Heat stability of allyl isothiocyanate and phenyl isothiocyanate complexed with randomly methylated β-cyclodextrin

Inclusion complexation reactions between isothiocyanates (ITCs), namely, allyl isothiocyanate (AITC) and phenyl isothiocyanate (PITC), and randomly methylated β-cyclodextrin (RM-β-CD) displayed “A_P” type solubility isotherms, indicating continuous increases in the stoichiometry of the inclusion complexes. The complex powders were prepared by spray drying and their heat stability was studied by thermal analysis and heat treatment in paraffin. The solid-state dissociation reactions were satisfactorily described by the unimolecular decay law. The apparent activation energy of dissociation, E_D, was determined to be 48 and 58 kJ/mol, respectively, for the AITC/RM-β-CD and PITC/RM-β-CD complexes, while the apparent activation energies of decrease of ITCs in paraffin, E_P for the pure AITC and PITC, and the corresponding inclusion complexes were 26, 48, 44, and 54 kJ/mol, respectively, suggesting volatility suppression and physical stabilisation by inclusion complexation as the plausible mode of stabilisation of the ITCs in paraffin. A compensation effect in the kinetics of depletion of the ITCs in paraffin was established.     

Properties of cholesterol-reduced butter made with beta-cyclodextrin and added evening primrose oil and phytosterols

The present study was carried out to examine changes in the chemical and sensory properties of butter in which the cholesterol was reduced and to which evening primrose oil (EPO) and phytosterols were added. Crosslinked β-cyclodextrin (β-CD) made from adipic acid was used, and approximately 90% of the cholesterol was removed. The color measurement values “L” and “a” were significantly different between the control (butter with no β-CD treatment and no added EPO and phytosterols) and treatment A (butter treated with 10% crosslinked β-CD); however, the color values for “L” and “a” were similar. The color value “b” in treatment B (butter treated with 10% crosslinked β-CD and 5% phytosterols and 3% EPO added) was significantly higher than in the other treatments. The thiobarbituric acid value of treatment B was significantly higher than that of the control and treatment A. Scores for hardness, elasticity, and cohesiveness were significantly lower in the control than in treatment A. Differences in sensory characteristics did not result from the β-CD treatment but from the addition of EPO and phytosterols. In microscopic examinations, no noticeable differences were found among the treatments, and a smooth texture and a fine, uniform crystalline structure were observed. Results indicated that about 90% of the cholesterol was reduced by crosslinked β-CD and that the β-CD treatment itself did not adversely influence the chemical and sensory properties of the butter. However, the addition of EPO and phytosterols to the butter appeared to impair its sensory properties, especially in terms of rancidity and overall acceptability.     

Comparison tests of hydroxylpropyl β-cyclodextrin (HPβ-CD) and β-cyclodextrin (β-CD) on retrogradation of rice amylose

Retarding effect of hydroxypropyl β-cyclodextrin (HPβ-CD) and β-cyclodextrin (β-CD) on the retrogradation of rice amylose was compared and studied. Size-exclusion high performance liquid chromatography (SE-HPLC) revealed that the degree of amylose retrogradation was reduced more by HPβ-CD than by β-CD. HPβ-CD preferentially interacted with amylose to form the potential amylose-HPβ-CD complex and lessened the stored amylose decrease in molecular weight (Mw). Differential scanning calorimeter (DSC) detected the potential complex formation as well. DSC data was analyzed using Avrami equation. Results showed that HPβ-CD significantly decreased the constant rate (k) and increased the Avrami exponent (n). The X-ray diffraction (XRD) patterns of retrograded amylose were further analyzed. It indicated that HPβ-CD caused an existence of intermediate type (V + B) and retarded the transformation from V- to B-type during the storage of the prepared amylose sample.     

Hydroxypropyl β-cyclodextrin improving multiple stresses tolerance of Lactococcus lactis subsp. lactis

L. lactis is known as industrial starter in the fermentation of dairy and meat products, and it plays an important role in human health as an edible probiotic. During industrial production, L. lactis often experiences different stresses that delay the growth and decrease the survival in some serious conditions. In this study, the protective effects of hydroxypropyl β-cyclodextrin (HP β-CD) on L. lactis under multiple stresses were investigated. The microbial cells were treated with different stresses including heat, NaCl, cold, and H₂O₂ stresses, and the results were showed by measuring the OD₆₀₀ or spot plating method. The growth and tolerance were improved when HP β-CD was added during different stress conditions, better than that of trehalose. Besides, the scanning electron microscopic and fluorescence spectrum studies showed that HP β-CD could combine with L. lactis to protect the cell structure, suggesting that HP β-CD may act as a protective agent of L. lactis. Therefore, HP β-CD could be considered as a potential protective agent to be applied in food industry, and its protective mechanism on L. lactis still needs further investigation.     

