Publications by authors named "Alfonso Larqué-Saavedra"

12 Publications

  • Page 1 of 1

Sensory attributes, physicochemical and antioxidant characteristics, and protein profile of wild prickly pear fruits (O. macrocentra Engelm., O. phaeacantha Engelm., and O. engelmannii Salm-Dyck ex Engelmann.) and commercial prickly pear fruits (O. ficus-indica (L.) Mill.).

Food Res Int 2021 Feb 24;140:109909. Epub 2020 Nov 24.

Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo Envolvente del Pronaf y Estocolmo s/n, C.P. 32310 Ciudad Juárez, Chihuahua, Mexico. Electronic address:

Mexico presents the highest richness of Opuntia Mill. species. These species are an important economic factor for the country, and source of nutrients, bioactive compounds, pigments, and nutraceuticals which can be of interest for the food and pharmaceutical industry. However, there are some wild Opuntia species in the Chihuahua desert, that have not been analyzed to establish their properties and potential use. The aim of study was to evaluate the sensory, physicochemical and protein profile in wild prickly pear fruits (O. macrocentra Engelm. (OM), O. phaeacantha Engelm. (OP), and O. engelmannii Salm-Dyck ex Engelmann. (OE)) from Samalayuca, Chihuahua and compare them with two commercial prickly pear fruits (O. ficus-indica (L.) Mill. (green-OFG, red-OFR). The sensory profile of wild species was characterized by highest color, odor, and sour taste compared to the commercial fruits. Pulp, peel, and seeds from wild prickly pear fruits showed lower pH, and higher titratable total acidity, total phenolic compounds, total flavonoids, antioxidant capacity, protein, lipids, ash, carbohydrates (only peel), and crude fiber content than commercial Opuntia species. Furthermore, O. engelmannii showed a tendency to present the highest betacyanins, betaxanthins, and betalains contents. A total of 181, 122, 113, 183 and 140 different proteins were identified in OM, OP, OE, OFG, OFR species, respectively. All species showed the highest enrichment in three main pathways such as amino acids biosynthesis, glycolysis (dark)/gluconeogenesis (light), and the citric acid cycle. The wild prickly pear fruits of this study showed important nutritional, protein, and antioxidant properties with biological interest, and can be a potential source of functional ingredients and nutraceuticals.
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http://dx.doi.org/10.1016/j.foodres.2020.109909DOI Listing
February 2021

Sw. (Ramón): An Alternative to Improve the Nutritional Properties and Functional Potential of the Wheat Flour Tortilla.

Foods 2019 Nov 24;8(12). Epub 2019 Nov 24.

Instituto de Ciencias Biomédicas, Universidad Autónoma de Ciudad Juárez, Anillo Envolvente del Pronaf y Estocolmo s/n, C.P. Ciudad Juárez, Chihuahua 32310, Mexico.

The wheat flour tortilla (WFT) is a Mexican food product widely consumed in the world, despite lacking fiber and micronutrients. Ramón seed flour (RSF) is an underutilized natural resource rich in fiber, minerals and bioactive compounds that can be used to improve properties of starchy foods, such as WFT. The study evaluated the impact of partial replacement of wheat flour with RSF on the physicochemical, sensory, rheological and nutritional properties and antioxidant capacity (AC) of RSF-containing flour tortilla (RFT). Results indicated that RFT (25% RSF) had higher dietary fiber (4.5 times) and mineral (8.8%; potassium 42.8%, copper 33%) content than WFT. Two sensory attributes were significantly different between RTF and WFT, color intensity and rollability. RFT was soft and it was accepted by the consumer. Phenolic compounds (PC) and AC were higher in RFT (11.7 times, 33%-50%, respectively) than WFT. PC identification by ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) showed that phenolic acids esterified with quinic acid, such as chlorogenic and other caffeoyl and coumaroyl derivatives were the major PC identified in RSF, resveratrol was also detected. These results show that RSF can be used as an ingredient to improve nutritional and antioxidant properties of traditional foods, such as the WFT.
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http://dx.doi.org/10.3390/foods8120613DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6963599PMC
November 2019

Consolidated Bioprocess for Bioethanol Production from Raw Flour of Seeds Using the Native Strain of Bm-2.

Microorganisms 2019 Oct 23;7(11). Epub 2019 Oct 23.

Renewable Energy Department, Yucatan Center for Scientific Research, Merida 97302, Mexico.

