scholarly journals Exopolysaccharide Production and Ropy Phenotype Are Determined by Two Gene Clusters in Putative Probiotic Strain Lactobacillus paraplantarum BGCG11

2014 ◽  
Vol 81 (4) ◽  
pp. 1387-1396 ◽  
Author(s):  
Milica Zivkovic ◽  
Marija Miljkovic ◽  
Patricia Ruas-Madiedo ◽  
Ivana Strahinic ◽  
Maja Tolinacki ◽  
...  
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Gene Cluster ◽  
Probiotic Strain ◽  
Gene Clusters ◽  
Size Exclusion ◽  
High Molecular Weight Fraction ◽  
Transposase Gene ◽  
Content Type ◽  
Lactobacillus Paraplantarum

ABSTRACTLactobacillus paraplantarumBGCG11, a putative probiotic strain isolated from a soft, white, artisanal cheese, produces a high-molecular-weight heteropolysaccharide, exopolysaccharide (EPS)-CG11, responsible for the ropy phenotype and immunomodulatory activity of the strain. In this study, a 26.4-kb region originating from the pCG1 plasmid, previously shown to be responsible for the production of EPS-CG11 and a ropy phenotype, was cloned, sequenced, and functionally characterized. In this region 16 putative open reading frames (ORFs), encoding enzymes for the production of EPS-CG11, were organized in specific loci involved in the biosynthesis of the repeat unit, polymerization, export, regulation, and chain length determination. Interestingly, downstream of theepsgene cluster, a putative transposase gene was identified, followed by an additionalrfbgene cluster containing therfbACBDgenes, the ones most probably responsible for dTDP-l-rhamnose biosynthesis. The functional analysis showed that the production of the high-molecular-weight fraction of EPS-CG11 was absent in two knockout mutants, one in theepsand the other in therfbgene cluster, as confirmed by size exclusion chromatography analysis. Therefore, bothepsandrfbgenes clusters are prerequisites for the production of high-molecular-weight EPS-CG11 and for the ropy phenotype of strainL. paraplantarumBGCG11.

ACTH-like bioactivity and immunoactivity in fetal lamb pituitaries at 0.65 and 0.95 gestation

1996 ◽  
Vol 8 (1) ◽  
pp. 195 ◽  
Author(s):  
MA Fora ◽  
NK Valego ◽  
MO Lively ◽  
MI Castro ◽  
JC Rose
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Late Gestation ◽  
Size Exclusion ◽  
Molecular Forms ◽  
Low Molecular Weight ◽  
High Molecular Weight Fraction ◽  
Crude Extracts ◽  
Molecular Weight Peak ◽  
Ovine Fetal

We wished to determine if the concentration of bioactive ACTH-like activity increased during development and if there was heterogeneity in ovine fetal anterior pituitary ACTH activity as measured by bioassay and radioimmunoassay (RIA). We obtained anterior pituitaries from eight sheep fetuses (four at 0.65 and four at 0.95 gestation; term 145 +/- 5 days) and extracted and homogenized them in ice-cold 5N acetic acid, 0.3% phenylmethanesulfonyl fluoride (PMSF) and 0.2% BSA. Fractionation of each pituitary extract was performed by size-exclusion chromatography using Sepadex G-50. The ACTH-like immunoactivity (ALI) profile for each pituitary showed two well-defined peaks. One eluted with human ACTH1-39 and the other eluted with the high molecular weight fraction in the void volume. Four fractions from the first peak representing the high molecular weight forms of ACTH activity and four fractions from the second peak representing the low molecular weight forms of ACTH activity were pooled separately. These two pools were subjected to reverse-phase chromatography (RPC) on a C-8 column using a linear gradient of 70% acetonitrile in 0.8% trifluoroacetic acid over a 60 min period. Based upon the RIA, the high molecular weight forms of ACTH from the G-50 column were resolved into three main fractions, one eluting similar to the standard ACTH1-39 and the remaining two eluting after that. The low molecular weight forms of ACTH from the G-50 column were resolved into three peaks, before, with, and after the standard. We used collagenase-dispersed rat adrenal cells to test the ACTH-like bioactivity (ALB) of the crude extracts and of the different fractions obtained from the RPC of the high and low molecular weight material. The concentration of ACTH-like bioactivity in the crude extracts was similar at the two stages of gestation. However, there was a trend for the low molecular weight peak to have more peptide eluting with human ACTH1-39 and higher ratios of ALB/ALI than did the high molecular weight peak. These results suggest that multiple ACTH molecular forms with different ALB/ALI ratios are present in the ovine fetal pituitary and that there is no selective increase in ACTH1-39 concentration in the fetal pituitary in late gestation.

