Interaction Of Nisin Research Articles

Fucoidan as a carrier of antimicrobial peptide: Preparation and characterization of nisin-loaded particles

Fucoidan is an anionic sulphated polysaccharide found in brown seaweed. Due to its various biological activities, biodegradability, and biocompatibility, the application of fucoidan is increasing in various fields. The research was aimed to use fucoidan for nisin encapsulation. Nisin-loaded fucoidan particles were prepared by the complexation method in the pH range of 4.0–7.0. The interaction of nisin and fucoidan was confirmed by FT-IR spectroscopy, dynamic light scattering method, thermogravimetric and differential scanning calorimetry analysis. The stability of the particles was analysed under storage at 4 °C. After one month of storage, the hydrodynamic diameter and zeta potential of particles changed insignificantly. Encapsulated nisin preserved its biological activity. The antimicrobial activity was tested against Gram-positive bacteria Bacillus subtilis and Listeria innocua, and Gram-negative bacteria Escherichia coli and Salmonella typhimurium. Nisin-loaded-fucoidan particles exhibited antibacterial activity against Gram-positive bacteria B. subtilis and L. innocua comparable to free nisin. The application of fucoidan for the encapsulation of nisin is promising. A new nisin formulation could be used in the food industry for biopreservation.

Effect of the interaction of nisin Z with various polysaccharides on its antibacterial activity

The effect of various polysaccharides commonly used as thickening or gelling agents in foods on the antibacterial activity of nisin Z was investigated. Addition of 1 g.L−1 sodium alginate in tryptic soy broth resulted in a four-fold increase of minimal inhibitory (MIC) and bactericidal (MBC) concentrations of nisin Z against Listeria innocua ATCC 33090 and Kocuria rhizophila ATCC 9341, while the addition of citrus pectin or various exopolysaccharides (Leuconostoc mesenteroides P35 exopolysaccharides, low (9–11 kDa) or medium (60–76 kDa) molecular weight dextran from Ln. mesenteroides and 2 kefirans (extracted from kefir grains provided by Crokfun or Kefiralia)) did not increase MIC and MBC of nisin Z against these bacteria. Zeta potential determination at pH 7 of the nisin Z, bacteria and polysaccharides allowed us to observe that only nisin Z had a slightly positive zeta potential, while the bacteria, sodium alginate and highly methoxylated citrus pectin zeta potentials were lower than −20 mV and zeta potentials of all exopolysaccharides were between 0 and - 10 mV. It was thus proposed that anionic sodium alginate, which is the only polysaccharide having a lower zeta potential than both bacteria, forms complexes stabilized by electrostatic interactions with oppositely charged nisin Z, thereby limiting the quantity of “free” nisin Z interacting with nisin-susceptible bacteria. This hypothesis was substantiated by the observation of nisin Z - sodium alginate aggregation at pH 7 and the estimation of the apparent affinity of sodium alginate for “free” nisin Z and immobilized nisin Z by partition experiments and by surface plasmon resonance analysis, respectively. This study thus provides useful information for food formulation regarding the effect of different polysaccharides on nisin Z antibacterial activity and some understanding of how interactions between nisin Z, polysaccharides, and nisin Z - sensitive bacteria are explanatory.

Spectroscopic aspects on the interaction of nisin with serum albumin: thermodynamic and kinetic studies.

Nisin is a bacteriocin produced by Streptococcus and Lactococcus species and has antimicrobial activity against other bacteria. Nisin omits the need to use chemical preservatives in food due to its biological preserving properties. In the present in vitro study, we investigated nisin interaction with bovine serum albumin (BSA) using fluorescence spectroscopy and surface plasmon resonance (SPR) analysis to obtain information about the mechanisms of BSA complex formation with nisin. The BSA fluorescence intensity values gradually diminished with rising nisin concentration. The BSA fluorescence quenching analysis indicated that a combined quenching mechanism plays the main role. Finally, the Kb values were reduced with increasing temperature, which is demonstrative of nisin-BSA complex stability decrease at high temperatures. The negative values of ΔH° and ΔS° showed that hydrogen bonds and van der Waals forces are the foremost binding force between BSA and nisin. Meanwhile, the negative values of ΔG° demonstrated the exothermic and random nature of the reaction process. The results of the SPR verified the gained results through the fluorescence spectroscopy investigation, which denoted that the BSA affinity to nisin diminished upon increasing temperature. Overall, fluorescence spectroscopy and SPR results showed that the BSA interaction with nisin decreased with rising temperatures.

