scholarly journals Cloning and Heterologous Expression of Plantaricin ZJ5, a Novel Bacteriocin from Lactobacillus plantarum ZJ5, in Escherichia coli

2018 ◽  
Vol 13 (12) ◽  
pp. 1934578X1801301
Author(s):  
Dafeng Song ◽  
Ping Li ◽  
Qing Gu
Keyword(s):  
Escherichia Coli ◽  
Fusion Protein ◽  
Lactobacillus Plantarum ◽  
Micrococcus Luteus ◽  
Biologically Active ◽  
Biological Method ◽  
Lac Operon ◽  
E Coli ◽  
First Time ◽  
Soluble Material

A novel bacteriocin, plantaricin ZJ5 (PZJ5) was yielded from Lactobacillus plantarum ZJ5, cloned, and produced in Escherichia coli BL21 (DE3) pLys. The PZJ5 structural gene was fused with a Trx tag, and cloned into the pET32a plasmid under the control of the inducible lac operon. Induction was performed with isopropyl-β-D-thiogalactopyranoside (IPTG), with subsequent overexpression of the fusion protein, followed by purification to homogeneity via His affinity chromatography. Recombinant E. coli produced greater quantities of PZJ5 than L. plantarum ZJ5, and PZJ5 in E. coli was expressed in the form of soluble material. Biologically active PZJ5 was recovered by cleaving the purified fusion protein using enterokinase. The released PZJ5 demonstrated antibacterial activity against Micrococcus luteus. In this study, an inexpensive biological method using a Trx fusion system was presented, and for the first time, bacteriocin PZJ5 was expressed and purified in E. coli.

scholarly journals Characterization of a Shiga Toxin 2e-Converting Bacteriophage from an Escherichia coli Strain of Human Origin

2000 ◽  
Vol 68 (9) ◽  
pp. 4850-4855 ◽  
Author(s):  
Maite Muniesa ◽  
Jürgen Recktenwald ◽  
Martina Bielaszewska ◽  
Helge Karch ◽  
Herbert Schmidt
Keyword(s):  
Escherichia Coli ◽  
Shiga Toxin ◽  
Laboratory Strain ◽  
Coli Strain ◽  
Biologically Active ◽  
Trna Genes ◽  
Dependent Manner ◽  
E Coli ◽  
Lysis Cassette ◽  
First Time

ABSTRACT An infectious Shiga toxin (Stx) 2e-converting bacteriophage (φP27) was isolated from Stx2e-producing Escherichia coliONT:H− isolate 2771/97 originating from a patient with diarrhea. The phage could be transduced to E. colilaboratory strain DH5α, and we could show that lysogens were able to produce biologically active toxin in a recA-dependent manner. By DNA sequence analysis of a 6,388-bp HindIII restriction fragment of φP27, we demonstrated that thestx 2e gene was located directly downstream ofileZ and argO tRNA genes. Although no analogue of an antiterminator Q encoding gene was present on this fragment, a lysis cassette comprising two holin genes which are related to the holin genes of Pseudomonas aeruginosa phage φCTX and a gene homologous to the endolysin gene gp19 of phage PS3 were detected. The results of our study demonstrated for the first time that Stx2e can be encoded in the genome of an infectious bacteriophage.

EFECTO INHIBITORIO DE EXTRACTOS DE Vitex mollis Kunth CONTRA BACTERIAS Y ESPECIES DE Fusarium DE IMPORTANCIA HUMANA Y AGRÍCOLA

2018 ◽  
Vol 41 (4) ◽  
pp. 353-363
Author(s):  
Alberto J. Valencia-Botin ◽  
Melesio Gutiérrez-Lomelí ◽  
Juan A. Morales-Del-Río ◽  
Pedro J. Guerrero-Medina ◽  
Miguel A. Robles-García ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Salmonella Enterica ◽  
Micrococcus Luteus ◽  
E Coli

