Novel intervention strategies such as food grade antimicrobials are used to enhance food safety. Nisin is a widely used naturally occurring antimicrobial effective against many pathogenic and spoilage microorganisms. However, its antimicrobial efficacy in food matrices is much reduced because of interactions with food components. Novel delivery systems may overcome this problem by protecting nisin in capsules and releasing it in a controlled manner. The overall goal of this research was to develop delivery systems of nisin to improve its long-term antimicrobial effectiveness. The first objective was to develop a low-cost method to extract nisin from a commercial preparation containing ~2.5% nisin. The best extraction yield was observed at 6 mg solids/mL 50% v/v ethanol. The nisin extract, after adjusting to 70% v/v ethanol, was dissolved with 2% zein (corn prolamins) and different amounts of Tween 20, glycerol, and thymol (another naturally occurring antimicrobial) for spray drying, a practical encapsulation method. Spray-dried capsules were characterized for release profiles of nisin at different pH conditions. Spray drying inlet temperature was first studied at 75 to 120°C, and the greatest sustained release of nisin was observed for capsules produced at 105°C, which was used in rest of this study. The impacts of Tween 20 and glycerol supplemented in spray-drying formulations were studied next. Addition of either 0.05% Tween 20 or glycerol in the spray-drying solution reduced the burst release of nisin at pH 6.0. Capsules with a higher amount of Tween 20 showed more complete release of nisin at pH 8.0, while glycerol had no apparent impact. The most sustained release of nisin at pH 6.0 was observed for capsules produced with both thymol and glycerol. Various capsules were then studied for their ability to inhibit the growth of Listeria monocytogenes at pH 6.0 during incubation at 30°C. Un-encapsulated nisin and thymol showed inhibition only for 12 h, while capsules with nisin and thymol containing either low or medium level of glycerol inhibited L. monocytogenes for >96 h. Our antimicrobial delivery systems, based on food grade ingredients and practical processes, have potential for practical application to enhance microbial safety and extend the shelf-life of foods.

Thesis (MSc Food Sc)--University of Stellenbosch, 2011. / Includes bibliography. / ENGLISH ABSTRACT: Maize is an important crop for both human and animal consumption. Maize kernel texture (kernel hardness) is an important quality trait for many sectors in the South African maize industry, where a harder texture is desired. Both total protein content and the main storage proteins, zein, have been associated with kernel texture. The zein profiles of South African white maize hybrids, from a breeding program, grown at three localities together with their respective inbred parent lines were evaluated to determine the difference in zein expression. For only the hybrids, total protein content, zein content and degree of hardness (kernel texture) was determined to establish possible relationships. Zein consists of four main classes, α-, β-, γ-, and δ-zein, which can further be divided into sub-classes. Zein was characterised using matrix-assisted laser desorption ionisation time-offlight mass spectrometry (MALDI-TOF MS) after optimisation of the zein extraction and matrix preparation procedures. Two matrices [2-(4-hydroxyphenylazo)benzoic acid (HABA) and α- cyano-4-hydroxy-cinammic acid (CHCA)] and three pH levels (<1.5, 1.7 and 2.9) for matrix solutions were investigated. Two solvent concentrations (50% and 70% acetonitrile (ACN) were also investigated. Parallel to this investigation, a zein extraction procedure was optimised. Zein was extracted from non-defatted (NDF) and defatted (DF) maize meal at ambient temperature and 60°C and meal with different particle size distributions. Particle size of maize meal did not influence the quality of spectra. Regardless of matrix conditions used, zein extracted at 60°C from NDF meal produced spectra with a low signal-to-noise (S/N) ratio. When only HABA was included (pH<1.5), not all γ-zein sub-classes were observed. This was also true for zein extracted at ambient temperature from NDF and DF meal. Good S/N ratios for all zein classes were obtained when zein was extracted from NDF meal at ambient temperature using a matrix solution containing both matrices in 70% ACN at pH 2.9. MALDI-TOF MS provided accurate detection and good profile resolution for zein characterisation. Spectral profiles of hybrids were a combination of their respective inbred parent lines. Zein