Discovery of the Potentiator of the Pore-Forming Ability of Lantibiotic Nisin: Perspectives for Anticancer Therapy.

This study was focused on the action of lantibiotic nisin on the phospholipid membranes. Nisin did not produce ion-permeable pores in the membranes composed of DOPC or DOPE. The introduction of DOPS into bilayer lipid composition led to a decrease in the threshold detergent concentration of nisin. An addition of nisin to DOPG- and TOCL-enriched bilayers caused the formation of well-defined ion pores of various conductances. The transmembrane macroscopic current increased with the second power of the lantibiotic aqueous concentration, suggesting that the dimer of nisin was at least involved in the formation of conductive subunit. The pore-forming ability of lantibiotic decreased in the series: DOPC/TOCL ≈ DOPE/TOCL >> DOPC/DOPG ≥ DOPE/DOPG. The preferential interaction of nisin to cardiolipin-enriched bilayers might explain its antitumor activity by pore-formation in mitochondrial membranes. Small natural molecules, phloretin and capsaicin, were found to potentiate the membrane activity of nisin in the TOCL-containing membranes. The effect was referred to as changes in the membrane boundary potential at the adsorption of small molecules. We concluded that the compounds diminishing the membrane boundary potential should be considered as the potentiator of the nisin pore-forming ability that can be used to develop innovative formulations for anticancer therapy.

An investigation of bacteriocin nisin anti-cancer effects and FZD7 protein interactions in liver cancer cells

The bacteriocin, nisin, produced by Lactococcus and Streptococcus species during fermentation, is widely used for bio preservatives in a wide variety of foods. Liver cancer has a high mortality rate and is the fourth leading cause of cancer-related deaths worldwide. Recently, researchers have shown the anti-cancer effects of nisin through in vitro and in vivo studies. This study aimed to investigate the effect of nisin on liver cancer cell lines, which represented two subgroups of the disease model. Nisin exhibited significant growth inhibition and apoptosis in both cell lines, HuH-7, and SNU182. Drug resistance is the main problem in liver cancer and the epithelial-to-mesenchymal transition has a role in the development of drug resistance in hepatocellular carcinoma. The expression of EMT transcription factors ZEB1, SNAI1, and TWIST1 were analyzed depending on nisin treatment, TWIST1 expression was down-regulated after nisin treatment compared to the untreated SNU182 and HuH-7 cell lines. Besides, due to the reported correlation between the overexpression of Frizzled (FZD) proteins, specifically FZD7, in primary hepatocellular carcinomas (HCCs), molecular docking was assessed for Nisin A in the binding site of FZD7. Results confirmed that Nisin A was able to form important hydrogen bonding with key residues. This research not only determined the role of nisin in different liver cancer cell models but it also provided the first result of FZD7 and nisin interaction.

Synergistic antibacterial activity and mechanism of action of nisin/carvacrol combination against Staphylococcus aureus and their application in the infecting pasteurized milk

Synergistic antibacterial effect is a promising way to overcome the challenge of microbial contamination in food. In this study, we detected the synergistic interactions of nisin and carvacrol. The MIC of nisin and carvacrol against S. aureus were 60 and 125 μg/mL, respectively. The FICI and FBCI were 0.28125 and 0.09375, which suggested that the nisin/carvacrol combination presented synergistic antibacterial effect against S. aureus. The antibacterial activity of nisin/carvacrol combination was much higher than their individuals and the dose of antibacterials was obviously reduced. The combination could completely kill S. aureus within 8 h, accelerate the destruction of cell membrane, and inhibit formation of biofilm. Under the intervention of nisin, more CAR could enter cell to hunt intracellular targets, leading to an increase in intracellular antibacterial level. Besides, in the storage of pasteurized milk, the combinational treatment successfully inhibited microbial reproduction at 25 °C and 4 °C. Thus, the combination of nisin and carvacrol was a potential synergistic strategy for food preservation.

