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Fermentation behaviour and volatile compound production by agave and grape must yeasts in high sugar Agave tequilana and grape must fermentations.

Few studies have been performed on the characterization of yeasts involved in the production of agave distilled beverages and their individual fermentation properties. In this study, a comparison and evaluation of yeasts of different origins in the tequila and wine industries were carried out for technological traits. Fermentations were carried out in high (300 g l(-1)) and low (30 g l(-1)) sugar concentrations of Agave tequilana juice, in musts obtained from Fiano (white) and Aglianico (red) grapes and in YPD medium (with 270 g l(-1) of glucose added) as a control. Grape yeasts exhibited a reduced performance in high-sugar agave fermentation, while both agave and grape yeasts showed similar fermentation behaviour in grape musts. Production levels of volatile compounds by grape and agave yeasts differed in both fermentations.

Agave↗

Irritant contact dermatitis caused by needle-like calcium oxalate crystals, raphides, in Agave tequilana among workers in tequila distilleries and agave plantations.

It was found that needle-like calcium oxalate crystals, raphides, are found abundantly in all tissues of Agave tequilana plants; thus, 1 droplet (0.03 ml) of juice pressed from leaves contains 100-150 crystals, 30-500 microm in length, sharpened at both ends. In tequila distilleries, 5/6 of the workers who handle the agave stems have experienced the characteristic irritation. In contrast, only 1/3 of workers in agave plantations who harvest agave plants, complain of the irritation. It is confirmed that all the irritation suffered in both distilleries and plantations takes place at bodily locations where the plants come into contact with the worker's skin in the course of their work.

Agricultural Workers' Diseases↗

Monoxenic production of the entomopathogenic nematode Steinernema carpocapsae using culture media containing agave juice (aguamiel) from Mexican maguey-pulquero (Agave spp). Effects of the contents of nitrogen, carbohydrates and fat on infective juvenile production.

The production of infective juvenile stages (IJ) of the entomopathogenic nematode Steinernema carpocapsae in the presence of its symbiotic bacterium Xenorhabdus nematophilus was carried out in orbitally agitated bottles. Four complex culture media (M1-M4) were used, containing from 8% to 28% (by vol.) agave juice (aguamiel) from Mexican maguey-pulquero (Agave spp) as the main carbohydrate source. After 20 days of fermentation, a maximum viable IJ concentration of 249,000 IJ/ml and an initial nematode population multiplication factor of x620 were achieved when medium M4 was used (aguamiel concentration in this medium was 28% by vol.). M4 medium contained (w/v): 0.3% total nitrogen, 3.2% total carbohydrates and 3.0% total fat. According to the results obtained, total carbohydrates concentration appeared to be of great importance in obtaining high IJ concentrations.

Agave↗

Constituents of Agave americana and Agave barbadensis.

An investigation of Agave americana and Agave barbadensis resulted in the isolation of a new homoisoflavanoid, 7-hydroxy-3-(4-methoxybenzyl)-chroman (3), together with known compounds 7-hydroxy-3-(4-methoxybenzyl)-chroman-4-one (1), 5,7-dihydroxy-3-(4-methoxybenzyl)-chroman-4-one (2), cantalasaponin-1 (4), and 2-hydroxy-butanedioic acid-1-methyl ester (5).

Agave↗

Purification and characterization of the endocellular beta-glucosidase of a new strain of Candida entomophila isolated from fermenting agave (Agave sp.) juice.

A yeast strain isolated in the laboratory from fermenting agave (Agave sp.) juice was studied and classified as Candida entomophila. The beta-glucosidase of this yeast was purified by ion-exchange chromatography and gel filtration. Its molecular mass estimated by gel filtration was 400 kDa. The oligomeric structure was determined following treatment of the purified enzyme with SDS. Its optimum pH was between 5 and 6, and its optimum temperature was 60 degrees C. The enzyme was active against soluble glucosides with (1-->3)-beta, (1-->4)-beta and (1-->4)-alpha linkage configuration, and it possesses (1-->6)-alpha-arabinofuranosidase activity. It is competitively inhibited by glucose and by D-gluconic acid lactone. The enzyme was constitutive and a glucosyltransferase activity is observed in the presence of ethanol. Since the glycosides present in wines and fruit juices represent a potential source of aromatic flavour, the possible use of the yeast glucosidase for the liberation of the bound aroma is discussed.

