Cerein Research Articles

Bacteriocins from the rhizosphere microbiome - from an agriculture perspective.

Bacteria produce and excrete a versatile and dynamic suit of compounds to defend against microbial competitors and mediate local population dynamics. These include a wide range of broad-spectrum non-ribosomally synthesized antibiotics, lytic enzymes, metabolic by-products, proteinaceous exotoxins, and ribosomally produced antimicrobial peptides (bacteriocins). Most bacteria produce at least one bacteriocin. Bacteriocins are of interest in the food industry as natural preservatives and in the probiotics industry, leading to extensive studies on lactic acid bacteria (colicin produced by Escherichia coli is a model bacteriocin). Recent studies have projected use of bacteriocins in veterinary medicine and in agriculture, as biostimulants of plant growth and development and as biocontrol agents. For example, bacteriocins such as Cerein 8A, Bac-GM17, putidacin, Bac 14B, amylocyclicin have been studied for their mechanisms of anti-microbial activity. Bac IH7 promotes tomato and musk melon plant growth. Thuricin 17 (Th17) is the only bacteriocin studied extensively for plant growth promotion, including at the molecular level. Th17 functions as a bacterial signal compound, promoting plant growth in legumes and non-legumes. In Arabidopsis thaliana and Glycine max Th17 increased phytohormones IAA and SA at 24 h post treatment. At the proteome level Th17 treatment of 3-week-old A. thaliana rosettes led to >2-fold changes in activation of the carbon and energy metabolism pathway proteins, 24 h post treatment. At 250 mM NaCl stress, the control plants under osmotic-shock shut down most of carbon-metabolism and activated energy-metabolism and antioxidant pathways. Th17 treated plants, at 250 mM NaCl, retained meaningful levels of the light harvesting complex, photosystems I and II proteins and energy and antioxidant pathways were activated, so that rosettes could better withstand the salt stress. In Glycine max, Th17 helped seeds germinate in the presence of NaCl stress, and was most effective at 100 mM NaCl. The 48 h post germination proteome suggested efficient and speedier partitioning of storage proteins, activation of carbon, nitrogen and energy metabolisms in Th17 treated seeds both under optimal and 100 mM NaCl. This review focuses on the bacteriocins produced by plant-rhizosphere colonizers and plant-pathogenic bacteria, that might have uses in agriculture, veterinary, and human medicine.

Extraction of the antimicrobial peptide cerein 8A by aqueous two-phase systems and aqueous two-phase micellar systems

Cerein 8A is an antimicrobial peptide with potential application against food spoilage and pathogenic bacteria. The partitioning of cerein 8A was investigated in two liquid–liquid extraction systems that are considered promising for bioseparation and purification purposes. Aqueous two-phase systems (ATPSs) were prepared with polyethylene glycol (PEG) and inorganic salts, and the addition of NaCl was investigated in this system. The best results concerning partition coefficients (K b) were obtained with PEG + ammonium sulphate, and K b value significantly increases when NaCl was added. Cerein 8A was effectively extracted into the micelle-rich phase in a 4% Triton X-114 medium. Recovery yield was higher for ATPS compared to micellar systems. Cerein 8A can be isolated from a crude suspension containing the bioactive molecule by ATPSs. Successful implementation of peptide partitioning represents an important step towards developing a low-cost effective separation method for cerein 8A.

Inhibition of Salmonella Enteritidis by cerein 8A, EDTA and sodium lactate

The ability of the bacteriocin cerein 8A to inhibit Salmonella Enteritidis in combination with EDTA and sodium lactate was investigated. Salmonella Enteritidis was incubated with combinations of cerein 8A (3200 AU/mL) and EDTA (20, 50, 100 mmol/L) or sodium lactate (200 mmol/L). All treatments caused a significant reduction in the OD 600 values of Salmonella Enteritidis cultures. The addition of cerein 8A plus EDTA resulted in higher inhibition in comparison with the bacteriocin alone; the greater the concentration of EDTA, the greater the inhibitory effect. The combination of cerein 8A plus 100 mmol/L EDTA results in a more efficient treatment to reduce the number of viable cells of Salmonella Enteritidis. The combination of cerein 8A plus sodium lactate also showed significant inhibition of the indicator organism. Transmission electron microscopy showed damaged cell walls and loss of protoplasmic material in treated cells. The cells of Salmonella Enteritidis treated with cerein 8A plus EDTA appeared more injured. The bacteriocin cerein 8A may be useful to inhibit Gram-negative bacteria, with enhanced effect in combination with chelating agents. Control of Salmonella Enteritidis, a Gram-negative bacterium constantly linked to food outbreaks, addresses an important aspect of food safety.

Kinetics and thermodynamics of thermal inactivation of the antimicrobial peptide cerein 8A

Optimization of thermal processes relies on adequate degradation kinetic models to warrant food safety and quality. The knowledge on thermal inactivation kinetics of bacteriocins is necessary to allow their adequate use as natural biopreservatives in the food industry. In this work, the kinetics of thermal inactivation was studied for the previously characterized bacteriocin cerein 8A. Samples of cerein 8A were treated at different time–temperature combinations in the range of 0–30 min and 70–82 °C and the thermodynamic and kinetic parameters for bacteriocin inactivation were calculated. Results showed that inactivation followed a first-order reaction with k-values between 0.059 min −1 and 0.235 min −1. D- and k-values decreased and increased, respectively, with increasing temperature, indicating a faster bacteriocin inactivation at higher temperatures. Results suggest that cerein 8A is a relatively thermostable bacteriocin with a z-value of 21.98 °C and E a of 105.7 kJ mol −1.

