Antioxidants and Bioactive Compounds in Food: Critical Review of Issues and Prospects.

This review paper gives an insight into the effective delivery mechanisms for health-promoting substances and highlights the challenges of using antioxidants and bioactives in foods. The selection criteria for choosing bioactives and their extraction in bioavailable form with their adequate incorporation techniques and delivery mechanisms are covered. Moreover, an overview of existing methods for determination of bioactivity is given. The importance of scientifically evaluating the effects of foods or food components on consumer health before making claims about the healthiness is aligned. Finally, a scientific perspective on how to respond to the booming demand for health-promoting products is given, and we acknowledge that despite the work done, there are still many challenges that need to be overcome.

Microencapsulation of Hibiscus sabdariffa L. Calyx Anthocyanins with Yeast Hulls.

Yeast hulls, due to their specific thin mannoprotein layer and high content of ß-glucan, constitute a promising material to stabilise the colour of anthocyanins. This study evaluates the potential of yeast hulls on the freeze-drying encapsulation of anthocyanins-rich extract from Hibiscus sabdariffa L. calyx with comparison to maltodextrin microcapsules. The moisture content (5.28-16.38%), water activity (< 0.039-0.307) and hygroscopicity (17.50-25.99 g/100 g) of obtained powders were evaluated. The stability of encapsulated anthocyanins, monitored through the total anthocyanin content, was evaluated with the pH differential method immediately after production and after a 10-week storage under different conditions of temperature (5 or 37 °C), humidity (45 or 85% RH), in presence or absence of light. The colour parameters (a, b*, L*, C*, H°, ΔE*) of powders were measured. The results indicated that yeast-hulls showed a good ability to protect anthocyanin against the influence of temperature, light, moisture compared to freeze-dried anthocyanins-rich extracts (p < 0.05). Yeast hulls protected anthocyanin better than maltodextrin under high humidity conditions (p < 0.05).

Screening of lactic acid bacteria for their potential use as aromatic starters in fermented vegetables.

Lactic acid fermentation is a traditional process to preserve foods and to modify their organoleptic properties. This process is generally conducted in a spontaneous way, allowing indigenous lactic acid bacteria (LAB) of the matrix and of the environment to compete and grow. The aim of this study was to better characterise LAB strains ability to modify aroma profiles in fruit and vegetable matrices, by focusing on two key enzymatic activities: ß-glucosidase and alcohol dehydrogenase (ADH). Firstly, 200 LAB isolated from Cambodian and Vietnamese fermented foods were screened for their ß-glucosidase activity and duplicate isolates identified through RAPD-PCR analysis were discarded. Thereby, 40 strains were found positive for ß-glucosidase using p-nitrophenyl-ß-D-glucopyranoside as substrate. Among them, 14 displayed an activity greater than 10 nmol/min/mg dry cell. Thirteen were identified as Lactiplantibacillus (L.) plantarum and one as L. pentosus. Secondly, four strains of different phenotypes for ß-glucosidase activity were tested for ADH activity. The highest reduction ability for hexanal and (E)-2-hexenal was obtained for Limosilactobacillus (L.) fermentum V013-1A for which no ß-glucosidase activity was detectable. The three other strains (L. plantarum C022-2B, C022-3B, and V0023-4B2) exhibited a lower reduction ability and only for hexanal. Thirdly, mashed tomatoes were fermented with these four strains individually to evaluate their ability to release volatile compounds from the tomato precursors. Fifty-eight volatile compounds were identified and quantified by HS-SPME/GC-MS. Untreated tomatoes were rich in aldehydes. The tomatoes fermented with L. plantarum strains were rich in ketones whereas those with L. fermentum were rich in alcohols. However, for the generation of terpenoids that provide flower and fruit flavours, our screening of ß-glucosidase activity was not able to explain the differences among the strains. For ADH activity, L. fermentum exhibited a high activity in fermentation as most of the target aldehydes and ketones disappeared and were replaced by their corresponding alcohols. The L. plantarum strains exhibited a lower activity but with an important substrate-selectivity diversity. A better knowledge of the functionality of each LAB strain in the food matrix will permit to predict and shape the aroma profiles of fermented food.

Cultivable microbial ecology and aromatic profile of "mothers" for Vino cotto wine production.

