Application of qPCR for multicopper oxidase gene (MCO) in biogenic amines degradation by Lactobacillus casei
Language English Country England, Great Britain Media print-electronic
Document type Journal Article
PubMed
32539976
DOI
10.1016/j.fm.2020.103550
PII: S0740-0020(20)30139-8
Knihovny.cz E-resources
- Keywords
- Biogenic amines degradation, Histamine, Lactobacillus casei, Primers, qPCR,
- MeSH
- Bacterial Proteins genetics metabolism MeSH
- Biogenic Amines analysis metabolism MeSH
- Cysteine analysis metabolism MeSH
- Glyceraldehyde-3-Phosphate Dehydrogenases genetics MeSH
- Culture Media chemistry MeSH
- Real-Time Polymerase Chain Reaction MeSH
- Ascorbic Acid analysis metabolism MeSH
- Lacticaseibacillus casei enzymology genetics growth & development metabolism MeSH
- Lactobacillus enzymology genetics growth & development metabolism MeSH
- Milk chemistry MeSH
- Oxidoreductases genetics metabolism MeSH
- Gene Expression Regulation, Bacterial MeSH
- Chromatography, High Pressure Liquid MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Bacterial Proteins MeSH
- Biogenic Amines MeSH
- Cysteine MeSH
- Glyceraldehyde-3-Phosphate Dehydrogenases MeSH
- Culture Media MeSH
- Ascorbic Acid MeSH
- Oxidoreductases 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.
References provided by Crossref.org