Spent coffee grounds as feedstock for the production of biosurfactants and the improved recovery of melanoidins.

Spent coffee grounds (SCG) are wastes generated in high amounts worldwide. Their composition makes them a promising feedstock for biotechnological processes. Here we show that the production of the biosurfactant surfactin by submerged culture of a Bacillus subtilis strain growing on SCG is possible, reaching concentrations up to 8.8 mg/L when using SCG at 8.3 g/L in the medium. In addition, we report a synergy between the production of surfactin and the recovery of melanoidins, an added-value compound already present in SCG. More specifically, the concentration of melanoidins in the culture medium increased between 2.1 and 2.5 times thanks to the presence of the B. subtilis in the culture. Furthermore, we have observed a strong interaction between surfactin and melanoidin aggregates through dynamic light scattering measurements, and that both of them can be co-purified with an acid precipitation. We have also characterized the interfacial and antioxidant properties of the cell-free supernatant and surfactin extract, as well as the distribution of the congeners of the biosurfactant. Altogether, this work describes a promising approach to obtain biosurfactants and antioxidant molecules in a single operation, which can be used to design several new formulations of interest for bioremediation, amendment of soils, food and cosmetics.

Seeds as Potential Sources of Phenolic Compounds and Minerals for the Indian Population.

Seeds are major sources of nutrients and bioactive compounds for human beings. In this work, the chemical composition and physicochemical properties of 155 Indian seeds (belonging to 49 families) are reported. Moisture and ash were measured with reference protocols from AOAC; total polyphenols and flavonoids were measured with spectrophotometric methods after extraction with organic solvents, and mineral elements were determined by X-ray fluorescence spectrophotometry. Total phenolic compounds, flavonoids and mineral contents (Al, Ba, Ca, Cl, Co, Cr, Cu, Fe, K, Mg, Mn, Mo, Na, P, Rb, S, Sr, Ti, V and Zn) were found to vary in the ranges 182−5000, 110−4465 and 687−7904 mg/100 g (DW), respectively. Noticeably, polyphenol contents higher than 2750 mg/100 g were observed in 18 seeds. In addition, mineral contents >5000 mg/100 g were detected in the seeds from Cuminum cyminum, Foeniculum vulgare, Commiphora wightii, Parkia javanica, Putranjiva roxburghii, Santalum album and Strychnos potatorum. Botanical and taxonomical variations in the proximate characteristics of the examined seeds are also discussed.

A useful and simple tool to evaluate and compare the intake of total dietary polyphenols in different populations.

OBJECTIVE: Polyphenols are antioxidant compounds with an impact on different health factors. Thus, it is important to have precise tools to estimate the intake of polyphenols. This study focuses on the development of an intuitive tool to estimating the intake of dietary total polyphenols. DESIGN: The tool was developed in a spreadsheet to improve accessibility and use. It is divided into six different meals for each of the 7 d with a similar format to 24-h diet recalls. The total polyphenol values of 302 foods were included and the possibility of own values. SETTING: Framework of the European project Stance4Health, Granada, Spain. PARTICIPANTS: This tool was tested on 90 participants in different stages of life (girls, women and pregnant women). Ages ranged from 10 to 35 years. RESULTS: The total polyphenol intake obtained was of 1790 ± 629 mg polyphenols/d. The highest consumption of polyphenols was observed in pregnant women (2064 mg/d). Polyphenols intake during the weekend was lower for the three groups compared to the days of the week. The results were comparable with those of other studies. CONCLUSIONS: The current tool allows the estimation of the total intake of polyphenols in the diet in a fast and easy way. The tool will be used as a basis for a future mobile application.

Dietary Fatty Acids Sustain the Growth of the Human Gut Microbiota.

