Isolation and identification of metallotolerant bacteria with a potential biotechnological application.

Mining has led to severe environmental pollution in countries with exhaustive mining production and inadequate industrial waste regulation. Microorganisms in contaminated sites, like mine tailings, have adapted to high concentrations of heavy metals, developing the capacity of reducing or removing them from these environments. Therefore, it is essential to thoroughly characterize bacteria present in these sites to find different ways of bioremediation. In this regard, in this study, an enrichment and isolation procedure were performed to isolate bacteria with lower nutritional requirements and high tolerance to Cu(II) and Fe(II) from two Sonoran River basin mining tails. Two Staphylococcus species and a Microbacterium ginsengisoli strain were isolated and identified from the San Felipe de Jesús mining tail. Also, three strains were isolated from the Nacozari de García mining tail: Burkholderia cenocepacia, Sphingomonas sp. and Staphylococcus warneri. Significant microbiological differences were found between the two sites. All these species exhibited tolerance up to 300 mg/L for Cu (II)-Fe (II) solutions, indicating their capacity to grow in these conditions. Moreover, a consortium of isolated bacteria was immobilized in two different biocomposites and the biocomposite with larger pore size achieved greater bacterial immobilization showcasing the potential of these bacteria in biotechnological applications.

Use of Coffee Bean Bagasse Extracts in the Brewing of Craft Beers: Optimization and Antioxidant Capacity.

Coffee bean bagasse is one of the main by-products generated by industrial coffee production. This by-product is rich in bioactive compounds such as caffeine, caffeic and chlorogenic acid, and other phenols. The aims of this work are to optimize the extraction conditions of phenolic compounds present in coffee bean bagasse and incorporate them into stout-style craft beers, as well as to determine their effect on the phenol content and antioxidant capacity. The optimal conditions for extraction were 30% ethanol, 30 °C temperature, 17.5 mL of solvent per gram of dry sample, and 30 min of sonication time. These conditions presented a total phenol content of 115.42 ± 1.04 mg GAE/g dry weight (DW), in addition to an antioxidant capacity of 39.64 ± 2.65 µMol TE/g DW in DPPH• and 55.51 ± 6.66 µMol TE/g DW for FRAP. Caffeine, caffeic and chlorogenic acids, and other minor compounds were quantified using HPLC-DAD. The coffee bean bagasse extracts were added to the stout craft beer and increased the concentration of phenolic compounds and antioxidant capacity of the beer. This work is the first report of the use of this by-product added to beers.

Determination of Chemical Constituents and Antioxidant Activities of Leaves and Stems from Jatropha cinerea (Ortega) Müll. Arg and Jatropha cordata (Ortega) Müll. Arg.

Jatropha species have been shown to be an important source of secondary metabolites with different biological effects. Jatropha cinerea (Ortega) Müll. Arg and Jatropha cordata (Ortega) Müll. Arg are distributed in the Northwestern region of Mexico, are adapted to extreme weather conditions and are widely used (stems, leaves, and sap) in traditional medicine. The aim of the present study was to carry out the phytochemical characterization and the evaluation of the antioxidant activity in methanolic extracts of stems and leaves from J. cinerea and J. cordata. The compounds present in the extracts of both species were characterized by ESI-IT-MS/MS and quantified by HPLC-DAD. The results showed that the stem extracts of both species are rich in phenolic acids, while the leaf extracts are rich in flavonoids. Some of the main compounds found were gallic acid, gentisic acid, 3,4-Dihydroxybenzoic acid, vitexin, isovitexin, and catechol. Both species showed high concentrations of phenols and total flavonoids and antioxidant activity. J.cordata showed the highest antioxidant capacity and the highest concentration of phenolic compounds. Overall, both Jatropha species are a natural source of antioxidant compounds with potential biotechnological uses.

Pecan (Carya illinoinensis (Wagenh.) K. Koch) Nut Shell as an Accessible Polyphenol Source for Active Packaging and Food Colorant Stabilization.

