A novel kinetic model for a cocoa waste fermentation to ethanol reaction and its experimental validation.

A non-segregated kinetic model is proposed to describe a fermentation process of agro-industrial residues derived via cocoa (mucilage juice) by Pichia kudriavzevii. The novel proposed hybrid model is based on a multiple coupling reaction mechanisms (structured) to describe the kinetics of substrate consumption, biomass, carbon dioxide, and ethanol, coupled to an unstructured model for the activity enzyme. The parameters of the kinetic model are estimated by non-linear least-squares curve fitting using the Marquardt-Levenberg algorithm. In addition, numerical simulations were compared with the experimental data via residual graphs. The effectiveness of the model was statistically evaluated using dimensionless efficiency coefficients under different initial conditions. A global sensitivity analysis was applied (Fisher's information matrix). The experimental results of the batch reactor showed a maximum ethanol concentration of 29 g/L, with a yield of 0.48 g-ethanol/g-glucose and a productivity of 0.30 g/L h. The method determined that the cell formation coefficient and the specific substrate consumption rate (θ1 and θ2) directly influence most of the states of our system. The proposed scheme is particularly suitable to assist in the rational design of cell factory properties or fermentation processes because it can represent the complex biochemistry in more detail and under different initial experimental conditions; the above reveals that the generated model is robust and can be considered for control and optimization purposes.

Antifungal activity screening of fractions from Annona cherimola Mill. leaf extract against Fusarium oxysporum.

The antifungal effect of ethanolic extract fractions of Annona cherimola leaves against the mycelial growth of Fusarium oxysporum was studied. The ethanolic crude extract was solvent partitioned and the ethyl acetate phase was fractionated by column or preparative thin-layer chromatography. All fractions were developed on TLC and analyzed for acetogenins (ACG) with Kedde reagent. The antifungal effect assays were carried out in vitro by the diffusion method on PDA plates. The ethanolic extract of A. cherimola leaves was highly active against F. oxysporum growth; subfractions obtained from the antifungal screening had a significant effect (p < 0.05) on the F. oxysporum growth parameters. The screening showed that as the purification steps progressed, the inhibition of mycelial growth increased. Six bioactive ACG (Annomolon-B, 34-epi annomolon B, almunequin, cherimoline 1, cherimoline 2, and isocherimoline 1) were identified by LC-QTOF-MS/MS. These findings suggested that bioactive ACG from A. cherimola leaves could be an alternative resource of a promising botanical fungicide to control plant diseases caused by F. oxysporum.

Angiotensin-I converting enzyme inhibitory activity of Amaranthus hypochondriacus seed protein hydrolysates produced with lactic bacteria and their peptidomic profiles.

The aim of this work was to determine the in vitro antihypertensive activities of lactobacillus (L. plantarum and L. helveticus) prepared amaranth protein hydrolysates, to determine the contribution of zinc, and to identify peptides. Depending on the bacteria species and the duration of the hydrolysis, up to 45.9% inhibition of angiotensin converting enzyme (ACE) was obtained. Size separation of the most active hydrolysates to yield < 1, <3-1, <3, <10-3 and < 10 kDa fractions enhanced ACE inhibition by 2-fold. A mixed mechanism of inhibition is proposed due to low correlation of ACE and zinc chelation. Thirty-six peptides were identified in the fractions using tandem mass spectrometry. A bioinformatic analysis showed the presence of encrypted fragments such as GVSEE or VNVDDPSK with known ACE-inhibitory properties. In conclusion, lactic acid bacteria proteases released peptides from amaranth proteins with ACE-inhibitory properties that were related to the presence of peptides with known or predicted ACE-inhibitor motifs.

Human Health Risk Associated with the Consumption of Aflatoxins in Popcorn.

INTRODUCTION: Aflatoxins are secondary metabolites produced mainly by the molds Aspergillus flavus, A. parasiticus and A. nomius, and they contaminate cereals, dry fruits, oilseeds and spices. Aflatoxins have harmful effects in animals and humans, inducing vomiting, diarrhea, hepatitis, cirrhosis, immunosuppression, miscarriages, mutagenic and teratogenic effects, resulting in different cancers. Popcorn (Zea mays everta) is a cereal susceptible to aflatoxin contamination, and there are no reports about the risk of its consumption. PURPOSE: A study on the incidence and consumption of aflatoxins in popcorn marketed in the city of Veracruz, Mexico was conducted and evaluated to carry out a risk assessment for human health. METHODS: To obtain popcorn, a random sampling in 30 places was done. Frequency of consumption was obtained with informed consent of participants of 253 surveys that considered gender (56% women and 44% men), age (13 less than 18 years, 218 older than 18 years and 22 older than 60 years) and the average body weight, which was 65.5 kg for women and 72.7 kg for men. RESULTS: Aflatoxins were found in 47% of the 30 samples. The estimated daily consumption among women was 21 g of popcorn daily with 2.8 ng kg-1 body weight aflatoxin B1 (AFB1) and 18.29 ng kg-1 body weight total aflatoxins, and for men, the values were 3.0 ng kg-1 body weight AFB1, and 16.0 ng kg-1 body weight of total AF; 1 ng kg-1 body weight is recommended as the tolerance limit by the JECFA (2001). CONCLUSION: The highest liver cancer risk was detected in men population under 18 years of age, with 0.137 cases in 100,000 persons. The results show that 9.5% of the consumers of AFB1-contaminated popcorn are at risk, and 52.2% are at risk for total aflatoxin exposure. Popcorn is accessible to children with lower weight, increasing the risk.