While a substantial amount of dietary fats escape absorption in the human small intestine and reach the colon, the ability of resident microbiota to utilize these dietary fats for growth has not been investigated in detail. In this study, we used an in vitro multivessel simulator system of the human colon to reveal that the human gut microbiota is able to utilize typically consumed dietary fatty acids to sustain growth. Gut microbiota adapted quickly to a macronutrient switch from a balanced Western diet-type medium to its variant lacking carbohydrates and proteins. We defined specific genera that increased in their abundances on the fats-only medium, including Alistipes, Bilophila, and several genera of the class Gammaproteobacteria In contrast, the abundances of well-known glycan and protein degraders, including Bacteroides, Clostridium, and Roseburia spp., were reduced under such conditions. The predicted prevalences of microbial genes coding for fatty acid degradation enzymes and anaerobic respiratory reductases were significantly increased in the fats-only environment, whereas the abundance of glycan degradation genes was diminished. These changes also resulted in lower microbial production of short-chain fatty acids and antioxidants. Our findings provide justification for the previously observed alterations in gut microbiota observed in human and animal studies of high-fat diets.IMPORTANCE Increased intake of fats in many developed countries has raised awareness of potentially harmful and beneficial effects of high fat consumption on human health. Some dietary fats escape digestion in the small intestine and reach the colon where they can be metabolized by gut microbiota. We show that human gut microbes are able to maintain a complex community when supplied with dietary fatty acids as the only nutrient and carbon sources. Such fatty acid-based growth leads to lower production of short-chain fatty acids and antioxidants by community members, which potentially have negative health consequences on the host.

Development of a food composition database of different food contaminants CONT11 and estimation of dietary exposure in children of southern Spain.

Increasing food security is one of the Sustainable Development Goals. One of the main risks in food is the increase in food contaminants. Processing methods, such as the addition of additives or heat treatment, influence contaminant generation and increase their levels in food. The aim of the present study was to create a database using a methodology similar to that of food composition databases but with a focus on potential food contaminants. CONT11 collects information on 11 contaminants: hydroxymethyl-2-furfural, pyrraline, Amadori compounds, furosine, acrylamide, furan, polycyclic aromatic hydrocarbons, benzopyrene, nitrates, nitrites and nitrosamines. This is collected for more than 220 foods obtained from 35 different data sources. A food frequency questionnaire validated for use with children was used to validate the database. Contaminant intake and exposure in 114 children aged 10-11 years were estimated. Outcomes were within the range of values described by other studies, confirming the usefulness of CONT11. This database will allow nutrition researchers to go a step further in assessing dietary exposure to some food components and the association of this with disease, whilst also informing strategies to reduce exposure.

Profiles for identifying problematic dietary habits in a sample of recreational Spanish cyclists and triathletes.

There is a lack of sufficient information on the dietary intake and nutritional supplementation of recreational endurance athletes throughout the year. The present observational study sought to assess the dietary intake and nutritional supplementation habits of recreational cyclists and triathletes from Spain. 4,037 cyclists and triathletes completed self-report measures. Nutritional profiles were developed and differences were examined according to sporting discipline and gender. Differences between groups were compared using the Mann-Whitney U or chi-squared test. Next, micro- and macro-nutrients were grouped according to whether or not guideline intake amounts were met. The clustering of dietary habits was then examined via K-means cluster analysis. Triathletes took more supplements than cyclists (X2 = 36.489; p value = .000) and females took more supplements than males (X2 = 5.920; p value = .017). Females and triathletes reported greater protein and CHO consumption than males and cyclists, respectively. Triathletes also reported a higher consumption of total fat, MUFA, PUFA, EPA, DHA and fibre. Females and triathletes tended to consume more vitamins and minerals than males and cyclists, respectively. Two main dietary habit clusters emerged which may be used to inform nutritional interventions targeting recreational athletes not meeting nutritional requirements. There is an imbalance in the main nutrients making up the diet of recreational Spanish athletes, characterised by insufficient CHO and excessive protein.

Does the pelleting process affect the nutritive value of a pre-starter diet for suckling piglets? Ex vivo studies on mineral absorption.

