Protein Produced by Bacillus subtilis ATCC21332 in the Presence of Cymbopogon flexuosus Essential Oil

Proteins levels produced by bacteria may be increased in stressful surroundings, such as in the presence of antibiotics. It appears that many antimicrobial agents or antibiotics, when used at low concentrations, have in common the ability to activate or repress gene transcription, which is distinct from their inhibitory effect. There have been comparatively few studies on the potential of antibiotics or natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. Therefore, this study was focusing on the effect of essential oil fromCymbopogon flexuosusin regulating proteins production byBacillus subtilisATCC21332. The Minimum Inhibition Concentration (MIC) of theC. flexuosusessential oil onB. subtiliswas determined by using microdilution assay, resulting 1.76mg/ml. The bacteria cells were further exposed to theC. flexuosusessential oil at concentration of 0.01 MIC for 72 h. The proteins were then isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile showed that a band with approximate size of 30 kDa was appeared for the treated bacteria withC. flexuosusessential oil. Thus,B. subtilisATCC21332 in stressful condition with the presence ofC. flexuosusessential oils at low concentration could induce the protein production. The isolated protein also showed antimicrobial activity against selected Gram-positive and Gram-negative bacteria.

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Cymbopogon nardus Essential Oil as Protein Inducer in Bacillus subtilis ATCC21332

Malaysian Journal of Science Health & Technology ◽

10.33102/mjosht.v1i1.18 ◽

2018 ◽

Vol 1 (1) ◽

Author(s):

Hairul Shahril Muhamad ◽

Ismatul Nurul Asyikin Ismail ◽

Nabilah Ahmad Alhadi ◽

Salina Mat Radzi ◽

Maryam Mohamed Rehan ◽

...

Keyword(s):

Antimicrobial Activity ◽

Bacillus Subtilis ◽

Essential Oil ◽

Protein Production ◽

Antimicrobial Agents ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Extracellular Proteins ◽

Specific Chemical ◽

Cymbopogon Nardus

Protein production by bacteria might be increased in stressful conditions such as in the presence of antimicrobial agents. Many studies have proven that antibiotics or antimicrobial agents at low concentration are able to activate or repress gene transcription process in bacteria. However, there have been comparatively few studies on the potential of natural compounds in nature as a specific chemical signal that can trigger a variety of biological functions. An attempt was made to study the effect of essential oil from Cymbopogon nardus in regulating protein production by Bacillus subtilis ATCC21332. The bacterial cells were further exposed to the C. nardus essential oil at concentration of 0.02 % for 48 h at 37°C. The intracellular proteins were then isolated and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Proteins profile showed that a band with approximate size of 180 kDa appeared for the treated bacteria with C. nardus essential oil. An alignment of peptide sequences to the NCBI BLAST database revealed that B. subtilis ATCC21332 in stressful condition tend to produce intracellular protein recognized as respiratory nitrate reductase ? subunit enzyme. Besides, the extracellular proteins secreted by B. subtilis ATCC21332 after being subjected to 0.02% of C. nardus essential oil for 48 and 72 h at 30°C, were further analyzed on antimicrobial activity. The extracellular proteins secreted by B. subtilis ATCC21332 prior to enhancing with 0.02 % C. nardus essential oil at 30°C for 72 h exhibited antimicrobial activity towards two strains of bacteria, which are Bacillus cereus and Escherichia coli.

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Combination Effects of Cymbopogon sp. Essential Oil on Selected Bacteria

Malaysian Journal of Science Health & Technology ◽

10.33102/mjosht.v1i1.8 ◽

2018 ◽

Vol 1 (1) ◽

Author(s):

Nur Aishah Abdul Wahab ◽

Hairul Shahril Muhamad ◽

Nabilah Ahmad Alhadi ◽

Salina Mat Radzi ◽

Maryam Mohamed Rehan ◽

...

Keyword(s):

Escherichia Coli ◽

Staphylococcus Aureus ◽

Bacillus Subtilis ◽

Essential Oils ◽

Staphylococcus Epidermidis ◽

Antimicrobial Agents ◽

Synergistic Effects ◽

Combination Effects ◽

Cymbopogon Flexuosus ◽

Combination Study

Combination effects between Cymbopogon flexuosus and Cymbopogon nardus essential oils were studied to determine whether the combination could emerge as better and more powerful antimicrobial agents against six selected bacteria includes Bacillus subtilis, Escherichia coli, Salmonella typhimurium, Serratia marcescens, Staphylococcus aureus, and Staphylococcus epidermidis. This combination study exhibited 40.67% additive, 28.67% antagonistic, 16.00% indifferent and 14.66% synergistic effects. C. flexuosus and C. nardus essential oils in combination showed a high inhibitory activity against S. aureus with 16% synergistic, 64% additive and 20% indifferent effects.