Release of Allyl Isothiocyanate from Mustard Seed Meal Powder

Allyl isothiocyanate (AITC) is a wide‐spectrum antimicrobial compound found in mustard seeds, produced when their tissues are disrupted. The formation of AITC in mustard seed is mediated by the myrosinase enzyme which catalyzes the release of volatile AITC from a glucosinolate—sinigrin. Since water is a substrate in the reaction, humidity from the air can be used to activate the release of AITC from mustard seed. In this study, defatted and partially defatted mustard seed meals were ground into powders with particle size ranging from 5 to 300 μm. The mustard seed meal powder (MSMP) samples were enclosed within hermetically sealed glass jars wherein the headspace air was adjusted to 85% or 100% relative humidity at 5, 20, or 35 °C. Data from gas chromatography analysis showed that AITC release rate and amount increased with increasing relative humidity and temperature. Moreover, the release rate can be manipulated by particle size and lipid content of the MSMP samples. The amount of AITC released ranged from 2 to 17 mg/g MSMP within 24 h under the experimental conditions tested. In view of the antimicrobial properties of AITC, the mustard meal powder may be used as a natural antimicrobial material for extending the shelf life of food products.     

Chemical and antimicrobial analyses of essential oil of Lippia origanoides H.B.K

Lippia origanoides H.B.K. (Verbenaceae) is a plant known in Oriximiná (Brazil) as “Salva-de-Marajó”. Its leaves are widely used as of a spice in cooking and in traditional medicine. The chemical composition of the essential oil obtained from its leaves, analyzed by GC and GC/MS, showed a high content of oxygenated monoterpenes (66.0%), carvacrol (38.6%) and thymol (18.5%) being the major constituents. Considering that previous studies on the same plant species showed carvacrol as a trace or absent compound, we propose the existence of a new chemotype for this species. A high carvacrol content in the essential oil determines the plant’s suitability for the preparation of oregano condiment. The antimicrobial activity of this essential oil was determined by the drop diffusion method, showing highly significant inhibition zones for all microorganisms tested.     

Effects of the origins of Botrytis cinerea on earthy aromas from grape broth media further inoculated with Penicillium expansum

Earthy “off” aromas from wine and grape juice are highly detrimental to the production of quality grape products. These volatile compounds are produced on grapes by Botrytis cinerea, Penicillium expansum and/or a combination of P. expansum and B. cinerea strains. B. cinerea strains were isolated from different (a) vineyards in Spain and Portugal, (b) grape varieties (c) bunches (i.e., sound and botrytized) and (d) positions in the botrytized bunch (i.e., interior or exterior). A novel Headspace-Phase Microextraction (SPME) followed by Gas Chromatrography/Mass Spectrometry (GC-MS) dedicated to analyze geosmin, methylisoborneol (MIB), 1-octen-3-ol, fenchone and fenchol in grape broth medium was used. Approximately 50% of the B. cinerea strains induced detectable geosmin. One strain accumulated significant amounts of anisoles, demonstrating that this contamination might already occur in the vineyard. Strains from the interior of Cainho grape bunches induced more geosmin and hence it may be possible to reduce this volatile in wine by avoiding using these grapes in case of B. cinerea attack.     

Effect of hydrostatic pressure and antimicrobials on survival of Listeria monocytogenes and enterohaemorrhagic Escherichia coli in beef

The effects of marinades on Listeria monocytogenes and enterohaemorrhagic Escherichia coli were investigated in pressure treated beef steaks. Meat was treated with 600 MPa or 450 MPa. Marinades did not enhance pressure inactivation of E. coli in beef steaks and marinades prevented pressure-induced sublethal injury in L. monocytogenes. Membrane-active essential oils carvacrol and thymol, and thiol-reactive allyl-isothiocyanate (AITC) and cinnamaldehyde were selected to investigate potential synergistic activity of clean label antimicrobials with pressure. Carvacrol accelerated pressure inactivation of E. coli in beef steaks; however, carvacrol increased pressure resistance of E. coli in buffer, and had no effect on survival of E. coli in ground beef. Thymol had no effect in either buffer or meat. AITC and cinnamaldehyde exhibited synergistic activity with pressure on E. coli in buffer; however, cinnamaldehyde did not affect survival of E. coli after pressure treatment of meat. Synergistic inactivation of AITC with pressure was observed only at concentrations that are negatively affect meat quality. AITC and carvacrol may be practically applied for enhancing the bacterial inactivation and extending the shelf life of beef steaks.     

