Physicochemical properties and bioactivity of nisin-containing cross-linked hydroxypropylmethylcellulose films

Cross-linked hydroxypropylmethylcellulose (HPMC) cast films with citric acid as polycarboxylic cross-linker were elaborated to study the effect of cross-linking level on various properties. Increased amounts of cross-linking agent were not connected to statistically different tensile strength and Young's modulus. Whatever the cross-linking level of the film was, the ultimate elongation parameter decreased by approximately 60% compared to the HMPC control film. Moisture sorption isotherms and water contact angle meter showed that the effect of cross-linking degree tends to reduce the hygroscopic and hydrophilic characteristics of films. In addition, to control bacteria growth on food surfaces, the antimicrobial activity of both 98% cross-linked HPMC-nisin and control HPMC-nisin films was tested on Micrococcus luteus. Despite the incorporation of a significant content of nisin, cross-linked HPMC-nisin films were completely inactive on the microbial strain compared to the HPMC-nisin control films. Cross-linking conditions likely either denatured the nisin or irreversibly bound nisin to the cross-linked HPMC. However, nisin adsorbed into films made from previously cross-linked HPMC maintained its activity.     

Antimicrobial and physicochemical properties of chitosan-HPMC-based films

To prepare composite films from biopolymers with anti-listerial activity and moisture barrier properties, the antimicrobial efficiency of chitosan-hydroxy propyl methyl cellulose (HPMC) films, chitosan-HPMC films associated with lipid, and chitosan-HPMC films chemically modified by cross-linking were evaluated. In addition, the physicochemical properties of composite films were evaluated to determine their potential for food applications. The incorporation of stearic acid into the composite chitosan-HPMC film formulation decreased water sensitivity such as initial solubility in water and water drop angle. Thus, cross-linking of composite chitosan-HPMC, using citric acid as the cross-linking agent, led to a 40% reduction in solubility in water. The water vapor transfer rate of HPMC film, approximately 270 g x m(-2) x day(-1) x atm(-1), was improved by incorporating chitosan and was further reduced 40% by the addition of stearic acid and/or cross-linking. Anti-listerial activity of films was determined on solid medium by a numeration technique. Chitosan-HPMC-based films, with and without stearic acid, inhibited the growth of Listeria monocytogenes completely. On the other hand, a loss of antimicrobial activity after chemical cross-linking modification was observed. FTIR and 13C NMR analyses were then conducted in order to study a potential chemical modification of biopolymers such as a chemical reaction with the amino group of chitosan. To complete the study, the mechanical properties of composite films were determined from tensile strength assays.     

Water sensitivity, antimicrobial, and physicochemical analyses of edible films based on HPMC and/or chitosan

Several properties of chitosan films associated or not with hydroxypropylmethylcellulose polymer (HPMC) and HPMC films incorporating or not nisin and/or milk fat were studied. Nisin addition at a level of 250 microg mL-1 and likewise chitosan at 1% (w/v) concentration were efficient for total inhibiting Aspergillus niger and Kocuria rhizophila food deterioration microorganisms. HPMC and chitosan films were transparent, whereas nisin and/or fat incorporation induced a 2-fold lightness parameter increase and, consequently, involved more white films. Measurements of tensile strength, as well as ultimate elongation, showed that chitosan and HPMC initial films were elastic and flexible. High thermal treatments and additive incorporation induced less elastic and more plastic films. Water vapor transmission as far as total water desorption rates suggested that chitosan films were slightly sensitive to water. Water transfer was decreased by <60% as compared with other biopolymer films. Regarding its hydrophobic property, the capacity of fat to improve film water barrier was very limited.     

