21. Determination of biogenic amine-forming potential of naturally occuring lactic acid bacteria (LAB) strains isolated from the grapes of Vitis vinifera cv. Refošk grape varietyJelena Topić, Lorena Butinar, Martina Bergant Marušič, Dorota Korte, Branka Mozetič Vodopivec, 2020, published scientific conference contribution abstract Abstract: Biogenic amines are naturally occurring compounds that have been reported in variety of food, such as fish, meat, cheese, vegetables, and wines. When biogenic amines are formed by microorganisms such as lactic acid bacteria (LAB) they may have adverse effect on human health. The most common biogenic amines found in foodstuff are histamine, tyramine, putrescine and cadaverine. LAB are used as starters in the production of several foods and beverages. In wines they are used in malolactic fermentation, where the conversion of malic acid to lactic acid occurs. The aim of this work was to determine biogenic amine-forming capacity of indigenous LAB strains isolated from grapes which were PCR positive and screening medium positive for biogenic amines. Three methods for the confirmation of LAB biogenic amine-forming potential were developed and compared: HPLC-DAD and TLC methods both using derivatization agent dansyl chloride and enzymatic method using horseradish peroxidase (HRP) and diamine oxidase (DAO) enzymes. The applicability of methods was tested on the three known biogenic amine-producing LAB strains: 0006, 9809, 9906 (IOEB, France). The limit of detection (LOD) of the methods was <1 mg/L for HPLC-DAD, 5 mg/L for TLC and 25 mg/L for enzymatic method. Although the LODs between the proposed methods differed, all of the three methods confirmed BA-forming potential of the three strains. Furthermore, the proposed methods were applied for the analysis of three indigenous strains: MKBT-282 (BA+), MKBT-307 (BA-) and MKBT-325 (BA+). The positive or negative BA-forming potential was confirmed with all of the three proposed methods. Keywords: lactic acid bacteria, biogenic amines, histamine, grape Published in RUNG: 10.07.2020; Views: 4056; Downloads: 0 This document has many files! More... |
22. Implementation of high performance liquid chromatography coupled to thermal lens spectrometry (HPLC‑TLS) for quantification of pyranoanthocyanins during fermentation of Pinot Noir grapesJelena Topič Božič, Lorena Butinar, Natka Ćurko, Karin Kovačević Ganić, Branka Mozetič Vodopivec, Dorota Korte, Mladen Franko, 2020, original scientific article Abstract: In this work high performance liquid chromatography coupled to thermal lens spectrometry (HPLC-TLS) was applied for monitoring of vinylphenolic pyranoanthocyanins formation during the fermentation of Pinot Noir wines. Vinylphenolic pyranoanthocyanins are wine pigments, present in low concentrations, but very important for wine colour stability.
Fermentation process was conducted with four different yeast strains, used as starters, either in sequential fermentation of non-Saccharomyces with S. cerevisiae yeast or as single fermentation with S. cerevisiae yeasts in order to test the applicability of developed method for monitoring of selected compounds in real wine fermentation experiments. The
developed HPLC-TLS method showed higher sensitivity compared to HPLC coupled to diode array detection (DAD) technique for particular wine colour compounds. Obtained limits of detection (LODs), were 6- and 22-times lower in comparison to HPLC–DAD in gradient and isocratic elution mode, respectively, whereas limits of quantification (LOQs)
5 and 18-times lower. Lower LODs enabled earlier observation of vinylphenolic pyranoanthocyanins formation during fermentation (already at day 7) in the case of HPLC-TLS method in gradient mode, while by using HPLC–DAD in gradient elution mode the formation of vinylphenolic pyranoanthocyanins was noticed only after 12 days of fermentation. Keywords: Thermal lens spectrometry (TLS), High performance liquid chromatography (HPLC), Pyranoanthocyanins, Wine, Yeasts Published in RUNG: 18.06.2020; Views: 5054; Downloads: 0 This document has many files! More... |