The influence of electron beam irradiation of antimicrobial-coated LDPE/polyamide films on antimicrobial activity and film properties

We evaluated the effects of ionizing radiation (1-3 kGy) and incorporation of antimicrobials on the functional properties of low-density polyethylene (LDPE)/polyamide films. We established the antimicrobial effectiveness of several coatings of FDA-approved antimicrobial compounds including sorbic acid, carvacrol, trans-cinnamaldehyde, thymol and rosemary oleoresin using selected food pathogen surrogates. The antimicrobial coatings were applied to one side of the LDPE films and dried. Films were irradiated using a 10 MeV linear electron beam accelerator at room temperature. All films showed inhibition zones in an agar diffusion test against Listeria innocua ATCC 33090 and Escherichia coli ATCC 884. In the liquid culture test, the antimicrobials significantly (p?0.05) reduced the specific growth rate of L. innocua by 3.8-8.5%, and decreased final cell concentration of both strains by 5.7-14.6% and 7.2-16.8%, respectively. All active compounds retained the antimicrobial activity when exposed to 1-3 kGy. Neither the presence of active compound nor dose affected the film's tensile strength and toughness. Additionally, films became more ductile and had improved moisture barrier functionality. Film's oxygen permeability was not affected by either treatment. Results are an initial step toward the development of self-sterile active packaging materials for use in combination with irradiation treatment of foods.     

Kinetic model of apple juice enzymatic browning in the presence of cyclodextrins: The use of maltosyl-β-cyclodextrin as secondary antioxidant

Enzymatic browning reactions limit the commercial shelf life of apple juice, so that colour preservation during storage is one of the main objectives of fruit processors. In this paper, the colour of fresh apple juice was evaluated in the presence of different types of cyclodextrins (CDs) (α-CD, β-CD, γ-CD and maltosyl-β-CD), compounds that bind or complex polyphenol oxidase substrates. The effectiveness of CDs as browning inhibitors was determined as the difference between the colours observed in the CD-treated sample and the controls, using the colour space CIE-L^∗, a^∗, b^∗ system. Although the effect of CDs on apple juice enzymatic browning has been studied, the action mechanism involved remains a subject of controversy. In this work, we have kinetically modelled apple juice enzymatic browning in the absence and presence of maltosyl-β-CD. The complexation constant between the mixtures of diphenols present in apple juice and maltosyl-β-CD was calculated (K_c = 4.09 mM⁻¹). Different concentrations of maltosyl-β-CD modified the evolution of lightness (L^∗), total colour (ΔE^∗) and hue angle (H^∗) because of the higher affinity constant it shows for the compounds responsible for apple juice browning than do α-CD, β-CD and γ-CD. Moreover, in this paper we show that maltosyl-β-CD can enhance the ability of ascorbic acid to prevent the enzymatic browning due to the protective effect of maltosyl-β-CD against ascorbic acid oxidation. Hence, maltosyl-β-CD seems to act as a “secondary antioxidant”, reducing apple juice browning and enhancing the naturally occurring antioxidant capacity of a food.     

Complexation of allyl isothiocyanate with β-cyclodextrin and its derivatives and molecular microcapsule of allyl isothiocyanate in β-cyclodextrin

Complexation reactions of allyl isothiocyanate (AITC) that is the major component of horseradish oil, with β-cyclodextrin (β-CD), mono[2-O-(2-hydroxypropyl)]-β-CD (2-HP-β-CD), mono[2-O-(2-hydroxyethyl)]-β-CD (2-HE-β-CD), and heptakis(2,6-di-methyl)-β-CD (DM-β-CD) were investigated using spectrophotometry with methyl orange (MO) as a molecular probe. The effects of temperature, ionic strength and concentration of methanol on the equilibria were determined. The results suggest that the affinity between β-CD with its derivatives and AITC was the most important driving force to form the inclusion complexes. The association constants of CDs with AITC (K) decreased with the increasing of temperature from 36.3 to 24.5 l/mol and increased with the increasing ionic strength. The thermodynamic measurements showed that the inclusive process was enthalpic favored. In addition, the molecular microcapsule of AITC in β-CD was prepared by suspension method. The content of AITC in the microcapsules was 43.8 mg/g and their stabilities were obviously improved because of Van der Waals forces and the steric hindrance.     