Fatty acid effect on hydroxypropyl methylcellulose-beeswax edible film properties and postharvest quality of coated 'Ortanique' mandarins

The objective of this work was to investigate the effect of fatty acid (FA) type and content on mechanical properties, water vapor permeability and oxygen permeability of hydroxypropyl methycellulose (HPMC)-beeswax (BW) stand-alone edible films. The effect of these films formed as coatings on the postharvest quality of 'Ortanique' mandarins was also studied. Selected FAs were stearic acid (SA), palmitic acid (PA), and oleic acid (OA), using BW/FA ratios of 1:0.5 and 1:0.2 (w/w). HPMCBW coatings reduced weight and firmness loss of 'Ortanique' mandarins, without compromising flavor quality compared to uncoated mandarins. Coatings containing OA provided the best weight loss control at both concentrations tested; however, when the BW/OA ratio was 1:0.5, the coatings increased fruit internal CO2, ethanol, and acetaldehyde contents of 'Ortanique' mandarins, therefore reducing flavor compared to the rest of the coatings studied. Although barrier and mechanical properties might be used to understand coating performance, differences observed between film oxygen permeability and coating permeability indicate that permeance should be measured on the coated fruit.     

硬脂酸-胱氨酸-大豆分离蛋白复合膜的机械特性和吸湿性研究

以硬脂酸作为增塑剂，胱氨酸作为交联剂制备具有一定力学性能和良好抗湿性能的大豆分离蛋白复合膜。将膜放在25℃，相对湿度为50%的干燥器中平衡两天，用质构仪测定膜的抗拉强度(TS),延伸率(E(%))。在水分活度a_w为0.10～0.90的范围内研究了复合膜在25℃的吸湿特性。吸湿速率和吸湿等温线数据分别拟合到Peleg's 方程和GAB(Guggenheim-Anderson-de Boer)模型。结果表明：大豆蛋白复合膜的TS、延伸率E(%)以及吸湿速率随着硬脂酸和胱氨酸的添加比率显著地变化。硬脂酸和胱氨酸的最佳添加比率为40∶60(w/w)(每升蛋白质溶液中加入10 g混合添加剂)，此时，大豆蛋白膜的强度比原来提高2倍，并且有最佳的吸湿速率。吸湿数据和GAB 模型有很高的拟合度，拟合系数最高达0.99。     

Synergistic antimicrobial activities of natural essential oils with chitosan films

The synergistic antimicrobial activities of three natural essential oils (i.e., clove bud oil, cinnamon oil, and star anise oil) with chitosan films were investigated. Cinnamon oil had the best antimicrobial activity among three oils against Escherichia coli , Staphylococcus aureus , Aspergillus oryzae , and Penicillium digitatum . The chitosan solution exhibited good inhibitory effects on the above bacteria except the fungi, whereas chitosan film had no remarkable antimicrobial activity. The cinnamon oil-chitosan film exhibited a synergetic effect by enhancing the antimicrobial activities of the oil, which might be related to the constant release of the oil. The cinnamon oil-chitosan film had also better antimicrobial activity than the clove bud oil-chitosan film. The results also showed that the compatibility of cinnamon oil with chitosan in film formation was better than that of the clove bud oil with chitosan. However, the incorporated oils modified the mechanical strengths, water vapor transmission rate, moisture content, and solubility of the chitosan film. Furthermore, chemical reaction took place between cinnamon oil and chitosan, whereas phase separation occurred between clove bud oil and chitosan.     

纳米SiO2对PVA基复合涂膜包装材料成膜透湿性能的影响

为改善聚乙烯醇（PVA）涂膜的阻湿保鲜效能特性,该文采用添加纳米SiO2对PVA基复合涂膜包装材料进行改性,应用响应曲面方法研究SiO2、硬脂酸、戊二醛对三元复合涂膜材料成膜效能特性的影响及交互作用。结果表明：加入纳米SiO2改性可有效提高PVA基复合涂膜包装材料的阻水、阻湿性能,优化组成膜透湿率8.18 g/(m2·d)比对照组（不添加纳米SiO2的PVA复合膜）降低26.61%（p<0.05）;硬脂酸、戊二醛对复合涂膜材料成膜透湿率的影响与纳米SiO2存在显著的交互作用,每100 mL 0.05 g/mL的PVA溶液中纳米SiO2添加量在小于0.05 g范围内随着SiO2含量增大,复合涂膜材料成膜透湿率随硬脂酸、戊二醛的比例增加而降低,阻湿性能提高。     