23. Synthesis of pyranoanthocyanins from Pinot Noir grape skin extract using fermentation with high pyranoanthocyanin producing yeasts and model wine storage as potential approaches in the production of stable natural food colorantsJelena Topić, Natka Ćurko, Karin Kovačević Ganić, Lorena Butinar, Alen Albreht, Irena Vovk, Dorota Korte, Branka Mozetič Vodopivec, 2020, original scientific article Keywords: Pyranoanthocyanins, Anthocyanins, Food colorants, Yeasts, Fermentation Published in RUNG: 03.04.2020; Views: 4989; Downloads: 0 This document has many files! More... |
24. The impact of Saccharomyces and non-Saccharomyces yeasts on wine colour: a laboratory study of vinylphenolic pyranoanthocyanin formation and anthocyanin cell wall adsorptionJelena Topić, Lorena Butinar, Alen Albreht, Irena Vovk, Dorota Korte, Branka Mozetič Vodopivec, 2020, original scientific article Keywords: Non-Saccharomyces yeast, Hydroxycinnamate decarboxylase activity, Sequential fermentation, Vinylphenolic pyranoanthocyanins, Anthocyanin adsorption Published in RUNG: 23.01.2020; Views: 4248; Downloads: 0 This document has many files! More... |
25. Occurence of biogenic amine - producing lactic acid bacteria in Refošk grape and wineJelena Topić Božič, Jan Reščič, Martina Bergant Marušič, Darja Lisjak, Branka Mozetič Vodopivec, Lorena Butinar, 2019, published scientific conference contribution abstract Abstract: Lactic acid bacteria (LAB) are naturally present in grapes, musts and wines. During malolactic fermentation, besides the beneficial conversion of L-malic to L-lactic acid, LAB can form also other compounds like biogenic amines (BAs). BAs are formed through decarboxylation of corresponding amino acids, through the action of enzymes. Consumption of food containing BAs can lead to food poisoning such as histamine poisoning. Histamine, the most studied BA, is known to cause headaches, oedema, vomiting.
In the last years, BAs associated pathways have been described as strain dependent and not as species dependent. Because of the high variability of microorganisms to decarboxylase amino acids, the detection of bacteria that have the possibility to transform precursor amino acid into BAs is very important in order to estimate the risk of BAs accumulation in wines. Since in previous years during project AGROTUR I we detected the higher BA content in some wines, we decided to focus on the presence of BA-producing LAB in Refošk grapes and wines. Therefore, during AGROTUR II project LAB collection of autochthonous lactic acid bacteria was set-up by isolating LAB from Refošk grapes and wines, originated from the grape growing cross-border region Karst (Slovenia). Over 600 isolates were obtained using MRS medium with added cycloheximide and 2% tomato juice. In parallel, also selective enrichment was performed using MRS medium described above with additionally added 5% ethanol. BA-producing LAB were primarily detected in microtiter-plate format using the decarboxylase screening medium with added amino acids (histidine, lysine, ornithine or tyrosine), which enable us to identify histamine-, cadaverine-, ornithine and tyrosine-producing LAB. Selected representatives from each BA-producing LAB groups were further on confirmed with chromatographic analysis (HPLC) and molecular methods (multiplex PCR method). BA-producing LAB were in majority represented by thyramin-producers, thereafter followed cadaverine-producers, with putrescine and histamine producers being the least presented. This screening of the BA-producing LAB is also incorporated in our on-going accurate selection of LAB starters for potential production of Refošk wines. Keywords: biogenic amines, lactic acid bacteria, refošk, grape, wine Published in RUNG: 22.01.2020; Views: 4352; Downloads: 0 This document has many files! More... |
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28. Implementation of liquid chromatography coupled to thermal lens spectrometry (HPLC-TLC) in studies of yeats' role in formation of stable pigments in Pinot Noir winesBranka Mozetič Vodopivec, Jelena Topić, Lorena Butinar, Natka Ćurko, Karin Kovačević Ganić, Dorota Korte, Mladen Franko, 2019, published scientific conference contribution abstract Keywords: thermal lens spectrometry, pigments, Pinot Noir wines Published in RUNG: 16.07.2019; Views: 4802; Downloads: 0 This document has many files! More... |