proteins were quantified using reverse-phase high performance liquid chromatography (RP-HPLC). Total protein content was determined and kernel texture assessed using a particle size (PSI) method. Significant (P<0.01) differences were observed for total protein content and degree of hardness between certain hybrids as well as average values for localities. Correlations and principal component analysis indicated that hybrids with a higher protein and total zein content had a harder kernel texture. The β- and γ-zein contents also correlated positively with kernel hardness. However, when expressed as percentage of total zein the opposite was observed. Collectively the α-zeins and most individual α-zeins correlated positively with hardness. Scanning electron microscopy micrographs showed differences between the floury endosperm of harder and softer maize kernels; illustrating starch types (amylose and amylopectin) should also be analysed in future hardness studies. The correlations obtained were not strong (r<0.6) and the variation explained by first two principal components was low. Other maize constituents, more hardness measurements should be considered in future studies to improve this. / AFRIKAANSE OPSOMMING: Mielies is a belangrike gewas vir beide mens- en dierlike inname. Mieliepittekstuur (pithardheid) is ‘n belangrike kwaliteitseienskap vir baie sektore in die Suid Afrikaanse mielieindustrie, waar ‘n harder tekstuur verlang word. Beide totale proteïeninhoud en die hoof opbergingsproteïen, zein, is al geassosieer met pittekstuur. Die zein profiele van Suid- Afrikaanse witmielie basters, van ‘n teel program, wat by drie lokaliteite verbou is sowel as hul onderskeie ingeteelde ouerlyne is geevalueer om verskille in zein uitdrukking te bepaal. Die totale proteïeninhoud, zeininhoud en graad van hardheid is bepaal om verhoudings vas te stel. Zein bestaan uit vier hoof klasse, α-, β-, γ-, en δ-zein, wat verder onderverdeel word in subklasse. Zein is gekarakteriseer met matriks-ondersteunende laser desorpsie ionisasie tyd-vanvlug massa spektrometrie (MBLDI-TVV MS) na die zein ekstraksie en matriks voorbereidingprosedures geoptimaliseer is. Twee matrikse [2-(4-hydroksiephenylazo)benzoë suur (HABA) en α-cyano-4-hydroksie-kaneelsuur (CHCA)] en drie pH vlakke (<1.5, 1.7 and 2.9) vir matriksoplossings was ondersoek. Twee oplossingkonsentrasies [50% and 70% asetonitriel (ACN)] is ook ondersoek. Zein ekstraksie kondisies is ook geoptimiseer. Zein is geëkstraheer van nie-ontvette (NOV) en ontvette (OV) mieliemeel by omgewings temperatuur en 60°C. Die partikelgrootte van die mieliemeel het nie die kwaliteit van spektra beïnvloed nie. Ongeag watter matrikskondisies gebruik is het zein wat van NGV meel by 60°C ge-ekstraheer is, spektra met ‘n lae sein-tot-geraas (S/G) verhouding geproduseer. As die HABA alleenlik gebruik (by pH<1.5) is, is nie alle γ-zein klasse waargeneem nie. Dit was ook waar vir zein wat by omgewings temperatuur van NGV en OV mieliemeel ge-ekstraheer is. Goeie S/G verhoudings is waargeneem met zein, ge-ekstraheer van NGV mieliemeel by omringende temperatuur, met die bebruik van beide matrikse in 70% ACN by pH 2.9. (MBLDI-TVV MS) het akkurate en goeie resolusie van profiele vir zein karateriseering verksaf. Spektrale profiele van basters was ’n kombinasie van hul onderskeie ouer lyne. Zeinproteïene is met omgekeerde-fase hoë prestasie vloeistofchromatografie gekwantifiseer. Totale proteïeninhoud is bepaal en die hardheidsgraad was bepaal met ‘n partikelgroote indeks (PGI) metode. Beduidende verskille tussen proteïeninhoud en hardheidsgraad is waargeneem tussen sekere basters sowel as gemiddelde waardes by die verskeie lokalieteite. Korrelasies en hoofkomponent analise het aangedui dat basters met ‘n hoër proteïeninhoud en totale zein ‘n harder tekstuur gehad het. Die β- en γ-zeininhoud het ook positief gekorreleer met hardheid. Alhoewel, wanneer die proteïene as persentasie van totale zein uitgedruk is, is die teenoorgestelde waargeneem. Gesamentelik het α-zein asook die meeste indivduele α-zeins positief gekorreleer met hardheid. Skandeer elektron mikroskopie mikrograwe het verskille tussen die meelerige endosperm van harder en sagter pitte aangedui; meer proteïenliggame was aanwesig in die harder pitte en die stysel was digter gepak. Dus moet stysel tipes ook in ag geneem word in toekomstige hardheidsstudies. Korrelasies wat verkry is, was nie hoog (r<0.6) nie en die variasie verduidelik deur die eerste twee hoofkomponente was laag. Ander miielie samestellende dele as ook meer hardheidsmetings moet in toekomstige studies in ag geneem word om dit te verbeter.