Synergy between high-pressure, temperature and ascorbic acid on the inactivation of Bacillus cereus

B. cereus strain (CECF 148) was used as a model system in the study of the behaviour of bacillus under high pressure, at temperatures over and below 0 °C and with ascorbic acid added to the culture. Three different assays were carried out in the present experiment. The first assay was performed to observe how B. cereus reacted to pressure shift freezing (PSF) treatment at different subzero temperatures (−8, −12, −20 and −17 °C) and pressures (120, 150, 210 and 350 MPa) in their vegetative form. In the second assay, we observed how different concentrations of ascorbic acid (1, 2, 5 and 20 mM) added to the growing brought decreased B. cereus on its vegetative form. Finally, we tried to inactivate the vegetative and spore form of B. cereus under pressure of 210 MPa at room (20 °C) (HP) and at subzero (−20 °C) (PSF) temperatures, in presence of ascorbic acid (20 mM), added to the growing culture (TSB). The results confirmed that pressures of 210 and 350 MPa at low temperatures (−20 and −17 °C) in the PSF treatment were not enough to inactivate bacillus and only about 10% of B. cereus at the assayed conditions (350 MPa at −17 °C) lost its growth capacity. The presence of ascorbic acid reduced the amount of B. cereus. The initial amount of B. cereus in the vegetative form was 10⁸ to 10⁹ cfu/mL. After HP (210 MPa at 20 °C) and PSF (210 MPa at −20 °C) treatments, the amount of B. cereus decreased by 4 and 2 logarithmic units, respectively. However, in both treatments, the presence of ascorbic acid (20 mM concentration) reduced the B. cereus growth capacity in about 5 logarithmic units. The presence of ascorbic acid in the spore form decreased the amount of B. cereus only by 2 logarithmic units, but without the antioxidant, the values remained close to control. The present research is a contribution to elicit the safety standards of food treated by high pressure.     

Enhanced antimicrobial activity of nisin in combination with allyl isothiocyanate against Listeria monocytogenes,Staphylococcus aureus,Salmonella Typhimurium and Shigella boydii

This study was designed to evaluate the synergistic antimicrobial effect of nisin and allyl isothiocyanate (AITC) against Listeria monocytogenes, Staphylococcus aureus, Salmonella Typhimurium and Shigella boydii. The synergistic interactions between nisin and AITC were observed against all foodborne pathogens, showing the fractional inhibitory concentrations <1. The populations of L. monocytogenes and S. aureus at the combined treatment of nisin and AITC were decreased to below 1 log CFU mL^?1 after 10‐h incubation at 37 °C. The changes in fatty acid profiles of all strains were substantially influenced by nisin alone and the combined treatment of nisin and AITC. A good agreement was observed among cell viability, membrane permeability and depolarisation activity in response to nisin and AITC. The results suggest that nisin and AITC as synergistic inhibitors could be an effective approach to achieve satisfactory antimicrobial activity against a wide range of foodborne pathogens.     

Functional properties and in vitro trypsin digestibility of red kidney bean (Phaseolus vulgaris L.) protein isolate: Effect of high-pressure treatment

The effects of high-pressure (HP) treatment at 200–600 MPa, prior to freeze-drying, on some functional properties and in vitro trypsin digestibility of vicilin-rich red kidney bean (Phaseolus vulgaris L.) protein isolate (KPI) were investigated. Surface hydrophobicity and free sulfhydryl (SH) and disulfide bond (SS) contents were also evaluated. HP treatment resulted in gradual unfolding of protein structure, as evidenced by gradual increases in fluorescence strength and SS formation from SH groups, and decrease in denaturation enthalpy change. The protein solubility of KPI was significantly improved at pressures of 400 MPa or higher, possibly due to formation of soluble aggregate from insoluble precipitate. HP treatment at 200 and 400 MPa significantly increased emulsifying activity index (EAI) and emulsion stability index (ESI); however, EAI was significantly decreased at 600 MPa (relative to untreated KPI). The thermal stability of the vicilin component was not affected by HP treatment. Additionally, in vitro trypsin digestibility of KPI was decreased only at a pressure above 200 MPa and for long incubation time (e.g., 120 min). The data suggest that some physiochemical and functional properties of vicilin-rich kidney proteins can be improved by means of high-pressure treatment.     

