Paenibacillus Research Articles

In Vitro Antimicrobial Activity Screening of Bacteria Endophytic to Ethnomedicinal Plant Rauvolfia serpentina (L.) Benth. ex. Kurz

Introduction: Microbial endophytes colonizing internal tissues of living plants provide benefits to their host by promoting plant growth and protection against microbial infectious through the production of wide ranges of metabolites. Members of such endophytic community harbouring medicinally important plants are known to synthesize several antimicrobial compounds. The present study aims to explore bacterial endophytes of ethnomedicinal plant Rauvolfia serpentina (Apocynaceae) for producing novel antimicrobial metabolites of pharmaceutical and biotechnological importance. Materials and Methods: The culturable bacterial endophytic diversity of R. serpentina (L.) Benth. ex. Kurz. has been screened for producing antimicrobial compounds following cross-streak and agar well diffusion assay methods against several test microbial strains. The bioactive compound was isolated and partially purified from the cell-free culture filtrate following chromatographic methods. Results: The endophytes revealed low colonization frequency and isolation rates in the root and stem respectively. In vitro antimicrobial screening of 12 phenotypically distinguishable endophytes resulted in the selection of a potent antimicrobial isolate RAU 305 identified as Pseudomonas aeruginosa RAU 305 (Genbank accession number KR816098). Cell-free culture filtrate of RAU 305 showed broad spectrum of antimicrobial activity by inhibiting Paenibacillus, Micrococcus, Arthrobacter, Rhodobacter, Mycobacterium, Bacillus, Escherichia, Staphylococcus, Klebsiella, Aspergillus, Colletotrichum and Pythium. The antimicrobially active component extracted in butanol and chloroform was partially purified by column chromatography followed by a preparative thin layer chromatography and the homogeneity of the compound was confirmed using different solvent systems. Conclusions: More detailed characterization and identification of the active component is essential to explore the metabolic potential of this endophytic bacterium in future.

Erratum to Cytotoxic Effects and Production of Cytokines Induced by the Endophytic Paenibacillus polymyxa RNC-D In Vitro.

CORRIGENDUM Corrigendum to Manuscript entitled Cytotoxic Effects and Production of Cytokines Induced by the Endophytic Paenibacillus polymyxa RNC-D In Vitro by Débora M. Neris1,3,*, Genoveva L.F. Luna2,3, Joice M. de Almeida Rodolpho1,3, Ana C. Urbaczek4, Ricardo de Oliveira Correia3, Karina A. Feitosa1,3, Nadja F.G. Serrano2,3, Cristina P. de Sousa2,3 and Fernanda de Freitas Anibal1,2,3. Curr. Pharm. Biotechnol., 2017; 18(9), 758-768. I hereby informed that all the authors agree with the inclusion of the name of the funding agency (FAPESP - Fundação de Amparo à Pesquisa do Estado de São Paulo) and the process number (2018/12846-5) to the acknowledgements section of the manuscript.

Paenialvin A-D, four peptide antibiotics produced by Paenibacillus alvei DSM 29.

Four peptide antibiotics, named paenialvin A-D, were isolated from Paenibacillus alvei DSM 29. Mass spectrum analysis determined the molecular masses of paenialvin A-D to be 1891, 1875, 1877, and 1923 Da, respectively. Tandem mass spectra and nuclear magnetic resonance (NMR) were used to elucidate their chemical structures. Paenialvin A-D showed antimicrobial activity against most strains that were tested, including methicillin-resistant Staphalococcus aureus, Staphylococcus aureus, Bacillus subtilis, Loktanella hongkongensis, Escherichia coli, and Pseudomonas aeruginosa. In particular, the minimum inhibitory concentration of paenialvins against Staphalococcus aureus reached 0.8-3.2 μg/mL. Although they were cytotoxic against HeLa cells at a concentration of 50 μg/mL, the lack of hemolysis by paenialvins confirmed that they are potential candidates for anti-tumor drugs.

Effect of Carbon Source in the Culture Medium on Composition and Sorption Properties of Extracellular Polysaccharides of Paenibacillus polymyxa

Effect of Carbon Source in the Culture Medium on Composition and Sorption Properties of Extracellular Polysaccharides of Paenibacillus polymyxa

Medium optimization for nitrogen fixer Paenibacillus sp. 1-49.

