Metabolic Pathway Reconstruction Indicates the Presence of Important Medicinal Compounds in Coffea Such as L-DOPA.

The use of transcriptomic data to make inferences about plant metabolomes is a useful tool to help the discovery of important compounds in the available biodiversity. To unveil previously undiscovered metabolites of Coffea, of phytotherapeutic and economic value, we employed 24 RNAseq libraries. These libraries were sequenced from leaves exposed to a diverse range of environmental conditions. Subsequently, the data were meticulously processed to create models of putative metabolic networks, which shed light on the production of potential natural compounds of significant interest. Then, we selected one of the predicted compounds, the L-3,4-dihydroxyphenylalanine (L-DOPA), to be analyzed by LC-MS/MS using three biological replicates of flowers, leaves, and fruits from Coffea arabica and Coffea canephora. We were able to identify metabolic pathways responsible for producing several compounds of economic importance. One of the identified pathways involved in isoquinoline alkaloid biosynthesis was found to be active and producing L-DOPA, which is a common product of POLYPHENOL OXIDASES (PPOs, EC 1.14.18.1 and EC 1.10.3.1). We show that coffee plants are a natural source of L-DOPA, a widely used medicine for treatment of the human neurodegenerative condition called Parkinson's disease. In addition, dozens of other compounds with medicinal significance were predicted as potential natural coffee products. By further refining analytical chemistry techniques, it will be possible to enhance the characterization of coffee metabolites, enabling a deeper understanding of their properties and potential applications in medicine.

Production and characterization of a new distillate obtained from fermentation of wet processing coffee by-products.

Coffee is one of the most important commodities worldwide. The industrial processing of coffee cherries generates a considerable volume of by-products such as wastewater, coffee pulp, mucilage, and husk. These by-products have sugars and nutrients that can be converted into value-added products via microbial action. In this study, for the first time, we evaluated the potential of coffee pulp and coffee wastewater as substrate for alcoholic fermentation produce a distilled beverage. The must composed by dry or wet coffee pulp and coffee wastewater added of commercial sucrose or sugarcane molasses was fermented by S. cerevisiae. After a screening step, a larger fermentation was carried out with the wet pulp added of sucrose due to its higher alcoholic fermentation efficiency. The distilled beverage contained 38% (v/v) ethanol and 0.2 g/L of acetic acid. The contaminants furfural, hydroxymethylfurfural and ethyl carbamate were below detection level. Among the 48 volatile compounds detected, the majority (21) were ethyl esters usually associated with floral and sweet aromas. Ethyl decanoate (996.88 µg/L) and ethyl dodecanoate (1088.09 µg/L) were the most abundant esters. Coffee spirit presented taste acceptance of 80% and sugarcane spirit, 70%. The tasters indicated an aroma acceptance of 86% for the coffee spirit and 78% for the sugarcane spirit. The results of this work demonstrate the potential for using coffee by-products to produce a good quality distilled beverage. Considering our results, especially sensorial analysis, we can infer that the produced coffee beverage represents a new alternative for adding value to the coffee production chain.

Identification of volatile compounds, quantification of glycerol and trace elements in distilled spirits produced in Mozambique.

Distilled spirits, in addition to satisfying the legal requirements with respect to its composition, must possess a sensorial quality capable of satisfying its consumers. The present work describes the analysis of volatile compounds, trace elements and glycerol in thirteen samples of Mozambican spirits collected in different regions of the country. Extraction of volatile components was a accomplished using the headspace SPME technique, the quantification of glycerol was achieved by UV/Vis spectroscopy and the determination of trace elements by atomic absorption spectrometry. Of the 35 volatile compounds identified, 19 were esters, of which ethyl hexanoate, ethyl octanoate, ethyl nonanoate, ethyl decanoate, ethyl laurate, ethyl hexadecanoate, ethyl tetradecanoate, ethyl benzenepropanoate, 3-methylbutyl acetate and isoamyl alcohol were the predominant compounds found in the different samples, contributing to the characteristics and aroma of the beverages. As for glycerol, concentrations below 5 mg L-1 were observed in all the samples studied. The samples were found to be contaminated by metals such as copper, lead and zinc. In addition, the results provided information on the quality of Mozambican spirits regarding some aspects related to the determination of metals and glycerol. This information might stimulate producers to be more attentive to the conditions of production and the risks involved in achieving good quality spirits.