Orientador: José Francisco Lopes Filho / Banca: Elisângela Corradini / Banca: José Antônio Gomes Vieira / Resumo: O propósito deste estudo foi verificar alterações na estrutura e em algumas propriedades funcionais de biofilmes produzidos com zeína adicionados de argilas modificadas. As argilas modificadas Cloisite® 15A e Cloisite® 30B foram adicionadas na solução filmogênica sob agitação nas proporções de 1, 2 e 4% em relação à massa de zeína e os filmes formados pelo método "casting". Ácido oléico e glicerol foram utilizados como agentes plasticizantes. Para a caracterização da microestrutura dos biofilmes foram realizadas as análises de Microscopia Eletrônica de Varredura (MEV), Ótica (MO) e Espectroscopia no Infravermelho (FTIR) e as propriedades funcionais avaliadas foram Opacidade, Solubilidade e Permeabilidade ao Vapor d'Água. Por meio das imagens obtidas na MEV observou-se que os acréscimos de argilas nos filmes resultaram na formação de uma microestrutura heterogênea, ou seja, estrutura de fase separada chamada de microcompósito ou tactóide, uma vez que as argilas não dispersaram no filme, o que proporcionou a formação de agregados. No entanto, por meio das imagens obtidas com a MO, observou-se uma distribuição uniforme dos glóbulos de lipídeos na matriz protéica, para todos os filmes. Em concordância com a característica da microestrutura dos filmes elaborados identificados na MEV, o FTIR foi capaz de identificar número de onda referente às argilas, indicando que a mesma não estabeleceu estruturação com a proteína, confirmando a formação de uma estrutura de fase separada tipo tactoide. Todos os filmes elaborados apresentaram maior opacidade quando comparados com o polietileno em razão da presença de pigmentos carotenóides. Pequenas diferenças na espessura das amostras e a quantidade de argila adicionada não influenciaram na transparência dos materiais. Os testes... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The purpose of this study was to evaluate changes in the structure and some functional properties of zein edible films added with modified clays. The modified clays Cloisite® 15A and Cloisite® 30B were added in the solution under agitation in the proportions of 1, 2 and 4% based on the weigh of zein. The films were formed pouring the solution on a surface to dry by casting method. Oleic acid and glycerol were used as plasticizer agents. Analyses of Scanning Electron Microscopy (SEM), Optical Microscopy (MO) and Infrared Spectroscopy (FTIR) were performed to characterize the microstructure of biofilms. The functional properties evaluated were opacity, solubility and permeability of water vapor. SEM images showed that the additions of clays in the films resulted in the formation of a heterogeneous microstructure, i.e. the phase separated structure called microcomposite or tactoids, since the clays were not dispersed in the film, which forms points of aggregates. However, the images obtained with MO showed an even distribution of lipids globules in the protein matrix, for all films. In agreement with the characteristic of the microstructure of the films showed by SEM, FTIR analyses pointed out the absence of wave number related to the clays, indicating that it was not established a really binding of the clays with the protein molecule. This confirms the formation of a material with phase separated, tactoid type. All the films prepared showed higher opacity when compared with polyethylene because of the presence of carotenoid pigments. Little differences in thickness of the samples and the amount of clay added did not influence the transparency of materials. Statistical mean tests of water solubility of the samples showed no differences among those films added with clay and the control. Water... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2016-06-02T19:02:43Z (GMT). No. of bitstreams: 1 5710.pdf: 9081274 bytes, checksum: 235a22e72d32a0486237aee2952f268e (MD5) Previous issue date: 2013-11-27 / Financiadora de Estudos e Projetos / Brazil is a major producer of fruit, but the losses related to postharvest handling are considerable high, and it is important to develop methods to reduce them. Analysis techniques were used is the application of edible coatings in order to increase the shelf life of the fruit. This study evaluated the use of zein based films, storage proteins from corn, with the addition of plasticizers and cellulose nanofibers, for covering pears and apples. The zein are hydrophobic proteins and can form a barrier against moisture loss and gas exchange, thus reducing the rate of fruit respiration and consequently increasing it`s shelf life. The filmogenic solutions tested were obtained using 4% zein with addition of different concentrations of cellulose nanofibers and plasticizers. The solutions were applied on apples and pears by immersion. Fruits were evaluated for weight loss, appearance and texture, where it was shown that the optimal concentration was 4% + 0.1% zein cellulose nanofibers + 0.5% oleic acid, as obtained an increase of about 30 days the shelf life of fruit in addition there was better preservation of fruits in natural color and texture compared to other concentrations. For this we use techniques: colorimetry, texturometer and monitoring of mass loss. For characterization of the films were used analysis techniques and contact angle atomic force microscopy, where we also observe that the best result for these two parameters was the movie 4% zein + 0.1% cellulose nanofibers and 0.5% oleic acid, because the films were less hydrophilic and more homogeneous. To evaluate the toxicity of produced film an experiment was conducted with male Wistar rats that were divided into two groups: the first group was fed with feed coated with filmogenic solution, and the second group received a diet without coating. During the experiment they documented the intake of food and water, and collected all excrement. After the experiment various organs were collected for analysis. The results of this experiment indicated that the solution filmogenic showed no toxic effect on the mice. The results of the experiments with the fruit specified the solutions were more efficient for the preservation of fruit quality in the study. / O Brasil é um grande