Paenibacillus sp. 1-49 is a nitrogen-fixing bacterium that has the potential as a fertilizer. However, it grows poorly (OD600≤1) in both mineral medium and rich medium. To achieve high-yield biomass, we optimized fermentation medium for Paenibacillus sp. 1-49. Plackett-Burman Design and Central Composite Design in response surface methodology were used to optimize medium composition. The optimal fermentation medium contained:(per liter) 36.22 g Sucrose, 5.31 g Tryptone, 10.92 g Yeast Extract, 0.51 g MgSO4, 3.5 g NaCl, 0.02 g Na2MoO4 and 0.02 g FeSO4. The maximum OD600 of 10.280±0.009 was obtained in shake flask fermentation, reached 94.6% of the predicted value. We succeeded in using a response surface methodology to optimize the fermentation medium for Paenibacillus sp. 1-49. Our results will be useful for largescale fermentation of this strain and other Paenibacillus spp.

Solution structure of chitosan-binding module 2 derived from chitosanase/glucanase from Paenibacillus sp. IK-5

Solution structure of chitosan-binding module 2 derived from chitosanase/glucanase from Paenibacillus sp. IK-5

Intestinal microbiota of ornamental fish Carassius auratus

Intestinal microbiota is an essential component for any organism life, because it directly affects nutrient assimilation, growth and health processes. However, in many animals and in case of fish is unknown how Intestinal microbiota is formed, which species are dominant and which one has capacity probiotic for use in aquaculture. The goal of this study was to establish the bacterial load that dominates the back intestine of Carassius auratus. A batch of 200 healthyjuvenilefishes were obtained from an ornamental fish farm in Mexico City. The fish were maintained in two culture beakers of 100 L during 15 day to acclimatization to ensure any sign of injury disease. To extract the back of gastrointestinal tract, which was rinse several times with distilled water for food and feces residues elimination. The sample was inoculated in 9 mL of sterile saline solution and from this, three dilutions in a 1:10 relation were made, inoculating 0.1 mL from each dilution in agar plates MRS, BHI and TCBS, and were incubated at 27oC for 24 hours. Subsequently a colony forming units (CFU mL-1) count was made, with the help of a Quebec counter type. Colonies were purified through successive inoculations. The molecular identification was made out by sequencing the gene RNAr 16S using Wizard Genomic. Molecular identification showed that the back region of the intestinal tract of C. auratus was dominated by phylum Proteobacteria and Firmicutes represented by the genus Bacillus , Vibrio , Vagococcus , Brevibacillus , Aeromonas , Pseudomonas , Shewanella , Enterococcus , Paenibacillus and Morganella. In relation to bacterial abundance by specie it was established that Bacillus sp. and Pseudomonas stutzeri were the most abundant reaching CFU mL -1 counts of 210 and 167 respectively, followed by Paenibacillus lactis with 103 CFU mL -1 and Bacillus cereus with 100 CFU mL -1 while Enterococcus eurekensis was the least abundant with an average value of 6 CFU mL -1 .

Low-Voltage Producing Microbial Fuel Cell Constructs Using Biofilm-Forming Marine Bacteria

Biofilm-forming marine bacterial isolates Paenibacillus lautus NE3B01, Pseudomonas mendocina NR802, Stenotrophomonas acidaminiphila NCW702 and Pseudomonas pseudoalcaligenes NP103 in microbial fuel cell (MFC) were investigated for low-voltage power generation. Biofilm formation by the isolates was evaluated by glass tube assay, microtitre plate assay and fluorescence microscopy. A dual chamber MFC of 2 litre capacity was constructed for low-voltage power generation and current output. Two chambers were internally connected by salt bridge and externally the circuit was connected with copper wires which were joined to the electrodes at the two ends and to the multimeter. Maximum current was generated when the salt bridge was constructed using 1 M KCl for all the four bacterial isolates. With Paenibacillus lautus NE3B01, a maximum voltage of 727.5 ± 13.4 mV in 6 h with 7 g/l of glucose as the sole source of carbon was recorded. However, Pseudomonas mendocina NR802 MFC was the most stable in terms of potential generation among all the isolates used for MFC studies. The experimental data for current and voltage showed that the biofilm-forming marine bacterial isolates are useful in MFC technology.

