradine capsules were very similar at all dissolution media. Besides, the pharmacokinetic parameters such as $AVC_t,\;C_{max}\;and\;T_{max}$ were calculated and ANOVA test was utilized for the statistical analysis of the parameters using logarithmically transformed $AVC_t\;and\;C_{max}$ and untransformed $T_{max}$. The results showed that the differences in $AVC_t,\;C_{max}\;and\;T_{max}$ between two capsules based on the Cefradine Yuhan were -2.87%, -0.96% and -4.85%, respectively. There were no sequence effects between two capsules in these parameters. The 90% confidence intervals using logarithmically transformed data were within the acceptance range of 1og(0.8) to log(1.25) $(e.g.,\;log(0.93){\sim}log(1.02)\;and\;log(0.88){\sim}log(1.13)\;for \;AVC_t\;and\;C_{max},\;respectively)$. The 90% confidence interval using untransformed data was within ${\pm}20%$$(e.g., \;-17.54{\sim}7.78\;for\;T_{max})$. All parameters met the criteria of KFDA guideline for bioequivalence, indicating that Broadcef capsule is bioequivalent to Cefradine Yuhan capsule.
The nutritive value of 4 straws, obtained after thrashing of seeds from fodder crops, was assessed as complete feed for ruminants. Sixteen male Murrah buffaloes (liveweight 365.8${\pm}$19.5 kg), were divided into 4 equal groups and offered ad lib. straw of either Trifolium resupinatum, Trifolium alexandrium, Medicago sativa or Lolium perenne, supplemented with minerals and vitamin A, for 40 days in a completely randomized design. Simultaneously, each straw was offered to 3 rumen fistulated male buffaloes in order to assess the biochemical changes in the rumen. Compared to other straws M. sativa straw had higher (p<0.05) organic matter (OM), crude protein (CP), acid-detergent fiber (ADF) and cellulose content. L .perenne had the highest (p<0.05) hemicellulose and lowest (p<0.05) CP and acid-detergent lignin (ADL) content. T. resupinatum had the lowest concentration of cell wall constituents (CWC). The digestibility of nutrients of T. resupinatum and L. perenne straw was similar, but higher (p<0.05) than that of other straws. M.sativa straw showed highest (p<0.05) digestibility of CP. The highest OM digestibility of T. resupinatum and CP digestibility of M. sativa were responsible for highest (p<0.05) total volatile fatty acids and trichloroacetic acid precipitable nitrogen in the strained rumen liquor. The digestible crude protein (DCP) was highest (p<0.05) in M. sativa followed by that in T. alexandrium. The total purine derivatives excreted in urine varied from 0.22-0.32 mmol/kg $W^{.75}/d$. The efficiency of microbial protein synthesis indicated that OM of straws of M. sativa and that of T. alexandrium was used more (p<0.05) efficiently. The microbial protein synthesized was highest in T. resupinatum, but statistically similar to other groups. The values for N-retention and apparent biological value were highest for L. perenne, though comparable with that of M. sativa and T. alexandrium. The available metabolizable energy (ME) was highest (p<0.05) in T. resupinatum followed by that in L. perenne and lowest in M. sativa. It was concluded that all the straws, supplemented with minerals and vitamin A, could be fed exclusively to adult ruminants with no adverse affect, as animals were able to maintain body weight (372${\pm}$20.1 kg).
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