Water and moisture susceptibility of chitosan and paper-based materials: structure-property relationships

Environmentally friendly and potentially bioactive food packaging based on chitosan-coated papers were elaborated. The morphology and the microstructure of these new materials were characterized by infrared spectroscopy and scanning electron microscopy. These observations suggested that the chitosan penetrated deeply into the paper, embedding the cellulose fibers, instead of forming a layer as expected. Through the barrier properties against moisture, the liquid water sensitivity, and NMR-relaxometry measurements, the water interactions were evaluated on the chitosan films and the chitosan-coated papers. They revealed that the coating by a chitosan film forming solution improved the paper moisture barrier properties but the surface hydrophilicity remained high. Relaxometry studies showed that, due to its hydrophilic character, chitosan controlled the interaction with water, despite the very low amount of deposit. On the other hand, the mechanical properties of papers were unmodified by the chitosan coating, which did not fundamentally affect the solid structure of the papers.     

纳米SiO2对聚乙烯醇基复合涂膜包装材料成膜透湿性能的影响

聚乙烯醇（PVA）由于其亲水性能而影响其在食品包装上的应用。本研究采用添加纳米SiO2对聚乙烯醇基复合涂膜包装材料进行改性,利用响应曲面方法研究SiO2、硬脂酸、戊二醛对三元复合涂膜材料成膜效能特性的影响及交互作用。结果表明：加入纳米SiO2可有效提高聚乙烯醇基复合涂膜包装材料的阻水阻湿性能,优化组成膜透湿率（8.18 g•m-2•d-1）比对照组（不添加纳米SiO2的PVA复合膜）降低26.61%（p<0.05）。硬脂酸、戊二醛对复合涂膜材料成膜的透湿率的影响与纳米SiO2存在显著的交互作用,每100 mL 5%（W/VH2O）的PVA溶液中纳米SiO2添加量在小于0.05 g范围内随着其含量增大,复合涂膜材料成膜透湿率随硬脂酸、戊二醛的比例增加而降低,阻水阻湿性能提高。     

Compositional and moisture content effects on the biodegradability of zein/ethylcellulose films

The effect of moisture content and film composition on biodegradability is the focus of this study. Flexible films were first characterized for the effect on water sorption isotherms of relative humidity, temperature, zein content, and the addition of the plasticizers stearic acid, poly(ethylene glycol), or etoxylated ricine oil. Zein/ethylcellulose (EC) mixture films had a behavior between that for pure zein and EC films, which had the lowest water sorption. For films with plasticizer, the lowest water sorption at 25 degrees C was observed for those with stearic acid. Biodegradability of zein/EC films, evaluated using bacterial cultures selected for their zein proteolytic activity and isolated from a local solid waste landfill and a lagoon, showed no plasticizer effect even though its effect on moisture content was significant. Large differences were observed at different film zein concentration with the highest biodegradability for 100% zein. However, biodegradability did not mimic the water sorption behavior of zein/EC mixture films.     

Using ATR-FTIR spectroscopy to design active antimicrobial food packaging structures based on high molecular weight chitosan polysaccharide