BACKGROUND: The effects of pelleting on the extent of the Maillard reaction (MR) and on calcium, magnesium and zinc solubility and absorption were analysed in a conventional pre-starter diet for suckling piglets. Development was tested measuring colour, absorbance (280/420 nm), fluorescence, residual free lysine, furosine, hydroxymethylfurfural (HMF) and furfural contents before and after pelleting. Fluorescence, absorbance and mineral solubility were also measured after in vitro digestion of diets. The effects on mineral absorption were tested using Caco-2 cells. RESULTS: MR indexes confirmed the development of the reaction during the pelleting of this particular diet compared with the meal diet. The CIE-Lab colour parameters showed a decrease in luminosity (L*) and progress of the colour to the red zone (a*) in the pelleted diet. A 36% decrease in free lysine content was observed. Significant correlations were observed between fluorescence intensity and furosine levels, HMF and furfural. The pelleting process did not modify calcium and magnesium solubility after in vitro digestion, but soluble zinc increased. The efficiency of calcium and zinc transport across Caco-2 cell monolayers was greater in the pelleted diet. CONCLUSIONS: Evidence of MR development is shown, resulting in various nutritional consequences. Optimisation of pelleting could result in a better formulation of diets for feedstuffs.

Effect of Food Thermal Processing on the Composition of the Gut Microbiota.

Cooking modifies food composition due to chemical reactions. Additionally, food composition shapes the human gut microbiota. Thus, the objective of this research was to unravel the effect of different food cooking methods on the structure and functionality of the gut microbiota. Common culinary techniques were applied to five foods, which were submitted to in vitro digestion-fermentation. Furosine, 5-(hydroxymethyl)furfural, and furfural were used as Maillard reaction indicators to control the heat treatment. Short-chain fatty acids production was quantified as indicator of healthy metabolic output. Gut microbial community structure was analyzed through 16S rRNA. Both food composition and cooking methods modified the microbiota composition and released short-chain fatty acids. In general, intense cooking technologies (roasting and grilling) increased the abundance of beneficial bacteria like Ruminococcus spp. or Bifidobacterium spp. compared to milder treatments (boiling). However, for some foods (banana or bread), intense cooking decreased the levels of healthy bacteria.

Effect of coffee melanoidin on human hepatoma HepG2 cells. Protection against oxidative stress induced by tert-butylhydroperoxide.

Soluble high-molecular weight fraction (named melanoidin) from coffee brew was isolated by ultrafiltration, subsequently digested by simulating a gastric plus pancreatic digestive condition and partly characterized by CZE, gel-filtration and browning. The objective of the present study was to investigate the potential protective effect of the coffee melanoidin submitted to gastrointestinal digestion on cell viability (lactate dehydrogenase leakage) and redox status of cultured human hepatoma HepG2 cells submitted to oxidative stress induced by tert-butylhydroperoxide (t-BOOH). Concentration of reduced glutathione (GSH) and malondialdehyde (MDA), generation of reactive oxygen species (ROS) and activity of antioxidant enzymes glutathione peroxidase (GPx) and reductase (GR) were used as markers of cellular oxidative status. Pretreatment of cultured HepG2 cells with 0.5-10 microg/mL digested coffee melanoidin (DCM) for 2 or 20 h completely prevented the increase in cell damage and GR and partly prevented the decrease of GSH and the increase of MDA and GPx evoked by t-BOOH in HepG2 cells. In contrast, increased ROS generation induced by t-BOOH was not prevented when cells were pretreated with DCM. The results show that treatment of HepG2 cells with concentrations of DCM within the expected physiological range confers the cells a significant protection against an oxidative insult.

Angiotensin-I converting enzyme inhibitory activity of coffee melanoidins.

