Isolation and Characterization of Bacillus cereus Phage vB_BceP-DLc1 Reveals the Largest Member of the Φ29-Like Phages

Bacillus phage φ29 and its relatives have been considered as one of the most important model organisms for DNA replication, transcription, morphogenesis, DNA packaging studies, and nanotechnology applications. Here, we isolated and characterized a new member of the φ29-like phage, named Bacillus cereus phage vB_BceP-DLc1. This phage, with a unique inserted gene cluster, has the largest genome among known φ29-like phages. DLc1 can use the surface carbohydrate structures of the host cell as receptors and only infects the most related B. cereus strains, showing high host-specificity. The adsorption rate constant and life cycle of DLc1 under experimental conditions were also determined. Not only stable under temperatures below 55 °C and pH range from 5 to 11, the new phage also showed tolerance to high concentrations of NaCl, 75% ethanol, chloroform, and mechanical vortex, which is preferable for practical use in the food and pharmaceutical industries.

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Isolation and Characterization of A Bacillus Phage from Equine Carcass Disposal Site

Defence Life Science Journal ◽

10.14429/dlsj.4.13082 ◽

2018 ◽

Vol 4 (1) ◽

pp. 61-66

Author(s):

Taruna Anand ◽

Nitin Virmani ◽

Rajesh Kumar Vaid ◽

Bidhan Chandra Bera ◽

Umang Ahlawat ◽

...

Keyword(s):

Bacillus Cereus ◽

Disposal Site ◽

Host Bacterium ◽

Bacillus Sp ◽

Carcass Disposal ◽

Isolation And Characterization ◽

Bacillus Phage ◽

Group Member

Bacteriophages play an important role in bacterial control in natural niche however a little is known about Bacillus sp. phages prevailing in cadaver affected soils. In the current study, the Bacillus sp. phage was isolated from the equine cadaver disposal site and characterised to gain an insight into the issue of role of phages in biological dynamics of manure thus formed over years. Firstly, the host bacterium was isolated and identified as Bacillus cereus group member as assessed by phylogenetic analysis and secondly it’s corresponding phage from same soil sample was also enriched and characterised. The phage (VTCCBPA38) was found to belong to family Myoviridae and was active within the temperature range of 4°C - 45°C. As assessed by biological sensitivity by spot test, the phage was active against 6/19 (31.6 %) Bacilli tested including Bacillus cereus from goat mastitis. Thus the phagemay find potential use in biocontrol of diseases caused by Bacillus sp. Furthermore, this report is valuable as the first study for investigation of Bacillus sp. phage in carcass burial sites.

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Isolation and Characterization of Plasminogen Activator from Pig Leucocytes

Thrombosis and Haemostasis ◽

10.1055/s-0038-1649147 ◽

1974 ◽

Vol 31 (01) ◽

pp. 072-085 ◽

Cited By ~ 5

Author(s):

M Kopitar ◽

M Stegnar ◽

B Accetto ◽

D Lebez

Keyword(s):

Molecular Weight ◽

Plasminogen Activator ◽

Gel Electrophoresis ◽

Gel Filtration ◽

Ph Optimum ◽

Isolation And Characterization ◽

Ph Range ◽

Inhibition Studies ◽

Final Purification

SummaryPlasminogen activator was isolated from disrupted pig leucocytes by the aid of DEAE chromatography, gel filtration on Sephadex G-100 and final purification on CM cellulose, or by preparative gel electrophoresis.Isolated plasminogen activator corresponds No. 3 band of the starting sample of leucocyte cells (that is composed from 10 gel electrophoretic bands).pH optimum was found to be in pH range 8.0–8.5 and the highest pH stability is between pH range 5.0–8.0.Inhibition studies of isolated plasminogen activator were performed with EACA, AMCHA, PAMBA and Trasylol, using Anson and Astrup method. By Astrup method 100% inhibition was found with EACA and Trasylol and 30% with AMCHA. PAMBA gave 60% inhibition already at concentration 10–3 M/ml. Molecular weight of plasminogen activator was determined by gel filtration on Sephadex G-100. The value obtained from 4 different samples was found to be 28000–30500.

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Adsorption Studies on Magnetic Nanoparticles Functionalized with Silver to Remove Nitrates from Waters

Water ◽

10.3390/w13131757 ◽

2021 ◽

Vol 13 (13) ◽

pp. 1757

Author(s):

Yesica Vicente-Martínez ◽

Manuel Caravaca ◽

Antonio Soto-Meca ◽

Miguel Ángel Martín-Pereira ◽

María del Carmen García-Onsurbe

Keyword(s):