ATR-FTIR spectroscopy has been used in this study to characterize the molecular mechanisms and kinetic processes that take place when a chitosonium acetate thin coating is put in contact with water solutions, Staphylococcus aureus solutions, microbial nutrient solutions, and with a high water activity TSA hydrogel medium to simulate the effect of direct contact with high moisture foods such as fresh meats, fish, and seafood products or beverages. The results of this work suggest that the biocide carboxylate groups that form when chitosan is cast from acetic acid solutions are being continuously evaporated from the formed film in the form of acetic acid (mechanism I) in the presence of environmental humidity, rendering weak biocide film systems. On the other hand, upon direct contact of the cast chitosonium acetate film with liquid water, water solutions, or the high moisture TSA hydrogel, a positive rapid migration, with a diffusion coefficient faster than 3.7 x 10(-12) m2/s, of protonated glucosamine water soluble molecular fractions (mechanism II) takes place from the film into the liquid phase, yielding strong antimicrobial performance and leaving in the remaining cast film only the non-water soluble chitosan fractions. Finally, this study describes a refined spectroscopic methodology to predict the antimicrobial properties of chitosan and gives insight into the capacity of chitosan as a natural biocide agent.     

Antibacterial activity of coffee extracts and selected coffee chemical compounds against enterobacteria

The in vitro antimicrobial activity of commercial coffee extracts and chemical compounds was investigated on nine strains of enterobacteria. The antimicrobial activity investigated by the disc diffusion method was observed in both the extracts and tested chemical compounds. Even though pH, color, and the contents of trigonelline, caffeine, and chlorogenic acids differed significantly among the coffee extracts, no significant differences were observed in their antimicrobial activity. Caffeic acid and trigonelline showed similar inhibitory effect against the growth of the microorganisms. Caffeine, chlorogenic acid, and protocatechuic acid showed particularly strong effect against Serratia marcescens and Enterobacter cloacae. The IC(50) and IC(90) for the compounds determined by the microtiter plate method indicated that trigonelline, caffeine, and protocatechuic acids are potential natural antimicrobial agents against Salmonella enterica. The concentrations of caffeine found in coffee extracts are enough to warrant 50% of the antimicrobial effect against S. enterica, which is relevant to human safety.     

Incorporation of a high concentration of mineral or vitamin into chitosan-based films

Mineral or vitamin E was incorporated into chitosan-based films: 10-200% (w/w chitosan) Gluconal Cal (GC), a mixture of calcium gluconate and lactate; 5-20% zinc lactate (ZL); and 5-20% alpha-tocopheryl acetate (VE) with acetylated monoglyceride (AM). The functionality of film-forming solutions and dried films was analyzed with standard procedures, and mathematical equations were developed to coordinate selected film functionality with the type and concentration of incorporated mineral or vitamin E. GC incorporation significantly increased pH and decreased viscosity of film-forming solutions, but not the addition of ZL or VE. The water barrier property of the films was improved by increasing the concentration of mineral or vitamin E in the film matrix. The tensile strength of the films was more significantly affected by GC or VE addition than film elongation, puncture strength, and puncture deformation. While a major endothermic peak around 200 degrees C was observed in DSC thermograms of chitosan-based films, only 200% GC incorporation altered this endothermic peak. This study demonstrated the capability of chitosan-based film matrix to carry a high concentration of mineral or vitamin E. Such films may be used for wrapping or coating to enhance the nutritional value of foods.     

Kombucha fermentation and its antimicrobial activity

Kombucha was prepared in a tea broth (0.5% w/v) supplemented with sucrose (10% w/v) by using a commercially available starter culture. The pH decreased steadily from 5 to 2.5 during the fermentation while the weight of the "tea fungus" and the OD of the tea broth increased through 4 days of the fermentation and remained fairly constant thereafter. The counts of acetic acid-producing bacteria and yeasts in the broth increased up to 4 days of fermentation and decreased afterward. The antimicrobial activity of Kombucha was investigated against a number of pathogenic microorganisms. Staphylococcus aureus, Shigella sonnei, Escherichia coli, Aeromonas hydrophila, Yersinia enterolitica, Pseudomonas aeruginosa, Enterobacter cloacae, Staphylococcus epidermis, Campylobacter jejuni, Salmonella enteritidis, Salmonella typhimurium, Bacillus cereus, Helicobacterpylori, and Listeria monocytogenes were found to be sensitive to Kombucha. According to the literature on Kombucha, acetic acid is considered to be responsible for the inhibitory effect toward a number of microbes tested, and this is also valid in the present study. However, in this study, Kombucha proved to exert antimicrobial activities against E. coli, Sh. sonnei, Sal. typhimurium, Sal. enteritidis, and Cm. jejuni, even at neutral pH and after thermal denaturation. This finding suggests the presence of antimicrobial compounds other than acetic acid and large proteins in Kombucha.     