Studies on the Functionality of Newly Synthesized Fibrinogen after Treatment of Acute Myocardial Infarction with Streptokinase, Increase in the Rate of Fibrinopeptide Release

1993 ◽  
Vol 70 (06) ◽  
pp. 0978-0983 ◽  
Author(s):  
Edelmiro Regano ◽  
Virtudes Vila ◽  
Justo Aznar ◽  
Victoria Lacueva ◽  
Vicenta Martinez ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Acute Myocardial Infarction ◽  
Molecular Weight ◽  
Steady State ◽  
High Molecular Weight ◽  
Coronary Arteries ◽  
Risk Index ◽  
Weight Fraction ◽  
High Molecular Weight Fraction ◽  
Fibrinopeptide A

SummaryIn 15 patients with acute myocardial infarction who received 1,500,000 U of streptokinase, the gradual appearance of newly synthesized fibrinogen and the fibrinopeptide release during the first 35 h after SK treatment were evaluated. At 5 h the fibrinogen circulating in plasma was observed as the high molecular weight fraction (HMW-Fg). The concentration of HMW-Fg increased continuously, and at 20 h reached values higher than those obtained from normal plasma. HMW-Fg represented about 95% of the total fibrinogen during the first 35 h. The degree of phosphorylation of patient fibrinogen increased from 30% before treatment to 65% during the first 5 h, and then slowly declined to 50% at 35 h.The early rates of fibrinopeptide A (FPA) and phosphorylated fibrinopeptide A (FPAp) release are higher in patient fibrinogen than in isolated normal HMW-Fg and normal fibrinogen after thrombin addition. The early rate of fibrinopeptide B (FPB) release is the same for the three fibrinogen groups. However, the late rate of FPB release is higher in patient fibrinogen than in normal HMW-Fg and normal fibrinogen. Therefore, the newly synthesized fibrinogen clots faster than fibrinogen in the normal steady state.In two of the 15 patients who had occluded coronary arteries after SK treatment the HMW-Fg and FPAp levels increased as compared with the 13 patients who had patent coronary arteries.These results provide some support for the idea that an increased synthesis of fibrinogen in circulation may result in a procoagulant tendency. If this is so, the HMW-Fg and FPAp content may serve as a risk index for thrombosis.

scholarly journals ‘Tanned’ gelatin spheres and granules for exclusion chromatography

1968 ◽  
Vol 108 (4) ◽  
pp. 641-646 ◽  
Author(s):  
A. Polson ◽  
W. Katz
Keyword(s):  
Molecular Weight ◽  
Polyethylene Glycol ◽  
High Molecular Weight ◽  
Weight Fraction ◽  
High Flow ◽  
High Molecular Weight Fraction ◽  
Flow Rates ◽  
Protein Molecules ◽  
Exclusion Chromatography