Effect of the essential oils from Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on the synthesis of ochratoxin A by Aspergillus niger and Aspergillus carbonarius isolated from tropical wine grapes.

The influence of essential oils (EOs) extracted from the leaves of Melaleuca alternifolia, Melaleuca quinquenervia and Backhousia citriodora on ochratoxin A (OTA) synthesis by fungi was studied. The extraction of EOs was performed by hydrodistillation (Clevenger apparatus) over a 2-h period and subsequently analyzed by GC-MS and GC-FID. The toxigenic activity of the essential oils (31.25; 15.62 and 7.81 µg mL-1) was evaluated by inhibiting the production of OTA by Aspergillus niger and Aspergillus carbonarius in Czapek agar medium culture. The quantification of the toxin was performed by HPLC. The production of OTA was dependent on the fungal species, incubation temperature (15 and 25 °C) and the presence of the essential oils. In tests carried out at 15 °C, the oils caused a reduction in OTA synthesis that ranged from 57.60 to 76.93% and from 54.78 to 98.68% for the fungal species A. carbonarius and A. niger, respectively. At 25 °C, reductions ranged from the 38.66 to 75.93% and from 17.94 to 71.79% for the respective fungi. The study concluded that natural products could be potential biocontrol agents against OTA contamination in food.

Correlation of the presence of acrolein with higher alcohols, glycerol, and acidity in cachaças.

Acrolein is a toxic aldehyde that can be present in various beverages, such as cachaça and other distilled spirits from sugarcane. The objective of this work was to detect and quantify acrolein in samples of cachaça produced by different processes in all regions of Brazil and to evaluate the possible routes of formation of this contaminant from the correlation with other secondary compounds present in the beverage using principal component analysis. Approximately 27.0% of the samples analyzed were outside the limit established by Brazilian legislation for this contaminant, with an average acrolein concentration of 14.01 mg 100 mL-1 anhydrous alcohol (aa). In the other samples, the average concentration was 0.97 mg 100 mL-1 aa. After selecting the variables that most closely correlated with the presence of acrolein in beverages, a positive correlation was found with the presence of butan-2-ol, propan-1-ol and volatile acids, and a slight correlation with the presence of phenolic compounds. Therefore, the presence of acrolein in cachaça can be associated with contamination of the fermentation must by bacteria of the genus Lactobacillus, as a result of the chemical degradation and enzymatic conversion of the glycerol produced during fermentation.

Kinetic, thermodynamic and physical-chemical study of the removal of copper from cachaça using coconut fibers.

The use of coconut fiber as a copper adsorbent in cachaça was optimised, and changes in the chemical quality of the beverage were observed by analytical techniques. The influence of the adsorbent mass and copper concentration parameters was investigated using the Central Composite Design, and the optimum condition was obtained through the use of Response Surface Methodology. The equilibrium of the adsorption reaction was obtained within 120 min. The kinetic data were better adjusted for the Elovich model (Elovich and Zhabrova 1939) and the isotherm data for the Sips model (Sips 1948). The maximum adsorption capacity was 1.38 mg g-1. Slight changes in the physicochemical quality of the beverage were observed, including a decrease in volatile acidity and alcohol content. Copper levels in cachaça decreased from 8.57 mg L-1 to 4.97 mg L-1 after adsorption. Therefore, the use of biomass as a metal ion adsorbent in beverages can be successfully employed.

Production and characterization of a new distilled beverage from green coffee seed residue.