Synergistic antimicrobial interactions of nisin A with biopolymers and solubilising agents for oral drug delivery

In order to develop bacteriocins, like the lantibiotic nisin A, into effective alternatives to existing antibiotics, their biophysical and physicochemical properties must first be assessed, from solubility, to susceptibility and absorption. It has been well established that formulation strategies at early drug development stages can be crucial for successful outcomes during preclinical and clinical phases of development, particularly for molecules with challenging physicochemical properties. This work elucidates the physicochemical challenges of nisin A in terms of its susceptibility to digestive enzymes like pepsin, pancreatin and proteinase K and its poor solubility at physiological pHs. Low solution concentrations, below the minimum inhibitory concentration against Staphylococcus aureus, were obtained in phosphate buffered saline (PBS, pH 7.4) and in fasted state simulated intestinal fluid (FaSSIF, pH 6.5), while higher solubilities at more acidic pH’s such as in a KCl/HCl buffer (pH 2) and in fasted state simulated gastric fluid (FaSSGF, pH 1.6) are observed. Tween® 80 (0.01% v/v) significantly increased the solution concentration of nisin A in PBS (pH 7.4, 24 hr). Pancreatin doubled nisin A’s solution concentration at pH 7.4 (PBS) but reduced its’ inhibitory activity to ∼ 20%, and pepsin almost completely degraded nisin (after 24 hr), but retained activity at biologically relevant exposure times (∼15 min). Harnessing synergism between nisin A and either glycol chitosan or ε-poly lysine, combined with the solubilizing effect of Tween®, increased the antimicrobial activity of nisin A six fold in an in vitro oral administration model.

Nisin-Loaded Ulvan Particles: Preparation and Characterization.

Nisin is an attractive alternative to chemical preservatives in the food industry. It is a cationic peptide of 34 amino acid residues that exhibits antimicrobial activity against Gram-positive bacteria. To ensure nisin stability in food matrices, new nisin-loaded ulvan particles were developed by the complexation method. The interaction of nisin with ulvan was demonstrated by FT-IR spectroscopy and differential scanning calorimetry. The encapsulation efficiency was calculated at different pH values within the range of 4.0–7.0 and was found to have the highest value at pH 7.0. The size and surface charge of particles fabricated at different concentrations of nisin and pH values were determined. Nisin-loaded ulvan particles exhibited antimicrobial activity against Gram-positive bacteria comparable to that of free nisin. Therefore, the developed complexes have the potential for application as biopreservatives in the food industry. For the first time, the potential of ulvan as a carrier of antimicrobial agent nisin was demonstrated.

Dual-Propelled Lanbiotic Based Janus Micromotors for Selective Inactivation of Bacterial Biofilms.

Graphene oxide/PtNPs/Fe2 O3 "dual-propelled" catalytic and fuel-free rotary actuated magnetic Janus micromotors modified with the lanbiotic Nisin are used for highly selective capture/inactivation of gram-positive bacteria units and biofilms. Specific interaction of Nisin with the Lipid II unit of Staphylococcus Aureus bacteria in connection with the enhanced micromotor movement and generated fluid flow result in a 2-fold increase of the capture/killing ability (both in bubble and magnetic propulsion modes) as compared with free peptide and static counterparts. The high stability of Nisin along with the high towing force of the micromotors allow for efficient operation in untreated raw media (tap water, juice and serum) and even in blood and in flowing blood in magnetic mode. The high selectivity of the approach is illustrated by the dramatically lower interaction with gram-negative bacteria (Escherichia Coli). The double-propulsion (catalytic or fuel-free magnetic) mode of the micromotors and the high biocompatibility holds considerable promise to design micromotors with tailored lanbiotics that can response to the changes that make the bacteria resistant in a myriad of clinical, environmental remediation or food safety applications.

Dual‐Propelled Lanbiotic Based Janus Micromotors for Selective Inactivation of Bacterial Biofilms

AbstractGraphene oxide/PtNPs/Fe2O3 “dual‐propelled” catalytic and fuel‐free rotary actuated magnetic Janus micromotors modified with the lanbiotic Nisin are used for highly selective capture/inactivation of gram‐positive bacteria units and biofilms. Specific interaction of Nisin with the Lipid II unit of Staphylococcus Aureus bacteria in connection with the enhanced micromotor movement and generated fluid flow result in a 2‐fold increase of the capture/killing ability (both in bubble and magnetic propulsion modes) as compared with free peptide and static counterparts. The high stability of Nisin along with the high towing force of the micromotors allow for efficient operation in untreated raw media (tap water, juice and serum) and even in blood and in flowing blood in magnetic mode. The high selectivity of the approach is illustrated by the dramatically lower interaction with gram‐negative bacteria (Escherichia Coli). The double‐propulsion (catalytic or fuel‐free magnetic) mode of the micromotors and the high biocompatibility holds considerable promise to design micromotors with tailored lanbiotics that can response to the changes that make the bacteria resistant in a myriad of clinical, environmental remediation or food safety applications.