Consolidated bioprocessing (CBP), which integrates biological pretreatment, enzyme production, saccharification, and fermentation, is a promising operational strategy for cost-effective ethanol production from biomass. In this study, the use of a native strain of (Bm-2) was evaluated for bioethanol production from in a CBP. The raw seed flour obtained from the ramon tree contained 61% of starch, indicating its potential as a raw material for bioethanol production. Quantitative assays revealed that the Bm-2 strain produced the amylase enzyme with activity of 193.85 U/mL. The Bm-2 strain showed high tolerance to ethanol stress and was capable of directly producing ethanol from raw flour at a concentration of 13 g/L, with a production yield of 123.4 mL/kg flour. This study demonstrates the potential of Bm-2 for starch-based ethanol production in a consolidated bioprocess to be implemented in the biofuel industry. The residual biomass after fermentation showed an average protein content of 22.5%, suggesting that it could also be considered as a valuable biorefinery co-product for animal feeding.
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http://dx.doi.org/10.3390/microorganisms7110483DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6920830PMC
October 2019

Genetic diversity of Clavispora lusitaniae isolated from Agave fourcroydes Lem, as revealed by DNA fingerprinting.

J Microbiol 2015 Jan 4;53(1):14-20. Epub 2015 Jan 4.

Laboratorio GeMBio, Centro de Investigación Científica de Yucatán A.C., Calle 43 # 130, Col. Chuburná de Hidalgo, Mérida, Yucatán, 97200, México.

This study characterized Clavispora lusitaniae strains isolated from different stages of the processing and early fermentation of a henequen (Agave fourcroydes) spirit produced in Yucatan, Mexico using a molecular technique. Sixteen strains identified based on morphological features, obtained from different substrates, were typed molecularly. Nine different versions of the divergent D1/D2 domain of the large-subunit ribosomal DNA sequence were identified among the C. lusitaniae strains. The greatest degree of polymorphism was found in the 90-bp structural motif of the D2 domain. The MSP-PCR technique was able to differentiate 100% of the isolates. This study provides significant insight into the genetic diversity of the mycobiota present during the henequen fermentation process, especially that of C. lusitaniae, for which only a few studies in plants have been published. The applied MSP-PCR markers were very efficient in revealing olymorphisms between isolates of this species.
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http://dx.doi.org/10.1007/s12275-015-4373-4DOI Listing
January 2015

Understanding the physiological responses of a tropical crop (Capsicum chinense Jacq.) at high temperature.

PLoS One 2014 3;9(11):e111402. Epub 2014 Nov 3.

Unidad de Recursos Naturales, Centro de Investigación Científica de Yucatán, Mérida, Yucatán, México.

Temperature is one of the main environmental factors involved in global warming and has been found to have a direct effect on plants. However, few studies have investigated the effect of higher temperature on tropical crops. We therefore performed an experiment with a tropical crop of Habanero pepper (Capsicum Chinense Jacq.). Three growth chambers were used, each with 30 Habanero pepper plants. Chambers were maintained at a diurnal maximum air temperature (DMT) of 30 (chamber 1), 35 (chamber 2) and 40°C (chamber 3). Each contained plants from seedling to fruiting stage. Physiological response to variation in DMT was evaluated for each stage over the course of five months. The results showed that both leaf area and dry mass of Habanero pepper plants did not exhibit significant differences in juvenile and flowering phenophases. However, in the fruiting stage, the leaf area and dry mass of plants grown at 40°C DMT were 51 and 58% lower than plants at 30°C DMT respectively. Meanwhile, an increase in diurnal air temperature raised both stomatal conductance and transpiration rate, causing an increase in temperature deficit (air temperature - leaf temperature). Thus, leaf temperature decreased by 5°C, allowing a higher CO2 assimilation rate in plants at diurnal maximum air temperature (40°C). However, in CO2 measurements when leaf temperature was set at 40°C, physiological parameters decreased due to an increase in stomatal limitation. We conclude that the thermal optimum range in a tropical crop such as Habanero pepper is between 30 and 35°C (leaf temperature, not air temperature). In this range, gas exchange through stomata is probably optimal. Also, the air temperature-leaf temperature relationship helps to explain how temperature keeps the major physiological processes of Habanero pepper healthy under experimental conditions.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0111402PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4218724PMC
August 2015

Hydrolysis of Agave fourcroydes Lemaire (henequen) leaf juice and fermentation with Kluyveromyces marxianus for ethanol production.

BMC Biotechnol 2014 Feb 14;14:14. Epub 2014 Feb 14.

Centro de Investigación Científica de Yucatán AC, Calle 43 No, 130 Col, Chuburná de Hidalgo, Mérida, Yucatán 97200, Mexico.