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The isolation of surface array proteins from bacteria

Canadian Journal of Biochemistry and Cell Biology ◽

10.1139/o84-152 ◽

1984 ◽

Vol 62 (11) ◽

pp. 1181-1189 ◽

Cited By ~ 42

Author(s):

S. F. Koval ◽

R. G. E. Murray

Keyword(s):

Molecular Weight ◽

Protein Conformation ◽

Gel Electrophoresis ◽

Guanidine Hydrochloride ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Structural Features ◽

Proteolytic Degradation ◽

Low Concentrations ◽

Single Polypeptide

The methods used for the isolation of regularly structured (RS) surface array proteins of a range of prokaryotes are described. Most RS proteins can be selectively solubilized from envelope preparations with low concentrations of urea or guanidine hydrochloride. Sodium dodecyl sulfate – polyacrylamide gel electrophoresis analysis of the protein extracts shows that most RS arrays are composed of a single polypeptide that may contain carbohydrate. The molecular weight of the proteins varies from 41 000 to 200 000. Possible reasons for the presence of more than one polypeptide in RS protein preparations are discussed, as well as the evidence for proteolytic degradation of some RS proteins during isolation. Structural features of the RS proteins are described and the importance of protein conformation to assembly of the arrays is indicated.

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The purification of a novel amylase from Bacillus subtilis and its inhibition by wheat proteins

Biochemical Journal ◽

10.1042/bj2090561 ◽

1983 ◽

Vol 209 (2) ◽

pp. 561-564 ◽

Cited By ~ 12

Author(s):

A R Orlando ◽

P Ade ◽

D Di Maggio ◽

C Fanelli ◽

L Vittozzi

Keyword(s):

Bacillus Subtilis ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Alpha Amylase ◽

Wheat Protein ◽

Wheat Proteins ◽

Maximal Inhibition ◽

Molecular Weights ◽

Amylase Inhibitors

A new alpha-amylase (EC 3.2.1.1) from Bacillus subtilis was purified by affinity chromatography. The molecular weight of the purified enzyme, estimated from sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, was 93000, which is very different from the molecular weights of two well-characterized amylases from B. subtilis. Electrofocusing showed an isoelectric point of 5. Amylase shows a broad maximum of activity between pH 6 and 7; maximal inhibition of enzyme by wheat-protein alpha-amylase inhibitors is displayed at pH 7.

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Proteomic Analysis of the Spore Coats of Bacillus subtilis and Bacillus anthracis

Journal of Bacteriology ◽

10.1128/jb.185.4.1443-1454.2003 ◽

2003 ◽

Vol 185 (4) ◽

pp. 1443-1454 ◽

Cited By ~ 135

Author(s):

Erh-Min Lai ◽

Nikhil D. Phadke ◽

Maureen T. Kachman ◽

Rebecca Giorno ◽

Santiago Vazquez ◽

...

Keyword(s):

Bacillus Subtilis ◽

Bacillus Anthracis ◽

Proteomic Analysis ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Detection Methods ◽

Coat Proteins ◽

Bona Fide ◽

Spore Proteins ◽

Spore Coat Proteins

ABSTRACT The outermost proteinaceous layer of bacterial spores, called the coat, is critical for spore survival, germination, and, for pathogenic spores, disease. To identify novel spore coat proteins, we have carried out a preliminary proteomic analysis of Bacillus subtilis and Bacillus anthracis spores, using a combination of standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis separation and improved two-dimensional electrophoretic separations, followed by matrix-assisted laser desorption ionization-time of flight and/or dual mass spectrometry. We identified 38 B. subtilis spore proteins, 12 of which are known coat proteins. We propose that, of the novel proteins, YtaA, YvdP, and YnzH are bona fide coat proteins, and we have renamed them CotI, CotQ, and CotU, respectively. In addition, we initiated a study of coat proteins in B. anthracis and identified 11 spore proteins, 6 of which are candidate coat or exosporium proteins. We also queried the unfinished B. anthracis genome for potential coat proteins. Our analysis suggests that the B. subtilis and B. anthracis coats have roughly similar numbers of proteins and that a core group of coat protein species is shared between these organisms, including the major morphogenetic proteins. Nonetheless, a significant number of coat proteins are probably unique to each species. These results should accelerate efforts to develop B. anthracis detection methods and understand the ecological role of the coat.