The aim of the present study was to assess the cultivable microbiota of "mothers" of Vino cotto collected from production of different years 1890, 1895, 1920, 1975, 2008. A total of 73 yeasts and 81 bacteria were isolated. Starmerella lactis-condensi, Starmerella bacillaris, Hanseniaspora uvarum, Saccharomyces cerevisiae, Hanseniaspora guillermondi and Metschnikowia pulcherrima were identified. Bacteria isolates belonged to lactic acid bacteria (Lactiplantibacillus plantarum and Pediococcus pentosaceus) and acetic acid bacteria (Gluconobacter oxydans). Remarkable biodiversity was observed for Starm. bacillaris, as well as L. plantarum and G. oxydans. Organic acids and volatile compounds were also determined. Malic and succinic acids were the main ones with values ranging from 8.49 g/L to 11.76 g/L and from 4.15 g/L to 7.73 g/L respectively, while citric acid was present at low concentrations (<0.2 g/L) in all samples. Esters and higher alcohols were the main volatile compounds detected followed by alkanes. This study permits to better understand the microbial communities associated to this product and could be considered a starting point for the definition of tailored starter cultures to improve the quality of Vino cotto preserving its typical traits.

Behind the Myth of the Fruit of Heaven, a Critical Review on Gac (Momordica cochinchinensis Spreng.) Contribution to Nutrition.

Gac, Momordica cochinchinensis (Lour.) Spreng. belongs to the Cucurbitaceae family. It is more considered as a super fruit. The demand for this plant is growing in countries where its reputation is high, including traditional countries of gac culture and countries fond of super fruits and food supplements. In these latter countries, the industrial strategy aims at producing high added value in food supplements or nutritional rich preparations. However, when marketing is not the driving force and claims have to be related to scientific data, the situation of gac is less "heavenly", mainly because its most remarkable properties are in the field of micronutrients. These latter components are indeed very important for health but their supplementation on healthy populations brings no significant advantage. This paper proposes to review aspects important for the nutritional reputation of this plant: where it comes from, how it is cultured to have an optimal nutritional composition, what is its composition and how it can impact health of consumers, in which products it is used and what are the regulations to use it in different markets. One important goal of this review is to give a critical and scientific approach to confirm data on this fruit, which has been promoted by marketing departments injecting so many wrong and unverified information. Missing data will be highlighted and potential positive applications are proposed all along the text.

Prospects for Food Fermentation in South-East Asia, Topics From the Tropical Fermentation and Biotechnology Network at the End of the AsiFood Erasmus+Project.

Fermentation has been used for centuries to produce food in South-East Asia and some foods of this region are famous in the whole world. However, in the twenty first century, issues like food safety and quality must be addressed in a world changing from local business to globalization. In Western countries, the answer to these questions has been made through hygienisation, generalization of the use of starters, specialization of agriculture and use of long-distance transportation. This may have resulted in a loss in the taste and typicity of the products, in an extensive use of antibiotics and other chemicals and eventually, in a loss in the confidence of consumers to the products. The challenges awaiting fermentation in South-East Asia are thus to improve safety and quality in a sustainable system producing tasty and typical fermented products and valorising by-products. At the end of the "AsiFood Erasmus+ project" (www.asifood.org), the goal of this paper is to present and discuss these challenges as addressed by the Tropical Fermentation Network, a group of researchers from universities, research centers and companies in Asia and Europe. This paper presents current actions and prospects on hygienic, environmental, sensorial and nutritional qualities of traditional fermented food including screening of functional bacteria and starters, food safety strategies, research for new antimicrobial compounds, development of more sustainable fermentations and valorisation of by-products. A specificity of this network is also the multidisciplinary approach dealing with microbiology, food, chemical, sensorial, and genetic analyses, biotechnology, food supply chain, consumers and ethnology.

Aroma compounds production by solid state fermentation, importance of in situ gas-phase recovery systems.

Flavour and fragrance compounds are extremely important for food, feed, cosmetic and pharmaceutical industries. In the last decades, due to the consumer's increased trend towards natural products, a great interest in natural aroma compounds has arisen to the detriment of chemically synthesised ones. Recently, solid state fermentation (SSF) has been applied in the production of many metabolites. Aroma compounds can be produced by SSF with a higher yield compared to submerged fermentation (SmF). In SSF processes, aroma compounds can be produced in the solid matrix or in the headspace, but they can be lost or stripped when aeration is required. This review focuses on the production of aroma compounds by SSF processes with a special highlight on in situ systems to recover the volatiles released in the gaseous phase and stripped due to aeration. Following a brief presentation of specificities of SSF processes concerning the choice of microorganisms and the solid matrix used for the production of aroma compounds, bioreactor aspects, factors affecting production of aroma compounds and in situ gas phase aroma recovery systems in aerated SSF bioreactors are discussed.