Background: Carbon sources for biofuel production are wide-ranging and their availability depends on the climate and soil conditions of the land where the production chain is located. Henequen (Agave fourcroydes Lem.) is cultivated in Yucatán, Mexico to produce natural fibers from the leaves, and a juice containing fructans is produced during this process. Fructans can be hydrolyzed to fructose and glucose and metabolized into ethanol by appropriate yeasts. In Mexico, different Agave species provide the carbon source for (distilled and non-distilled) alcoholic beverage production using the stem of the plant, whilst the leaves are discarded. In this work, we investigated the effect of thermal acid and enzymatic hydrolysis of the juice on the amount of reducing sugars released. Growth curves were generated with the yeasts Saccharomyces cerevisiae and Kluyveromyces marxianus and fermentations were then carried out with Kluyveromyces marxianus to determine alcohol yields.

Results: With thermal acid hydrolysis, the greatest increase in reducing sugars (82.6%) was obtained using 5% H2SO4 at 100°C with a 30 min reaction time. Statistically similar results can be obtained using the same acid concentration at a lower temperature and with a shorter reaction time (60°C, 15 min), or by using 1% H2SO4 at 100°C with a 30 min reaction time. In the case of enzymatic hydrolysis, the use of 5.75, 11.47 and 22.82 U of enzyme did not produce significant differences in the increase in reducing sugars. Although both hydrolysis processes obtained similar results, the difference was observed after fermentation. Ethanol yields were 50.3 ± 4 and 80.04 ± 5.29% of the theoretical yield respectively.

Conclusions: Final reducing sugars concentrations obtained with both thermal acid and enzymatic hydrolysis were similar. Saccharomyces cerevisiae, a good ethanol producer, did not grow in the hydrolysates. Only Kluyveromyces marxianus was able to grow in them, giving a higher ethanol yield with the enzymatic hydrolysate. The leaves account for a non-negligible weight of the total agave plant biomass, so this work complements the knowledge already developed on agave fermentations by making it possible to produce ethanol from almost the entire plant (stem and leaves).
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http://dx.doi.org/10.1186/1472-6750-14-14DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3932041PMC
February 2014

PCR-based detection and characterization of the fungal pathogens Colletotrichum gloeosporioides and Colletotrichum capsici causing anthracnose in papaya (Carica papaya l.) in the Yucatan peninsula.

Mol Biotechnol 2008 Nov 1;40(3):293-8. Epub 2008 Aug 1.

Laboratorio GeMBio, Centro de Investigación Científica de Yucatán, Calle 43 # 130, Col. Chuburná de Hidalgo, Merida, Yucatan 97200, Mexico.

Colletotrichum gloeosporioides is the common causal agent of anthracnose in papaya (Carica papaya L.) fruits, and infection by this fungal pathogen results in severe post-harvest losses. In the Yucatán peninsula (Mexico) a different Colletotrichum species was isolated from papaya fruits with atypical anthracnose lesions. The DNAs from a variety of Colletotrichum isolates producing typical and atypical lesions, respectively, were amplified by PCR with C.gloeosporioides-specific primers. All isolates from typical anthracnose lesions yielded a 450 bp PCR product, but DNAs from isolates with atypical lesions failed to produce an amplification product. For further characterization, the rDNA 5.8S-ITS region was amplified by PCR and processed for sequencing and RFLP analysis, respectively, to verify the identity of the papaya anthracnose pathogens. The results revealed unequivocally the existence of two Colletotrichum species causing anthracnose lesions on papaya fruits: C. gloeosporioides and C. capsici. PCR-RFLP using the restriction endonuclease MspI reliably reproduced restriction patterns specific for C. capsici or C. gloeosporioides. The generation of RFLP patterns by MspI (or AluI or RsaI) is a rapid, accurate, and unequivocal method for the detection and differentiation of these two Colletotrichum species.
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http://dx.doi.org/10.1007/s12033-008-9093-0DOI Listing
November 2008

Ethanol production from henequen (Agave fourcroydes Lem.) juice and molasses by a mixture of two yeasts.

Bioresour Technol 2008 Dec 3;99(18):9036-9. Epub 2008 Jun 3.

Centro de Investigación Científica de Yucatán, Chuburná de Hidalgo, Mérida 97200, Yucatán, Mexico.

In the fermentation process of henequen (Agave fourcroydes Lem.) leaf juice, complemented with industrial molasses, the use of an inoculum comprising two yeasts: Kluyveromyces marxianus (isolated from the henequen plant) and Saccharomyces cerevisiae (commercial strain) was studied. An ethanol production of 5.22+/-1.087% v/v was obtained. Contrary to expected, a decrease on ethanol production was observed with the use of the K. marxianus strain. The best results were obtained when a mixture of 25% K. marxianus and 75% S. cerevisiae or S. cerevisiae alone were used with an initial inoculum concentration of 3x10(7)cellmL(-1). Furthermore, it was possible to detect a final concentration of approximately 2-4gL(-1) of reducing sugars that are not metabolized by the yeasts for the ethanol production. These results show that although the use of a mixture of yeasts can be of interest for the production of alcoholic beverages, it can be the opposite in the case of ethanol production for industrial purposes where manipulation of two strains can raise the production costs.
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http://dx.doi.org/10.1016/j.biortech.2008.04.063DOI Listing
December 2008

Molecular characterization of Kluyveromyces marxianus strains isolated from Agave fourcroydes (Lem.) in Yucatan, Mexico.