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T lymphocyte-derived differentiation-inducing factor inhibits proliferation of leukemic and normal hemopoietic cells

Blood ◽

10.1182/blood.v68.6.1333.1333 ◽

1986 ◽

Vol 68 (6) ◽

pp. 1333-1338 ◽

Cited By ~ 8

Author(s):

U Gullberg ◽

E Nilsson ◽

MG Sarngadharan ◽

I Olsson

Keyword(s):

Cell Line ◽

T Lymphocyte ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Inhibitory Effect ◽

Exchange Chromatography ◽

Leukemia Cells ◽

Gel Chromatography ◽

Wild Type ◽

Clonogenic Growth

Abstract A differentiation-inducing factor (DIF) for the promyelocytic HL-60 cell line is constitutively produced by the malignant T lymphocyte line HUT-102. DIF was highly purified from HUT-102-conditioned media by means of diethylaminoethanol (DEAE)-chromatography, gel chromatography, and high-resolution, ion-exchange chromatography on a MonoQ column and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). In addition to inducing differentiation of wild-type HL-60 cells, resulting in secondary inhibition of growth, DIF, at a tenfold lower concentration, inhibited the growth of some clones of the monoblastic U- 937 cell line as well as that of subclones of HL-60. The latter effect was most likely a primary growth inhibition and not secondary to differentiation; 50% inhibition of clonogenic growth in agar was seen at approximately 1.0 pmol/L of DIF. In addition, the clonogenic growth of fresh leukemia cells from 10 of 12 patients with acute myeloid leukemia (AML) was inhibited with 50% inhibition at approximately 10 pmol/L of DIF. The growth of normal granulocyte-macrophage colonies was inhibited at a similar concentration, whereas early erythroid colonies were much more resistant. DIF and interferon-gamma (gamma-IFN) were shown to be separate molecules inasmuch as a neutralizing antibody for gamma-IFN did not abolish the DIF effect. The differentiation effect on wild-type HL-60 and the proliferation inhibitory effect on leukemic and normal myeloid cells cochromatographed through all purification steps suggest that both activities are exhibited by identical polypeptides. DIF may have a role in regulating normal hemopoiesis. The growth inhibitory effect of DIF and the ability to induce differentiation of some leukemia cells may suggest a clinical utility in the treatment of leukemia.

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Studies on the action of dexamethasone on prostaglandin production by freshly dispersed amnion cells

Acta Endocrinologica ◽

10.1530/acta.0.1280563 ◽

1993 ◽

Vol 128 (6) ◽

pp. 563-567 ◽

Cited By ~ 3

Author(s):

William Gibb ◽

Robert Breton

Keyword(s):

Arachidonic Acid ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Inhibitory Effect ◽

Arachidonic Acid Release ◽

Dexamethasone Treatment ◽

Glucocorticoid Treatment ◽

Prostaglandin Production ◽

Acid Release ◽

Dispersed Cells

The human amnion may be an important source of prostaglandins (PGs) during pregnancy and possibly labor. Glucocorticoids stimulate PG production in confluent amnion cell cultures, but in freshly dispersed cells they inhibit PG production. The purpose of the present study was to determine if this inhibitory effect occurred at the level of arachidonic acid release from lipids. Cells were labeled with radioactive arachidonate and the release of radioactivity was measured in the presence or absence of dexamethasone. No significant effect of dexamethasone treatment was observed. The possibility that glucocorticoid treatment inhibited the release of arachidonate from a specific species of phospholipid was also examined. However, no difference was found in the distribution of arachidonate between lipids isolated from glucocorticoid-treated and untreated cells. Furthermore, dexamethasone treatment did not alter the lipocortin 1 and 2 content of dispersed cells, determined following sodium dodecyl sulfate polyacrylamide gel electrophoresis and immunoblotting. These studies indicate that the inhibition of prostaglandin production by dispersed amnion cells by glucocorticoids most likely occurs at a point distal to arachidonic acid release.

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Isolation, heterologous expression, and purification of a novel antifungal protein from Bacillus subtilis strain Z-14

Microbial Cell Factories ◽

10.1186/s12934-020-01475-1 ◽

2020 ◽

Vol 19 (1) ◽

Author(s):

Xuechao Zhang ◽

Xiaojun Guo ◽

Cuihong Wu ◽

Chengcui Li ◽

Dongdong Zhang ◽

...

Keyword(s):

Bacillus Subtilis ◽

Antifungal Activity ◽

Pichia Pastoris ◽

Fungal Infections ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Exchange Chromatography ◽