Effect of Transglutaminase on Protein Electrophoretic Pattern of Rice,Soybean, and Rice‐Soybean Blends

The interactions taking place in composite dough containing rice flour and soybean proteins (5% w/w) in the presence of transglutaminase, an enzyme with cross‐linking activity, were studied using different electrophoretic analyses. The interaction between rice proteins and soybean proteins was intensified by the formation of new intermolecular covalent bonds catalyzed by transglutaminase and the indirect formation of disulfide bonds among proteins. The main protein fractions involved in those interactions were both β‐conglycinin and glycinin of soybean and the glutelins of the rice flour, although albumins and globulins were also cross‐linked. The addition of soybean proteins to rice flour improves the amino acid balance and they also might play an important role on the rice dough properties because soybean proteins interact with rice proteins, yielding protein aggregates of high molecular weight.     

Structural investigation of edible zein films/coatings and directly determining their thickness by FT-Raman spectroscopy

Near-infrared Fourier transform Raman (FT-Raman) spectroscopy was employed to study the molecular structure of edible zein films/coatings, which were fabricated directly from zein protein. The secondary structure of zein protein was mainly in alpha-helix and remained unaltered during film formation as evidenced by the vibrational modes of amide I at 1656 cm(-1) and amide III at 1274 cm(-1). Raman results indicated that hydrophobic interaction played an important role in the formation of zein film and disulfide bonding might be responsible for the structural stability of zein protein during film formation. To enhance its antimicrobial property, an antimicrobial zein film was manufactured by incorporating zein protein with benzoic acid whose structure was then characterized by FT-Raman. It showed that physical entrapment or hydrophobic interaction was crucial to the incorporation of benzoic acid with zein protein, and the secondary structure of the antimicrobial film was still maintained in alpha-helical form. In addition, FT-Raman exhibits its preference in directly determining the thickness of zein films/coatings. By correlating the Raman intensity ratio of nu(1003) to nu(84) (I(1003/84)) versus the thickness of zein film, a linear relationship with high coefficient (R(2) = 0.9927) was obtained, which was then used pragmatically to determine the thickness of zein coatings on apple. It showed that the FT-Raman result (thickness = 0.27 +/- 0.01 mm) was consistent with that of classical micrometric measurement (thickness = 0.28 +/- 0.02 mm). Consequently, FT-Raman provides a direct, simple, and reagent-free method to characterize the structure and the thickness of zein films/coatings.     

天然花青素提取物与壳聚糖明胶复合膜的制备和表征

为了开发天然的抗氧化活性包装材料,以紫甘蓝、黑米、玫瑰、蓝莓为原料制备天然花青素提取物与壳聚糖明胶的复合膜,比较分析了不同天然花青素提取物对复合膜的物理、机械、抗氧化活性及形貌结构的影响。结果表明：天然花青素提取物的加入,增加了膜的厚度,显著（P<0.05）影响膜的含水率、水溶性及外观形貌。壳聚糖明胶复合膜的水蒸汽透过率（water vapor permeability,WVP）为10.69×10-11 g/(m·s·Pa)。玫瑰花青素提取物的加入使得WVP值降低,而其他花青素提取物的加入使得WVP值增大。玫瑰复合膜的拉伸强度最大,达到27.03 MPa,断裂伸长率最小,黑米花青素提取物可增加复合膜的延展性,断裂伸长率最大为57.67%。傅里叶红外光谱表明天然花青素提取物的羟基基团与壳聚糖的氨基基团产生相互作用。扫描电镜结果表明花青素提取物影响微观结构,而且生物相容性较好。加入天然花青素提取物后,复合膜抗氧化活性均显著（P<0.05）提高,且玫瑰复合膜有着较高的抗氧化活性,1,1-二苯基-2-苦基肼（DPPH）自由基清除能力达到95.47%。结果表明：玫瑰花青素提取物更有利于开发阻湿性能好,水溶性低,抗拉伸和抗氧化活性高的包装材料,具有良好的应用前景。     