超高压促进β-环糊精向大米颗粒内部的有效渗入

文中探究了超高压对β-环糊精在大米颗粒中的渗入情况。在粳米浸泡液中添加2%β-环糊精在60℃温水中浸泡30 min,随后在超高压(100～500 MPa)下浸泡20 min,最后将浸泡后的大米蒸汽催熟并焖饭制成米饭。采用高效液相法定量检测浸泡米液残留的β-环糊精含量,研究发现随着压强的增大,浸泡液中β-环糊精的含量显著降低,表示渗入米粒的β-环糊精更多。通过β-环糊精包埋1,8-二羟基蒽醌荧光剂来示踪β-环糊精在超高压浸泡过程中向米粒内部的渗入,用激光共聚焦显微镜观察常压处理和超高压处理米饭切片的荧光分布。实验发现超高压能有效地促进β-环糊精渗透到米粒内部。     

Active Packaging of Fresh Chicken Breast,with Allyl Isothiocyanate (AITC) in Combination with Modified Atmosphere Packaging (MAP) to Control the Growth of Pathogens

ABSTRACT: Listeria monocytogenes and Salmonella typhimurium are major bacterial pathogens associated with poultry products. Ally isothiocyanate (AITC), a natural antimicrobial compound, is reportedly effective against these pathogenic organisms. A device was designed for the controlled release of AITC with modified atmosphere packaging (MAP), and then evaluated for its ability to control the growth of L. monocytogenes and S. typhimurium on raw chicken breast during refrigerated storage. In order to obtain controlled release during the test period, a glass vial was filled with AITC and triglyceride. It was then sealed using high-density polyethylene film. The release of AITC was controlled by the concentration (mole fraction) of AITC in the triglyceride and by the AITC vapor permeability through the film. The fresh chicken samples were inoculated with one or the other of the pathogens at 10⁴ CFU/g, and the packages (with and without AITC-controlled release device) were flushed with ambient air or 30% CO₂/70% N₂ before sealing, and then stored at 4 °C for up to 21 d. The maximum reduction in MAP plus AITC (compared to MAP alone) was 0.77 log CFU/g for L. monocytogenes and 1.3 log CFU/g for S. typhimurium. The color of the chicken breast meat was affected by the concentration of AITC. Overall, a release rate of 0.6 μg/h of AITC was found to not affect the color, whereas at 1.2 μg/h of AITC the surface of the chicken was discolored.     

Effect of hydroxypropyl β-cyclodextrin on physical properties and transition parameters of amylose–lipid complexes of native and acetylated starches

The effect of hydroxpropyl β-cyclodextrin (HPβ-CD) on physical properties and digestibility of wheat, potato, waxy maize and high-amylose maize starches before and after acetylation was studied. Effect of HPβ-CD on amylose–lipid complexes in native and acetylated potato starches synthesized using α-lysophosphatidylcholine was also studied. Acetylation increased swelling factor, amylose leaching, peak viscosity and susceptibility to α-amylase hydrolysis, but decreased gelatinization temperature and enthalpy and gel hardness in all starches. HPβ-CD markedly increased swelling factor and amylose leaching in native and acetylated wheat starches but had little or no impact on other starches. Wheat starch gelatinization enthalpy decreased in the presence of HPβ-CD but gelatinization temperature of all the starches was slightly increased. HPβ-CD had no influence on enzymatic hydrolysis. Melting enthalpy of amylose–lipid complex in both native and acetylated wheat starches was decreased by HPβ-CD. Acetylation also decreased the melting enthalpy of amylose–lipid complex in wheat starch. Similar trend of thermal transitions was observed in the presence of HPβ-CD for the amylose–lipid complexes synthesized in potato starch. Acetylation reduces the complex formation ability of the amylose polymer. Similar to gelatinization, acetylation widened the melting temperature range of amylose–lipid complexes while shifting it to a lower temperature. Higher swelling and amylose leaching, and decreased gelatinization temperature and enthalpy resulting from acetylation of wheat starch is consistent with its influence on starch hydration. Similar effects resulting from the inclusion of HPβ-CD were consistent with the disruption of amylose–lipid complex by HPβ-CD which promotes granular hydration.     