Herein, the antioxidant and food stabilizing properties of a pecan nut shell (PNS) hydroalcoholic extract (PNSE) are reported. Chemical degradation of PNSE demonstrated the presence of condensed tannins as the main phenolic components. PNSE showed remarkable antioxidant properties in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay (EC50 = 0.004 mg/mL). PNSE was initially tested as an inhibitor of mushroom tyrosinase, exhibiting a quite low IC50 value (0.055 mg/mL) against the enzyme diphenolase activity, suggesting its use in enzymatic browning inhibition. The anthocyanin stabilization properties were evaluated under accelerated aging conditions of both pure pigments and commercial fruit juices, and PNSE was found to be effective at concentrations (0.05 mg/mL) at which well-known stabilizers such as chlorogenic and ferulic acids proved to fail. PNSE also performed well in the stabilization of spray-dried anthocyanins for use as a food colorant, increasing the half-life of blackberry anthocyanins up to 20%. In order to explore the possibility of using PNSE as a functional additive for active packaging, polylactic acid (PLA) films containing PNSE were prepared by solvent casting, and no substantial alteration of the mechanical properties was found on addition of the extract up to 10% w/w. The films showed remarkable antioxidant properties (DDPH reduction >60% with a 3% w/w loading, at a dose of 1 mg/mL in the DPPH solution) and delayed the onset of browning of apple smoothies (ca. 30% inhibition with a 10% w/w loading). These results highlight the exploitation of PNS as a low-cost polyphenol source for food industry applications.

A novel process of the isolation of nitrifying bacteria and their development in two different natural lab-scale packed-bed bioreactors for trichloroethylene bioremediation.

Trichloroethylene (TCE) is a carcinogenic compound that is commonly present in groundwater and has been detected in drinking water sources for Mexican towns in the Mexico-US border area. Nitrifying bacteria, such as Nitrosomonas europaea, have been shown to be capable of degrading halogenated compounds, including TCE, but it is difficult to obtain high cell concentrations of these bacteria. The aim of the present study was to generate biomass of a nitrifying bacterial consortium from the sludge of an urban wastewater treatment plant (WWTP) and evaluate its capacity to biodegrade TCE in two different natural lab-scaled packed bed bioreactors. The consortium was isolated by a novel method using a continuous stirred-tank bioreactor inoculated with activated sludge from the Domos WWTP located in Cd. Obregón, Sonora, Mexico. The bioreactor was fed with specific media to cultivate ammonia-oxidizing bacteria at a dilution rate near the maximum specific growth rate reported for Nitrosomonas europaea. Optical density and suspended solids measurements were performed to determine the culture biomass production, and the presence of inorganic nitrogen species was determined by spectrophotometry. The presence of nitrifying ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) was confirmed by PCR amplification, and biofilm formation was observed by scanning electron microscopy. Batch-scale experiments confirmed the biodegradative activity of the isolated consortium, which was subsequently fixed in an inorganic carrier as zeolite and a synthetic carrier such as polyurethane to both be used as lab-scale packed-bed bioreactors, with up to 58.63% and 62.7% of TCE biodegradation achieved, respectively, demonstrating a possible alternative for TCE bioremediation in environmental and engineering systems.

Lignin and holocellulose from pecan nutshell as reinforcing fillers in poly (lactic acid) biocomposites.

Lignocellulose from agro-food biowaste represents a valuable source of cost-effective structural fillers for wholly renewable polymer composites. In this work, pecan (Carya illinoinensis) nutshell (NS) fiber and its structural components, holocellulose (HC) and acid insoluble lignin (AIL), were isolated, characterized and used as reinforcing fillers to manufacture poly(lactic acid) (PLA) based biocomposites. Thermal, morphological and mechanical properties of the prepared materials were analyzed. NS and HC acted as heterogeneous nucleating agents, potentially able to control PLA physical aging. Moreover, they significantly enhanced the viscoelastic response of PLA, mainly restricting the melt molecular mobility due to hydrodynamic effects and the formation of a three-dimensional particulate network. Flexural tests demonstrated that HC induced a 25% increase in modulus compared to the plain polymer. AIL, conversely, conferred higher ductility to the PLA matrix producing an increase in stress and strain at break of 55% and 65%, respectively. Finally, all the biocomposites showed lower resilience with respect to plain PLA due to the lack of chemical adhesion between filler and matrix. These results emphasize the potential of NS as a source of reinforcing filler in polymer-based biocomposites.

