4.5 Article

Detection and relative quantification of amine oxidase gene (yobN) in Bacillus subtilis: application of real-time quantitative PCR

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出版社

SPRINGER INDIA
DOI: 10.1007/s13197-021-05090-9

关键词

Biogenic amines degradation; Histamine; Real-time PCR; Primers

资金

  1. Grant Agency of the Czech Republic (GACR) [17-09594S]
  2. Internal Grant Agency of the Tomas Bata University in Zlin [IGA/FT/2020/009]

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Degradation of undesirable biogenic amines in food by microorganisms, particularly Bacillus subtilis, has been proven effective in eliminating their toxicity. However, the degradation ability of B. subtilis strains may be limited by sporulation.
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 design two sets of primers for the detection and quantification of the amine oxidase gene (yobN) and endogenous (housekeeping) gene (gyrB) in Bacillus subtilis. Moreover, these sets can be used for relative quantification of yobN by real-time PCR (qPCR). We also tested the degradation of BAs by three bacterial strains (B. subtilis strains: IB1a, CCM 2216, CCM 2267) in a mineral medium over a two-day period. Their degradation abilities were verified by high performance liquid chromatography with UV detection (HPLC/UV). According to the results, two strains significantly (P < 0.05) reduced histamine, tyramine, putrescine, and cadaverine. Moreover, our results indicate that the degradation ability of B. subtilis strains could be limited by sporulation because the gene encoding amine oxidase (yobN) is no longer expressed in the spores.

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