Melanoidins formed at the last stage of the Maillard reaction have been pointed out to possess certain functional properties. Potential antihypertensive activity of food melanoidins (coffee, beer, and sweet-wine) has been evaluated according to in-vitro ACE-inhibitory activity. Precision of the assay (3.2% of coefficient of variation, n = 10) for melanoidins is similar to those reported of well-known antihypertensive peptides. Assay was applied on food melanoidins obtained from coffee (three roasting degrees), beer, and sweet-wine. All samples showed in-vitro ACE-inhibitory activity. The activity in coffee melanoidins was significantly higher at more severe heating conditions. These experiments demonstrate that food melanoidins could inhibit ACE activity. In-vitro ACE-inhibitory activity of coffee melanoidins is likely located within the melanoidin structure. But ACE-inhibitory activity is also partly due to the low-molecular-weight compound nonchemically bound to the melanoidin structure, then melanoidins can act as carrier-protecting agents. These compounds could be naturally phenolic compounds present in the green beans or intermediary Maillard reaction products with antihypertensive activity.

Effect of in vitro enzymatic digestion on antioxidant activity of coffee melanoidins and fractions.

Traditionally antioxidant activity of melanoidins has only been evaluated in food for implication in shelf life but gastrointestinal digestion is necessary to study their potential bioactivity. In addition, the biological fate of melanoidins has been stressed during the past decade since they did not behave as inert substances. In the present paper a soluble coffee melanoidin isolated from brewed coffee after ultrafiltration with a 10 kDa cutoff membrane was treated ionically and enzymatically collecting the respective high and low molecular weight fractions. Antioxidant activity of these fractions was evaluated with five well-described assays (DPPH, ABTS, ORAC, HOSC, and FRAP) that were previously setup in a plate reader based automatized analysis. Low molecular weight compounds released from melanoidin after gastrointestinal digestion exerted the highest antioxidant activity, even higher than compounds bound ionically to melanoidins. Gastrointestinal digestion is able to modify coffee melanoidins to some extent, as hypothesized from their absolute antioxidant activities. Two options are plausible: by modifying/releasing the ionically bound compounds and/or by genesis of new more active structures from the melanoidin skeleton after enzymatic treatment.

New Method To Estimate Total Polyphenol Excretion: Comparison of Fast Blue BB versus Folin-Ciocalteu Performance in Urine.

Polyphenols are bioactive substances of vegetal origin with a significant impact on human health. The assessment of polyphenol intake and excretion is therefore important. The Folin-Ciocalteu (F-C) method is the reference assay to measure polyphenols in foods as well as their excretion in urine. However, many substances can influence the method, making it necessary to conduct a prior cleanup using solid-phase extraction (SPE) cartridges. In this paper, we demonstrate the use of the Fast Blue BB reagent (FBBB) as a new tool to measure the excretion of polyphenols in urine. Contrary to F-C, FBBB showed no interference in urine, negating the time-consuming and costly SPE cleanup. In addition, it showed excellent linearity (r2 = 0.9997), with a recovery of 96.4% and a precision of 1.86-2.11%. The FBBB method was validated to measure the excretion of polyphenols in spot urine samples from Spanish children, showing a good correlation between polyphenol intake and excretion.

Relationship between HMF intake and SMF formation in vivo: An animal and human study.

SCOPE: 5-Hydroxymethylfurfural (HMF) is a furanic compound produced in heat-processed foods by nonenzymatic browning reactions. HMF has been demonstrated to be hepato- and nephrotoxic in animals with a link to its metabolite 5-sulfooxymethylfurfural (SMF). To date little is known about either the formation of SMF from ingested HMF or the formation of DNA adducts in animals or human beings. METHODS AND RESULTS: To assess SMF in vivo formation, we first performed a study in mice treated with high/low doses of oral HMF. We found increased concentrations of SMF in plasma and DNA SMF-adducts in leukocytes, hepatic tissue, and kidneys by means of LC-MS/MS, but no spatial formation in such tissues was observed by MALDI-MS imaging technology due to low sensitivity. In a second experiment, we measured the exposure to HMF in a Spanish preadolescent population. We analyzed the concentration of HMF metabolites (plasma, urine) and measured, for the first time, the presence of SMF in plasma and DNA SMF-adducts in leukocytes. CONCLUSION: This study provides the first evidence that oral HMF is readily transformed into SMF in vivo, giving rise to the formation of DNA adducts in a direct relation with HMF intake, both in animals and human beings.