Candida↗

Timing and rate of speciation in Agave (Agavaceae).

The Agave (Agavaceae) are keystone species of semiarid to arid regions where the geographic center of origin is Mexico but whose populations spread from the southwestern U.S. through Central America, the Caribbean, and into northern South America. Our analyses indicate that Agave is a young genus, between 7.8 and 10.1 million years old, and yet it harbors the most species of any genera in the family. Of the eight genera in the family, Agave is paraphyletic with respect to three of them, and these four genera are often grouped into a genus termed Agave sensu lato, which harbors 208 of the 293 recognized species in the Agavaceae. In this article, we examine the phylogenetic limits of Agave sensu lato and present analyses elucidating the origin and rate of speciation in the group. These analyses lead to some new insights into the phylogenetic limits of Agave, indicate an estimated age of the family between 20 and 26 million years and an age of the Agave sensu lato of </=10 million years. Furthermore, we estimate a high mean per-lineage rate of diversification for the genus and find that rates of speciation were significantly elevated between 8 and 6 million years ago and then again between 3 and 2.5 million years ago. We discuss the potential for both monocarpy and the evolution of a generalist pollination system, largely dependent on nectarivorous bat species, as possible driving factors in the radiation of the group.

Agave↗

Utilization of by-products from the tequila industry. Part 2: Potential value of Agave tequilana Weber azul leaves.

The leaves of the agave plant are left in the field after harvesting the heads for tequila production. Different types of agave leaves were isolated, classified, and their content in the total plant determined. The usable fractions were collected and their properties determined. Of the total wet weight of the agave plant, 54% corresponds to the agave head, 32% corresponds to materials which could be usable for sugar and fiber production which leaves 14% of the wet plant without apparent utility. The fractions with higher total reducing sugars (TRS) content were the fresh fraction of partially dry leaves stuck to the head and the leaf bases with a TRS content of 16.1% and 13.1%, respectively. The highest TRS concentration (16-28%) is in the agave head which is used for tequila production. The leaves are 90-120 cm long and 8-12 cm wide and contain fiber bundles that are 23-52 cm long and 0.6-13 mm wide. The ultimate fiber length is approximately 1.6 mm with an average width of 25 microns. There are several types of leaf fibers that can be utilized depending on what part of the plant they come from and what product is desired. Agave leaf fibers were pulped using a soda pulping process and the pulp was hand formed into test sheets. Test sheets made from pulped agave leaf fibers had a breaking length comparable to paper made from both pine and eucalyptus fibers, but the tear index and burst index were lower than the other two papers.

Alcoholic Beverages↗

Pulp and paper from blue agave waste from tequila production.

Pulping of blue agave waste, from the production of tequila, was evaluated by both chemical and biomechanical pulping processes. Two conventional and two organosolv systems were used to pulp the agave waste under a standard set of conditions. The soda-ethanol process was superior in terms of delignification and pulp properties in comparison to the soda and ethanol organosolv processes for pulping of agave waste; however, the kraft process gave the best strength properties. In general, the strength of the agave waste pulps was rather poor in comparison to wood and other agro-based pulps; however, the tear strength was relatively high. This result is typical of poorly bonded sheets and may be due to the coarseness of the agave fibers and/or loss of hemicelluloses in the steaming process for the tequila production. Fungal treatment of the agave waste with Ceriporiopsis subvermispora reduced the energy consumption for mechanical refining but gave biomechanical pulps with inferior strength properties. The blue agave chemical pulps should be suitable for blending with softwood kraft pulps for publication grade paper.

Alcoholic Beverages↗

Utilization of byproducts from the tequila industry: part 1: agave bagasse as a raw material for animal feeding and fiberboard production.

Agave bagasse was successfully separated into fractions that were used in sheep feeding trials. Agave bagasse can be substituted for corn stubble in the sheep's diet which resulted in improved weight gain. Agave bagasse was also processed into long and short fiber fractions with a hammermill and fiberboards of medium and high specific gravities being produced. Medium specific gravity agave fiberboards had moisture and mechanical properties comparable to medium specific gravity fiberboards made using aspen fiber. All high specific gravity agave fiberboards made from short or long fibers were stronger in bending than the ANSI standard for hardboards.