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30. Characterization of lactic bacteria for biogenic amine formationJelena Topic, Lorena Butinar, Martina Bergant Marušič, Dorota Korte, Branka Mozetič Vodopivec, 2018, independent scientific component part or a chapter in a monograph Abstract: Biogenic amines are compounds present in many different foods and beverages (wine, beer, dairy products, fermented vegetables and soy products, fish, etc.). Their presence in foodstuff is a result of a microbial action during storage and ageing. The most important are histamine, tryptamine, β-phenylethylamine and tryptamine, which can induce undesirable physiological effects in humans. They are formed through decarboxylation of corresponding amino acids, through the action of enzymes. Consumption of food containing biogenic amines can lead to food poisoning such as histamine poisoning. Histamine, the most studied biogenic amine, is known to cause headaches, oedema, vomiting, etc. [1]–[4]. Monitoring of the content of biogenic amines in foods is of concern for public health in their relation to the food safety, food spoilage and food intolerance. Because microorganisms are used in food productions as starters and biopreservers, characterization of microorganisms for their ability to produce biogenic amines is equally important. Lactic acid bacteria are often used as biopreservers as they can produce antimicrobial metabolites and antifungal peptides. Some strains can also produce undesirable biogenic amines [5]. In order to use lactic acid bacteria as starters or biopreservers, the selection of strains that would not produce biogenic amines is necessary.
When considering studies of biogenic amines in foods, focus should be on developing new or improving analysis methods for biogenic amines detection. Secondly, the connections between microorganisms capable of producing biogenic amines and the content of biogenic amines in foods should be investigated [3]. The most widely technique used for quantification of biogenic amines in foodstuff is liquid chromatography, Alternatively to chromatographic techniques, other techniques such as enzymatic biosensors, ELISA and flow-injection analysis have also been employed. Sensors are interesting due to the fact that they do not require special instrumentations, and there is no need for sample clean-up and derivatization, which are the main drawback of chromatographic methods [4]. Detection of biogenic amines producing lactic bacteria is important due to the concerns for public health and there is a need for the early and rapid detection of such microorganisms. Most of the methods that are used for screening involved the measurement of amino acid-decarboxylase activity, although there were been some methods reported that used differential media and pH indicators. Nowadays, molecular methods are replacing culture methods. Molecular approaches are used to determine the presence or absence of genes responsible for biogenic amines formation. The main advantages of DNA hybridization and PCR methods are speed, simplicity, sensitivity and specificity as they allow detection of targeted genes. Culture independent methods which are based on PCR techniques are now regarded as most suitable methods for screening isolates [5].
[1] A. R. Shalaby, “Significance of biogenic amines to food safety and human health,” Food Res. Int., vol. 29, no. 7, pp. 675–690, Oct. 1996.
[2] J. M. Landete, S. Ferrer, and I. Pardo, “Biogenic amine production by lactic acid bacteria, acetic bacteria and yeast isolated from wine,” Food Control, vol. 18, pp. 1569–1574, 2007.
[3] F. B. Erim, “Recent analytical approaches to the analysis of biogenic amines in food samples,” TrAC - Trends in Analytical Chemistry, vol. 52. pp. 239–247, 2013.
[4] J. L. Ordóñez, A. M. Troncoso, M. D. C. García-Parrilla, and R. M. Callejón, “Recent trends in the determination of biogenic amines in fermented beverages – A review,” Analytica Chimica Acta, vol. 939. pp. 10–25, 2016.
[5] R. M. Elsanhoty and M. F. Ramadan, “Genetic screening of biogenic amines production capacity from some lactic acid bacteria strains,” Food Control, vol. 68, pp. 220–228, Oct. 2016. Keywords: lactic bacteria, biogenic amines Published in RUNG: 13.12.2018; Views: 4680; Downloads: 0 This document has many files! More... |