produtor de frutas, porém as perdas relacionadas ao manuseio póscolheita são consideráveis, sendo importante o desenvolvimento de metodologias que as reduzam. Uma técnica que tem sido usada é aplicação de revestimentos comestíveis com o objetivo aumentar o tempo de prateleira do fruto. O presente trabalho avaliou o uso de filmes a base de zeína, proteínas de reserva do milho, com a adição de plastificantes e nanofibras de celulose, para recobrimento de peras e maçãs. As zeína são proteínas hidrofóbicas e podem formar uma barreira contra a perda de umidade e as trocas gasosas, reduzindo assim a taxa de respiração dos frutos e consequentemente aumentando o seu tempo de prateleira. As soluções filmogênicas testadas foram obtidas utilizando-se 4% de zeína com adição de diversas concentrações de nanofibras de celulose e plastificantes. As soluções foram aplicadas sobre peras e maçãs por imersão. As frutas foram avaliadas quanto a perda de massa, aparência e textura. Para isso utilizamos as técnicas de: colorimetria, texturômetro e acompanhamento da perda de massa, onde foi mostrado que a melhor concentração foi a de 4% de zeína + 0,1% de nanofibras de celulose + 0,5% de ácido oleico, pois obtivemos um aumento de cerca de 30 dias no tempo de prateleira das frutas, além disso, houve uma melhor conservação na coloração natural das frutas e na textura em relação as outras concentrações. Para caracterização dos filmes foram utilizadas as técnicas de análise de ângulo de contato e microscopia de força atômica, onde também podemos observar que o melhor resultado para esses dois parâmetros foi o filme de 4% de zeína + 0,1% de nanofibras de celulose e 0,5% de ácido oleico, pois os filmes ficaram menos hidrofílicos e mais homogêneos. Para avaliar a toxicidade do filme produzido foi feito um experimento com ratos machos da linhagem Wistar, divididos em dois grupos, sendo que o primeiro grupo foi alimentado com ração revestida com solução filmogênica, e o segundo grupo recebeu a ração sem revestimento. Durante o experimento foram compilados o consumo de ração e água e coletadas todas as excreções. Após o experimento foram coletados vários órgãos para análise.

This experimental work investigated the development and testing of a controlled release system for methionine. Methionine is one of the limiting amino acids for the milk production in dairy cows. The quantities of methionine which reach the small intestine are affected by the bacteria in the rumen which utilize methionine. A controlled release system which will offer a protective barrier for methionine may ensure that the methionine reaches the small intestine in sufficient quantities. The work involved the development of a coating around methionine crystals, which would act as a barrier, protecting it from the rumen conditions. Zein and kafirin proteins from maize and sorghum, respectively, were used as the principal coating components for the controlled release system. Two different approaches were used in the development of the controlled release system. First, the zein and kafirin proteins were tested for their ability to act as barriers for the controlled release of methionine, and second, zein and kafirin microparticles were used as the controlled release agents. Relatively successful, laboratory-scale methods were developed for coating the methionine with the proteins and the microparticles. Protein coatings were made by addition of methionine crystals to acid-dissolved proteins which led to the formation of a protein/methionine matrix. For coating the methionine with microparticles, glacial acetic acid was used to fuse microparticles around the methionine crystals. Dissolution assays were performed to test the release of methionine from the coatings under simulated rumen conditions. Both the zein and kafirin and microparticle coatings exhibited a barrier effect for methionine. The barrier effects of these coatings were influenced by several factors. Increasing the proportion of the coating agents led to improved barrier properties. However, this only occurred until a certain proportion of coating agent was present (50%), after which the barrier properties no longer increased. Heat treatment of the coatings also increased the barrier properties of the coatings. This may be due to the formation of disulphide cross-links being formed during the application of heat. When a simple extrusion method was used to form the coatings, the barrier properties also improved in comparison to those coatings which were not formed using extrusion. When producing the microparticles, it was found that only the laboratory extracted kafirin preparation with 85% (db) protein formed microparticles. It was hypothesized that microparticle formation might be related to the purity of the protein preparations. Scanning electron microscopy of the coatings after the dissolution tests and pepsin digestion revealed pores on the surface of the coating. These were probably where the methionine leached from the coating into the dissolution medium. The protein coatings did act as partial barriers, extending the release of methionine. From the release curves of methionine from the coatings, it could be seen that a sustained release of methionine occurred over a period of time, rather than a controlled release of methionine at a certain time. The aim of the application was thus only partially achieved as a complete protective barrier for methionine was not obtained from the protein coatings. No significant difference between the barrier properties of the coatings prepared from the proteins themselves and the microparticles were found. However, when based on equal protein purity the kafirin protein coatings showed the most effective barrier properties. Further research regarding kafirin coatings as a controlled release agent is recommended based on the results of the above named calculation. This research would entail investigating various coating technologies and methods. / Dissertation (MSc)--University of Pretoria, 2011. / Food Science / unrestricted

