• Title/Summary/Keyword: osmotolerance

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Reduced Susceptibility of a Model Saccharomyces cerevisiae Biofilm to Osmotic Upshifts

  • Jirku Vlacimir;Jan Masak;Alena Cejkova
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.17-20
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    • 2001
  • Whole-cell attachment by covalent linkage, thereby simulating natural and specific attachments, improves the osmotolerance of Saccharomyces cerevisiae cells. The enhanced osmoresistance is correlated with a decrease in the intercellular concentration of trehalose and accompanied by membrane compositional changed. The results obtained indicate that yeast cell-support (physical) contact is sensed and responded to.

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FUNGAL EXTRACELLULAR POLYSACCHARIDES INVOLVED IN RECYCLING OF METABOLITES AND OSMOTOLERANCE OF PENICILLIUM FELLUTANUM : APPLICATION OF $^{13}$ C-NMR SPECTROSCOPY FOR THE STUDY ON FUNGAL PHYSIOLOGY AND METABOLISM

  • Park, Yong-Il;Gander, John.-E.
    • Proceedings of the Korean Society for Applied Microbiology Conference
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    • 2000.04a
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    • pp.208-213
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    • 2000
  • Penicillium fellutanum produces a phosphorylated, choline-containing extracellular peptido-polysaccharide, peptidophosphogalactomannan (pPxGM) (8). The $\^$13/C-methyl labeled pPxGM ([methyl-$\^$13/C]pPxGM) was prepared from the cultures supplemented with L-[methyl-$\^$13/C]methionine or [2-$\^$13/C]glycine and was used as a probe to monitor the fate of phosphocholine in this polymer. Addition of purified [methyl-$\^$l3/C]pPxGM to growing cultures in low phosphate medium resulted in the disappearance of [methyl-$\^$13/C]phosphocholine and -N,N'-dimethyl-phosphoethanolamine from the added [methyl-$\^$13/C]pPxGM. Two $\^$l3/C-methyl-enriched cytoplasmic solutes, choline-O-sulfate and glycine betaine, were found in mycelial extracts, suggesting that phosphocholine-containing extracellular pPxGM of P.fellutanum is a precursor of intracellular choline-O-sulfate and glycine betaine and thus of phosphatydilcholine (l0). $\^$13/C-Methyl-labeled cells grown in 3 M NaCl-containing medium showed 2.6- and 22-fold more accumulation of $\^$13/C-methyl labeled choline-O-sulfate and glycine betaine, respectively, originated from the extracellular [$\^$13/C-methyl]pPxGM than those grown without added NaCl. The results suggest that, in addition to glycerol and erythritol, glycine betaine and choline-O-sulfate and thus choline are also osmoprotectants and hence that pPxGM is involved in osmotolerance of this fungus (11). Taken collectively, the $\^$l3/C- and $\^$31/P-NMR analyses of cytosolic solute pools and structural modulation of extracellular pPxGM corresponding to environmental stimuli in P. fellutanum, provided evidence that pPxGM is involved in cellular choline metabolism, osmotolerance, and recycling of metabolites.

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Expression of Bacillus subtilis proBA Genes and Reduction of Feedback Inhibition of Proline Synthesis Increases Proline Production and Confers Osmotolerance in Transgenic Arabidopsis

  • Chen, Mingqing;Wei, Hongbo;Cao, JunWei;Liu, Ruijie;Wang, Youliang;Zheng, Congyi
    • BMB Reports
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    • v.40 no.3
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    • pp.396-403
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    • 2007
  • Proline accumulation has been shown to correlate with tolerance to drought and salt stresses in plants. We attempt to introduce the wild-type, mutant, and fusion proBA genes derived from Bacillus subtilis into Arabidopsis thaliana under the control of a strong promoter cauliflower mosaic virus 35S (CaMV35S). The transgenic plants produced higher level of free proline than control and the overproduction of proline resulted in the increased tolerance to osmotic stress in transgenic plants. Besides, the mutation in proBA genes, which were proved to lead $\alpha$-glutamyl kinase ($\alpha$-GK) reduces sensitivity to the end-product inhibition and the fusion of proB and proA also result in increasing proline production and confer osmotolerance in transgenic lines.