Antimicrobial edible apple films inactivate antibiotic resistant and susceptible Campylobacter jejuni strains on chicken breast

Abstract: Campylobacter jejuni is the leading cause of bacterial diarrheal illness worldwide. Many strains are now becoming multidrug resistant. Apple‐based edible films containing carvacrol and cinnamaldehyde were evaluated for bactericidal activity against antibiotic resistant and susceptible C. jejuni strains on chicken. Retail chicken breast samples inoculated with D28a and H2a (resistant strains) and A24a (a sensitive strain) were wrapped in apple films containing cinnamaldehyde or carvacrol at 0.5%, 1.5%, and 3% concentrations, and then incubated at 4 or 23 °C for 72 h. Immediately after wrapping and at 72 h, samples were plated for enumeration of viable C. jejuni. The antimicrobial films exhibited dose‐ and temperature‐dependent bactericidal activity against all strains. Films with ≥1.5% cinnamaldehyde reduced populations of all strains to below detection at 23 °C at 72 h. At 4 °C with cinnamaldehyde, reductions were variable for all strains, ranging from 0.2 to 2.5 logs and 1.8 to 6.0 logs at 1.5% and 3.0%, respectively. Films with 3% carvacrol reduced populations of A24a and H2a to below detection, and D28a by 2.4 logs at 23 °C and 72 h. A 0.5‐log reduction was observed for both A24a and D28a, and 0.9 logs for H2a at 4 °C at 3% carvacrol. Reductions ranged from 1.1 to 1.9 logs and 0.4 to 1.2 logs with 1.5% and 0.5% carvacrol at 23 °C, respectively. The films with cinnamaldehyde were more effective than carvacrol films. Reductions at 23 °C were greater than those at 4 °C. Our results showed that antimicrobial apple films have the potential to reduce C. jejuni on chicken and therefore, the risk of campylobacteriosis. Possible mechanisms of antimicrobial effects are discussed. Practical Application: Apple antimicrobial films could potentially be used in retail food packaging to reduce C. jejuni commonly present on food.     

Preparation of active antimicrobial methyl cellulose/carvacrol/montmorillonite nanocomposite films and investigation of carvacrol release

Methyl cellulose (MC)/carvacrol (CRV)/montmorillonite (MMT) nanocomposite films were prepared to obtain active antimicrobial packaging materials. The characterization of film samples by X-ray diffraction and transmission electron microscopy showed that composite films were of nano structures. CRV addition to the MC film and MC/MMT nanocomposite films led to a decrease in the thickness and opacity values of them, whereas MMT addition to the film matrix caused an increase in these values. Thermal stability of films slightly increased with increasing MMT concentration in film matrix. CRV release from films was investigated at different temperatures for 30 days. An increase in the MMT concentration matrix caused a decrease in CRV release at 25.0 ± 0.5 °C and in 60 ± 4% relative humidity (RH). CRV release increased with temperature at a constant RH. The antimicrobial activities of films were tested against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) by the microatmosphere method and these organisms were completely inhibited on the nutrient broth/bacteriological agar medium when film samples containing 11.1 ± 0.2 mg CRV were present. MC/CRV film and MC/CRV/MMT-60% nanocomposite films on sausage reduced E. coli and S. aureus counts by 0.9 and 0.7 log cfu/mL, respectively, compared to the control film. The amount of CRV release from developed antimicrobial films can be controlled by MMT concentration within the film matrix and by the storage temperature of film.     

Physical and antimicrobial properties of Gelidium corneum/nano-clay composite film containing grapefruit seed extract or thymol

To improve the physical properties of Gelidium corneum (GC) film, Cloisite Na⁺ and 30B were incorporated into the preparation of the film. X-ray diffraction patterns of the films indicated that a degree of exfoliation and intercalation are formed depending on the type of nano-clays and its concentration and the film structure affects the physical properties of the GC/nano-composite films. Tensile strength (TS) of the GC film was increased by the addition of nano-clays. GC film had a TS of 19.59 MPa for the control, while the GC film having 3% Cloisite Na⁺ or 30B had TS of 27.37 and 26.40 MPa, respectively. Elongation at break and water vapor permeability were also improved by the addition of 3% nano-clays. The physical properties of the film were not improved by the addition of GSE or thymol, but the additions did inhibit the growth of Escherichia coli O157:H7 and Listeria monocytogenes. The results suggest that the nano-composite films containing GSE or thymol may extend the shelf life of food.