The only treatment for coeliac disease, a common autoimmune disorder, is life-long adherence to a gluten-free diet. However, the replacement of wheat gluten, a key structural and functional component in bread, poses a major technological challenge for food scientists. The use of non-wheat cereal proteins, as alternatives to gluten, shows much promise in gluten-free bread making. Literature has shown that when zein, the maize prolamin protein, is subjected to wet heat above its glass transition temperature (Tg), the protein becomes viscoelastic, rubbery and dough-like. Gluten-like fibrils are visible, which form complex protein networks similar to those found in wheat dough. The resulting zein dough has viscoelastic characteristics and can be successfully used with hydrocolloids to produce gluten-free bread. This project examined the influence of wet heat treatment and dilute organic acids (lactic acid and acetic acid) on the dough-making quality of non-wheat cereal proteins, such as kafirin and zein. Zein was the only non-wheat cereal protein to show any physical change when it was subjected to wet heat treatments, forming a dough-like substance. Acidification of the zein dough prepared at 40°C with concentrations of 0.7, 1.3 and 5.4% (v/v) organic acid in distilled water solutions, showed that the higher the concentration of acid used, the greater its effect on the dough's rheological properties. Tensile tests using a Keifer rig on zein dough showed that as the concentration of organic acid was increased from 0.7 to 1.3 and to 5.4% (v/v) the dough become softer and increasingly more extensible. The dough also exhibited less resistance to extension and reduced elasticity. CLSM revealed that the zein doughs contained a protein network, made up of fine protein fibrils, which became smoother and more homogenous as the concentration of acid was increased. Although SDS-PAGE revealed that no oligomerization took place with acid addition, iv FTIR showed that zein dough prepared with distilled water at 40°C had elevated levels of β-sheets. When organic acids were added in increasing levels, corresponding increases in the quantities of α-helices in the protein were observed. Alveography showed that zein-based doughs prepared with dilute organic acids retained gases well and that the concentration of dilute organic acids influenced dough distensibility (biaxial extensibility) and stability (the ability of the dough to retain gas). Low concentration of acids (0.7 and 1.3%) increased dough stability to levels similar to that of strong wheat flour, 103 mm H2O, but higher concentrations of acids (5.4%) led to a marked reduction in dough stability. Thus, by increasing zein dough functionality to such an extent, the apparent usefulness of the doughs and their ability to retain gases produced during fermentation is reduced. Simple distensibility tests on zein doughs showed that added organic acids promoted ‘clumping’ of the fine protein fibrils in the dough network into pronounced fibres. This would account for the decreased dough stability when high levels (5.4%) of organic acids were used. Baking trials with zein doughs were not successful as adequate leavening was impossible without an acid-tolerant leavening agent. It is believed that dilute organic acids influence the rheological properties of zein dough by creating a positively charged environment, in which the protein is partially solubilized. The higher the level of organic acid used, the greater the positive net charge and the more pronounced the effect on the protein network structure. Organic acids could also improve fluidity of the zein dough by acting as plasticizers. From this work it can be seen that although a protein network is present in all zein-based doughs, the ability of this network to retain gases is dependant on the level of organic acids present. The functional properties of zein-doughs made with low levels of organic acids (0.7 and 1.3%) shows potential in the production of gluten-free bread for individuals suffering from coeliac disease. / Dissertation (MSc)--University of Pretoria, 2013. / gm2014 / Food Science / unrestricted

Identifying and detecting health hazards in food products, especially contaminants and toxic substances such as allergens, food toxins and agricultural residues from pesticides, remains a challenge. Increasing demand for food products and growing health consciousness necessitate rapid and accurate measurements which can be easily conducted on-site without long measurement times and high costs. Due to their ease of use, accuracy sample preparation and rapidity, biosensors have started to outcompete time-consuming lab-scale analytical devices. However, as the use of biosensors increase, a concern of the amount of plastics and synthetic polymers used in the fabrication of these biosensors rises. In this dissertation, new ways to create biodegradable and eco-friendly plant-based SERS biosensor platforms from corn protein, zein, are presented. Its higher hydrophobicity and film forming capability make zein a very suitable biopolymer for fabricating biosensors. In the first part of this dissertation, chemical crosslinking was tested to improve the surface hydrophobicity, surface roughness (using AFM), mechanical properties, kinetics of gelation and film formation of zein films, and as a result zein-film based SERS platforms with fewer defects could be fabricated. In the second part, the detection sensitivity of the zein film-based SERS platforms was increased with metallic nanoparticle decoration (gold, silver or silver-shelled-gold). The addition of all three types of nanoparticles significantly increased the SERS enhancement factors of the platforms, with silver-shelled-gold nanoparticles giving the highest enhancement factor of 10⁵. In the last part of this thesis, a novel approach was tested, where electrospun zein nanofibers decorated with metallic nanoparticles were used as a SERS biosensor platform. Due to their higher surface area-to-volume ratios, electrospun zein nanofibers gave a higher SERS enhancement factor (10⁶). This enhancement factor enabled the detection of acrylamide, a food carcinogen, with a 10⁴ times lower detection limit than nanophotonic based nanoimprinted zein, acrylamide sensor platform. Overall, this dissertation successfully shows the fabrication of biodegradable and eco-friendly SERS sensor platforms that have comparable detection sensitivities to those of non-biodegradable ones.

