0000000000003355
AUTHOR
J.j. Mateo
Biotechnological characterisation of exocellular proteases produced by enologicalHanseniasporaisolates
Summary Twenty-six enological Hanseniaspora isolates were identified by ITS PCR-RFLP and D1/D2 domain of 26S rDNA sequencing and assayed for exocellular protease production. Based on qualitative data, six isolates, belonging to H. guilliermondii, H. valbyensis and H. occidentalis species, were selected to continue the study. Analytical procedure was optimised, and protease activities were quantified and characterised on the basis of different biotechnological factors. Protease activity was quite glucose, fructose and ethanol content independent, but divalent cation affects activity; these data support that they were aspartic proteases. The effect of 2-mercaptoethanol suggests the importance…
Characterisation of Hanseniaspora isolates with potential aroma-enhancing properties in Muscat wines
The sensorial characteristics of the wines produced with Muscat grapes are related to the level of terpene alcohols, so an improvement of such a level is expected as a result of hydrolytic processes conducted by Hanseniaspora. The aim of this work was to select and identify new strains for this purpose. In a second step, the strains were assayed to evaluate their oenological abilities. H. uvarum H107 and H. vineae G26 and P38 were proven to be good candidates to be used in commercial vinification processes to enhance wine properties. Wine inoculated with yeasts showed an increase in the level of aromatic compounds.
Critical study of and improvements in chromatographic methods for the analysis of type B trichothecenes
Various analytical methods used in the analysis of type B trichothecenes (deoxynivalenol, nivalenol, 3- and 15-acetyldeoxynivalenol) in cereals were compared and optimised in this work. These methods use either GC-electron-capture detection (ECD) of trimethylsilyl, trifluoroacetyl and heptafluorobutyryl derivatives or HPLC with UV or photodiode array detection of analytes. A new HPLC procedure using fluorescence detection prior derivatisation with coumarin-3-carbonyl chloride has been also tested. Five extraction solvents and two solid-phase extraction cartridges (silica, Florisil) plus a especial clean-up column (MycoSep 225) were compared in order to obtain the best recovery of the mycoto…
Enzyme contribution of non-Saccharomyces yeasts to wine production
The fermentation of grape must to produce wine is a biologically complex process, carried on by yeasts and malolactic bacteria. The yeasts present in spontaneous fermentation may be divided into two groups, the Saccharomyces yeasts, particularly S. cerevisiae, and the non-Saccharomyces yeasts which include members of the genera Rhodotorula, Pichia, Candida, Debaryomyces, Metschtnikowia, Hansenula and Hanseniaspora. S. cerevisiae yeasts are able to convert sugar into ethanol and CO2 via fermentation. They have been used for thousands of years by mankind for the production of fermented beverages and foods, including wine. Their enzymes provide interesting wine organoleptic characteristics. β-…
Comparison of extraction and clean-up procedures for analysis of zearalenone in corn, rice and wheat grains by high-performance liquid chromatography with photodiode array and fluorescence detection.