Encapsulation of Hibiscus sabdariffa L. anthocyanins as natural colours in yeast.

Hibiscus sabdariffa extracts, a rich source of anthocyanin, were subjected to encapsulation in yeast cells. An encapsulation yield (EY) of 208 µg/100 mg of cells and an encapsulation efficiency (EE) of 27%, were reached after optimisation of the ratios (0.5 g wet yeast cells for 5 ml of anthocyanin extracts at 1 g·L-1) and with 10% of ethanol. The storage stability of encapsulated pigments was investigated in water and buffer pH 1.5 at 5 & 37 °C for 10 days and 90 °C for 30 min. The percentage of loss of colour was determined by colourimetry assays. The microparticles made of yeast with or without heat treatment exhibited different protecting effects (P < 0.01). At 37 °C, the percentage of loss of colour in water was of 2.5% for heat-treated and 36.5% for non-treated yeast microparticles, suggesting that yeast enzymes would be responsible for the loss of anthocyanin during storage. These results are confirmed by the percentage of loss of colour which was far lower in conditions of low enzymatic activity: 3.1% at 5 °C for non-heat-treated cells in water. The pH of solvent had also an important effect on the degradation of encapsulated anthocyanin; in buffer at pH 1.5 and 37 °C with the non-heat-treated cells, the degradation decreased strongly to 9.4% compared with 36.5% in water. These results show that yeast cells are a good mean of encapsulation of pigments for a colouring purpose and that they provide anthocyanins a good protection as long as their enzymes are inactivated.

Fluorescence Lifetime and UV-Vis Spectroscopy to Evaluate the Interactions Between Quercetin and Its Yeast Microcapsule.

Quercetin is a fragile bioactive compound. Several works have tried to preserve it by encapsulation but the form of encapsulation (mono- or supra-molecular structure, tautomeric form), though important for stability and bioavailability, remains unknown. The present work aims at developing a fluorescence lifetime technique to evaluate the structure of quercetin during encapsulation in a vector capsule that has already proven efficiency, yeast cells. Molecular stabilization was observed during a 4-month storage period. The time-correlated single-photon counting (TCSPC) technique was used to evaluate the interaction between quercetin molecules and the yeast capsule. The various tautomeric forms, as identified by UV-Vis spectroscopy, result in various lifetimes in TCSPC, although they varied also with the buffer environment. Quercetin in buffer exhibited a three-to-four longer long-time after 24 h (changing from 6-7 to 18-23 ns), suggesting an aggregation of molecules. In yeast microcapsules, the long-time population exhibited a longer lifetime (around 27 ns) from the beginning and concerned about 20% of molecules compared to dispersed quercetin. This shows that lifetime analysis can show the monomolecular instability of quercetin in buffer and the presence of interactions between quercetin molecules and their microcapsules.

Molecule structural factors influencing the loading of flavoring compounds in a natural-preformed capsule: Yeast cells.

Yeast cells are efficient microcapsules for the encapsulation of flavoring compounds. However, as they are preformed capsules, they have to be loaded with the active. Encapsulation efficiency is to a certain level correlated with LogP. In this study, the effect of structural factors on the encapsulation of amphiphilic flavors was investigated. Homological series of carboxylic acids, ethyl esters, lactones, alcohols and ketones were encapsulated into the yeast Yarrowia lipolytica. Although, in a single homological series, the length of the molecule and thus the LogP were correlated with encapsulation efficiency (EY%), big differences were observable between series. For instance, carboxylic acids and lactones exhibited EY% around 45%-55%, respectively, for compounds bigger than C8 and C6, respectively, whereas ethyl esters reached only about 15-20% for C10 compounds. For a group of various C6-compounds, EY% varied from 4% for hexanal to 45% for hexanoic acid although the LogP of the two compounds was almost similar at 1.9. In total our results point out the importance of the level of polarity and localization of the polar part of the compound in addition to the global hydrophobicity of the molecule. They will be of importance to optimize the encapsulation of mixtures of compounds.

Conidiation of Penicillium camemberti in submerged liquid cultures is dependent on the nitrogen source.