Magnetic Nanoparticles ◽

Nitrate Content ◽

Adsorption Efficiency ◽

Experimental Conditions ◽

Dependent Behavior ◽

Before And After ◽

Naoh Solution ◽

High Concentrations ◽

Interference Studies

This paper presents a novel procedure for the treatment of contaminated water with high concentrations of nitrates, which are considered as one of the main causes of the eutrophication phenomena. For this purpose, magnetic nanoparticles functionalized with silver (Fe3O4@AgNPs) were synthesized and used as an adsorbent of nitrates. Experimental conditions, including the pH, adsorbent and adsorbate dose, temperature and contact time, were analyzed to obtain the highest adsorption efficiency for different concentration of nitrates in water. A maximum removal efficiency of 100% was reached for 2, 5, 10 and 50 mg/L of nitrate at pH = 5, room temperature, and 50, 100, 250 and 500 µL of Fe3O4@AgNPs, respectively. The characterization of the adsorbent, before and after adsorption, was performed by energy dispersive X-ray spectroscopy, scanning electron microscopy, Brunauer-Emmett-Teller analysis and Fourier-transform infrared spectroscopy. Nitrates can be desorbed, and the adsorbent can be reused using 500 µL of NaOH solution 0.01 M, remaining unchanged for the first three cycles, and exhibiting 90% adsorption efficiency after three regenerations. A deep study on equilibrium isotherms reveals a pH-dependent behavior, characterized by Langmuir and Freundlich models at pH = 5 and pH = 1, respectively. Thermodynamic studies were consistent with physicochemical adsorption for all experiments but showed a change from endothermic to exothermic behavior as the temperature increases. Interference studies of other ions commonly present in water were carried out, enabling this procedure as very selective for nitrate ions. In addition, the method was applied to real samples of seawater, showing its ability to eliminate the total nitrate content in eutrophized waters.

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Screening a Strain of Klebsiella sp. O852 and the Optimization of Fermentation Conditions for Trans-Dihydrocarvone Production

Molecules ◽

10.3390/molecules26092432 ◽

2021 ◽

Vol 26 (9) ◽

pp. 2432

Author(s):

Li Chen ◽

Lu-Lu Zhang ◽

Jing-Nan Ren ◽

Xiao Li ◽

Gang Fan ◽

...

Keyword(s):

Ethanol Concentration ◽

Flavor Compounds ◽

Accession Number ◽

Reaction Parameters ◽

Isolation And Characterization ◽

Biotransformation Process ◽

Pharmaceutical Industries ◽

Final Ethanol Concentration ◽

Optimization Of Fermentation

Flavors and fragrances have high commercial value in the food, cosmetic, chemical and pharmaceutical industries. It is interesting to investigate the isolation and characterization of new microorganisms with the ability to produce flavor compounds. In this study, a new strain of Klebsiella sp. O852 (accession number CCTCC M2020509) was isolated from decayed navel orange (Citrus sinensis (L.) Osbeck), which was proved to be capable of converting limonene to trans-dihydrocarvone. Besides, the optimization of various reaction parameters to enhance the trans-dihydrocarvone production in shake flask was performed for Klebsiella sp. O852. The results showed that the yield of trans-dihydrocarvone reached up to 1 058 mg/L when Klebsiella sp. O852 was incubated using LB-M medium for 4 h at 36 °C and 150 rpm, and the biotransformation process was monitored for 36 h after adding 1680 mg/L limonene/ethanol (final ethanol concentration of 0.8% (v/v)). The content of trans-dihydrocarvone increased 16 times after optimization. This study provided a basis and reference for producing trans-dihydrocarvone by biotransformation.

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Incidence and characterisation of Bacillus cereus bacteriophages from Thua Nao, a Thai fermented soybean product

BioMolecular Concepts ◽

10.1515/bmc-2021-0009 ◽

2021 ◽

Vol 12 (1) ◽

pp. 85-93

Author(s):

Wallapat Phongtang ◽

Ekachai Chukeatirote

Keyword(s):

Bacillus Cereus ◽

Food Industry ◽

Pathogenic Bacteria ◽

Morphological Analysis ◽

Food Poisoning ◽

Fermented Soybean ◽

Foodborne Pathogenic Bacteria ◽

Ph Range ◽

Potential Use

Abstract Bacillus cereus is considered to be an important food poisoning agent causing diarrhea and vomiting. In this study, the occurrence of B. cereus bacteriophages in Thai fermented soybean products (Thua Nao) was studied using five B. cereus sensu lato indicator strains (four B. cereus strains and one B. thuringiensis strain). In a total of 26 Thua Nao samples, there were only two bacteriophages namely BaceFT01 and BaceCM02 exhibiting lytic activity against B. cereus. Morphological analysis revealed that these two bacteriophages belonged to the Myoviridae. Both phages were specific to B. cereus and not able to lyse other tested bacteria including B. licheniformis and B. subtilis. The two phages were able to survive in a pH range between 5 and 12. However, both phages were inactive either by treatment of 50°C for 2 h or exposure of UV for 2 h. It should be noted that both phages were chloroform-insensitive, however. This is the first report describing the presence of bacteriophages in Thua Nao products. The characterization of these two phages is expected to be useful in the food industry for an alternative strategy including the potential use of the phages as a biocontrol candidate against foodborne pathogenic bacteria.

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Characterization of a New Intracellular Alginate Lyase with Metal Ions-Tolerant and pH-Stable Properties

Marine Drugs ◽

10.3390/md18080416 ◽

2020 ◽

Vol 18 (8) ◽

pp. 416

Author(s):

Yan Ma ◽

Jie Li ◽

Xin-Yue Zhang ◽

Hao-Dong Ni ◽

Feng-Biao Wang ◽

...