Animal Feed↗

Quantification of selected volatile constituents and anions in Mexican Agave spirits (Tequila, Mezcal, Sotol, Bacanora).

A large collection (n = 95) of Mexican Agave spirits with protected appellations of origin (Tequila, Mezcal, Sotol, and Bacanora) was analyzed using ion and gas chromatography. Because of their production from oxalate-containing plant material, all Agave spirits contained significant concentrations of oxalate (0.1-9.7 mg/L). The two Tequila categories ("100% Agave" and "mixed") showed differences in the methanol, 2-/3-methyl-1-butanol, and 2-phenylethanol concentrations with lower concentrations in the mixed category. Mezcal showed no significant differences in any of the evaluated parameters that would allow a classification. Sotol showed higher nitrate concentrations and lower 2-/3-methyl-1-butanol concentrations. Bacanora was characterized by exceptionally high acetaldehyde concentrations and a relatively low ethyl lactate content. The methanol content was the most problematic compound regarding the Mexican standards: two Tequilas (4%), five Sotols (31%), and six Bacanoras (46%) had levels above the maximum methanol content of 300 g/hL of alcohol. In conclusion, the composition of Mexican Agave spirits was found to vary over a relatively large range.

Agave↗

Water-soluble carbohydrates and fructan structure patterns from Agave and Dasylirion species.

Fructans, storage carbohydrates with beta-fructofuranosyl linkages, are found in approximately 15% of higher plants. The metabolic flexibility of those molecules allows them easily to polymerize and depolymerize to soluble carbohydrates according to plant development stage and environmental conditions. In this work, water-soluble carbohydrates, including fructan structure patterns, were compared among Agave and Dasylirion species grown in different environmental regions in Mexico. Fructans were the main storage carbohydrate present in Agave stems, in addition to other carbohydrates related to its metabolism, whereas Dasylirion spp. presented a different carbohydrate distribution. A good correlation of water-soluble carbohydrate content with climatic conditions was observed. Fructans in Agave and Dasylirion genera were found in the form of polydisperse molecules, where structural heterogeneity in the same plant was evidenced by methylation linkage analysis and chromatographic methods. Fructans from the studied species were classified into three groups depending on DP and linkage-type abundance. These storage carbohydrates share structural characteristics with fructans in plants that belong to the Asparagales members. Agave and Dasylirion fructans can be categorized as graminans and branched neo-fructans, which we have termed agavins.

Agave↗

Isolation and partial characterization of a protease from Agave americana variegata.

A new protease was isolated from an extract of leaves of Agave americana variegata. The protease (EC 3.4.-) was purified 565-fold with a yield of 39.5%. The 43.8 mg enzyme had a specific activity of 0.44 units/mg. According to electrophoretic, ultracentrifugal and other physical characterizations the enzyme was homogeneous. The enzyme had a MR of 57000, a S20,W-value of 4.37 S, a D20, W-value of 6.8-7.0 - 10(-7) cm2sec-1, a Stokes radius of 3.18 nm, a partial specific volume of 0.735 cm3g-1, a frictional ration of 1.25, a molecular absorbancy index at 280 nm of 5.773-10(4), an isoelectric point of 5.25 and contained 8-10% carbohydrate. The enzyme contained no cysteine. Agave protease could hydrolyze a variety of protein substrates although it did have a restricted specificity. It is not a sulphhydryl protease but seems to be an alkaline "serine" protease with an optimum pH of 7.8-8.0 Agave protease had marked esterolytic activity and with Cbz-Tyr-ONp had an apparent Michaelis constant of 0.0345 -10(-3) M and a V of 1.24 mol substrate/mol enzyme per sec. The enzyme did not need metal ions for optimal activity, monovalent cations did not influence its kinetic parameters, but it was inhibited by cobalt, pC1HgBzO- and TosPheCH2C1. With respect to its primary specificity, as well as its pH-dependence there was a resemblance with chymotrypsin, although the rate of hydrolysis of Agave protease is much lower.