Influence of Free Fatty Acids on Lipid Membrane-Nisin Interaction.

The influence of free fatty acids (FFAs) on the nisin–membrane interaction was investigated through micro-DSC and fluorescence spectroscopy. A simple but informative model membrane was prepared (5.7 DMPC:3.8 DPPS:0.5 DOPC molar ratio) by considering the presence of different phospholipid headgroups in charge and size and different phospholipid tails in length and unsaturation level, allowing the discrimination of the combined interaction of nisin and FFAs with the single phospholipid constituents. The effects of six FFAs on membrane stability were evaluated, namely two saturated FFAs (palmitic acid and stearic acid), two monounsaturated FFAs (cis-unsaturated oleic acid and trans-unsaturated elaidic acid) and two cis-polyunsaturated FFAs (ω-6 linoleic acid and ω-3 docosahexaenoic acid). The results permitted assessment of a thermodynamic picture of such interactions which indicates that the peptide–membrane interaction does not overlook the presence of FFAs within the lipid bilayer since both FFAs and nisin are able to selectively promote thermodynamic phase separations as well as a general lipid reorganization within the host membrane. Furthermore, the magnitude of the effects may be different depending on the FFA chemical structure as well as the membrane lipid composition.

A natural food preservative peptide nisin can interact with the SARS-CoV-2 spike protein receptor human ACE2

Nisin, a food-grade antimicrobial peptide produced by lactic acid bacteria has been examined for its probable interaction with the human ACE2 (hACE2) receptor, the site where spike protein of SARS-CoV-2 binds. Among the eight nisin variants examined, nisin H, nisin Z, nisin U and nisin A showed a significant binding affinity towards hACE2, higher than that of the RBD (receptor binding domain) of the SARS-CoV-2 spike protein. The molecular interaction of nisin with hACE2 was investigated by homology modeling and docking studies. Further, binding efficiency of the most potent nisin H was evaluated through the interaction of hACE2:nisin H complex with RBD (receptor-binding domain) of SARS-CoV-2 and that of hACE2:RBD complex with nisin H. Here, nisin H acted as a potential competitor of RBD to access the hACE2 receptor. The study unravels for the first time that a globally used food preservative, nisin has the potential to bind to hACE2.

Characterization, Release Profile and Antimicrobial Properties of Bioactive Polyvinyl Alcohol-Alyssum homolocarpum Seed Gum- Nisin Composite Film

A common goal of active packaging is to improve the shelf life, safety, or quality of packaged foods. The integrity of an active package must be remained in order to prevent the growth of microorganisms on the surface of food. Therefore, active polyvinyl alcohol-Alyssum homolocarpum seed gum (PVA-AHSG) composite films with different nisin concentrations (3000, 5000 and 10,000 IU) were prepared and their physico-chemical and antimicrobial properties were determined. Addition of nisin to the composite films increased their water vapor permeability (WVP), elongation at break (EB) and opacity, whereas their total color difference (ΔE), glass transition temperature (Tg), melting temperature (Tm), tensile strength (TS) and young modulus (YM) were decreased. Increasing the nisin concentration remarkably increased the chain mobility, interactions between polymers and water molecules and also the free volume of polymer matrix. The antimicrobial potential of film against L. monocytogene, S. aureus and E. coli as well as the release of nisin into phosphate buffer solution (pH 7.2) were investigated. Films containing nisin had inhibition effect against gram positive pathogens among which L. monocytogenes was the most sensitive bacterium. In liquid media, all films containing nisin prevented the growth of L. monocytogenes and S. aureus, but it was only the film with 10,000 IU nisin content which was able to control 100% of the microbial population during incubation time. Nisin release and diffusion coefficient (D) increased as its concentration increased in the film matrix due to the interaction of nisin with film polymer chains. Therefore, the resultant film had appropriate controlled release property and suitable microbial inhibitory against gram positive bacteria. Graphical Abstract Composite bioactive films based on polyvinyl alcohol-Alyssum homolocarpum seed gum blend and Nisin: Physiochemical characterization and antimicrobial properties. Monjazeb et al. (2017).