Inhibition of Listeria monocytogenes in dairy products using the bacteriocin-like peptide cerein 8A

The efficacy of the antimicrobial peptide cerein 8A to control the development of Listeria monocytogenes in milk and soft cheese was investigated. The addition of 160 AU ml − 1 cerein 8A to UHT milk resulted in a decrease of 3 log cycles in viable cells within the 14-day period at 4 °C. The viable counts of L. monocytogenes in pasteurized milk samples containing cerein 8A was lower than those observed in controls without bacteriocin. Addition of cerein 8A to Minas-type soft cheese caused a delay in the start of exponential growth phase, although similar counts were observed after day 6. When cerein 8A was used to control cheese surface contamination by L. monocytogenes, a decrease of 2 log cycles in viable counts of cerein-treated samples was observed during 30 days at 4 °C. This antimicrobial peptide shows potential use as a biopreservative for application in dairy products.

Cerein 8A production in soybean protein using response surface methodology

In this work a 2 3 factorial design was performed with the aim of optimizing the bacteriocin (cerein 8A) production by Bacillus cereus 8A, using response surface methodology (RSM). The bacteriocin production was first tested on different substrates, like soybean meal, soybean protein, fish meal, industrial fibrous soybean residue and cheese whey. Soybean protein was the selected medium to test the effect of three variables (temperature, initial pH and soybean protein concentration) by RSM. Statistical analysis of results showed that, in the range studied, the three variables have a significant effect on cerein 8A production. Response-surface data showed maximum bacteriocin production at initial pH between 6.5 and 9.0, temperature between 22 and 34 °C and soybean protein concentration higher than 20 g l −1. It was not possible to determine a relation between bacteriocin production and optimal conditions for bacterial growth in terms of pH and temperature, although increased bacteriocin yields and bacterial growth were obtained with higher soybean protein concentration.

Purification and partial chemical characterization of the antimicrobial peptide cerein 8A

To purify and to characterize the antimicrobial compound cerein 8A. Cerein 8A was isolated by ammonium sulfate precipitation, 1-butanol extraction and ion-exchange chromatography. Direct activity on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was observed. The purified substance corresponded to a 26 kDa peptide band. The native protein eluted at the void volume of Sephadex G-100, but within the included volume when a 1.5 mol l(-1) NaCl buffer was used, indicating that cerein 8A aggregates extracellularly. The antimicrobial activity was lost by treatment with proteases and heat. The ultraviolet spectrum was typical of a polypeptide and the infrared spectrum indicates that the peptide contains acyl group(s) in its structure. Intact Bacillus cereus spores were sensitive to cerein 8A at 1600 AU ml(-1). Cerein 8A show distinct properties from other antimicrobial peptides of B. cereus, and has a significant inhibitory effect on spores. The characterization of a substance active against important pathogens addresses an important aspect of food safety.

Antibacterial activity of cerein 8A, a bacteriocin-like peptide produced by Bacillus cereus.

The mode of action of cerein 8A, a bacteriocin produced by the soil bacterium Bacillus cereus 8A, was investigated. The effect of cerein 8A was tested against Listeria monocytogenes and a bactericidal effect at 400 arbitrary units (AU)/ml was observed. In addition, cerein 8A was bactericidal against Bacillus cereus at 200 AU/ml, and inhibited the growth of Escherichia coli and Salmonella Enteritidis. Stronger inhibition of these gram-negative bacteria was achieved when the chelating agent EDTA was added together with bacteriocin. The effect of cerein 8A on B. cereus and L. monocytogenes was also investigated by Fourier transform infrared spectroscopy (FTIR). Treated cells had an important frequency increase at 2920 cm-1 and a decrease at 1400 cm-1, corresponding to assignments of fatty acids. Transmission electron microscopy showed damaged cell walls and loss of protoplasmic material. These results suggest that the mode of action of cerein 8A is to interfere with cell membranes and the cell wall.

Influence of media and temperature on bacteriocin production by Bacillus cereus 8A during batch cultivation.

Cerein 8A is a bacteriocin produced by the soil bacterium Bacillus cereus 8A, isolated from native woodlands of Brazil. The influence of temperature and media on the growth of B. cereus 8A and the production of this bacteriocin was studied during batch cultivation. Maximum activity was detected by cultivation in brain/heart infusion broth, reaching 3200 activity units ml(-1). Bacteriocin was also produced in peptone, MRS, Mueller-Hinton and nutrient broth, while no activity was observed during cultivation in thioglycollate or tryptic soy broth. Temperature had a strong influence on bacteriocin production, which was higher at 30 degrees C than at 25 degrees C. An important decrease in bacteriocin activity was observed at 37 degrees C. The relationship between growth and specific production rates, as a function of the temperature, showed different kinetics of production and there were several peaks in the specific production rates during growth. Bacteriocin was produced at the stationary phase, indicating it is synthesized as a secondary metabolite.