Mol Biotechnol 2007 Nov 10;37(3):181-6. Epub 2007 Jul 10.

Laboratorio GeMBio, Centro de Investigación Científica de Yucatán, Calle 43 # 130, Col. Chuburná de Hidalgo, Merida, Yucatan, Mexico,

The molecular characterization of 14 strains of Kluyveromyces marxianus isolated from Agave fourcroydes (Lem.) in Yucatan, Mexico, was performed by AP-PCR analysis, PCR-RFLP of 5.8S-ITS, and complete NTS regions. A sequence analysis of the D1/D2 domain of the 26S rDNA was also carried out in six selected strains. The AP-PCR approach had the highest discrimination power for the molecular characterization of new henequen K. marxianus strains. PCR-RFLP of 5.8S-ITS regions did not reveal polymorphisms in this group of strains. The restriction enzyme digestion analysis of NTS region enables the separation among strains which coincides with ascospore shape groups. The molecular tools used in this article may be useful to confirm a preliminary screen of yeasts isolated from henequen without the use of growth characteristics or morpho-physiological tests.
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http://dx.doi.org/10.1007/s12033-007-0036-yDOI Listing
November 2007

Responses of transformed Catharanthus roseus roots to femtomolar concentrations of salicylic acid.

Plant Physiol Biochem 2007 Jun-Jul;45(6-7):501-7. Epub 2007 Apr 14.

Unidad de Bioquímica y Biología Molecular de Plantas, Centro de Investigación Científica de Yucatán, A. C., Calle 43 No. 130, Col. Chuburná de Hidalgo, CP 97200, Mérida, Yucatán, Mexico.

Catharanthus roseus transformed roots were cultured in the presence of salicylic acid (SA) at concentrations between 0.1 fM and 100 pM and the effect on root growth was evaluated. Significant morphological changes in the lateral roots were recorded on day two in the SA treatment. Presence of SA increased root cap size and caused the appearance of lateral roots closer to the root tip. The bioassay was sensitive enough to allow testing of low concentrations of other growth regulators that may affect root morphology and physiology.
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http://dx.doi.org/10.1016/j.plaphy.2007.04.003DOI Listing
October 2007

A rapid and simple method for DNA extraction from yeasts and fungi isolated from Agave fourcroydes.

Mol Biotechnol 2006 May;33(1):67-70

Laboratorio GeMBio, Centro de Investigación Científica de Yucatán, Calle 43 # 130, Col. Chuburná de Hidalgo, Mérida 97200, Yucatán, México.

A simple and easy protocol for extracting high-quality DNA from different yeast and filamentous fungal species is described. This method involves two important steps: first, the disruption of cell walls by mechanical means and freezing; and second, the extraction, isolation, and precipitation of genomic DNA. The absorbance ratios (A(260)/A(280)) obtained ranged from 1.6 to 2.0. The main objective of this procedure is to extract pure DNA from yeast and filamentous fungi, including those with high contents of proteins, polysaccharides, and other complex compounds in their cell walls. The yield and quality of the DNAs obtained were suitable for micro/minisatellite primer-polymerase chain reaction (MSP-PCR) fingerprinting as well as for the sequence of the D1/D2 domain of the 26S rDNA.
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http://dx.doi.org/10.1385/MB:33:1:67DOI Listing
May 2006

A fast, simple, and reliable high-yielding method for DNA extraction from different plant species.

Mol Biotechnol 2005 Oct;31(2):137-9

Laboratorio GeMBio, Centro de Investigación Científica de Yucatán, Calle 43, #130, Col. Chuburná de Hidalgo, Mérida 97200, Yucatán, México.

Genetic studies and pathogen detection in plants using molecular methods require the isolation of DNA from a large number of samples in a short time span. A rapid and versatile protocol for extracting high-quality DNA from different plant species is described. This method yields from 1 to 2 mg of DNA per gram of tissue. The absorbance ratios (A260/A280) obtained ranged from 1.6 to 2.0. A minimal presence of contaminating metabolites (as polymerase chain reaction [PCR] inhibitors) in samples and a considerable savings in reagents are characteristics of this protocol, as well as the low cost of the analysis per sample. The quality of the DNA was suitable for PCR amplification.
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http://dx.doi.org/10.1385/MB:31:2:137DOI Listing
October 2005