OBJECTIVE: To study the ability of a commercial Penicillium camemberti strain, used for Camembert type cheese ripening, to produce conidia during growth in liquid culture (LC), in media containing different sources of nitrogen as, industrially, conidia are produced by growth at the surface of a solid state culture because conidiation in stirred submerged aerobic LC is not known. RESULTS: In complex media containing peptic digest of meat, hyphae ends did not differentiate into phialides and conidia. Contrarily, in a synthetic media containing KNO3 as sole nitrogen source, hyphae ends differentiated into phialides producing 0.5 × 10(7) conidia/ml. Conidia produced in LC were 25 % less hydrophobic than conidia produced in solid culture, and this correlates with a seven-times-lower expression of the gene rodA encoding hydrophobin RodA in the mycelium grown in LC. CONCLUSION: Conidiation of P. camembertii is stimulated in iquid medium containing KNO3 as sole source of nitrogen and therefore opens up opportunities for using liquid medium in commercial productions.

Effect of heat-processing on the antioxidant and prooxidant activities of ß-carotene from natural and synthetic origins on red blood cells.

Extraction of bioactives is a cause of structural changes in these molecules. In this work, the bioactivity of commercial natural ß-carotenes, one softly extracted without heat-assistance from Momordica cochinchinensis (BCG), one conventionally extracted from another natural source (BCC), and a synthetic one (BCS), was assessed during an additional heat-treatment mimicking formulation. Their antioxidant activities were evaluated after heat-treatment at different concentrations through hemolysis of horse red blood cells. The thermal 15-cis-isomerization of ß-carotene, characterized by DAD-HPLC, resulted in a 2.5- to 4.8-fold increase in the anti-hemolytic effect but this was undetected in chemical assay, at 4 µM. At 100 µM, BCC lost its antioxidant properties and became pro-oxidant. This effect might be caused by long-chain-oxidized-products of BCC. Results demonstrated that a short heat-treatment improves the bioactivity of ß-carotene but longer treatments made BCC prooxidant, showing that samples that underwent drastic extraction processes could not tolerate additional steps for functional food production.

Production of conidia of Penicillium camemberti in liquid medium through microcycles of conidiation.

Microcycle conidiation is a survival mechanism of fungi encountering unfavorable conditions. In this phenomenon, asexual spores germinate secondary spores directly without formation of mycelium. As Penicillium camemberti conidia have the ability to produce conidiophores after germination in liquid culture induced by a thermal stress (18 and 30 °C), our work has aimed at producing conidia through this mean. Incubation at 18 and 30 °C increased the swelling of conidia and their proportion thereby producing conidiophores. Our results showed that the microcycle of conidiation can produce 5 × 10(8) conidia ml(-1) after 7 days at 18 °C of culture. The activity of these conidia was checked through culture on a solid medium. Conidia produced by microcycle conidiation formed a normal mycelium on the surface of solid media and 25 % could still germinate after 5 months of storage.

Physico-chemical state influences in vitro release profile of curcumin from pectin beads.

Curcumin is a polyphenolic compound with diverse effects interesting to develop health benefit products but its formulation in functional foods or in food supplement is hampered by its poor water solubility and susceptibility to alkaline conditions, light, oxidation and heat. Encapsulation of curcumin could be a mean to overcome these difficulties. In this paper, curcumin was encapsulated by ionotropic gelation method in low methoxyl pectin beads associated with different surfactants: Solutol(®), Transcutol(®) and sodium caseinate. After encapsulation, physico-chemical properties of encapsulated curcumin such as its solubility, physical state, tautomeric forms and encapsulation efficiency as well as encapsulation yield were characterized. In vitro dissolution of curcumin from beads displayed different kinetic profiles according to bead composition due to different matrix network. As Solutol(®) was a good solvent for curcumin, the drug was present into amorphous form in these beads inducing a rapid release of curcumin in the simulated digestive fluids. In contrast, drug release was slower from sodium caseinate beads since curcumin was not totally dissolved during the manufacturing process. Moreover, the FLIM studies showed that a part of curcumin was encapsulated in caseinate micelles and that 34% of this drug was in keto form which may delay the curcumin release. The Transcutol beads showed also a slow drug release because of the low curcumin solubility and the high density of the matrix.

Isomerization and increase in the antioxidant properties of lycopene from Momordica cochinchinensis (gac) by moderate heat treatment with UV-Vis spectra as a marker.