Enhanced growth of cabbage seedlings by a <i>Paenibacillus</i> isolate in the presence of <i>Xanthomonas campestris</i> pv <i>campestris</i>

Paenibacillus spp are rhizobacteria that can promote plant growth through a range of mechanisms A New Zealand isolate of Paenibacillus P16 has reduced the incidence of black rot caused by Xanthamonas campestris pv campestris (Xcc) in brassicas To investigate if this response was provided through plant growth promotion isolate P16 was coapplied with Xcc as a seed treatment In the presence of Xcc P16treated seedlings had significantly greater root length leaf area and root and shoot dry weight compared to the positive control (Xcc alone) There were no significant differences in plant growth parameters between P16treated seedlings in the absence of the pathogen and the negative control (seeds without Xcc or P16) Isolate P16 enabled plants to survive and grow normally by preventing disease development; the mechanism of disease suppression requires further investigation

Occurrence and characterization of Paenibacillus sp. isolated from rabbits.

Occurrence and characterization of Paenibacillus sp. isolated from rabbits.

Intestine-specific expression of the β-glucanase in mice

Guan, L.-Z., Xi, Q.-Y., Sun, Y.-P., Wang, J.-L., Zhou, J.-Y., Shu, G., Jiang, Q.-Y. and Zhang, Y.-L. 2014. Intestine-specific expression of the β-glucanase in mice. Can. J. Anim. Sci. 94: 287-293. The β-glucanase gene (GLU, from Paenibacillus polymyxa CP7) was cloned into a specific expression plasmid (MUC2-GLU-LV). Transgenic mice were prepared by microinjection. Polymerase chain reaction and Southern blot analysis of genomic DNA extracted from the tail tissue of transgenic mice showed that the mice carried the β-glucanase gene. Northern blot analysis indicated that β-glucanase was specifically expressed in the intestine of the transgenic mice. The β-glucanase activity in the intestinal contents was found to be 1.23±0.32 U mL-1. The crude protein, crude fat digestibility of transgenic mice were increased by 9.32 and 5.09% (P<0.05), respectively, compared with that of the non-transgenic mice, while moisture in feces was reduced by 12.16% (P<0.05). These results suggest that the expression of β-glucanase i...

Preliminary studies regarding antioxidant and antimicrobial capacity for different types of Romanian honeys.

Different types of Romanian honeys were evaluated for their antioxidant and antibacterial activity. Total polyphenols, total fl avonoids and two colorimetric methods for radical scavenging activity and the total antioxidant power showed different amounts of biologically active compounds in tested honey samples. Antibacterial ac tivity was tested against Escherichia coli and Paenibacillus larvae , the first a model organism for bacteria and other a pathogenic bacterium causing American foul brood disease in Apis melliefra . Heather and honeydew honey had the highest content of total polyphenols (127.35 and 123.28 mgGAE/100 g and), while sunflower honey together with honeydew honey show the highest content of total fl avonoids (64.25 and 65.98 mgQE/100 g honey). The most effective in antibacterial activity for P.larvae was found in sunflower honey whereas multifloral honey showed highest activity against E.coli .

Isolation and characterization of heavy metal removing bacterial bioflocculants

Thirteen metal-tolerant bacteria capable of producing metal-removing bioflocculants were isolated from an industrial effluent sample. Pseudomonas sp. was found to be the pre-dominant species among the isolates (8 out of 13), followed by Herbaspirillium spp. (4) and one Paenibacillus sp. The flocculating activity of bioflocculants produced by these microorganisms was assayed using the kaolin clay. The heavy-metal-removal efficiency was determined using atomic absorption spectrometer before and after mixing the bioflocculant with the heavy metal solutions. Bioflocculants exhibit different flocculating abilities of removing kaolin clay in the presence of different heavy metals. Bioflocculants produced by Pseudomonas sp. CH9 possessed the highest flocculating activity (1.8) compared to the remaining bioflocculants. The flocculating activities of CH11 and CH13increased to 0.95 and 0.87 in the presence of Pb2+ and to 0.89 and 0.98 in the presence of Hg2+ respectively from 0.015 in the presence of Ca2+ in the standard kaolin clay assay. Up to 90% of Pb2+ was removed by Pseudomonas sp. CH8 bioflocculants. Seventy-eight percent of Hg2+ and 66% of Cd2+ was removed by Pseudomonas sp. CH6 andHerbaspirillium sp. CH13 bioflocculants respectively. Most of the bioflocculants demonstrated a higher percentage of heavy-metal removal at low concentrations. This study demonstrates that microbial bioflocculants have potential to be used as an alternative bioremedial tool for industrial effluents and wastewater treatments which are co-contaminated with heavy metals. Key words: Bioflocculant; heavy metals, Pseudomonas sp., Herbaspirillium sp.,Paenibacillus sp., Industrial effluent.