Actualmente existe la necesidad de hacer frente al problema de la resistencia a los antibióticos y al uso indiscriminado de fungicidas químicos en la agricultura. El objetivo de este trabajo fue evaluar el efecto inhibitorio de extractos acuosos, metanólicos, acetónicos y hexánicos de hoja y tallo de Vitex mollis Kunth (Lamiaceae) contra diferentes bacterias (Escherichia coli, Micrococcus luteus, Salmonella enterica y Staphylococcus aureus) y especies del hongo Fusarium (F. verticillioides, F. oxysporum, F. tapsinum y F. oxysporum f.sp. lycopersici) de importancia en la salud y en la agricultura, así como determinar su composición química general. Se determinaron las concentraciones inhibitorias mínimas (CIM) de todos los extractos por la técnica de microdilución, excepto del hexánico, que no presentó inhibición en las bacterias estudiadas. S. enterica fue la bacteria que mostró mayor sensibilidad al extracto metanólico de tallo (CIM = 28 μg mL-1), le siguieron M. luteus (CIM = 32 μg mL-1), S. aureus (CIM = 75 μg mL-1) y E. coli (CIM = 80 μg mL- 1). Los extractos metanólicos y acuosos de tallo presentaron mayor porcentaje de inhibición contra los diferentes tipos de Fusarium evaluados por el método de dilución en agar. Los extractos de V. mollis inhibieron a F. verticillioides entre 62 y 91 % con 120 μg mL-1 de extracto. El orden de las especies de hongos inhibidas por los extractos fue: F. verticillioides > F. oxysporum > F. tapsinum > F. oxysporum f.sp. lycopersici. La composición química de las especies se determinó mediante pruebas para fenoles, taninos, flavonoides, triterpenos, alcaloides, cumarinas y saponinas. Ninguno de los extractos presentó alcaloides y saponinas. Los fenoles (37.1 mg EAG/g muestra seca) y flavonoides (26.8 mg EQ/g muestra seca) fueron los compuestos mayoritarios en los extractos metanólicos y acuosos. En conclusión, se requieren cantidades muy pequeñas de extracto para la inhibición de bacterias y de Fusarium; por lo tanto, V. mollis puede ser considerada una fuente de metabolitos para este fin y en la agricultura como control alternativo dentro de un manejo integrado de enfermedades.

Iso-octenidine: Promising Octenidine Analogue with Improved Solubility

2020 ◽  
Vol 17 ◽  
Author(s):  
Igor K. Yakuschenko ◽  
Nataliya N. Pozdeeva ◽  
Viktoriya A. Mumyatova ◽  
Alexey A. Terentiev ◽  
Svyatoslav Ya. Gadomsky
Keyword(s):  
Escherichia Coli ◽  
Antibacterial Activity ◽  
Micrococcus Luteus ◽  
Octenidine Dihydrochloride ◽  
First Time

: Iso-octenidine, an isomer of octenidine dihydrochloride, was synthesized and studied for the first time. Isooctenidine was demonstrated to be 3-fold more soluble in water in comparison to original octenidine, and both substances had remarkably similar antibacterial activity (tested on Escherichia Coli and Micrococcus luteus).

Expression and partial purification of human pro-opiomelanocortin in Escherichia coli

1989 ◽  
Vol 3 (2) ◽  
pp. 105-112 ◽  
Author(s):  
T. S. Grewal ◽  
P. J. Lowry ◽  
D. Savva
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Fusion Protein ◽  
Western Blot ◽  
Blot Analysis ◽  
Expression Vectors ◽  
Partial Purification ◽  
Amino Acid Residues ◽  
E Coli ◽  
Dna Fragment

ABSTRACT A large portion of the human pro-opiomelanocortin (POMC) peptide corresponding to amino acid residues 59–241 has been cloned and expressed in Escherichia coli. A 1·0 kb DNA fragment encoding this peptide was cloned into the expression vectors pUC8 and pUR291. Plasmid pJMBG51 (a pUC8 recombinant) was found to direct the expression of a 24 kDa peptide. The recombinant pUR291 (pJMBG52) was shown to produce a β-galactosidase fusion protein of 140 kDa. Western blot analysis showed that both the 24 kDa and 140 kDa peptides are recognized by antibodies raised against POMC-derived peptides. The β-galactosidase fusion protein has been partially purified from crude E. coli cell lysates using affinity chromatography on p-aminobenzyl-1-thio-β-d-galactopyranoside agarose.

scholarly journals Antimicrobial activity of lactic acid bacteria isolated from garri on Escherichia coli strains isolated from clinical and environmental samples

2020 ◽  
Vol 12 (4) ◽  
pp. 357-365
Author(s):  
H.I. Atta ◽  
A. Gimba ◽  
T. Bamgbose
Keyword(s):  
Escherichia Coli ◽  
Lactic Acid ◽  
Lactic Acid Bacteria ◽  
Lactobacillus Plantarum ◽  
Lactobacillus Acidophilus ◽  
Plant Pathogens ◽  
Resistant Bacteria ◽  
Fermented Foods ◽  
Environmental Isolates ◽  
E Coli

