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Transcription activity of lactic acid bacterial proteolysis-related genes during cheese maturation

Pangallo, Domenico, Kraková, Lucia, Puškárová, Andrea, Šoltys, Katarína, Bučková, Maria, Koreňová, Janka, Budiš, Jaroslav, Kuchta, Tomáš
Food microbiology 2019 v.82 pp. 416-425
Lactobacillus, Lactococcus, aminopeptidases, cheese ripening, cheeses, complementary DNA, dipeptidyl-peptidase IV, ewe milk, genes, lactic acid bacteria, metabolism, metagenomics, milk, milk proteins, proteinases, proteolysis, ribosomal RNA, transcription (genetics), transcriptomics, transferases
The catabolism of milk protein in cheese is one way how the microorganisms influence the sensorial characteristics of the final product. In this investigation, we paid attention to four genes [prtP (cell-envelope proteinase gene), pepX (X-prolyl dipeptidyl aminopeptidase gene), pepN (aminopeptidase gene) and bcaT (branched chain aminotransferase gene)] responsible for the production of volatile aroma-active compounds from milk proteins by lactic acid bacteria (LAB). We studied the dynamics of these genes and their corresponding LAB host, during the maturation of a raw ewes’ milk-based cheese, using metagenomics and metatranscriptomics approaches. The transcriptome-oriented experiments included the analysis of total RNA (at three stages of cheese maturation) and also the construction of specific cDNA sub-libraries of the abovementioned genes. The proteolytic transcriptome analysis was supported by following the transcription activity of 16S rRNA gene and by metagenomic investigation. The combination of the described methods permitted to screen the dynamics of targeted genes throughout the cheese production. Lactococci were the major players in the LAB group, but the analysis provided also information on the role and properties of members of the genus Lactobacillus, such as Lb. rhamnosus, Lb. helveticus, Lb. pentosus, Lb. curvatus, Lb. parabuchneri, Lb. plantarum, Lb. brevis, Lb. delbrueckii, Lb. paracasei, Lb. fermentum and Lb. heilongjiangensis, proteolysis-related genes of which were active during cheese ripening.