Optimization of culture conditions for thermostable chitinase production by Paenibacillus sp. D1

A hyper-chitinase producing isolate Paenibacillus sp. D1 was obtained from common effluent treatment plant of seafood industries at Veraval (Gujarat, India). The isolate exhibited chitinase production over a wide temperature (25 - 45°C) and pH (6 - 9) range with maxima at 30°C in medium with initial pH 7.0. The crude chitinase had activity in broad pH (4 - 10) and temperature (30 - 60°C) range with optima at pH 5.0 and 50°C, respectively. The enzyme was highly thermostable with t1/2 of 36 to 60 h at 40 and 45°C. Crab shell chitin, urea and K2HPO4were identified as best carbon, nitrogen and phosphorous sources influencing chitinase production by Paenibacillus sp. D1. Addition of tween 80 and FeCl3 enhanced the chitinase production by 1.44 and 1.33 fold, respectively. Identification of essential nutrients affecting chitinase production by Paenibacillus sp. D1 would help to formulate a suitable medium for its production. Moreover, low cost chitin from crab shells can be used as carbon source for thermostable chitinase production by the isolate for industrial and agricultural applications. Key words: Paenibacillus, optimization, thermostable, chitinase.

Cloning and characterization of a novel mannanase from Paenibacillus sp. BME-14.

A mannanase gene (man26B) was obtained from a sea bacterium, Paenibacillus sp. BME-14, through the constructed genomic library and inverse PCR. The gene of man26B had an open reading frame of 1,428 bp that encoded a peptide of 475- amino acid residues with a calculated molecular mass of 53 kDa. Man26B possessed two domains, a carbohydrate binding module (CBM) belonging to family 6 and a family 26 catalytic domain (CD) of glycosyl hydrolases, which showed the highest homology to Cel44C of P. polymyxa (60% identity). The optimum pH and temperature for enzymatic activity of Man26B were 4.5 and 60 degrees C, respectively. The activity of Man26B was not affected by Mg(2+) and Co(2+), but was inhibited by Hg(2+), Ca(2+), Cu(2+), Mn(2+), K(+), Na(+), and beta-mercaptoethanol, and slightly enhanced by Pb(2+) and Zn(2+). EDTA did not affect the activity of Man26B, which indicates that it does not require divalent ions to function. Man26B showed a high specific activity for LBG and konjac glucomannan, with K(m), V(max), and k(cat) values of 3.80 mg/ml, 91.70 micromol/min/mg protein, and 77.08/s, respectively, being observed when LBG was the substrate. Furthermore, deletion of the CBM6 domain increased the enzyme stability while enabling it to retain 80% and 60% of its initial activity after treatment at 80 degrees C and 90 degrees C for 30 min, respectively. This finding will be useful in industrial applications of Man26B, because of the harsh circumstances associated with such processes.

Cloning, sequencing, and expression of the gene encoding a multidomain endo-beta-1,4-xylanase from Paenibacillus curdlanolyticus B-6, and characterization of the recombinant enzyme.

The nucleotide sequence of the Paenibacillus curdlanolyticus B-6 xyn10A gene, encoding a xylanase Xyn10A, consists of 3,828 nucleotides encoding a protein of 1,276 amino acids with a predicted molecular mass of 142,726 Da. Sequence analysis indicated that Xyn10A is a multidomain enzyme comprising nine domains in the following order: three family 22 carbohydrate-binding modules (CBMs), a family 10 catalytic domain of glycosyl hydrolases (xylanase), a family 9 CBM, a glycine-rich region, and three surface layer homology (SLH) domains. Xyn10A was purified from a recombinant Escherichia coli by a single step of affinity purification on cellulose. It could effectively hydrolyze agricultural wastes and pure insoluble xylans, especially low substituted insoluble xylan. The hydrolysis products were a series of short-chain xylooligosaccharides, indicating that the purified enzyme was an endo-beta-1,4-xylanase. Xyn10A bound to various insoluble polysaccharides including Avicel, alpha-cellulose, insoluble birchwood and oat spelt xylans, chitin, and starches, and the cell wall fragments of P. curdlanolyticus B-6, indicating that both the CBM and the SLH domains are fully functioning in the Xyn10A. Removal of the CBMs from Xyn10A strongly reduced the ability of plant cell wall hydrolysis. These results suggested that the CBMs of Xyn10A play an important role in the hydrolysis of plant cell walls.

