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Application of qPCR for multicopper oxidase gene (MCO) in biogenic amines degradation by Lactobacillus casei
H. Pištěková, P. Jančová, L. Berčíková, F. Buňka, I. Sokolová, T. Šopík, K. Maršálková, OMRP. Amaral, L. Buňková
Jazyk angličtina Země Velká Británie
Typ dokumentu časopisecké články
PubMed
32539976
DOI
10.1016/j.fm.2020.103550
Knihovny.cz E-zdroje
- MeSH
- bakteriální proteiny genetika metabolismus MeSH
- biogenní aminy analýza metabolismus MeSH
- cystein analýza metabolismus MeSH
- glyceraldehyd-3-fosfátdehydrogenasy genetika MeSH
- kultivační média chemie MeSH
- kvantitativní polymerázová řetězová reakce MeSH
- kyselina askorbová analýza metabolismus MeSH
- Lactobacillus casei enzymologie genetika růst a vývoj metabolismus MeSH
- Lactobacillus enzymologie genetika růst a vývoj metabolismus MeSH
- mléko chemie MeSH
- oxidoreduktasy genetika metabolismus MeSH
- regulace genové exprese u bakterií MeSH
- vysokoúčinná kapalinová chromatografie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Degradation of undesirable biogenic amines (BAs) in foodstuffs by microorganisms is considered one of the most effective ways of eliminating their toxicity. In this study, we designed two sets of primers for the detection and quantification of the multicopper oxidase gene (MCO), which encodes an enzyme involved in BAs degradation, and endogenous (glyceraldehyde-3-phosphate dehydrogenase) gene (GAPDH) in Lactobacillus casei group by real-time PCR (qPCR). We tested 15 Lactobacillus strains in the screening assays (thus, MCO gene possessing assay (PCR) and monitoring of BAs degradation by HPLC-UV), in which Lactobacillus casei CCDM 198 exhibited the best degradation abilities. For this strain, we monitored the expression of the target gene (MCO) in time (qPCR), the effect of redox treatments (cysteine, ascorbic acid) on the expression of the gene, and the ability to degrade BAs not only in a modified MRS medium (MRS/2) but also in a real food sample (milk). Moreover, decarboxylase activity (ability to form BAs) of this strain was excluded. According to the results, CCDM 198 significantly (P < 0.05) reduced BAs (putrescine, histamine, tyramine, cadaverine), up to 25% decline in 48 h. The highest level of relative expression of MCO (5.21 ± 0.14) was achieved in MRS/2 media with cysteine.
Citace poskytuje Crossref.org
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- $a Degradation of undesirable biogenic amines (BAs) in foodstuffs by microorganisms is considered one of the most effective ways of eliminating their toxicity. In this study, we designed two sets of primers for the detection and quantification of the multicopper oxidase gene (MCO), which encodes an enzyme involved in BAs degradation, and endogenous (glyceraldehyde-3-phosphate dehydrogenase) gene (GAPDH) in Lactobacillus casei group by real-time PCR (qPCR). We tested 15 Lactobacillus strains in the screening assays (thus, MCO gene possessing assay (PCR) and monitoring of BAs degradation by HPLC-UV), in which Lactobacillus casei CCDM 198 exhibited the best degradation abilities. For this strain, we monitored the expression of the target gene (MCO) in time (qPCR), the effect of redox treatments (cysteine, ascorbic acid) on the expression of the gene, and the ability to degrade BAs not only in a modified MRS medium (MRS/2) but also in a real food sample (milk). Moreover, decarboxylase activity (ability to form BAs) of this strain was excluded. According to the results, CCDM 198 significantly (P < 0.05) reduced BAs (putrescine, histamine, tyramine, cadaverine), up to 25% decline in 48 h. The highest level of relative expression of MCO (5.21 ± 0.14) was achieved in MRS/2 media with cysteine.
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