GC-MS study of changes in polar/mid-polar and volatile compounds in Persian lime (Citrus latifolia) during fruit growth.

BACKGROUND: Citrus fruits possess a high content of bioactive compounds whose changes during fruit maturation have not been studied in depth. Fruits were sampled from week 1, after fruit onset (7 days after flowering), to week 14. Volatile compounds isolated by headspace-solid-phase microextraction and polar extracts from all samples were analyzed by gas chromatography-mass spectrometry. RESULTS: The relative abundance of 107 identified metabolites allowed differences among samples at different stages of fruit growth to be established. Principal component analysis showed a clear discrimination among samples, and analysis of variance revealed significant differences in 94 out of the 107 metabolites. Among total volatiles, monoterpenes increased their relative abundance from 86% to 94% during fruit growth, d-limonene, γ-terpinene and ß-pinene being the most abundant; conversely, sesquiterpenes decreased from 11.5% to 2.8%, ß-bisabolene and α-bergamotene being the most concentrated. Sugars, in general, exhibited a gradual increase in abundance, reaching a maximum between weeks 9 and 12. Citric and malic acids, representing approximately 90% of the total identified carboxylic acids, reached a maximum concentration at commercial maturity (week 14). CONCLUSION: Of the 107 tentatively identified metabolites during Persian lime growth, sugars, carboxylic acids, and volatiles were those that experienced more significant changes and more clearly created differences among fruit growth stages. © 2018 Society of Chemical Industry.

Assessment of Aflatoxin M1 and M2 exposure risk through Oaxaca cheese consumption in southeastern Mexico.

The present study evaluated the exposure of Southeast Mexican population to Aflatoxin M1 (AFM1) and M2 (AFM2) through the consumption of Oaxaca cheese. The intake of Oaxaca cheese was assessed via a food 7-day dairy questionnaire (N = 1100, 2014 and 2015). Thirty Oaxaca cheeses were randomly sampled, and the origin of the samples was also investigated. AFM1 and AFM2 were quantified by HPLC-FD. The exposure was assessed through the combination of the Probabilistic Density Functions (probabilistic approach). The percentage of the population at risk was calculated through the population exceeding the toxicological reference values (TDI). The risk assessment revealed that the population at higher risk to AFM1 and AFM2 was the children, followed by the adolescents and adult women. To our knowledge, the present study is the first to assess the exposure risk of different age groups of a population to AFM1 and AFM2 through the consumption of cheese.

Targeted Analysis of the Concentration Changes of Phenolic Compounds in Persian Lime (Citrus latifolia) during Fruit Growth.

Citrus fruits possess a high content of phenolic compounds; however, few studies have focused on the changes occurring during fruit growth. In this study, the changes in the concentration of 20 flavonoids, 4 phenolic acids, and their biosynthetic precursors phenylalanine and tyrosine have been evaluated during fruit maturation (14 weeks). Extracts from all samples, obtained by ultrasound assistance, were analyzed by liquid chromatography coupled to tandem mass spectrometry with a triple quad system (LC-QqQ MS/MS). In general, the concentration of flavanones, which represented over 70% of the studied phenols, and flavones increased during fruit growth, reaching their maximum concentration around week 12. In general, flavanols and phenolic acids exhibited their maximum concentration at week 5 and then decreasing significantly during the rest of maturation. Phenylalanine and tyrosine showed a sinuous behavior during fruit growth. Partial least-squares showed a clear differentiation among fruits belonging to different maturation stages, coumaric acid derivatives being the most influential variables on the projection.

Purification and characterization of peroxidase from avocado (Persea americana Mill, cv. Hass).

