Non-targeted Metabolomics Analysis of Benzo（α）pyrene by Microorganisms in Kefir Grains

doi:10.13560/j.cnki.biotech.bull.1985.2021-1074

Abstract

Abstract:

In order to understand the mechanism of microbial action on the polycyclic aromatic hydrocarbon benzo（α）pyrene in the traditional fermenter Kefir grains from Xinjiang,and the characteristic changes in the metabolites of Kefir grains in fermented milk under the stress of 50 mg/L benzo（α）pyrene were investigated. Using non-targeted metabonomics technology,combined with principal component analysis and partial minor binary discriminant analysis,it was confirmed that phenylacetic acid,phenol,protocatechuic acid,diisopropyl phthalate and phthalic acid were intermediate metabolic characteristic products. A total of 56 metabolic pathways were annotated in the KEGG database,among which there were 3 key pathways with high enrichment of metabolites,high significance and related to amino acid metabolism. Intermediate metabolites cleaved the benzene ring under the action of a series of enzymes such as dehydrogenase,dioxygenase,and dehydroisomerase. Therefore,it is inferred that the presence of microbial strains in the Kefir grains may have degraded benzo（α）pyrene through the“naphthalene pathway”,the“phthalic acid pathway”and the“phenylalanine pathway”. Thus study may provide a certain theoretical basis for the action mechanism of microorganisms on benzo（α）pyrene,may propose possible biological control strategies for reducing the harms of benzo（α）pyrene to the human body,and may provide research basis for the rational development and application of kefir fermented dairy products in the food industry.

Key words: Kefir grains, benzo（α）pyrene, metabolomics, metabolic pathways

GULJAMAL·Aisa , XING Jun, LI An, ZHANG Rui. Non-targeted Metabolomics Analysis of Benzo（α）pyrene by Microorganisms in Kefir Grains[J]. Biotechnology Bulletin, 2022, 38(5): 123-135.

Figures/Tables 11

Fig. 1 Principal component analysis of the samples at diff-erent fermentation stages under benzo（α）pyrene stress A：Five green circles indicate five Kefir samples fermented for 0 h under benzo（α）pyrene stress. B：Five red circles indicate five Kefir samples fermented for 8 h under benzo（α）pyrene stress. C：Five orange circles indicate five Kefir samples fermented for 16 h under benzo（α）pyrene stress. D：Five red circles indicate five Kefir samples fermented for 24 h under benzo（α）pyrene stress. E：Yellow five circles indicate five Kefir samples fermented for 32 h under benzo（α）pyrene stress

Fig. 2 PLS-DA score map of samples at different fermen-tation stages under benzo（α）pyrene stress

Fig. 3 Distribution of the test statistic（Q2）of the OPLS-DA substitution test and the P value

Fig. 4 Stacked histogram of the percentages of metabolites in the samples at different fermentation stages under benzo（α）pyrene stress

Fig. 5 Percentage of metabolites playing a biological role in the samples of different fermentation stages under benzo（α）pyrene stress

Fig.6 Box plot of the differences in metabolites of the samples at different fermentation stages under benzo（α）pyrene stress *P<0.05,**P<0.01,***P<0.001

Fig.7 Importance map of PLS-DA metabolites in the samples at different fermentation stages under benzo（α）pyrene stress

Fig.8 Metabolic pathways with significant enrichment of differential metabolites in the samples at different fermentation stages under benzo（α）pyrene stress（ORA enrichment analysis）

Fig.9 Topological analysis of metabolic pathways with significant enrichment of different metabolites in the samples at different fermentation stages under benzo（α）pyrene stress

Fig. 10 Phenylalanine pathway

Fig. 11 Phthalic acid pathway

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