Simulated Gastrointestinal Digestion, Bioaccessibility and Antioxidant Capacity of Polyphenols from Red Chiltepin (Capsicum annuum L. Var. glabriusculum) Grown in Northwest Mexico.

Chiltepin, a wild chili mostly used in different traditional foods and traditional medicine in Northwest Mexico, represents a source of polyphenols. However, studies about the bioaccessibility of polyphenols as a parameter to measure the nutritional quality and bioefficacy of them in the fruit after consumption are scarce. Chiltepin showed phenolic acids and flavonoids contents between 387 and 65 µg/g, respectively. Nevertheless, these values decreased after the digestion process. Before digestion, gallic acid, 4-hydroxibenzoinc acid, chlorogenic acid, caffeic acid, p-coumaric acid, quercetin and luteolin were the main polyphenols found in chiltepin by HPLC-DAD and confirmed by FIA-ESI-IT-MS/MS. Gallic and chlorogenic acids were non-detected in the gastric phase, while only p-coumaric acid (5.35 ± 3.89 µg/g), quercetin (5.91 ± 0.92 µg/g) and luteolin (2.86 ± 0.62 µg/g) were found in the intestinal phase. The bioaccessibility of phenolic acids, flavonoids, and total polyphenols after the intestinal phase was around 24, 17 and 23%, respectively. Overall, results indicated that release of polyphenols from chiltepin fruit might be affected by the food matrix and gastrointestinal conditions due to the low bioaccessibility values observed.

Expression analysis of genes involved in the synthesis of oleic and linoleic acids in Jatropha cinerea seeds from Northwestern Mexico / Análise de expressão de genes envolvidos na síntese de ácidos oleico e linoleico em sementes de Jatropha cinerea do noroeste do México

ABSTRACT: Plants belonging to genus Jatropha has arisen interest because of its high oil content that could be used to produce biodiesel. It is also widely reported that the main fatty acids in Jatropha oilseed are oleic and linoleic acids. However, there are scarce studies related to native species of Jatropha from Northwestern Mexico which are adapted to arid conditions, and the expression of genes involved in fatty acid synthesis for these species is still unknown. Therefore, the aim of this study was to analyze the expression of five genes, ACP1, KASII, D9SD, FAD2-1 and FAD2-2, which are involved in the oleic and linoleic acids synthesis in mature wild-seeds of Jatropha cinerea, a native species from Sonoran Desert, using semi-quantitative RT-PCR. The J. cinerea seeds were randomly collected in Bahía de Kino, Sonora (México) which is a region characterized by its harsh environments such as saline soils and extreme temperature changes and J. curcas mature seeds from a non-toxic variety from Veracruz, Mexico were used as a reference. The RT-PCR analysis of three biological replicates were considered to ensure data consistent. Our analysis showed a higher expression of KASII and FAD2-1 genes in J. cinerea seeds compared to J. curcas, meanwhile the expression of ACP1, D9SD and FAD2-2 were higher in J. curcas. Furthermore, Actin and FAD2-1 genes sequences here obtained are the first reported for J. cinerea, thus providing information to develop further studies.