The demand for biodegradable packaging materials as an alternative to synthetic ones to reduce environmental cost has seen an increase in recent years. In addition, functionalizing the packaging film to provide specific advantages like antimicrobial properties has yet to be explored thoroughly. This study considers adding antimicrobial agents to improve the quality and safety of actively packaged fresh produce using an antimicrobial enzyme (lysozyme) immobilized on a biopolymer based packaging film (corn-zein). The developed packaging material is aimed as an active biodegradable packaging to reduce bacterial contamination on the surface of fresh organic produce, specifically tomatoes. The study uses glutaraldehyde and glyoxal as binding agents to immobilize the enzyme on the packaging film. The effect of concentration of glutaraldehyde and glyoxal on the controlled release of the enzyme was studied. Concentrations of 0.1 and 0.2 g/g lysozyme: cross linking agent had controlled release properties. However, concentrations of 0 or 0.05 are about 20-30% more effective at inactivating bacteria. Antimicrobial activity in the constructed zein films are also tested against selected pathogens (Salmonella Newport and Listeria monocytogenes). Developed zein based film is tested against inoculated tomatoes to determine the efficacy of the films in reducing the pathogen population. The inoculated tomatoes are stored at room temperature over a storage period of one week. The film was able to reduce Listeria monocytogenes population by three logs but was unable to reduce the population of Salmonella Newport. / Master of Science

Nanostructured materials play a vital role in almost all aspects of science and technology in the 21st century. The materials include nanoparticles, nanofilms, biological membranes etc. whose physicochemical properties are size-dependent. Thin films have wide range of applications in various branches of science. One of the efficient methods to form miniaturized structures for device applications is to fabricate nanostructured films on different substrates. Surfactant assembly on metallic and non-metallic surfaces based on self assembly and Langmuir-Blodgett technique offers a unique way to form thin films at molecular levels. The process of formation of unimolecular assemblies gives the flexibility of tuning the properties of underlying substrates for various applications including wetting characteristics, lubrication, passivation, mimicking biological phenomena etc. Towards this direction, self assembled monolayers (SAMs) of alkanethiols on gold and silver surfaces have been studied comprehensively for the past two decades. The reported literature on short chain length thiol-based monolayers is however, limited since the formation using conventional methods yield poor quality monolayers. The short chain length monolayers are useful in various applications like tribology, layer-by-layer assemblies, biosensors etc. Hence, it is essential to reproducibly form SAMs of various chain lengths and understand their properties. The present study is related to the formation of SAMs of alkanethiols and diselenides on gold and silver surfaces to form ordered and well-oriented monolayers. Monolayers of varying chain lengths (CH3(CH2)nSH where n = 3, 5, 7, 9, 11, 15) have been formed on gold and silver surfaces using different methods, (1) adsorption from neat thiols; (2) adsorption under electrochemical control and (3) adsorption from alcoholic solutions of the thiols. The characteristics features of the SAMs have been followed based on three different aspects, (i) structure and stability of the methylene groups (ii) interfacial characteristics involving the end group and the solvent and (iii) metal-head group interactions. The structure and stability of the monolayers have been followed based on vibrational spectroscopy and electrochemistry under different environment including thermal perturbations. The stability of the SAMs at different temperatures and subsequent changes associated with the orientation / packing has been monitored both in the dry state using reflection absorption infrared spectroscopy (RAIRS) and under electrochemical conditions using cyclic voltammetry and impedance analysis. Monolayers adsorbed from neat thiols show superior quality in terms of stability and structural arrangement. Short chain thiols with n = 3, 5, 7 show substantial stability when the adsorption is carried out from neat thiols. Figure 1 shows the RAIR spectra of hexanethiol SAM on gold adsorbed by three different procedures. Monolayers adsorbed under potential control behave very similar to the monolayers adsorbed from neat thiol as for as stability and structural orientation are concerned. Monolayers prepared using conventional methods of adsorption from alcoholic solutions are of inferior quality in terms of stability and arrangement especially for the short chain lengths. This is likely to be due to the fact that monolayers prepared using conventional methods may have intercalated solvent molecules within the monolayer assembly that degrade the integrity of the SAM leading to poor quality. The blocking characteristics of the monolayers for diffusing redox couple have been followed by determining the heterogeneous electron transfer rate constant using electrochemical techniques. The spectroscopic data and the electrochemical data follow similar trend indicating the superior quality of monolayer adsorbed from neat thiol in terms of stability as compared to conventionally prepared monolayers. Figure 1. RAIR spectra of hexanethiol-SAMs on Au(111) surface at 25C. The monolayers are formed by adsorption (A) from neat thiol (B) under potential control and (C) from alcoholic solution of the thiol. Wavenumber (cm-1) The interfacial characteristics of the monolayers (effect of end group functionality on the solvent properties) have been monitored on the basis of capacitance, contact angle and atomic force microscopy- measurements. Well-organized monolayers behave like good capacitors with relatively low values of double layer capacitance in presence of a liquid electrolyte as compared to the expected values based on known thickness and dielectric constant of the SAMs. This behavior can be explained by invoking the depletion of water density at the methyl terminated SAM-water interface where the solvent properties are different from that of bulk. Variation of one such property, dielectric constant, has been mapped using force measurement based on AFM. Dielectric constant of water changes from the bulk value of 78 to a low value as given in figure 2. This cross-over occurs within a span of 1-3 nm depending on the chain length of the thiol. Of the three procedures used, the ones based on the use of neat thiol and electrochemical adsorption result in well-oriented alkyl chains followed by highly oriented methyl terminal groups. This is responsible for the high hydrophobic nature of the interface and the subsequent observation of interfacial water properties. The SAMs prepared from ethanol fail to show the hydrophobic effects. Hydrophilic monolayers (NH2 terminated monolayers) fail to show depletion of water density at the interface indicating the importance of end group functionality in altering the interfacial characteristics of the monolayer. Figure 2. Spatial variation of dielectric permittivity of water at the hexanethiol SAM - water interface. The SAM is formed on gold (111) surface; (a) from ethanolic solution of the thiol (b) under electrochemical control (c) from neat thiol. The origin on the x-axis is the position of the methyl groups of SAM and the direction towards right side is in to the bulk water. The well-oriented SAMs have been used to follow the adsorption of a biopolymer. Zein protein is a prolamine of maize and is projected to be a biocompatible coating for food products and food containers. Hence, it is essential to prepare impermeable coatings of zein with different surface wetting properties. The adsorption of zein on highly ordered SAMs with hydrophobic or hydrophilic end group functionality has been studied and the orientation of the protein followed using spectroscopy, microscopy and electrochemistry. It is observed that zein shows higher affinity towards hydrophilic than hydrophobic surfaces with small foot print size on the Figure 3. Orientation of zein protein on hydrophilic and hydrophobic SAM as deciphered from the experimental data. hydrophilic surface resulting in large surface coverage. Figure 3 shows the schematics of zein deposits on hydrophilic and hydrophobic SAM surfaces determined based on spectroscopy, quartz crystal microbalance and electrochemical studies. The AFM shows cylindrical, rod-like and disc-like features of zein on hydrophilic surfaces that form the base units for the growth of cylindrical structures of zein. The published literature on the studies on SAMs on silver surfaces reveals that there is no consensus on the structure of the monolayers on silver. This may be due to the difficulty in getting pristine oxide-free surfaces in the case of silver and this is likely to affect the monolayer quality. Hence, it is decided to prepare SAMs of alkanethiols on silver and study their characteristics. Subtle differences between the monolayers adsorbed from neat thiol and from alcoholic solutions of thiols have been observed in terms of stability and permeability. Atomic force microscopic studies illustrate the presence of depletion of water at the SAM-aqueous interface. Diselenide-based monolayers have been formed on gold to understand the head group-substrate interactions on the monolayer properties. The disorder observed on short chain diselenide-based monolayers formed from alcoholic solutions can be eliminated by adsorption from neat compounds as described for the thiols. A preliminary account on the stability of SAMs under hydrodynamic conditions has been given based on rotating disc electrode voltammetry. It is observed that the SAMs get well-ordered when the electrode is rotated at a fast rate leading to the hypothesis that the monolayer assembly gets annealed as a function of the rotation rate. The thesis is planned as follows: Chapter 1 gives general introduction about organic thin films with particular emphasis on self-assembled monolayers on gold and silver, their characteristics in terms of stability, interfacial properties and adsorption behaviour. Chapter 2 deals with the experimental methodologies and schematics used for the preparation and characterization of the monolayers. Chapter 3 is on the contribution of alkyl spacer to the stability of the monolayers studied using spectroscopy and electrochemistry. Chapter 4 deals with the interfacial properties of the SAMs in presence of aqueous medium. In order to emphasize the importance of the terminal functional groups, adsorption of zein has been demonstrated on surfaces of controlled wettablity. Chapter 5 explains the formation and stability of monolayers of short and long chain alkyl diselenides on gold surfaces. Chapter 6 gives the structural and interfacial characteristics of alkanethiol monolayers on silver surfaces. The stability and subsequent changes of alkanethiol monolayers under hydrodynamic conditions has been discussed in the appendix section.(For fig pl refer pdf file.)