The aim of this work was the optimization of some procedures usually used in the analysis of zearalenone (ZEA) in corn and other cereals by high-performance liquid chromatography (HPLC) with photodiode array and/or fluorescence detection. The comparison of five extraction solvents is presented. Three solid-phase extraction cartridges (C-18, silica, Florisil) and immuno-affinity columns were also compared to obtain the best recovery of the mycotoxin with the minimal presence of co-extractives in the chromatograms. Mixtures of methanol-1% aqueous NaCl (80.20 or 60:40 v/v) were the best extraction solvents. Florisil provided higher recovery of ZEA than C-18, and silica proved unsuitable. The i…
Characterization of a β-glucosidase isolated from an alpeorujo strain of Candida adriatica
ABSTRACTA β-glucosidase-producing strain, Candida adriatica CECT13142, was isolated from olive oil wastes (alpeorujo) and identified by PCR/restriction fragment length polymorphism of the rDNA internal transcribed spacer and sequence analysis of the D1/D2 region of the 26S rRNA gene techniques. The enzyme was purified by sequential chromatography on DEAE-cellulose and Sephadex G-100. The relative molecular mass of the enzyme was estimated to be 50 kDa by SDS-PAGE. The hydrolytic activity of the β-glucosidase had an optimum pH of 8.2 and an optimum temperature of 40°C. The enzyme displayed high substrate specificity and high catalytic efficiency (Km 0.85 mM, Vmax 12.5 U/g of cells) for p-nit…
Characterization of an ethanol-tolerant 1,4-β-xylosidase produced byPichia membranifaciens
Aims: The purification and biochemical properties of the 1,4-β-xylosidase of an oenological yeast were investigated. Methods and Results: An ethanol-tolerant 1,4-β-xylosidase was purified from cultures of a strain of Pichia membranifaciens grown on xylan at 28°C. The enzyme was purified by sequential chromatography on DEAE cellulose and Sephadex G-100. The relative molecular mass of the enzyme was determined to be 50 kDa by SDS-PAGE. The activity of 1,4-β-xylosidase was optimum at pH 6·0 and at 35°C. The activity had a Km of 0·48 ± 0·06 mmol l−1 and a Vmax of 7·4 ± 0·1 μmol min−1 mg−1 protein for p-nitrophenyl-β-d-xylopyranoside. Conclusions: The enzyme characteristics (pH and thermal st…
Monoterpenes in grape juice and wines.
The importance of monoterpenes on varietal flavour of wines has been reviewed. These compounds were mainly found linked to sugar moieties in the grape juice and wines, showing no olfactive characteristics. In this way, mechanisms to liberate terpenes were studied, making a comparative study between acidic and enzymic hydrolysis of terpene glycosides. Finally, analytical techniques developed to study these compounds, in both free or glycosidically forms, and also to fractionate glycosidic precursors, have been discussed.
Differentiation of Penicillium griseofulvum Dierckx isolates by enzyme assays and by patulin and griseofulvin analyses
The production of patulin and griseofulvin by 49 different isolates of Penicillium griseofulvum Dierckx was analyzed by high-performance liquid chromatography. Eleven isolates were obtained from pistachio nuts, 37 were obtained from wheat seeds, and 1 was obtained from the American Type Culture Collection. Activities of 19 enzymes were also assayed by the API ZYM system. From these results it may be deduced that there are two different groups among the strains tested which cannot be distinguished by morphological and cultural characteristics. One group of isolates did not produce detectable amounts of patulin and griseofulvin when grown in sucrose-yeast extract and Wickerham media, while en…
Liquid chromatographic determination of toxigenic secondary metabolites produced by Fusarium strains.
Various liquid chromatographic methods used in the analysis of mycotoxins (zearalenone, trichothecenes and fumonisins) produced by Fusarium species were compared in this work. The results demonstrate the suitability of modern clean-up procedures employing multifunctional MycoSep and immunoaffinity columns although these methods are more expensive than conventional methodologies for clean-up. HPLC with both fluorescence and photodiode array detection is a suitable technique for the analysis of toxic secondary metabolites produced by Fusarium species; different derivatisation strategies have been studied to improve the sensitivity of the technique because of the low concentration of these met…
Aroma compounds in wine as influenced by apiculate yeasts
Aroma compounds of wines resulting from fermentation of sterile grape musts from Monastrell variety inoculated with pure and mixed cultures of apiculate and Saccharomyces yeasts, were isolated and analyzed by gas chromatography with flame ionization and mass spectrometry. Samples fermented with mixed cultures produced a higher concentration of selected compounds and higher total amounts of alcohols and acids, in contrast with wines produced with pure cultures of Saccharomyces spp. Apiculate yeasts are important in the chemical composition and quality of wine.