RESUMO: Plantas pertencentes ao gênero Jatropha têm despertado interesse devido ao seu alto teor de óleo que poderia ser usado para produzir biodiesel. Também é amplamente relatado que os principais ácidos graxos das oleaginosas da Jatropha são os ácidos oléico e linoleico. No entanto, existem poucos estudos relacionados a espécies nativas de pinhão-manso do noroeste do México que estão adaptados a condições áridas, e a expressão de genes envolvidos na síntese de ácidos graxos para essas espécies ainda é desconhecida. Portanto, o objetivo deste estudo foi analisar a expressão de cinco genes, ACP1, KASII, D9SD, FAD2-1 e FAD2-2, que estão envolvidos na síntese de ácidos oleico e linoleico em sementes selvagens maduras de Jatropha cinerea, espécies nativas do Deserto de Sonora, usando RT-PCR semi-quantitativa. Sementes de J. cinerea foram coletadas aleatoriamente em Bahía de Kino, Sonora (México), região caracterizada por ambientes agressivos como solos salinos e mudanças extremas de temperatura, e sementes de J. curcas maduras de uma variedade não-tóxica de Veracruz, México, usado como referência. Análise RT-PCR de três repetições biológicas foram consideradas para garantir dados consistentes. Nossa análise mostrou uma maior expressão dos genes KASII e FAD2-1 em sementes de J. cinerea comparado a J. curcas, enquanto a expressão de ACP1, D9SD e FAD2-2 foi maior em J. curcas. Além disso, as sequências dos genes Actin e FAD2-1 obtidas são as primeiras relatadas para J. cinerea, fornecendo, assim, informações para o desenvolvimento de novos estudos.

Drying and pickling on phenols, capsaicinoids, and free radical-scavenging activity in Anaheim and Jalapeño peppers / Secagem e conserva sobre os fenóis, capsaicinoides e neutralização de radicais livres em pimentas Anaheim e Jalapeño

ABSTRACT: The effects of sun-drying, air-drying and pickling processes on phenol and capsaicinoid contents, and free radical-scavenging activity [2,2-diphenyl-1-picrylhydrazyl (DPPH), and 2,2′-azino-bis 3-ethylbenzthiazoline-6-sulphonic acid (ABTS)] in Anaheim (red) and Jalapeño peppers were evaluated. Sun-drying process in Anaheim pepper caused the highest phenols retention (100%), and the free radical-scavenging activity (100%) when compared with air-drying (80%). Pickling process in Jalapeño pepper caused a moderate reduction on the phenol content (24%) and the radical-scavenging activity by DPPH (35%). Processes studied did not cause variations in the capsaicinoid fractions neither in its radical-scavenging activity. Results suggested that dried and pickled peppers are a good source of phenolics and capsaicinoids with antioxidant activity.

RESUMO: Os efeitos dos processos de secagem ao sol, secagem ao ar e conserva sobre o conteúdo dos fenóis e capsaicinoides, assim como a atividade de eliminação de radicais livres [2,2-difenil-1-picrilhidrazilo (DPPH), e ácido 2, 2'-azino-bis 3-etilbenztiazolino-6-sulfónico ABTS)] em pimentas Anaheim (vermelho) e Jalapeño foram avaliados. A secagem ao sol mostrou maior retenção de fenóis (100%) e atividade de eliminação dos radicais livres (100%) quando foi comparada com a secagem ao ar (80%). O processo de conserva mostrou uma redução moderada no teor de fenóis (24%) e na capacidade de remoção de radicais (35%). Os processos estudados não causaram alterações nas frações capsaicinoides nem na seu atividade de eliminação de radicais. Os resultados sugerem que as pimentas secas e em conserva são uma boa fonte de fenóis e capsaicinoides com atividade antioxidante.

Development of an antioxidant biomaterial by promoting the deglycosylation of rutin to isoquercetin and quercetin.

Quercetin-3-O-rutinoside (rutin), quercetin-3-O-glucoside (isoquercetin) and quercetin have shown antioxidant, cytoprotective, vasoprotective, antiproliferative and antiinflammatory properties. The aim of this work was to determine the conversion of rutin to isoquercetin and quercetin during the production of poly(l-lactic acid) films with potential to deliver these flavonoids toward tissues, pharmaceuticals or food matrices. Three poly(l-lactic acid) formulations with 17.7, 39.6 and 39.1mg/g of rutin were prepared by the extrusion process. Processing temperatures (130-165°C) promoted the deglycosylation of rutin to produce isoquercetin and subsequently quercetin, identified by high performance liquid chromatography coupled to mass spectrometry. The effect of the process on the antioxidant activity of the films was determined by measuring the capacity to scavenge 2,2 diphenyl-1-picrylhydrazyl radicals. The material with the highest proportion of quercetin showed the highest antioxidant activity which could be used to produce delivering devices of the flavonoids to tissues, pharmaceuticals or food matrices.