Amino Acids↗

Pollination biology of two chiropterophilous agaves in Arizona.

I studied the pollination biology of two closely related species of agave, Agave palmeri and A. chrysantha (Agavaceae), which exhibit several chiropterophilous (bat-pollinated) traits. Floral studies, floral visitor observations, and pollination studies were conducted over four summers at six different sites to examine floral traits and determine the relative importance of diurnal vs. nocturnal pollinators. Agave chrysantha appears to have developed minor shifts in several floral characters that enhance diurnal pollination. Although floral shifts towards diurnal pollination were fewer in A. palmeri, stigmas were diurnally receptive and copious floral rewards were available in the morning, indicating that some adaptations exist to allow for multiple pollinators. Differences in fruit and seed set between naturally day- and night-pollinated umbels for both species were either not significant or significantly higher in day-pollinated plants. Bats were not important pollinators of A. chrysantha, and the mutualistic relationship between A. palmeri and the lesser long-nosed bat was found to be asymmetric. "Bat-adapted" floral traits appear to be flexible enough to respond to the climatic and pollinator unpredictability experienced by agaves at the northern edge of their distribution. This variability may be a more important factor affecting evolution of floral characters than a particular pollinator.

Journal Article↗

Characterization of three agave species by gas chromatography and solid-phase microextraction-gas chromatography-mass spectrometry.

Steam distillation (SD) extraction-solid-phase microextraction coupled to GC-MS was developed for the determination of terpenes and Bligh-Dyer extraction-derivatization coupled with GC for the determination of fatty acids such as ethyl esters were used. It was found that the three different Agave species have the same profile of fatty acids; the quantity of these compounds is different in each Agave variety. On the other hand, different terpenes were identified in the three Agave plants studied: nine in A. salmiana, eight in A. angustifolia and 32 in A. tequilana Weber var. azul.

Agave↗

Molecular structures of fructans from Agave tequilana Weber var. azul.

Agave plants utilize crassulacean acid metabolism (CAM) for CO(2) fixation. Fructans are the principal photosynthetic products generated by agave plants. These carbohydrates are fructose-bound polymers frequently with a single glucose moiety. Agave tequilana Weber var. azul is an economically important CAM species not only because it is the sole plant allowed for tequila production but because it is a potential source of prebiotics. Because of the large amounts of carbohydrates in A. tequilana, in this study the molecular structures of its fructans were determined by fructan derivatization for linkage analysis coupled with gas chromatography-mass spectrometry (GC-MS), nuclear magnetic resonance (NMR), and matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF-MS). Fructans were extracted from 8-year-old A. tequilana plants. The linkage types present in fructans from A. tequilana were determined by permethylation followed by reductive cleavage, acetylation, and finally GC-MS analysis. Analysis of the degree of polymerization (DP) estimated by (1)H NMR integration and (13)C NMR and confirmed by MALDI-TOF-MS showed a wide DP ranging from 3 to 29 units. All of the analyses performed demonstrated that fructans from A. tequilana consist of a complex mixture of fructooligosaccharides containing principally beta(2 --> 1) linkages, but also beta(2 --> 6) and branch moieties were observed. Finally, it can be stated that fructans from A. tequilana Weber var. azul are not an inulin type as previously thought.

Agave↗

The uses of AFLP for detecting DNA polymorphism, genotype identification and genetic diversity between yeasts isolated from Mexican agave-distilled beverages and from grape musts.

AIMS: The objectives were to determine the variability and to compare the genetic diversity obtained using amplified fragment length polymorphism (AFLP) markers in analyses of wine, tequila, mezcal, sotol and raicilla yeasts. METHODS AND RESULTS: A molecular characterization of yeasts isolated from Mexican agave musts, has been performed by AFLP marker analysis, using reference wine strains from Italian and South African regions. CONCLUSIONS: A direct co-relation between genetic profile, origin and fermentation process of strains was found especially in strains isolated from agave must. In addition, unique molecular markers were obtained for all the strains using six combination primers, confirming the discriminatory power of AFLP markers. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report of molecular characterization between yeasts isolated from different Mexican traditional agave-distilled beverages, which shows high genetic differences with respect to wine strains.

Agave↗