Screening of Hanseniaspora Strains for the Production of Enzymes with Potential Interest for Winemaking
Some non-Saccharomyces yeasts, including Hanseniaspora, participate in the first stages of wine fermentation. Besides their importance in the wine production process some of these yeasts have been described to be potential producers of hydrolytic enzymes to industrial level. In this work, we pretend to evaluate the technological abilities of the Hanseniaspora strains deposited in the Spanish Type Culture Collection (CECT). First of all, we considered verification of the correct identification of the strains using several miniaturized biochemical systems and molecular techniques (PCR, RFLP and sequencing of the ribosomal D1/D2 region). The results allowed us to verify the correct adscription…
Influence of the storage conditions on some physicochemical and mycological parameters of honey
The authors studied the changes in yeast and mould counts, pH, free and lactone acidity, colour and sugar composition undergone by honeys with water contents of 169, 188 and 220 g kg-1 on storage at refrigeration temperature (4–7°C), room temperature and 28°C for two years. Of the solid culture media assayed, honey agar and osmophilic medium fostered yeast growth while malt-extract agar and yeast-extract-malt-extract agar allowed for better development of filamentous fungi. The yeasts isolated belonged to the genera Saccharomyces, Schizosaccharomyces and Zygosaccharomyces, while filamentous moulds were of the genera Aspergillus, Penicillium, Fusarium and Alternaria. The honeys studied under…
Characterization of Gibberella fujikuroi complex isolates by fumonisin B-1 and B-2 analysis and by RAPD and restriction analysis of PCR-amplified internal transcribed spacers of ribosomal DNA
Summary Twenty nine isolates of Fusarium spp. (twenty four of them belonging to the Gibberella fujikuroi complex) isolated from banana and corn from different geographical regions were analyzed for their ability to produce fumonisins B 1 and B 2 and for genetic relatedness using random amplified polymorphic DNA (RAPD) and restriction analysis of PCR amplification products of the 5.8s ribosomal DNA-intervening internal transcribed spacer regions (ITS I-5.8S-ITS II). For RAPD analysis, six of twenty oligonucleotide primers were selected after testing with five Fusarium spp. isolates and used to characterize 24 additional isolates. DNA fragments from the 29 isolates of Fusarium spp., which wer…
Valorization of winery and oil mill wastes by microbial technologies
Agro-food industrial wastes represent a serious environmental problem in producing countries. In addition to the traditional irrigation with wastewaters, application of solids to the land, and the extraction of valuable compounds for the pharmaceutical, cosmetics and food industries, a microbiological approach is also available. In this review, the most promising microbiological processes for the valorization of wine and oil by-products are presented. Recently, there has been an upsurge in the exploitation of the waste materials generated by the wine and olive oil industries. Treatments by physical and chemical methods are generally expensive, and they do not often provide complete solution…
Sustainability of food industry wastes: a microbial approach
Abstract Agro-food industrial by-products signify a grave environmental problem. In addition to the conventional irrigation with wastewaters, the application of solids to the soil, and the extraction of worthwhile compounds for the pharmaceutical, cosmetics, and food industries, a microbiological strategy is also possible. In this review, the most promising microbiological methods for the valorization of wine and oil by-products are shown. Recently, there has been an upsurge in the exploitation of the waste materials generated by the wine and olive oil industries. Treatments by physical and chemical processes are generally costly, and they do not often provide entire solutions. Microbiologi…
Determination of type A trichothecenes by high-performance liquid chromatography with coumarin-3-carbonyl chloride derivatisation and fluorescence detection.
A method for the analysis of type A trichothecenes T-2 toxin, HT-2 toxin, neosolaniol and diacetoxyscirpenol by high-performance liquid chromatography with fluorescence detection using coumarin-3-carbonyl chloride has been developed. Different parameters concerning the analytical procedure such as stability of both the reagent and derivatised analytes, time and temperature of the derivatisation reaction, were studied and optimised. Three different clean-up procedures (solid-phase extraction with silica gel or C-18 cartridges, and liquid-liquid partition between toluene and dihydrogen phosphate buffer) were tested in order to remove the excess reagent peaks. The last procedure gave the best …