1. The preparation of tanned gelatin spheres and granules from high-molecular-weight gelatin is described. This material is comparatively hard, giving high flow rates, is insoluble in water at temperatures between 0° and 100° and is resistant to digestion by trypsin and chymotrypsin. The high-molecular-weight fraction of gelatin was prepared by precipitation with polyethylene glycol, and the spheres and granules prepared from this fraction were hardened and insolubilized by tanning with either formalin or chromium salts or both. 2. The spheres and granules were used successfully for the separation of protein molecules and other protein-aceous materials ranging in molecular weight from 200 to greater than 6000000. This gel exclusion material has several properties superior to those of other products used for similar purposes. Further, it was noticed that the porosity of the spheres differed considerably from that of the granules.

scholarly journals Low-Molecular-Weight Metabolites Secreted by Paenibacillus larvae as Potential Virulence Factors of American Foulbrood

2014 ◽  
Vol 80 (8) ◽  
pp. 2484-2492 ◽  
Author(s):  
Hedwig-Annabell Schild ◽  
Sebastian W. Fuchs ◽  
Helge B. Bode ◽  
Bernd Grünewald
Keyword(s):  
Molecular Weight ◽  
Virulence Factors ◽  
Gene Clusters ◽  
Paenibacillus Larvae ◽  
Economic Losses ◽  
Low Molecular Weight ◽  
American Foulbrood ◽  
Toxic Activity ◽  
Content Type

ABSTRACTThe spore-forming bacteriumPaenibacillus larvaecauses a severe and highly infective bee disease, American foulbrood (AFB). Despite the large economic losses induced by AFB, the virulence factors produced byP. larvaeare as yet unknown. To identify such virulence factors, we experimentally infected young, susceptible larvae of the honeybee,Apis mellifera carnica, with differentP. larvaeisolates. Honeybee larvae were rearedin vitroin 24-well plates in the laboratory after isolation from the brood comb. We identified genotype-specific differences in the etiopathology of AFB between the tested isolates ofP. larvae, which were revealed by differences in the median lethal times. Furthermore, we confirmed that extracts ofP. larvaecultures contain low-molecular-weight compounds, which are toxic to honeybee larvae. Our data indicate thatP. larvaesecretes metabolites into the medium with a potent honeybee toxic activity pointing to a novel pathogenic factor(s) ofP. larvae. Genome mining ofP. larvaesubsp.larvaeBRL-230010 led to the identification of several biosynthesis gene clusters putatively involved in natural product biosynthesis, highlighting the potential ofP. larvaeto produce such compounds.

scholarly journals Overproduction of Ristomycin A by Activation of a Silent Gene Cluster in Amycolatopsis japonicum MG417-CF17

2014 ◽  
Vol 58 (10) ◽  
pp. 6185-6196 ◽  
Author(s):  
Marius Spohn ◽  
Norbert Kirchner ◽  
Andreas Kulik ◽  
Angelika Jochim ◽  
Felix Wolf ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gene Cluster ◽  
High Performance ◽  
Pathogenic Bacteria ◽  
Genome Mining ◽  
Gene Clusters ◽  
Von Willebrand Disease ◽  
Biosynthesis Gene Cluster ◽  
Content Type ◽  
Bioinformatic Tools

ABSTRACTThe emergence of antibiotic-resistant pathogenic bacteria within the last decades is one reason for the urgent need for new antibacterial agents. A strategy to discover new anti-infective compounds is the evaluation of the genetic capacity of secondary metabolite producers and the activation of cryptic gene clusters (genome mining). One genus known for its potential to synthesize medically important products isAmycolatopsis. However,Amycolatopsis japonicumdoes not produce an antibiotic under standard laboratory conditions. In contrast to mostAmycolatopsisstrains,A. japonicumis genetically tractable with different methods. In order to activate a possible silent glycopeptide cluster, we introduced a gene encoding the transcriptional activator of balhimycin biosynthesis, thebbrgene fromAmycolatopsis balhimycina(bbrAba), intoA. japonicum. This resulted in the production of an antibiotically active compound. Following whole-genome sequencing ofA. japonicum, 29 cryptic gene clusters were identified by genome mining. One of these gene clusters is a putative glycopeptide biosynthesis gene cluster. Using bioinformatic tools, ristomycin (syn. ristocetin), a type III glycopeptide, which has antibacterial activity and which is used for the diagnosis of von Willebrand disease and Bernard-Soulier syndrome, was deduced as a possible product of the gene cluster. Chemical analyses by high-performance liquid chromatography and mass spectrometry (HPLC-MS), tandem mass spectrometry (MS/MS), and nuclear magnetic resonance (NMR) spectroscopy confirmed thein silicoprediction that the recombinantA. japonicum/pRM4-bbrAbasynthesizes ristomycin A.