Effect of high pressure on the antimicrobial activity and secondary structure of the bacteriocin nisin

Abstract Effect of high pressure (HP) treatment on the antimicrobial properties and the structure of nisin was evaluated. Nisin solutions at pH 2.8 or 6.1 were treated by HP at 500 MPa – 10 min – 20 °C and their antimicrobial potency was determined. It appeared that HP clearly impacted the antimicrobial activity of nisin, with respective activity loss of 22.5% and 49.9% at pH 2.8 and 6.1. Structural analysis of nisin by circular dichroism and Fourier transform-infrared spectroscopies revealed that the decrease of nisin antimicrobial activity was likely due to the unfolding of the protein induced by HP. A loss of nisin β-turns structure, particularly significant at neutral pH, was linked to the drastic drop in antimicrobial activity, as these structures are implicated in the nisin interaction with the bacterial membrane. Industrial relevance The combination of nisin and high pressure (HP) can be use at an industrial scale to inactivate bacteria. Nisin is allowed as a food additive (E234) and can be added at a final concentration ranging from 120 to 500 IU/g, depending on the product. In this work, we showed that HP can induce a significant reduction of nisin activity (-22.5% at pH 2.8 and -49.9% at pH 6.1). Therefore, this activity loss could be taken into account to manage the final nisin concentration in HP-treated food products.

Synergistic antimicrobial effects of nisin and p-Anisaldehyde on Staphylococcus aureus in pasteurized milk

Staphylococcus aureus (S. aureus) infections resistant to a variety of drugs have been increasingly documented in recent years. Drug resistance has appeared largely due to extensive use of antibiotics. Nisin has been widely used in food industry to prevent bacterial growth. However, nisin resistance has been found in several foodborne pathogenic bacteria. p-Anisaldehyde (AS), an extract from Pimpinella anisum seeds, is a very common digestive herb of north India. In this study, synergistic interactions of nisin combined with AS against S. aureus were observed. The synergy was confirmed by checkerboard microdilution method, with the fractional inhibitory concentration index values ranging from 0.25 to 0.375. The positive interactions were verified by the challenge tests in pasteurized milk and agar diffusion assays. In addition, when treatment with nisin or AS alone or in combination, an obvious loss of 260 nm absorbing materials was observed. And flow cytometry assay showed that nisin or AS alone or in combination caused bacterial membrane permeabilization. The LIVE/DEAD BacLight experiment, scanning electron microscope and transmission electron microscopy observation further confirmed that the compounds could kill bacteria via disrupting membranes. All results above verified that the combination of nisin and AS which showed antibacterial activity by a membrane damage mechanism could not only reduce drug resistance but also be used as a new promising naturally sourced food preservative.

Biodegradable and antimicrobial films based on poly(butylene adipate-co-terephthalate) electrospun fibers

Biodegradable poly(butylene adipate-co-terephthalate) (PBAT) films incorporated with different levels of the antimicrobial peptide nisin were developed using the electrospinning technique. The characterization included thermal, structural, morphological, mechanical and antimicrobial properties. Thermal analysis indicated good thermal stability of PBAT. Nisin incorporation seems to increase nanofiber stability. The PBAT/nisin fibers presented no significant differences in the melting temperature (124–125.4 °C) and the glass transition temperature. PBAT showed characteristic diffraction peaks of the crystal structure. PBAT fibers were uniformly distributed and nisin was well dispersed throughout the fiber. The samples showed similar mechanical properties, and the addition of nisin caused no significant changes in the values of tensile strength, although Young’s modulus tended to decrease with higher nisin levels. Antimicrobial fibers inhibited the Gram-positive bacterium Listeria monocytogenes. These results provided insights into the interaction of nisin and PBAT in nanofibers produced by the electrospinning technique and their application in the food packaging industry.