Relationship between acrylamide and thermal-processing indexes in commercial breakfast cereals: a survey of Spanish breakfast cereals.

Breakfast cereals are significant contributors to the daily intake of food-derived acrylamide in Western countries. Acrylamide was determined by LC-MS in 60 commercial breakfast cereals marketed in Spain. Several SPE cartridges were evaluated for clarification of the aqueous extract. LOQ was 62 microg/kg . Acrylamide content ranged from < 62-803 microg/kg (average 292 microg/kg , and median 258 microg/kg , with an estimated acrylamide intake from breakfast cereals of 2.68 microg acrylamide/person/day. According to the German concept of minimization, a signal value 450 microg/kg was calculated. Relationships among acrylamide and some parameters of the studied samples such as type of cereal, its physical form (puffed and flaked) or certain ingredients in the formulation (proteins and dietary fibre content) were also investigated. Wheat-based cereals contained significantly higher levels of acrylamide, as did samples with higher fibre or protein content. In addition, puffed breakfast cereals also contained significantly higher levels of acrylamide. There was no significant correlation between acrylamide levels and contents of 5-hydroxymethylfurfural, furosine or cereal browning.

Study on fluorescence of Maillard reaction compounds in breakfast cereals.

During the advanced stage of the Maillard reaction (MR) in food processing and cooking, Amadori rearrangement products undergo dehydration and fission and fluorescent substances are formed. Free and total (free + linked to the protein backbone) fluorescence (FIC) due to Maillard compounds in 60 commercial breakfast cereals was evaluated. Pronase was used for efficient release of linked fluorescent Maillard compounds from the protein backbone. Results were correlated with some heat-induced markers of the extent of the MR or sugar caramelisation during cereal processing, such as hydroxymethylfurfural, furfural, glucosilisomaltol and furosine. The effect of sample composition (dietary-fibre added, protein, etc.) on levels of FIC, expressed as fluorescence intensity (FI) per milligram of sample, is discussed. FIC is significantly correlated to the protein content of the sample and fluorescent Maillard compounds are mainly linked to the protein backbone. The ratio of total-FIC to free-FIC was 10.4-fold for corn-based, wheat-based and multicereal-based breakfast cereals but significantly higher in rice-based samples. Addition of dietary fibre or honey increased the FIC values. Data support the usefulness of FIC measurement as an unspecific heat-induced marker in breakfast cereals.

Application of a fast high-performance liquid chromatography method for simultaneous determination of furanic compounds and glucosylisomaltol in breakfast cereals.

The high-performance liquid chormatography method reported in the present paper provides a fast, low-cost, precise, and simple technique to analyze simultaneously 3 different indicators of nonenzymatic browning, i.e., hydroxymethylfurfural (HMF), furfural, and glucosylisomaltol (GIM), in breakfast cereals. These compounds were extracted in aqueous solution and separated on a reversed-phase C18 column with water-acetonitrile (95 + 5, v/v) mobile phase under isocratic conditions. Average recovery rates for HMF, furfural, and GIM were 99.1, 98.4, and 99.4%, respectively. The coefficients of variation were 3.67, 2.42, and 1.59% for HMF, furfural, and GIM, respectively. The detection limit was 0.01 mg/kg, and the quantitation limit was 0.05 mg/kg for the 3 studied compounds.

Antioxidant capacity, total phenols and color profile during the storage of selected plants used for infusion.