Proteinase K ◽

Antifungal Protein ◽

Subtilis Strain

Abstract Background Wheat sheath blight, a soil borne fungal disease caused by Rhizoctonia cerealis, is considered as one of the most serious threats to wheat worldwide. Bacillus subtilis Z-14 was isolated from soil sampled from a wheat rhizosphere and was confirmed to have strong antifungal activity against R. cerealis. Results An antifungal protein, termed F2, was isolated from the culture supernatant of Z-14 strain using precipitation with ammonium sulfate, anion exchange chromatography, and reverse phase chromatography. Purified F2 had a molecular mass of approximately 8 kDa, as assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis. Edman degradation was used to determine the amino acid sequence of the N-terminus, which was NH2ASGGTVGIYGANMRS. This sequence is identical to a hypothetical protein RBAM_004680 (YP_001420098.1) synthesized by B. amyloliquefaciens FZB42. The recombinant F2 protein (rF2) was heterologously expressed in the yeast host Pichia pastoris, purified using a Niaffinity column, and demonstrated significant antifungal activity against R. cerealis. The purified rF2 demonstrated broad spectrum antifungal activity against different varieties of fungi such as Fusarium oxysporum, Verticillium dahliae, Bipolaris papendorfii, and Fusarium proliferatum. rF2 was thermostable, retaining 91.5% of its activity when incubated for 30 min at 100 °C. Meanwhile, rF2 maintained its activity under treatment by proteinase K and trypsin and over a wide pH range from 5 to 10. Conclusions A novel antifungal protein, F2, was purified from biocontrol Bacillus subtilis Z-14 strain fermentation supernatant and heterologously expressed in Pichia pastoris to verify its antifungal activity against R. cerealis and the validity of the gene encoding F2. Considering its significant antifungal activity and stable characteristics, protein F2 presents an alternative compound to resist fungal infections caused by R. cerealis.

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Preliminary characterization of two thymus-dependent xenoantigens from mouse lymphocytes

Biochemical Journal ◽

10.1042/bj1630211 ◽

1977 ◽

Vol 163 (2) ◽

pp. 211-217 ◽

Cited By ~ 26

Author(s):

I S Trowbridge ◽

M Nilsen-Hamilton ◽

R T Hamilton ◽

M J Bevan

Keyword(s):

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Filtration ◽

Membrane Vesicles ◽

Sodium Deoxycholate ◽

Pea Lectin ◽

Preliminary Characterization ◽

Low Concentrations ◽

Membrane Bound

Preliminary characterization of two mouse thymus-dependent (T) lymphocyte xenoantigens, T25 and T200, which are selectively labelled by lactoperoxidase-catalysed iodination of T-cells, is described. Both molecules are membrane-bound glycoproteins. Fractionation of membrane vesicles prepared from BW5147 lymphoma cells by sedimentation through sucrose density gradients show that antigens T25 and T200 are in fractions enriched with plasma membrane. Moreover antigen T200 is partially degraded when viable cells are treated briefly with low concentrations of trypsin. Both molecules are efficiently solubilized in buffers containing sodium deoxycholate or Nonidet P-40, as measured by failure to sediment at 100000g for 60min. However, gel filtration on Sepharose 6B showed the presence of aggregated material in Nonidet P-40 extracts which was not found in deoxycholate-solubilized membranes. After solubilization in detergent, antigens T25 and T200 bind to, and may be specifically eluted from, columns of pea lectin--Sepharose or concanavalin A--Sepharose. Both molecules are heterogeneous when examined by polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate. As judged by its binding to columns of pea lectin, at least part of the heterogeneity of mouse thymocyte antigen T25 resides in its carbohydrate moiety.

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Characterization of Spores of Bacillus subtilis That Lack Most Coat Layers

Journal of Bacteriology ◽

10.1128/jb.00896-08 ◽

2008 ◽

Vol 190 (20) ◽

pp. 6741-6748 ◽

Cited By ~ 61

Author(s):

Sonali Ghosh ◽

Barbara Setlow ◽

Paul G. Wahome ◽

Ann E. Cowan ◽

Marco Plomp ◽

...

Keyword(s):

Bacillus Subtilis ◽

Dipicolinic Acid ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Great Majority ◽

Coat Proteins ◽

Expression Of Genes ◽

Genes Encoding ◽

Wet Heat ◽

Coat Material

ABSTRACT Spores of Bacillus subtilis have a thick outer layer of relatively insoluble protein called the coat, which protects spores against a number of treatments and may also play roles in spore germination. However, elucidation of precise roles of the coat in spore properties has been hampered by the inability to prepare spores lacking all or most coat material. In this work, we show that spores of a strain with mutations in both the cotE and gerE genes, which encode proteins involved in coat assembly and expression of genes encoding coat proteins, respectively, lack most extractable coat protein as seen by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, as well as the great majority of the coat as seen by atomic force microscopy. However, the cotE gerE spores did retain a thin layer of insoluble coat material that was most easily seen by microscopy following digestion of these spores with lysozyme. These severely coat-deficient spores germinated relatively normally with nutrients and even better with dodecylamine but not with a 1:1 chelate of Ca2+ and dipicolinic acid. These spores were also quite resistant to wet heat, to mechanical disruption, and to treatment with detergents at an elevated temperature and pH but were exquisitely sensitive to killing by sodium hypochlorite. These results provide new insight into the role of the coat layer in spore properties.

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