Abstract. The production of bacteriocins by lactic acid bacteria affords them the ability to inhibit the growth of bacteria; they are particularly important in the biocontrol of human and plant pathogens. Lactic acid bacteria have been frequently isolated from fermented foods due to the high acidity these foods contain. In this study, lactic acid bacteria were isolated from garri, a popular Nigerian staple food, which is fermented from cassava, and their antagonistic activity against clinical and environmental isolates of Escherichia coli was determined. The species of Lactobacillus isolated include: Lactobacillus plantarum (50%), Lactobacillus fermentum (20%), Lactobacillus acidophilus (20%), and Lactobacillus salivarius (10%). Growth inhibition of the strains of E.coli was observed in Lactobacillus plantarum that inhibited the growth of both. The clinical and environmental isolates of E. coli were inhibited by Lactobacillus plantarum, while Lactobacillus acidophilus showed activity against only the clinical isolate. The greatest zone of inhibition against the strains of E. coli was recorded by Lactobacillus acidophilus (22.7±1.53 mm). The bacteriocins produced by Lactobacillus species have a good potential in the biocontrol of pathogens, and should be the focus of further studies on antibiotic resistant bacteria.

scholarly journals CONSTRUCTION, EXPRESSION AND PURIFICATION OF RECOMBINANT PRE-MATURE PEPTIDE OF PLANTARICIN F FROM Lactobacillus plantarum S34 IN Escherichia coli

2015 ◽  
Vol 16 (1) ◽  
pp. 31
Author(s):  
Kusdianawati Kusdianawati ◽  
Apon Zaenal Mustopa ◽  
Suharsono Suharsono ◽  
Bugi Ratno Budiarto ◽  
Fatimah Fatimah ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Lactobacillus Plantarum ◽  
Proteolytic Enzymes ◽  
Leader Peptide ◽  
Human Consumption ◽  
Mature Peptide ◽  
Expression And Purification ◽  
Food Preservative ◽  
E Coli ◽  
Sds Page

Plantaricin is one of bacteriocins that have the potential to be used as food preservative. Plantaricin is safe for human consumption because it can be easily degraded by proteolytic enzymes. The objective of this study was to express and purify recombinant pre-mature peptide of plantaricin F from <em>Lactobacillus plantarum</em> S34 in <em>Escherichia coli</em>. Plantaricin gene-specific primer was used to obtain pln F structural gene amplicon from L. <em>plantarum</em> S34. This amplicon was cloned in pET32a vector and expressed in E. coli BL21 (DE3) pLysS. Pre-mature plantaricin F peptide was expressed as Histagged-fusion protein and separated by Co2+-chelating affinity chromatography. L. <em>plantarum</em> S34-derived pre-mature plantaricin F peptide fused with thioredoxin-(His)6tag had successfully been expressed in E. <em>coli</em> BL21 (DE3) pLysS using pET32a as an expression vector. The fused recombinant pln F as pre-mature state expressed had a molecular mass of +24 kDa, meanwhile the fused recombinant that contained only the leader peptide of pln F appeared as +20 kDa based on SDS-PAGE separations. The optimal production of fused recombinant pln F as soluble fraction was obtained when culture condition was added with 0.5 mM of IPTG and incubated at 22°C for 5 hours (OD~1). Furthermore, the expression of fused recombinant pln F as its pre-mature peptide pointed out that the pln F’s leader peptide could be proteolytically cleaved by a system in heterologous cells. Overall, heterologous pln F production as pre-mature peptide fused with thioredoxin-(His)6tag had been well established. From this research, we expect plantaricin F can be expressed and purified in E. coli.

scholarly journals Induction and control of the autolytic system of Escherichia coli

1982 ◽  
Vol 152 (1) ◽  
pp. 26-34
Author(s):  
M Leduc ◽  
R Kasra ◽  
J van Heijenoort
Keyword(s):  
Escherichia Coli ◽  
Fatty Acid ◽  
Acid Synthesis ◽  
Induction Method ◽  
E Coli ◽  
Escherichia Coli Cells ◽  
Carbonyl Cyanide ◽  
And Control ◽  
First Time ◽  
Induced Autolysis

Various methods of inducing autolysis of Escherichia coli cells were investigated, some being described here for the first time. For the autolysis of growing cells only induction methods interfering with the biosynthesis of peptidoglycan were taken into consideration, whereas with harvested cells autolysis was induced by rapid osmotic or EDTA shock treatments. The highest rates of autolysis were observed after induction by moenomycin, EDTA, or cephaloridine. The different autolyses examined shared certain common properties. In particular, regardless of the induction method used, more or less extensive peptidoglycan degradation was observed, and 10(-2) M Mg2+ efficiently inhibited the autolytic process. However, for other properties a distinction was made between methods used for growing cells and those used for harvested cells. Autolysis of growing cells required RNA, protein, and fatty acid synthesis. No such requirements were observed with shock-induced autolysis performed with harvested cells. Thus, the effects of Mg2+, rifampicin, chloramphenicol, and cerulenin clearly suggest that distinct factors are involved in the control of the autolytic system of E. Coli. Uncoupling agents such as sodium azide, 2,4-dinitrophenol, and carbonyl-cyanide-m-chlorophenyl hydrazone used at their usual inhibiting concentration had no effect on the cephaloridine or shock-induced autolysis.

