Isolation of aminopeptidase N genes of food associated propionibacteria and observation of their transcription in skim milk and acid whey.
In this study consensus oligonucleotides PN5/PN3 were designed by aligning the aminopeptidase N genes (pepN) of various actinobacteria and applied to the isolation of the pepN genes of dairy propionibacteria (PAB) and closely related species associated with food. This allowed sequencing of a pepN gene region from Propionibacterium jensenii LMG 16541. The sequence of this gene was completed by inverse PCR. Consensus primer pairs NU1/D1 and NU2/D1 were derived from the alignment of the new sequence with its homologues in Propionibacterium acnes and other actinobacteria; these were used to start sequencing of the pepN genes of Propionibacterium freudenreichii, Propionibacterium thoenii, Propionibacterium microaerophilum, Propionibacterium acidipropionici, Propioni bacterium cyclohexanicum and Propionibacterium microaerophilum. Reverse transcription coupled with PN5/PN3 and NU1/D1 PCR tests indicated that the pepN genes of P. jensenii and P. freudenreichii are expressed during growth in skim milk and acid whey.
Rossi
Franca
F
Dipartimento Scientifico e Tecnologico, Università degli Studi di Verona, Strada Le Grazie 15, 37134, Verona, Italy.
Busetto
Michela
M
Torriani
Sandra
S
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018826:CD13 Antigens / Q000235:genetics*; D002611:Cheese / Q000382:microbiology; D003470:Culture Media; D004269:DNA, Bacterial / Q000235:genetics; D005516:Food Microbiology; D005798:Genes, Bacterial; D008892:Milk / Q000382:microbiology; D008894:Milk Proteins; D008969:Molecular Sequence Data; D011424:Propionibacterium / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic
D003470:Culture Media; D004269:DNA, Bacterial; D008894:Milk Proteins; C024471:whey acidic proteins; D018826:CD13 Antigens