BACKGROUND: Avocado (Persea americana Mill, cv. Hass) fruit ranks tenth in terms of the most important products for Mexico. Avocado products are quite unstable due to the presence of oxidative enzymes such as polyphenol oxidase and peroxidase. The present study is to characterize the activity of purified avocado peroxidase from avocado in order to ascertain the biochemical and kinetic properties and their inhibition conditions. RESULTS: Purification was performed by Sephacryl S 200 HR gel filtration chromatography and its estimated molecular weight was 40 kDa. The zymogram showed an isoelectric point of 4.7. Six substrates were tested in order to ascertain the affinity of the enzyme for these substrates. The purified peroxidase was found to have low Km (0.296 mM) and high catalytic efficiency (2688 mM(-1) s(-1)) using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid), optimum activity being reached at 51°C, pH 3.8. The addition of dithiothreitol, ß-mercaptoethanol, ascorbic acid, sodium azide, L-cysteine and Tween-20 had high inhibitory effects, while metals ions such as Cu(+), Fe(2+) and Mn(2+) had weak inhibitory activity on purified avocado peroxidase. CONCLUSION: The purified avocado peroxidase exhibits high inhibition (Ki = 0.37 µM) with 1.97 µM n-propyl gallate using ABTS as substrate at 51°C, pH 3.8 for 10 min.

Relationship between fermentation index and other biochemical changes evaluated during the fermentation of Mexican cocoa (Theobroma cacao) beans.

BACKGROUND: During traditional cocoa processing, the end of fermentation is empirically determined by the workers; consequently, a high variability on the quality of fermented cocoa beans is observed. Some physicochemical properties (such as fermentation index) have been used to measure the degree of fermentation and changes in quality, but only after the fermentation process has concluded, using dried cocoa beans. This would suggest that it is necessary to establish a relationship between the chemical changes inside the cocoa bean and the fermentation conditions during the fermentation in order to standardize the process. RESULTS: Cocoa beans were traditionally fermented inside wooden boxes, sampled every 24 h and analyzed to evaluate fermentation changes in complete bean, cotyledon and dried beans. The value of the fermentation index suggested as the minimal adequate (≥1) was observed at 72 h in all bean parts analyzed. At this time, values of pH, spectral absorption, total protein hydrolysis and vicilin-class globulins of fermented beans suggested that they were well fermented. CONCLUSION: Since no difference was found between the types of samples, the pH value could be used as a first indicator of the end of the fermentation and confirmed by evaluation of the fermentation index using undried samples, during the process.

Spray-dried Bacillus thuringiensis Serovar israelensis formulations for control of Aedes aegypti larvae.

Suspensions containing 0.25 and 1.25 g/liter of Bacillus thuringiensis subsp. israelensis (Bti) spore-toxin complex were spray-dried by using maltodextrin DE-6, corn starch, and nixtamalized corn flour (25 g/liter) as materials to entrap active delta-endotoxin. The inlet air temperature of the drier was kept constant at 141 degrees C and the outlet temperature was maintained at 60 or 70 degrees C. The Probit analysis of the concentration-mortality response of third instars of Aedes aegypti (L.) larvae of the spray-dried products at 60 degrees C showed that LC50 values for maltodextrin DE-6 with 1 and 5% spore-toxin complex were 4 and 10% higher in toxicity, respectively, than that for the unformulated spore-toxin complex without drying. The LC50 value for corn starch with 1 and 5% of spore-toxin complex were also higher in toxicity (7 and 8% respectively). However, LC50 values for nixtamalized corn flour with one and 5% spore-toxin complex were 81 and 55% higher in toxicity, respectively. Dried products contain an a(w) < or = 0.7, suggesting that they are able to keep the products without microorganism growth for longer periods. The scanning electron microscope of Bti spray-dried formulations with nixtamalized corn flour showed smooth spherical particles entrapping the active ingredient. These results suggested that Bti spore-toxin complex formulated with maltodextrin DE-6, corn flour, and nixtamalized corn flour, and then spray-dried may increase larval feeding and thus increase activity against Ae. aegypti larvae.

Lactobacillus reuteri beta-galactosidase activity and low milk acidification ability.

Beta-galactosidase activity was studied as a possible cause of the low milk acidification ability observed in Lactobacillus reuteri NRRL 14171. Enzymatic activity was determined in MRS broth supplemented with either glucose or lactose and milk at the middle and final stage of the exponential phase, as well as at the stationary phase. Results were compared with beta-galactosidase activity in Lactobacillus casei NRRL-B1922, a strain that shows the milk acidification ability. The effects of the types of carbon and nitrogen sources were established by comparison of growth parameters (higher maximum cell concentration and specific growth rate) in broth culture and skim milk supplemented with 2% glucose or 1% casein peptone. In milk, L. reuteri showed higher beta-galactosidase activity in all growth phases compared with L. casei. Greater cell concentration maxima, specific growth rates, and acidification abilities were observed in L. reuteri when it was cultured in milk supplemented with 1% casein peptone compared with non-supplemented milk cultures. Results suggest that the poor milk acidification ability observed in L. reuteri may be more related to a weak proteolytic system than to deficient beta-galactosidase activity.