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1.
Occurence of biogenic amine - producing lactic acid bacteria in Refošk grape and wine
Jelena Topić Božič, Jan Reščič, Martina Bergant Marušič, Klemen Lisjak, Branka Mozetič Vodopivec, Lorena Butinar, 2019, objavljeni povzetek znanstvenega prispevka na konferenci

Opis: 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.
Najdeno v: ključnih besedah
Ključne besede: biogenic amines, lactic acid bacteria, refošk, grape, wine
Objavljeno: 22.01.2020; Ogledov: 1630; Prenosov: 0
.pdf Polno besedilo (185,72 KB)

2.
Determination of biogenic amine-forming potential of naturally occuring lactic acid bacteria (LAB) strains isolated from the grapes of Vitis vinifera cv. Refošk grape variety
Dorota Korte, Branka Mozetič Vodopivec, Lorena Butinar, Martina Bergant Marušič, Jelena Topić, 2020, objavljeni povzetek znanstvenega prispevka na konferenci

Opis: 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.
Najdeno v: ključnih besedah
Ključne besede: lactic acid bacteria, biogenic amines, histamine, grape
Objavljeno: 10.07.2020; Ogledov: 1266; Prenosov: 0
.pdf Polno besedilo (225,99 KB)

3.
Determination of biogenic amines formation by autochthonous lactic acid bacteria from ‘Refošk’ grapes using different analytical methods
Jelena Topić, Lorena Butinar, Martina Bergant Marušič, Dorota Korte, Branka Mozetič Vodopivec, 2022, izvirni znanstveni članek

Opis: Different analytical methods were tested and optimized for the determination of four biogenic amines (BA) histamine, putrescine, cadaverine and tyramine produced by grape-associated lactic acid bacteria (LAB). Autochthonous LAB were isolated from ‘Refošk’ grapes belonging to Slovenian-Italian Karst region as they represent a potential pool of beneficial LAB starter bacteria for improving the typicality and quality of wine. Six isolated strains were screened by multiplex PCR, of which four strains were positive for BA-forming genes (MKBT-49, MKBT-282, MKBT-568, MKBT-570). The production of BA was evaluated by thin-layer chromatography (TLC), high-performance liquid chromatography coupled to diode array detection (HPLC-DAD) and enzymatic method. The HPLC-DAD results showed that strain MKBT-49 (tyrdc+) had high tyramine production (386.6 ± 0.14 mg/L), which was confirmed by TLC. The ability to produce putrescine was confirmed in strain MKBT-282 by PCR, HPLC-DAD (16.4 ± 1.72 mg/L), and TLC. Histamine-producing ability was detected in strain MKBT-570, with a concentration below the limit of detection of the HPLC-DAD (<0.2 mg/L), while the other two methods were not sensitive enough for confirmation. This study shows that the production of BA can detected in native LAB and that relatively simple method such as TLC can be used effectively for the initial screening.
Najdeno v: ključnih besedah
Ključne besede: grapes, wine-like matrices, biogenic amines, autochthonous lactic acid bacteria, analytical methods
Objavljeno: 03.01.2022; Ogledov: 101; Prenosov: 1
URL Polno besedilo (0,00 KB)
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