Supplementation of hydroxypropyl methylcellulose into yeast leavened all-whole grain barley bread potentiates cholesterol-lowering effect

We investigated in Syrian Golden hamsters the biological impact and its underlying mechanism of single whole grain breads supplemented with 2-3% hydroxypropyl methylcellulose (HPMC), a semisynthetic viscous soluble dietary fiber (SDF) as a substitute for gluten. Hamsters were fed high-fat diets supplemented with 48-65% (w/w) differently ground, freeze-dried single grain breads including whole grain wheat, barley, barley supplemented with HPMC, debranned oat, and oat supplemented with HPMC which were compared to a diet containing microcrystalline cellulose (control). All single grain breads significantly lowered plasma LDL-cholesterol concentrations compared to the control. Enrichment with HPMC further lowered plasma and hepatic cholesterol concentrations. Despite the reduced molecular weight of naturally occurring soluble (1--->3),(1--->4)-β-d-glucan (β-glucan) caused by the bread-making process, whole grain barley breads downregulated hepatic expression of CYP7A1 and HMG-CoAR genes that are responsible for bile acid and cholesterol synthesis, suggesting a possible role of bioactive compounds such as short-chain fatty acids and phenolic compounds from barley bread. Barley bread enriched with HPMC downregulated expression of ABCG5 gene. Taken together, it appears that distinctive modulation of synthesis and excretion of hepatic cholesterol and bile acid contributes to the cholesterol-lowering properties of whole grain barley breads and breads enriched with HPMC. These data suggests that alternative whole grain breads supplemented with HPMC may provide consumers with a staple food that can assist in cholesterol management.     

New bioactive biomaterials based on quaternized chitosan

Chitosan was chemically modified to produce quaternary ammonium salts in order to improve its antimicrobial activity and physicochemical properties. Quaternization of N-alkyl chitosan derivatives was carried out using alkyl iodide to elaborate water-soluble cationic polyelectrolytes ( N, N, N-trimethylchitosan, TMC). TMC was characterized by (1)H NMR spectroscopy; the quaternization degree was determined from (1)H NMR spectra and by titration of iodide ion. The antibacterial activity of hydroxypropylcellulose (HPC) films or coatings associated with chitosan or TMC as biocide was evaluated against the growth of Listeria monocytogenes and Salmonella typhimurium. The HPC-chitosan and HPC-TMC coatings exhibited a total inhibition on solid medium of both bacterial strains. Experiments conducted in liquid medium showed that the inhibitory activity against the growth of Listeria innocua was improved after chemical modification. Moreover, physicochemical properties of films were evaluated to determine their potential for food applications. The addition of the antibacterial agents showed a significant impact on the moisture barrier and mechanical properties of HPC films.     

Heat treatment to modify the structural and physical properties of chitosan-based films

This work was focused on studying the changes undergone by heat-treated chitosan films with and without tannic acid addition by monitoring both microstructure and physical properties. Once the films were submitted to different heat treatments, they exhibited higher barrier properties as well as lower water uptake, solubility, and moisture content. These results were also confirmed through X-ray patterns, which changed from the hydrated to the anhydrous conformation, sharper FTIR peaks specifically associated with water, and shift of T(g) toward higher temperatures determined by DMA. Moreover, the modifications caused by the curing process at a molecular scale were observed at a structural level by using a TEM technique. FTIR evaluation granted new insights into the interactions between tannic acid and chitosan molecules, before and after the heat curing, especially due to the occurrence of new peaks and changes in the wavenumber region 1550-1750 cm(-1).