title: Proteome analysis of Streptococcus thermophilus grown in milk reveals pyruvate formate-lyase as the major upregulated protein.
abstract: We investigated the adaptation to milk of Streptococcus thermophilus LMG18311 using a proteomic approach. Two-dimensional electrophoresis of cytosolic proteins were performed after growth in M17 medium or in milk. A major modification of the proteome concerned proteins involved in the supply of amino acids, like the peptidase PepX, and several enzymes involved in amino acid biosynthesis. In parallel, we observed the upregulation of the synthesis of seven enzymes directly involved in the synthesis of purines, as well as formyl-tetrahydrofolate (THF) synthetase and serine hydroxy-methyl transferase, two enzymes responsible for the synthesis of compounds (THF and glycine, respectively) feeding the purine biosynthetic pathway. The analysis also revealed a massive increase in the synthesis of pyruvate formate-lyase (PFL), the enzyme which converts pyruvate into acetyl coenzyme A and formate. PFL has been essentially studied for its role in mixed-acid product formation in lactic acid bacteria during anaerobic fermentation. However, formate is an important methyl group donor for anabolic pathway through the formation of folate derivates. We hypothesized that PFL was involved in purine biosynthesis during growth in milk. We showed that PFL expression was regulated at the transcriptional level and that pfl transcription occurred during the exponential growth phase in milk. The complementation of milk with formate or purine bases was shown to reduce pfl expression, to suppress PFL synthesis, and to stimulate growth of S. thermophilus. These results show a novel regulatory mechanism controlling the synthesis of PFL and suggest an unrecognized physiological role for PFL as a formate supplier for anabolic purposes.
authors.affiliation: Unité Biologie des Bactéries Pathogénes à Gram Positif, Institut Pasteur, 25 rue du Dr. Roux, 75724 Paris, France.
authors.forename: Sylviane
authors.initials: S
authors.lastname: Derzelle
authors.forename: Alexander
authors.initials: A
authors.lastname: Bolotin
authors.forename: Michel-Yves
authors.initials: MY
authors.lastname: Mistou
authors.forename: Françoise
authors.initials: F
authors.lastname: Rul
mesh_terms: D000123:Acetyltransferases / Q000235:genetics*; D000818:Animals; D003593:Cytoplasm / Q000201:enzymology; D015180:Electrophoresis, Gel, Two-Dimensional; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006358:Hot Temperature; D007700:Kinetics; D008892:Milk / Q000382:microbiology*; D020543:Proteome; D012333:RNA, Messenger / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D048249:Streptococcus thermophilus / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development*; D014158:Transcription, Genetic
chemical_list: D020543:Proteome; D012333:RNA, Messenger; D000123:Acetyltransferases; C022560:formate C-acetyltransferase