scholarly journals Plasmid-Borne Type E Neurotoxin Gene Clusters in Clostridium botulinum Strains

2013 ◽  
Vol 79 (12) ◽  
pp. 3856-3859 ◽  
Author(s):  
Zhen Zhang ◽  
Hannamari Hintsa ◽  
Ying Chen ◽  
Hannu Korkeala ◽  
Miia Lindström
Keyword(s):  
Gene Cluster ◽  
Gel Electrophoresis ◽  
Clostridium Botulinum ◽  
Southern Hybridization ◽  
Gene Clusters ◽  
Pulsed Field Gel Electrophoresis ◽  
Content Type ◽  
Pulsed Field ◽  
Large Plasmids

ABSTRACTA collection of 36Clostridium botulinumtype E strains was examined by pulsed-field gel electrophoresis (PFGE) and Southern hybridization with probes targeted tobotEandorfX1in the neurotoxin gene cluster. Three strains were found to contain neurotoxin subtype E1 gene clusters in large plasmids of about 146 kb in size.

Some Osmotic Measurements of the Molecular Weight of a High Molecular-Weight Fraction of Rubber Hydrocarbon

1955 ◽  
Vol 28 (2) ◽  
pp. 504-507
Author(s):  
G. W. Drake
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Average Molecular Weight ◽  
Weight Fraction ◽  
Considerable Proportion ◽  
Prolonged Storage ◽  
High Molecular Weight Fraction ◽  
The United Kingdom ◽  
Molecular Weight Fractions ◽  
Temperate Climates

Abstract Fractionation of the rubber hydrocarbon in temperate climates has usually resulted in high molecular-weight fractions, with a molecular weight of the order of one million. Bloomfield has shown that fresh latex contains a considerable proportion of hydrocarbon having an intrinsic viscosity (η) of 10 or over and, therefore, a molecular weight of well over 106. The fractionation technique used by Bloomfield in Malaya has now been applied by the writer to smoked sheet and to F rubber, working in the United Kingdom. No very high molecular-weight fractions were found in the smoked sheet, but the F rubber yielded a fraction of (η)=7.3 and a number average molecular weight 6×106, determined osmometrically. The average molecular weight of natural rubber when freshly prepared is probably well over a million, and includes a substantial portion having a molecular weight of several millions. By the time smoked sheet has reached temperate climates, the high molecular-weight portion has probably been converted to gel. F rubber, presumably because of its different method of preparation, retains the major part of its high molecular-weight material during prolonged storage.

scholarly journals Important Late-Stage Symbiotic Role of theSinorhizobium melilotiExopolysaccharide Succinoglycan

2018 ◽  
Vol 200 (13) ◽  
pp. e00665-17 ◽  
Author(s):  
Markus F. F. Arnold ◽  
Jon Penterman ◽  
Mohammed Shabab ◽  
Esther J. Chen ◽  
Graham C. Walker
Keyword(s):  
Molecular Weight ◽  
High Molecular Weight ◽  
Late Stage ◽  
Sinorhizobium Meliloti ◽  
Antimicrobial Action ◽  
Protective Function ◽  
Content Type ◽  
Early Stages ◽  
Symbiotic Interactions ◽  
Legume Symbiosis