Keyword(s):

Metal Ion ◽

Brown Seaweed ◽

Alginate Lyase ◽

Industrial Applications ◽

Alginate Oligosaccharides ◽

Potential Applications ◽

Pharmaceutical Industries ◽

Ph Range ◽

Alginate Lyases

Alginate lyases play an important role in alginate oligosaccharides (AOS) preparation and brown seaweed processing. Many extracellular alginate lyases have been characterized to develop efficient degradation tools needed for industrial applications. However, few studies focusing on intracellular alginate lyases have been conducted. In this work, a novel intracellular alkaline alginate lyase Alyw202 from Vibrio sp. W2 was cloned, expressed and characterized. Secretory expression was performed in a food-grade host, Yarrowia lipolytica. Recombinant Alyw202 with a molecular weight of approximately 38.3 kDa exhibited the highest activity at 45 °C and more than 60% of the activity in a broad pH range of 3.0 to 10.0. Furthermore, Alyw202 showed remarkable metal ion-tolerance, NaCl independence and the capacity of degrading alginate into oligosaccharides of DP2-DP4. Due to the unique pH-stable and high salt-tolerant properties, Alyw202 has potential applications in the food and pharmaceutical industries.

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Isolation and Characterization of a Lectin-like Protein (SBLP) from the Dried Roots of Scutellaria baicalensis (Lamiaceae)

Natural Product Communications ◽

10.1177/1934578x1801301232 ◽

2018 ◽

Vol 13 (12) ◽

pp. 1934578X1801301

Author(s):

Huiqin Wang ◽

Guanzhen Gao ◽

Lijing Ke ◽

Jianwu Zhou ◽

Pingfan Rao

Keyword(s):

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Scutellaria Baicalensis ◽

Dependent Manner ◽

Scutellaria Baicalensis Georgi ◽

Isolation And Characterization ◽

Pas Staining ◽

Ph Range ◽

Dose Dependent

A novel lectin-like protein with MW 63.2 kDa, designated as SBLP, has been isolated and characterized from the dried roots of Scutellaria baicalensis Georgi (Lamiaceae). SBLP was purified by ammonium sulfate precipitation and anion exchange chromatography. It is a glycoprotein according to a PAS staining assay and consisting of protein (86.0%) and sugar (14.0%). Its N-terminal amino acid sequence was determined as GSAVGFLY by Edman degradation. SBLP showed hemagglutinating activity against human and rooster erythrocytes, which were stable below 60°C and in the pH range of 4 −10. Furthermore, SBLP was found to be stimulated by Ca2+, Na+, Ba2+, Zn2+ ions, which suggested it was a metal-dependent lectin. SBLP inhibited the growth of Fusarium oxysporum f.sp. lycopersici and Alternaria eichhorniae in the a dose-dependent manner, and suppressed the proliferation of HepG2 tumor cells with an IC50 of 1.00 μM. This is the first report of a lectin from Radix Scutellariae.

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Isolation and Characterization of NaCl-Sensitive Mutants of Caulobacter crescentus

Applied and Environmental Microbiology ◽

10.1128/aem.69.6.3029-3035.2003 ◽

2003 ◽

Vol 69 (6) ◽

pp. 3029-3035 ◽

Cited By ~ 18

Author(s):

Luiz Fernando G. Zuleta ◽

ValÃ¯Â¿Â½ria C. S. Italiani ◽

Marilis V. Marques

Keyword(s):

Caulobacter Crescentus ◽

Osmotic Shock ◽

Promoter Regions ◽

Isolation And Characterization ◽

Lipopolysaccharide Biosynthesis ◽

Severe Reduction ◽

Mutant Strains ◽

High Concentrations ◽

Severe Growth

ABSTRACT An attempt to characterize Caulobacter crescentus genes important for the response to high concentrations of NaCl was initiated by the isolation of mutants defective in survival in the presence of 85 mM NaCl. A transposon Tn5 library was screened, and five strains which contained different genes disrupted by the transposon were isolated. Three of the mutants had the Tn5 in genes involved in lipopolysaccharide biosynthesis, one had the Tn5 in the nhaA gene, which encodes a Na+/H+ antiporter, and one had the Tn5 in the ppiD gene, which encodes a peptidyl-prolyl cis-trans isomerase. All the mutant strains showed severe growth arrest in the presence of 85 mM NaCl, but only the nhaA mutant showed decreased viability under these conditions. All the mutants except the nhaA mutant showed a slightly reduced viability in the presence of 40 mM KCl, but all the strains showed a more severe reduction in viability in the presence of 150 mM sucrose, suggesting that they are defective in responding to osmotic shock. The promoter regions of each disrupted gene were cloned in lacZ reporter vectors, and the pattern of expression in response to NaCl and sucrose was determined; this showed that both agents induced ppiD and nhaA gene expression but did not induce the other genes. Furthermore, the ppiD gene was not induced by heat shock, indicating that it does not belong to the σ32 regulon, as opposed to what was observed for its Escherichia coli homolog.

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