Synergistic interactions of nisin in combination with cinnamaldehyde against Staphylococcus aureus in pasteurized milk

Staphylococcus aureus (S. aureus) is the leading cause of a broad range of human infections and associated with food poisoning worldwide. Nisin can be considered as a natural preservative and is widely used by the food industry. However, resistance of nisin has been reported in several species of bacteria. Identification of antibacterial agents that are synergistic with nisin is a method of minimising resistance to nisin. Cinnamaldehyde (CA), a well-studied essential oil isolated from cinnamon bark, has been reported to exhibit a broad spectrum of antimicrobial activity across a wide range of microorganisms. Previous studies have demonstrated that CA could be used in the food industry due to its active antibacterial activity and its safety. In this study, the synergistic antibacterial activity in vitro of nisin and CA against 13 food-borne isolates of S. aureus and S. aureus ATCC 29213 were evaluated by a checkerboard assay. The FICI values of nisin combined with CA ranged from 0.1875 to 0.375, which demonstrated a synergistic effect. The positive interactions were verified using the challenge tests in pasteurized milk and agar diffusion assays. The LIVE/DEAD BacLight experiment and scanning electron microscope (SEM) assay were also performed to distinguish the intact and damaged both of cell walls and membranes with the compounds, the combination group displayed stronger damage both of cell wall and cell membrane than when nisin or CA was used alone. These findings indicated that the combination of nisin and CA could not only be used as a promising naturally sourced food preservative but also to reduce the problem of bacterial resistance.

Effect of nisin and perilla oil combination against Listeria monocytogenes and Staphylococcus aureus in milk.

In the present study, in vitro interaction of nisin and perilla oil (PO) against 20 food-borne isolates of L. monocytogenes and S. aureus were assessed using a checkerboard microdilution method. Synergism was observed in tested strains with the fractional inhibitory concentration indexs (FICIs) ranges from 0.125-0.25 and 0.19-0.375, respectively. Scanning electron microscopy was carried out to investigate the effect of nisin and PO on the integrity of cell wall and membrane of L. monocytogenes and S. aureus. The results showed that nisin and PO were more effective in damaging cell wall and membrane in combination.

ارزیابی فاکتورهای موثر در تشکیل نانولیپوزوم های حاوی نایسین با استفاده از روش سطح پاسخ

Nisin has numerous applications as a natural preservative in foods, including dairy product, canned food, processed cheese and milk. Several studies demonstrated that proteolytic degradation and the interaction of nisin with food components might result in decreased its antimicrobial activity. Encapsulation of antimicrobial peptides into nanoliposomes may offer a potential alternative to protect antimicrobials, enhancing their efficacy and stability for food applications. In first stage of this research, Response Surface methodology was used for optimization of nanoliposomes produced by heating method. A central composite design (CCD) consisting of 18 experimental run with three independent variables: phospholipid concentration (2-30 mM), stirring speed (500-1360 rpm) and processing time (30-90 min) were used and their effects on size of nanliposome were evaluated.In the next stages, stability of nanoliposomes was investigated during 2 month. The optimum operating conditions obtained from the quadratic form of RSM model for particle size were phospholipids 30 (mM), stirring speed 930 (rpm) and process time 90 (min). The results of stability indicated that samples in the range of 400 to 500 nm were stable up to 2 month (P > 0/05) but samples larger than 500 nm were unstable during 2 month but stable up to 1 month(P> 0/05).

In Vitro synergy of nisin and coenzyme Q0 against Staphylococcus aureus

Nisin has been widely used as a natural preservative in foods including dairy products, but the emergence of the nisin-resistant strains could compromise its use to control food-borne pathogens. It has been reported that coenzyme Q (CoQ) is a powerful antioxidant, but a small number of studies have described the antibacterial activity of CoQ. One of the most abundant forms of CoQ is CoQ10. CoQ0 is chosen for this study, which is the water-soluble homolog of CoQ10. In the present study, the in vitro interaction of nisin and CoQ0 against 15 food-borne isolates of Staphylococcus aureus was assessed using a checkerboard microdilution method. Synergism was observed in strains tested, the FICI values ranging from 0.25 to 0.375. No antagonistic interaction between nisin and CoQ0 occurred. The positive interactions were verified confirmed using the time-killing test and agar diffusion assay. Our discovery of efficacy of combining nisin with CoQ0 might pave the way for a novel solution for spoilage problem in food industry and provide an alternative approach to overcome antimicrobial drug resistance.