This study evaluated green coffee seed residue (GCSR) as an alternative substrate for producing distilled beverages. Two proportions of GCSR, 10% and 20% (w/v), were fermented and distilled in a copper alembic still. The spirits were characterized by GC-FID, HS-SPME GC-MS, and sensory analysis by trained panelists. Most of the 62 identified volatile compounds were affected by the GCSR concentration. Total terpenes, higher alcohols, and acetals showed the highest concentrations in the 10% GCSR spirit. Esters, acetates, and aldehydes were most abundant in the 20% GCSR. In the sensory analysis, the 10% GCSR spirit was characterized by floral, dairy, and almond aromas, while the 20% GCSR spirit was embodied coffee, vegetable, hazelnut, cooked cabbage, and nut descriptors. The results demonstrate the potential of GCSR as a substrate for producing coffee spirits with chemical and sensory qualities, with the 10% GCSR being the better option for fermentation.

Effect of Fungicides on Growth and Ochratoxin A Production by Aspergillus carbonarius from Brazilian Wine Grapes.

Prevention in the field of mycotoxin-producing fungi is the most effective strategy for controlling the presence of mycotoxins in foods. Chemical fungicides are widely used to protect crops, so their implications on mycotoxin production need to be considered. Therefore, the aim of this study was to evaluate the effect in vitro and on grapes of five fungicides commonly used on grape cultures in Brazil on Aspergillus carbonarius growth and ochratoxin A (OTA) production. At the doses recommended by manufacturers, most fungicides significantly reduced A. carbonarius growth and OTA production in vitro, whereas this effect was influenced by the type of fungicide, dose, and temperature. Temperature was the main factor that influenced the effectiveness of fungicides. In general, at 15°C, fungicides showed the greatest reduction on fungal growth and OTA production. Fungicide effect on grapes was different to that on a semisynthetic grape medium. All fungicide doses were not effective at controlling A. carbonarius in grapes. Thus, the direct effect of fungicides on grapes must be studied to obtain a better approximation of field conditions. The results indicate that the use of fungicides at the doses recommended by manufacturers is better than the application at low doses. This study showed that at the lowest doses, where fungal growth is not inhibited, fungicides positively stimulate OTA production.

Effect of temperature, water activity, and pH on growth and production of ochratoxin A by Aspergillus niger and Aspergillus carbonarius from Brazilian grapes.

The growth of ochratoxigenic fungus and the presence of ochratoxin A (OTA) in grapes and their derivatives can be caused by a wide range of physical, chemical, and biological factors. The determination of interactions between these factors and fungal species from different climatic regions is important in designing models for minimizing the risk of OTA in wine and grape juice. This study evaluated the influence of temperature, water activity (aw), and pH on the development and production of OTA in a semisynthetic grape culture medium by Aspergillus carbonarius and Aspergillus niger strains. To analyze the growth conditions and production of OTA, an experimental design was conducted using response surface methodology as a tool to assess the effects of these abiotic variables on fungal behavior. A. carbonarius showed the highest growth at temperatures from 20 to 33°C, aw between 0.95 and 0.98, and pH levels between 5 and 6.5. Similarly, for A. niger, temperatures between 24 and 37°C, aw greater than 0.95, and pH levels between 4 and 6.5 were optimal. The greatest toxin concentrations for A. carbonarius and A. niger (10 µg/g and 7.0 µg/g, respectively) were found at 15°C, aw 0.99, and pH 5.35. The lowest pH was found to contribute to greater OTA production. These results show that the evaluated fungi are able to grow and produce OTA in a wide range of temperature, aw, and pH. However, the optimal conditions for toxin production are generally different from those optimal for fungal growth. The knowledge of optimal conditions for fungal growth and production of OTA, and of the stages of cultivation in which these conditions are optimal, allows a more precise assessment of the potential risk to health from consumption of products derived from grapes.