Momordica cochinchinensis (gac) is a plant rich in lycopene. This pigment tends to solubilize in oil and get damaged during extraction. The impact of heating on cis-isomerization of oil-free lycopene in hexane was studied at 50 and 80°C during 240min with UV-Vis spectrometry, DAD-HPLC and TEAC test. The initial all-trans-form isomerized to the 13-cis isomer more rapidly at 80°C. After this treatment, 16% of the lycopene compounds were in the 9-cis-form. This isomer triggered an increase in the antioxidant properties which was detectable from concentrations above 9% and resulted in a change from 2.4 to 3.7µmol Trolox equivalent. It is thus possible to increase the bioactivity of lycopene samples by controlling heating. The evolution of ratios calculated from the global UV-Vis spectrum was representative of cis-isomerization and spectrometry can thus be a simple way to evaluate the state of isomerization of lycopene solutions.

Encapsulation in a natural, preformed, multi-component and complex capsule: yeast cells.

From the first observation about 40 years ago that yeast cells were interesting protective structures that could be used in several industrial applications, processes have been developed enabling technologists to incorporate several compounds possessing different physico-chemical (hydrophobic/hydrophilic) properties. Technologists screened yeast diversity to choose strains possessing the best potential and modified their physiological state to increase the uptake capability and the envelope plasticity, for instance by increasing the amount of lipids. Physico-chemical treatments were also used to improve the uptake and decrease the yeast natural material impact on the final products. For example, yeast cells could be "emptied" of their plasmic material. Yeast cells can also be coated with an additional polymeric material to increase resistance to heat treatment or decrease material liberation.These capsules can be used for several applications including carbonless paper, perfuming tissues and drug targeting, but the main industrial application deals currently with flavour encapsulation, although encapsulation in yeast is also interesting for the global food industry trend for health products.This paper proposes to review the use of yeast as an encapsulation structure focusing particularly on the properties of the yeast capsule and their impact on loading, protection, targeting and release.

Preferential localization of Lactococcus lactis cells entrapped in a caseinate/alginate phase separated system.

This study aimed to entrap bioprotective lactic acid bacteria in a sodium caseinate/sodium alginate aqueous two-phase system. Phase diagram at pH=7 showed that sodium alginate and sodium caseinate were not miscible when their concentrations exceeded 1% (w/w) and 6% (w/w), respectively. The stability of the caseinate/alginate two-phase system was also checked at pH values of 6.0 and 5.5. Lactococcus lactis subsp. lactis LAB3 cells were added in a 4% (w/w) caseinate/1.5% (w/w) alginate two-phase system at pH=7. Fluorescence microscopy allowed to observe that the caseinate-rich phase formed droplets dispersed in a continuous alginate-rich phase. The distribution of bacteria in such a system was observed by epifluorescence microscopy: Lc. lactis LAB3 cells stained with Live/Dead(®) Baclight kit™ were located exclusively in the protein phase. Since zeta-potential measurements indicated that alginate, caseinate and bacterial cells all had an overall negative charge at pH 7, the preferential adhesion of LAB cells was assumed to be driven by hydrophobic effect or by depletion phenomena in such biopolymeric systems. Moreover, LAB cells viability was significantly higher in the ternary mixture obtained in the presence of both caseinate and alginate than in single alginate solution. Caseinate/alginate phase separated systems appeared thus well suited for Lc. lactis LAB3 cells entrapment.

Advanced fluorescence technologies help to resolve long-standing questions about microbial vitality.

Advances in fundamental physical and optical principles applied to novel fluorescence methods are currently resulting in rapid progress in cell biology and physiology. Instrumentation devised in pioneering laboratories is becoming commercially available, and study findings are now becoming accessible. The first results have concerned mainly higher eukaryotic cells but many more developments can be expected, especially in microbiology. Until now, some important problems of cell physiology have been difficult to investigate due to interactions between probes and cells, excretion of probes from cells and the inability to make in situ observations deep within the cell, within tissues and structures. These technologies will enable microbiologists to address these topics. This Review aims at introducing the limits of current physiology evaluation techniques, the principles of new fluorescence technologies and examples of their use in this field of research for evaluating the physiological state of cells in model media, biofilms or tissue environments. Perspectives on new imaging technologies, such as super-resolution imaging and non-linear highly sensitive Raman microscopy, are also discussed. This review also serves as a reference to those wishing to explore how fluorescence technologies can be used to understand basic cell physiology in microbial systems.