Many plants, like tea, are widely used for preparing herbal infusions. These plants have an interesting antioxidant capacity that may change after harvesting depending on the technological processing and the storage conditions. We determined the antioxidant capacity (ABTS, DPPH and FRAP methods), total phenolic content and color analysis (reflectance) of 36 plants traditionally consumed in Spain as infusion. Green tea was the most antioxidant herb, although oregano and lemon balm showed also a very high antioxidant capacity, as well as phenolic content. The antioxidant study after 3-month storage at different temperatures showed that up to a 50% of the total antioxidant capacity could be lost. Color analysis correlated with antioxidant capacity evolution, being a quick tool to control the storage conditions. Finally, our data confirm that the intake of one serving of plant infusion could release the equivalent of up to 1,500 µmol trolox, being a good source of antioxidants for the human diet.

Relationship between Glycation and Polyphenol Content and the Bioactivity of Selected Commercial Soy Milks.

Soy milk is a health-promoting beverage of which consumption is steadily expanding. Different bioactivities have been associated with soy products such as antioxidant capacity, anti-inflammatory properties, or decrease of cancer development risk. These activities have been related to the presence of several compounds, including polyphenols and serine protease inhibitors, although factors influencing such activities have been scarcely studied. In this study, we have determined the antioxidant capacity (ABTS and FRAP methods measured with the global antioxidant response, GAR protocol), total phenolic content, serine protease inhibitory activity, and presence of heat damage indicators in commercial soy milks. Polyphenols were primarily responsible for the antioxidant capacity of soy milks, increasing their concentration after digestion. Glycation under heat treatment might be responsible for decreasing protease inhibitory activities in soy milks. The results obtained support a role for furosine, a known marker of Maillard reaction and glycation, as a potential indicator to monitor both thermal treatment and effects on protease inhibitory activities in soy milk. The contribution of soy milk consumption to the daily intake of antioxidants and serine protease inhibitory activities is discussed.

Assessing the antioxidant activity of melanoidins from coffee brews by different antioxidant methods.

Antioxidant activity of instant coffees produced from the same green coffee beans roasted at three different degrees was analyzed. Coffee melanoidins were obtained by ultrafiltration (10 kDa cutoff) and subsequent diafiltration. Pure melanoidins were isolated from melanoidins after overnight incubation in 2 M NaCl and then ultrafiltered. Filtrates, corresponding to the low molecular weight (LMW) fraction noncovalently linked to the melanoidin skeleton, were also preserved. Antioxidant activity of coffee brews (CB), melanoidins (M), pure melanoidins (PM), and bounded melanoidin compounds (BMC) were tested using the 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and ferric reducing power (FRAP) methods. The correlation between the different methods was studied. The higher contribution of melanoidins to the total antioxidant activity of coffees was shown to be caused by the LMW compounds linked noncovalently to the melanoidin skeleton, as data from BMC confirmed. CB, M, and BMC fractions exert the highest antioxidant activity in aqueous media, whereas PM was not dependent on the reaction media. The highest correlation was found between DPPH and FRAP methods.

Contribution of melanoidins to the antioxidant capacity of the Spanish diet.

Most foods included within the western diet are subjected to heat processing. During such treatment, the Maillard reaction takes place, generating brown polymers known as melanoidins. Melanoidins could have health beneficial properties such as chemopreventive, antimicrobial or antioxidant capacity. Here we have established the content of melanoidins in thermally processed foods consumed regularly in the Spanish diet (chocolate, sweet wine, balsamic vinegar, beer, bread, breakfast cereals and biscuits) and their antioxidant capacity by different in vitro methodologies (ABTS, FRAP, DPPH, ORAC, HOSC). The mean intake of melanoidins is 12.2g/person/day within the Spanish diet, bread, pilsner beer and biscuits being the foods that contributed most. The most antioxidant melanoidins were those from coffee, followed by balsamic vinegar and sweet wine. Finally, the average intake of antioxidant capacity coming from melanoidins was 717 µmol Trolox/day, melanoidins from coffee, biscuits, pilsner beer and chocolate being those which possessed more antioxidant capacity.