scholarly journals A New 3-Prenyl-2-flavene and Other Extractives from Baphia massaiensis and Their Antimicrobial Activities

2018 ◽  
Vol 13 (4) ◽  
pp. 1934578X1801300 ◽  
Author(s):  
Ngonye Keroletswe ◽  
Runner R. T. Majinda ◽  
Ishmael B. Masesane
Keyword(s):  
Escherichia Coli ◽  
2D Nmr ◽  
Antimicrobial Activities ◽  
Spectroscopic Techniques ◽  
Good Activity ◽  
Gram Negative ◽  
E Coli ◽  
Chromatographic Techniques ◽  
Inhibition Zones ◽  
First Time

One new 3-prenyl-2-flavene, named baphiflavene A, 1, and eleven known compounds, 2-12, were isolated and reported for the first time from Baphia massaiensis using several chromatographic techniques. Their structures were elucidated using different spectroscopic techniques; 1D and 2D-NMR, HRMS, GC-MS, UV/Vis, FTIR and by comparison with literature data. The isolates were tested against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli and Candida albicans to establish their preliminary antimicrobial activities. The results revealed that compound 1 had moderate activities against both Gram positive ( B. subtilis and S. aureus) and Gram negative ( E. coli and P. aeruginosa) bacteria, and good activity against C. albicans with inhibition zones of 10–23 mm (compared to 19 mm for chloramphenicol and miconazole standards). To the best of our knowledge, this is the first phytochemical work reported on Baphia massaiensis.

scholarly journals Spread of NDM-5 and OXA-181 Carbapenemase-Producing Escherichia coli in Chad

2019 ◽  
Vol 63 (11) ◽  
Author(s):  
Oumar Ouchar Mahamat ◽  
Manon Lounnas ◽  
Mallorie Hide ◽  
Abelsalam Tidjani ◽  
Julio Benavides ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Healthy Volunteers ◽  
Resistance Genes ◽  
Sequence Type ◽  
Hospitalized Patients ◽  
Content Type ◽  
E Coli ◽  
First Time

ABSTRACT We detected for the first time blaNDM-5 and blaOXA-181 in Escherichia coli isolates from hospitalized patients and healthy volunteers in Chad. These resistance genes were located on IncX3 and IncF plasmids. Despite the large diversity of E. coli clones, the identified resistant intestinal isolates belonged mainly to the same sequence type.

scholarly journals The Transcriptional Factors MurR and Catabolite Activator Protein Regulate N-Acetylmuramic Acid Catabolism in Escherichia coli

2008 ◽  
Vol 190 (20) ◽  
pp. 6598-6608 ◽  
Author(s):  
Tina Jaeger ◽  
Christoph Mayer
Keyword(s):  
Escherichia Coli ◽  
Cell Wall ◽  
Sole Source ◽  
Lag Phase ◽  
Face To Face ◽  
E Coli ◽  
Catabolite Activator Protein ◽  
Activator Protein ◽  
High Level ◽  
First Time

ABSTRACT The MurNAc etherase MurQ of Escherichia coli is essential for the catabolism of the bacterial cell wall sugar N-acetylmuramic acid (MurNAc) obtained either from the environment or from the endogenous cell wall (i.e., recycling). High-level expression of murQ is required for growth on MurNAc as the sole source of carbon and energy, whereas constitutive low-level expression of murQ is sufficient for the recycling of peptidoglycan fragments continuously released from the cell wall during growth of the bacteria. Here we characterize for the first time the expression of murQ and its regulation by MurR, a member of the poorly characterized RpiR/AlsR family of transcriptional regulators. Deleting murR abolished the extensive lag phase observed for E. coli grown on MurNAc and enhanced murQ transcription some 20-fold. MurR forms a stable multimer (most likely a tetramer) and binds to two adjacent inverted repeats within an operator region. In this way MurR represses transcription from the murQ promoter and also interferes with its own transcription. MurNAc-6-phosphate, the substrate of MurQ, was identified as a specific inducer that weakens binding of MurR to the operator. Moreover, murQ transcription depends on the activation by cyclic AMP (cAMP)-catabolite activator protein (CAP) bound to a class I site upstream of the murQ promoter. murR and murQ are divergently orientated and expressed from nonoverlapping face-to-face (convergent) promoters, yielding transcripts that are complementary at their 5′ ends. As a consequence of this unusual promoter arrangement, cAMP-CAP also affects murR transcription, presumably by acting as a roadblock for RNA polymerase.

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