Osmotic-inducible Gene Expression using MudJ(Km.lac) Operon Fusion in Salmonella typhimurium (Salmonella typhimurium에서 MudJ(Km.lac) 오페론 융합을 이용한 삼투유도유전자의 발현)

  • 주성관;우영대;허연주;안정선;박용근
    • Korean Journal of Microbiology
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    • v.29 no.4
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    • pp.215-220
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    • 1991
  • MudJ(Km.lac) operon fusions were used in the identification of osmotic-inducible genetic(osi) loci in Salmonella typhimurium. Expression of osi::lacZ(osi5001, 5027) genes were dramatically induced 39-189 fold when the osmolarity was increased. Seven osm::lacZ genes were constituvely expressed under both low and high osmotic strength. The osi5001::lacZ fusion strains showed the enhanced osmotolerance and the reduced expression of the osi5001::lacZ in the presence of 1mM proline or betaine as osmoprotectants. Four osmotic inducible genetic loci were mapped into 36 (YK531), 44 (YK504), 57 (YK501) and 84 (YK528) map unit by testing the cotransduction frequency.

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Production of Mannitol by Lactobacillus sp. KY-107 (Lactobacillus sp. KY-107에 의한 Mannitol의 생산)

  • 윤종원;강선철류병호송승구
    • KSBB Journal
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    • v.11 no.3
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    • pp.374-379
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    • 1996
  • The production of extracellular mannitot by an efficient mannitol-producing bacterium, Lactobacillus sp. KY-107 was studied in shake flask culture using the modified MRS medium. Maximum mannitol production was obtained with fructose as the sole carbon source. Within 95 hours of incubation, a final concentration of 70g/L of mannitol from 100g/L fructose was obtained with an indicated yield of 86% based on fructose consumed. However, higher concentrations of fructose could not effectively be transformed to mannitol due to a lack of osmotolerance. The strain produced no other polyols such as glycerol and sorbitol as by-products. Yeast extract was best nitrogen source and high levels of inorganic phosphate up to 10g/L did not show any detrimental effect for mannitol formation. Manganese ion played important role in both cell growth and mannitol production. The optimum culture temperature and initial pH were $35^{\circ}C$ and 6-8, respectively.

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Proline Accumulation and P5CS ($\Delta^1$-pyrroline-5-carboxylate synthetase) Gene Expression in Response to Salt Stress in Zoysiagrasses

  • Lee, Dong-Joon;Hwang, Cheol-Ho
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.48 no.1
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    • pp.20-24
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    • 2003
  • Proline is known as an osmotrotectant to enhance tolerance against both salt and dehydration stresses. A P5CS ($\Delta^1$-pyrroline-5-carboxylate synthetase) plays a major role in regulation of synthesis of proline. An overexpression of the mothbean P5CS gene in transgenic tobacco plant increased the levels of proline and osmotolerance. In an attempt to look for the possibility to use content of proline as well as a level of P5CS gene expression as molecular markers for salt tolerance, the amounts of proline and transcript levels of P5CS were measured as functions of either concentration of NaCl or length of treatment period among different species of zoysiagrass. Hybridzoysia showed the highest level of proline ($329\mu\textrm{g}$/g.f.w.) among five different species of zoysiagrass at 250 mM NaCl in 24 hours. The level of P5CS transcript was also the highest in the hybridzoysia at 250 mM NaCl in 24 hours. The transcriptions of P5CS gene were induced at the rates of 1.2, 1.2, 1.8, and 1.8, upon treatment of 250 mM NaCl in Z. japonica, Z. matrella, Z. sinica and hybridzoysia respectively. Based on a correlation between the level of P5CS transcript and the proline content among different species of zoysiagrass, a comparative structural analysis of the gene for P5CS from either Z. sinica or hybridzoysia may lead to an understanding of mechanism for salt tolerance shown differently among zoysiagrasses.