LABORATORY DIAGNOSIS OPTIMIZING TESTS IN AMERICAN FULLBROAD

We examined 24 honey samples with sapling, and 4 of them(16,67%) contained Paenibacillus larvae. Bacteriological exam allowed us to isolate P. larvae after 3-5 days of inoculation period at 37oC. On solid medium (glucose agar with 1‰ yolk of an egg 1/5 and BHI agar with 0,2 mg/l thiamine). In case of lichid growth, we isolated P. larvae after two days. The study of biochemical properties of P. larvae strains isolated from the four samples on API 20E galery confirmed that this belong to Paenibacillus larvae species subsp. larvae. Serological exam, trought VITA AFB Diagnostic Kit established the quick diagnostic (aproximatly 10 minutes), at the four samples witch were positive t hrought the classical bacteriological techniques, w ith new perspectives in optimising the diagnostics in Ameri can foulbrood.

OPTIMIZATION OF HPLC METHOD FOR QUANTIFYING TETRACYCLINE RESIDUE IN HONEY

Tetracyclines are used for the treatment or prevention of American and European fouldbrood in bee colonies which are caused especially by two species of bacteria: Paenibacillus larvae larvae and Melissococcus pluton, respectively. In the present study a method for determination of residual tetracyclines in honey has been optimized and validated. It was used the reversed-phase high performance liquid chromatography (HPLC) with UV detection. Tetracyclines were extracted from honey with 0.01M sodium succinate buffer. For the clean up step, solid phase extraction using metal chelate resin was applied. Tetracyclines were eluted with buffer containing EDTA. Linearity, precision, recovery and sensitivity were satisfactory. The limit of detection was established at 5 µg/kg

Optimization of cultivation conditions for the production of chitinase from Paenibacillus sp CHE-N1

A bacterial strain capable of secreting high levels of chitinase was isolated from chitin-enriched soil and identified as Paenibacillus sp. CHE-N1. This strain, which shows high potential as an antagonist against fungi, was grown in a chitin-containing medium to evaluate its ability to produce chitinase. The statistical design method was used to determine the optimal chitinase-producing conditions. Factors, including carbon sources and nitrogen sources in the medium compositions, initial pH, and the temperature of fermentation, were investigated. The obtained optimal conditions were: CSCP 55.4g/L, peptone 1.8g/L, and (NH4)2SO4 1.6g/L, with an optimal initial pH and temperature of 8.5 and 34.3℃, respectively. Under these conditions, the optimal volumetric enzyme activity could reach 8.1U/mL, about a 4-fold increase as compared to results under basic conditions.

Vertical distribution of phosphorus and P-dissolving/decomposing bacteria in the sediment of Guanting reservoir

In this study, we determined pH, phosphorus content and the number of P-dis-solving/decomposing bacteria in relation to the depth in the sediment of Guanting reservoir in Beijing. The pH value was slightly increased from 8.0 in the surface to 8.5 in the bottle (69 cm in depth) of sediment. The highest P content, 1269 mg/kg, was detected in the 35 cm layer, followed by the 5 cm (993 mg/kg) and 69 cm (580 mg/kg) layers. The number of inorganic P-dissolving bacteria varied from 6×102 to 8×104 and the organic P-discomposing bacteria were from 1.9×103 to 6.3×104 per gram sediment in different depths, which were counted under 28℃and 20℃×pH7.5 and 8.5. The number of P-dissolving/discomposing bacteria was directly correlated to the P content in each layer of sediments. The analysis of P-dissolving/decomposing ability of bacteria showed that some of these bacteria were also capable of accumulating P in-tracellularly. The intracellular P-accumulation was more efficient at lower temperature; in contrast, the activity of P-dissolving/decomposing was stronger at higher temperature. So the content of dissolved P in water body, or quality of water, could be affected by the change of temperature via the regulation of bacterial activity. On the basis of 16S rDNA sequencing, the 13 efficient P-dissolving/decomposing bacteria were identified as Bacillus spp., Bacterium sp., Microbacterium sp., Paenibacillus sp. and Pseudomonas sp.