Antiproliferative and apoptotic activities of extracts of Asclepias subulata.

CONTEXT: Asclepias subulata Decne. (Apocynaceae) is a shrub used in the Mexican traditional medicine for the treatment of cancer. OBJECTIVE: The objective of this study was to evaluate the antiproliferative activity of methanol extract of aerial parts of A. subulata and its fractions against different cancer cell lines. Additionally, we analyzed the mechanism of action of the active fractions. MATERIALS AND METHODS: Methanol extract fractions were prepared by serial extraction with n-hexane, ethyl acetate, and ethanol. The antiproliferative activity of methanol extract and its fractions was evaluated, against several murine (M12.C3.F6, RAW 264.7, and L929) and human (HeLa, A549, PC-3, LS 180, and ARPE-19) cell lines by the MTT assay, using concentrations of 0.4-400 µg/mL for 48 h. Ethanol and residual fractions were separated using silica gel column. Apoptosis induction of cancer cells was evaluated by Annexin and JC-1 staining using flow cytometry. RESULTS: Methanol extract and its fractions showed antiproliferative activity against all human cancer cell lines tested. Methanol extract had the highest antiproliferative activity on A549 and HeLa cells (IC50 values < 0.4 and 8.7 µg/mL, respectively). Ethanol and residual fractions exerted significant antiproliferative effect on A549 (IC50 < 0.4 µg/mL) and PC3 cells (IC50 1.4 and 5.1 µg/mL). Apoptotic assays showed that CEF7, CEF9, CRF6, and CRF5 fractions induced mitochondrial depolarization in A549 cells, 70, 73, 77, and 80%, respectively. Those fractions triggered the apoptosis mitochondrial pathway. CONCLUSION: Our data show that A. subulata extracts have potent antiproliferative properties on human cancer cell lines. This plant should be considered an important source of potent anticancer compounds.

Chemical characterisation of kernels, kernel meals and oils from Jatropha cordata and Jatropha cardiophylla seeds.

BACKGROUND: Jatropha cordata and Jatropha cardiophylla are native to northwestern Mexico and are adapted to arid and semi-arid conditions (<500 mm of precipitation and temperatures from 8 to 45 °C). The aim of this study was to evaluate the chemical composition of J. cordata and J. cardiophylla kernels and oils as well as antinutrients in the defatted kernel meals of these species. RESULTS: Kernels of J. cordata and J. cardiophylla seeds analysed in this study were rich in crude protein (283 and 289 g kg(-1) respectively) and lipid (517 and 537 g kg(-1) respectively). The main fatty acids in J. cordata and J. cardiophylla oils were linoleic and oleic acids. High levels of trypsin inhibitor and phytates and low levels of saponins were present in the meals. The phorbol ester contents in J. cordata and J. cardiophylla kernel meals were 2.73 and 1.46 mg g(-1) respectively. CONCLUSION: For both J. cordata and J. cardiophylla it could be inferred that (a) the oil and kernel meal were toxic and the kernel meal could be used as livestock feed only after detoxification, (b) the oil could be used for non-alimentary purposes, i.e. biodiesel production, and (c) the seed or oil could be used for isolating various bioactive compounds for pharmaceutical and agricultural applications.

Antioxidant activity and diffusion of catechin and epicatechin from antioxidant active films made of poly(L-lactic acid).

Active membranes and food packaging containing antioxidants like catechin and epicatechin, combined with the use of materials made of biopolymers obtained from renewable sources, could create a novel alternative to reduce oxidation in food, pharmaceutical, and cosmetic products. Poly(94% L-lactic acid) films containing 1.28% catechin and 1.50% epicatechin were extruded in a pilot plant-scale extrusion machine. The diffusion kinetics of catechin and epicatechin into 95% ethanol at 20, 30, 40, and 50 °C and 50% ethanol at 40 °C displayed Fickian release behavior and diffusion coefficients between 0.5 and 50 × 10(-11) cm(2)/s. According to the Arrhenius equation, the energy of activation for the diffusion of catechin and epicatechin in the films was 110.43 and 98.92 kJ/mol, respectively. The antioxidant activity of the films was measured in methanol extracts containing 46.42 µg/mL of catechin and 57.52 µg/mL of epicatechin as 32.90 and 36.68% of scavenging the 2,2-diphenyl-1-picrylhydrazyl radical, respectively.