ABSTRACTSinorhizobium melilotienters into beneficial symbiotic interactions withMedicagospecies of legumes. Bacterial exopolysaccharides play critical signaling roles in infection thread initiation and growth during the early stages of root nodule formation. After endocytosis ofS. melilotiby plant cells in the developing nodule, plant-derived nodule-specific cysteine-rich (NCR) peptides mediate terminal differentiation of the bacteria into nitrogen-fixing bacteroids. Previous transcriptional studies showed that the intensively studied cationic peptide NCR247 induces expression of theexogenes that encode the proteins required for succinoglycan biosynthesis. In addition, genetic studies have shown that someexomutants exhibit increased sensitivity to the antimicrobial action of NCR247. Therefore, we investigated whether the symbiotically activeS. melilotiexopolysaccharide succinoglycan can protectS. melilotiagainst the antimicrobial activity of NCR247. We discovered that high-molecular-weight forms of succinoglycan have the ability to protectS. melilotifrom the antimicrobial action of the NCR247 peptide but low-molecular-weight forms of wild-type succinoglycan do not. The protective function of high-molecular-weight succinoglycan occurs via direct molecular interactions between anionic succinoglycan and the cationic NCR247 peptide, but this interaction is not chiral. Taken together, our observations suggest thatS. melilotiexopolysaccharides not only may be critical during early stages of nodule invasion but also are upregulated at a late stage of symbiosis to protect bacteria against the bactericidal action of cationic NCR peptides. Our findings represent an important step forward in fully understanding the complete set of exopolysaccharide functions during legume symbiosis.IMPORTANCESymbiotic interactions between rhizobia and legumes are economically important for global food production. The legume symbiosis also is a major part of the global nitrogen cycle and is an ideal model system to study host-microbe interactions. Signaling between legumes and rhizobia is essential to establish symbiosis, and understanding these signals is a major goal in the field. Exopolysaccharides are important in the symbiotic context because they are essential signaling molecules during early-stage symbiosis. In this study, we provide evidence suggesting that theSinorhizobium melilotiexopolysaccharide succinoglycan also protects the bacteria against the antimicrobial action of essential late-stage symbiosis plant peptides.

SiO2 modified graphene oxide hybrids for improving fretting wear performance of ultra-high molecular weight polyethylene

2021 ◽  
Vol ahead-of-print (ahead-of-print) ◽  
Author(s):  
Xiaocui Xin ◽  
Yunxia Wang ◽  
Zhaojie Meng ◽  
Fengyuan Yan
Keyword(s):  
Molecular Weight ◽  
Wear Resistance ◽  
High Molecular Weight ◽  
Design Methodology ◽  
Fretting Wear ◽  
Wear Performance ◽  
Content Type ◽  
Modified Graphene Oxide ◽  
Modified Graphene ◽  
Ultra High Molecular Weight

Purpose The purpose of this paper is to investigate the fretting wear performance of ultra-high-molecular-weight-polyethene (UHMWPE) with addition of GO and SiO2. Design/methodology/approach In this study, GO were synthesized and SiO2 nanoparticles were grafted onto GO. The effect of nanofiller on fretting wear performance of UHMWPE was investigated. Findings The results indicated that GO was successfully synthesized and SiO2 nanoparticles successfully grafted onto GO. Incorporation of GS was beneficial for the reduction in friction and the improvement in wear resistance of UHMWPE. GO was beneficial for reducing friction coefficient, while SiO2 was good for improving wear resistance. There existed a tribological synergistic effect between GO nanosheet and SiO2 nanoparticles. Research limitations/implications The hybrids of GS were promising nanofiller for improving the fretting wear performance of UHMWPE. Originality/value The main originality of the research is to reveal the effect of GO and SiO2 nanoparticles on fretting behavior of UHMWPE. The result indicated hybrids of GS were promising nanofiller for improving the fretting wear performance of UHMWPE.

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