Biological function of CpSlt2, an ortholog of the cell wall integrity (CWI) MAPK of Saccharomyces cerevisiae, in the chestnut blight fungus Cryphonectria parasitica

  • So, Kum-Kang;Ko, Yo-Han;Chun, Jeesun;Kim, Jung-Mi;Kim, Dae-Hyuk
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.11-11
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    • 2018
  • Cryphonectria parasitica, chestnut blight fungus, has a characteristic of decreasing pathogenicity when infected with Cryphonectria hypovirus 1. C. parasitica is known to be one of the most representative model systems used to observe the interaction between viruses, plants and fungi. The mitogen-activated protein kinase (MAPK) pathway, which is well conserved in various organisms ranging from yeast to humans, functions in relaying phosphorylation-dependent signals within MAPK cascades to diverse cellular functions involved in the regulation of pheromone, cell wall integrity, and osmotolerance in filamentous fungi. Several genes in the MAPK pathway were revealed to be regulated by hypovirus, or to be involved in pathogenicity in C. parasitica. Among these pathways, the CWI pathway has aroused interest because CpBck1, an ortholog of yeast Bck1 (a CWI MAPKKK), was previously reported to be involved in cell wall integrity and sectorization. Interestingly, sporadic sectorization was observed in the CpBck1 mutant and sectored phenotypes were stably inherited in the progeny that were successively transferred from sectored mycelia. In this study, we analyzed the biological function of CpSlt2, downstream gene of CpBck1, to confirm whether the sectorization phenomenon occurred in the specific single gene or cell wall integrity (CWI) pathway. As results, the CpSlt2-null mutant exhibited marked changes in colonial growth, near absence of conidiation and aerial hyphae, abnormal pigmentation, CWI-related phenotypic defects, and dramatically impaired virulence. As cultivation of the mutant strains progressed, the majority of the colonies showed sporadic sectorization and mycelia from the sectored area stably maintained the sectored phenotype. These results suggest that the unique sectorization is CWI pathway-specific, though the components in the same CWI pathway have common and specific functions.

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Characteristics of Mannitol Production by Leuconostoc sp. KY-002 (Leuconostoc sp. KY-002의 Mannitol 생산 특성)

  • 류병호;김동현윤종원
    • KSBB Journal
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    • v.11 no.6
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    • pp.636-641
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    • 1996
  • The production of extracellular mannitol by a new mannitol-producing bacterium, Leuconostoc sp. KY-002 was studied in shake flask cultures. The new isolate has a capability of utilizing fructose and sucrose for mannitol formation. Maximum mannitol production was obtained with fructose as the sole carbon source. Under the optimal culture conditions, within 70 hours of incubation, a final concentration of 26 g/L of mannitol from 50 g/L fructose was obtained with an indicated yield of 52% based on fructose consumed. However, higher concentrations of fructose ranging from 100 to 250 g/L could not effectively be transformed to mannitol due to a lack of osmotolerance. The strain produced no other polyols such as glycerol and sorbitol as by-products. Yeast extract was the best nitrogen source and high levels of inorganic phosphate up to 10 g/L promoted mannitol formation. Any mineral ions and salts did not play important role in both cell growth and mannitol production. Nicotinic acid enhanced mannitol production by 16%. The optimum culture temperature and initial pH were $35^{\circ}C$ and 6, respectively.

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Effect of Sigma Factor ${\sigma}^{B}$ on Biofilm Formation of Listeria monocytogenes in High Osmotic and Low Temperature Conditions (고삼투압 및 저온 조건에서 sigma factor ${\sigma}^{B}$가 Listeria monocytogenes biofilm 생성에 미치는 영향)

  • Park, Sang-Gyu;Park, Shin
    • Korean Journal of Food Science and Technology
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    • v.36 no.3
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    • pp.456-460
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    • 2004
  • Effects of sigma factor (${\sigma}^{B}$) on biofilm formation in Listeria monocytogenes 10403S and ${\sigma}^{B}-deficient$ sigB null mutant were studied under high osmotic and low temperature conditions. In brain heart infusion (BHI) medium containing 6% NaCl, wild type 10403S and ${\sigma}^{B}-deficient$sigB null mutant formed biofilms of $6.83{\pm}0.38\;and\;5.33{\pm}0.45\;log\;cfu/cm^{2}$, respectively. L. monocytogenes 10403S in BHI medium containing 6% NaCl formed 4.7 times larger biofilm than without NaCl. Culture of L. monocytogenes 10403S and sigB null mutant at $4^{\circ}C$ did not show any significant differences in biofilm formation. The results suggest biofilm formation is activated by ${\sigma}^{B}$ and NaCl, whereas not affected by low temperature stress in L. monocytogenes 10403S. More studies are necessary to determine biofilm formation mechanism in osmotolerant L. monocytogenes.