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O consumo de bolos com reduÃÃo calÃrica vem crescendo, mas tem apresentado desafios a serem superados na formaÃÃo da sua estrutura quando o aÃÃcar ou a gordura sÃo substituÃdos por adoÃantes, gomas, espessantes ou substitutos de gordura. O presente trabalho propÃs avaliar as caracterÃsticas internas de bolos com reduÃÃo calÃrica e a viabilidade de produÃÃo do mesmo com caracterÃsticas similares ao bolo convencional. Inicialmente, foram definidos o tempo de mistura da massa e a fonte de gordura a ser utilizada na formulaÃÃo, utilizando-se o volume especÃfico como parÃmetro. Em seguida, determinou-se a formulaÃÃo do bolo padrÃo a partir de um delineamento experimental do tipo fatorial simples, medindo-se volume especÃfico e a contagem de cÃlulas. A partir dessa formulaÃÃo, a sacarose e a gordura foram parcialmente substituÃdos para a produÃÃo de bolos com valor calÃrico reduzido. A substituiÃÃo do aÃÃcar foi feita proporcionalmente por uma mistura de goma xantana (1,5%) e sucralose (1%), enquanto a zeÃna foi usada para substituir a gordura. Nos bolos com reduÃÃo de aÃÃcar avaliou-se o volume especÃfico (VE), a contagem de cÃlulas (CC) dos bolos, a viscosidade aparente da massa e as suas propriedades tÃrmicas por meio de calorimetria diferencial de varredura (DSC). Nos bolos com reduÃÃo do teor de gordura, as anÃlises realizadas foram VE, CC e viscosidade aparente. Verificou-se que o bolo com maior VE foi aquele elaborado com gordura vegetal hidrogenada e 1 (um) minuto de mistura da massa. A formulaÃÃo padrÃo foi definida como tendo 155,88 e 28,78 partes de aÃÃcar e de gordura, respectivamente. à medida que o teor de aÃÃcar decresceu (10,00-52,17%) foram reduzidos o volume especÃfico (1,94-0,7 mL/g) e a contagem de cÃlulas (36,2 â 4,0 cÃl/cmÂ) do bolo e a viscosidade aparente da massa (337,56-631,40 cP). Com a reduÃÃo do teor de gordura, nÃo foram observadas diferenÃas significativas entre as amostras para VE, CC e viscosidade aparente, sendo viÃvel a produÃÃo do bolo com reduÃÃo calÃrica substituindo-se a gordura em atà 46,86%. Pelos resultados encontrados, foi observado que a substituiÃÃo do aÃÃcar contribuiu mais acentuadamente que a substituiÃÃo da gordura na formaÃÃo de defeitos na estrutura do bolo. Os termogramas das massas dos bolos padrÃo e com reduÃÃo de sacarose sugeriram que a presenÃa de sucralose pode reduzir a temperatura de gelatinizaÃÃo do amido, acelerando esse processo e causando uma compactaÃÃo da estrutura durante o assamento, favorecendo assim a coalescÃncia das bolhas dispersas na massa. / The consumption of cakes with calorie reduction is growing, but has presented challenges to be overcome in the formation of its structure when the sugar or fat are replaced by sweeteners, gums, thickeners or fat substitutes. The present study was to investigate the internal characteristics of cakes with calorie reduction and viability of producing the same pattern similar to conventional cake. Initially, we defined the mixing time and mass of fat source to be used in the formulation, using as parameter the specific volume. Next, we determined the pattern of the cake formulation from a factorial experimental design of the simple type, measuring the specific volume and cell count. From this formulation, sucrose and fat were partially substituted for the production of calorie reduced cake. Replacement of sugar was made in proportion of a mixture of xanthan gum (1.5%) and sucralose (1%), while the zein was used to replace fat. In cakes with sugar lowering evaluated the specific volume (VE), the cell count (CC) of the cakes, the apparent viscosity of the mass and its thermal properties by differential scanning calorimetry (DSC). In cakes with reduced fat content, the analyzes were VE, CC and apparent viscosity. It was found that the cake with increased VE was prepared with one hydrogenated vegetable fat and one (1) minute of mixing the dough. The standard formulation was defined as having 155.88 and 28.78 shares of sugar and fat, respectively. As the sugar content decreased (10.00 to 52.17%) were reduced specific volume (1.94 to 0.7 ml / g) and cell counts (from 36.2 to 4.0 cells / cm Â) the cake and the apparent viscosity of the mass (337.56 to 631.40 cP). By reducing the fat content, there were no significant differences between the samples for VE, CC and viscosity, and viable to produce cake with reduced calorie fat replacing up to 46.86%. For the results, it was observed that replacement of sugar contributed more sharply than the replacement of fat in the formation of defects in the structure of the cake. Thermograms of the mass of standard and cakes with reduced sucrose suggested that the presence of sucralose can reduce the starch gelatinization temperature, accelerating the process and causing a compression of the structure during baking, thereby facilitating the coalescing of bubbles dispersed in mass.