Efectos de una mezcla de estearina de palma y aceite de canola sobre los parámetros reológicos de la masa de trigo y características del pan / Effects of a palm stearin and canola oil mixture on the rheological parameters of wheat dough and bread / Efeitos de uma mistura de estearina de palma e óleo de canola sobre os parâmetros reológicos da massa de trigo e características do pão

Se estudiaron los efectos de la sustitución de manteca comercial por una mezcla de estearina de palma:aceite:canola 50:50, sobre las características reológicas de la masa de harina de trigo y del pan elaborado con esta masa. Se calcularon las correlaciones (r= 0,82; p<0,05) entre el trabajo de deformación (W) y la absorción de agua en el farinógrafo; y la absorción de agua y el volumen específico del pan (r= -0,55), el trabajo de deformación (W) y el volumen específico del pan (r= -0,54). Los resultados mostraron que la adición de grasa, disminuyó W, la resistencia al estiramiento (P) y la absorción de agua, y aumentó la estabilidad y el tiempo de mezclado. También se realizaron correlaciones entre el volumen del pan y la fuerza de compresión (textura) en el pan (r= -0,74). La mezcla estearina:canola 50:50 mostró un nivel mayor de ácidos grasos poliinsaturados, sin la presencia de ácidos grasos trans, proporcionando propiedades reológicas similares a las de una masa elaborada con manteca comercial, sin presentar diferencias del volumen y textura del pan. El estudio muestra que la grasa elaborada con mezcla de estearina de palma y aceite de canola es una alternativa viable para sustituir a las mantecas comerciales convencionales.

The effects of the substitution of the commercial shortening for palm stearin and canola oil mixture on the rheological properties of a wheat dough and bread were studied. The correlation coefficients between alveograph deformation work (W) and water absorption (WA) on the farinograph (r= 0.82; p<0.05), between WA and the specific volume (r= -0.55), and between W and specific volume (r= -0.54) were determined. The data show that the addition of fat decreases W, stretch resistance (P), and WA, while it increased the stability and mixing time. The correlation between loaf volume and compression force (texture) of bread (r= -0.74) was also determined. The palm stearin: canola oil 50:50 mixture had a greater level of polyunsaturated fatty acid than commercial shortening without the presence of trans fatty acids, while providing similar rheological properties, without differences in loaf volume and bread texture. The study shows that the fat elaborated with the palm stearin:canola oil mixture is a viable alternative to replace the conventional commercial shortening.

Estudaram-se os efeitos da substituição de banha comercial por uma mistura de estearina de palma:óleo:canola 50:50, sobre as características reológicas da massa de farinha de trigo e do pão elaborado com esta massa. Calcularam-se as correlações (r= 0,82; p<0,05) entre o trabalho de deformação (W) e a absorção de água no farinógrafo; e a absorção de água e o volume específico do pão (r= -0,55), o trabalho de deformação (W) e o volume específico do pão (r= -0,54). Os resultados mostraram que a adição de gordura, diminuiu W, a resistência ao estiramento (P) e a absorção de água, e aumentou a estabilidade e o tempo de mistura. Também se realizaram correlações entre o volume do pão e a força de compressão (textura) no pão (r= -0,74). A mistura estearina:canola 50:50 mostrou um nível maior de ácidos graxos poliinsaturados, sem a presença de ácidos graxos trans, proporcionando propriedades reológicas similares às de uma massa elaborada com banha comercial, sem apresentar diferenças no volume e textura do pão. O estudo mostra que a gordura elaborada com mistura de estearina de palma e óleo de canola é uma alternativa viável para substituir às banhas comerciais convencionais.