Skip to main content
NCBI home page
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1997 Jun;63(6):2213–2217. doi: 10.1128/aem.63.6.2213-2217.1997

Cloning and functional expression in Escherichia coli of the gene encoding the di- and tripeptide transport protein of Lactobacillus helveticus.

H Nakajima 1, A Hagting 1, E R Kunji 1, B Poolman 1, W N Konings 1
PMCID: PMC168514  PMID: 9172341

Abstract

The gene encoding the di- and tripeptide transport protein (DtpT) of Lactobacillus helveticus (DtpTLH) was cloned with the aid of the inverse PCR technique and used to complement the dipeptide transport-deficient and proline-auxotrophic Escherichia coli E1772. Functional expression of the peptide transporter was shown by the uptake of prolyl-[14C] alanine in whole cells and membrane vesicles. Peptide transport via DtpT in membrane vesicles is driven by the proton motive force. The system has specificity for di- and tripeptides but not for amino acids or tetrapeptides. The dtpTLH gene consists of 1,491 bp, which translates into a 497-amino-acid polypeptide. DtpTLH shows 34% identity to the di- and tripeptide transport protein of Lactococcus lactis and is also homologous to various peptide transporters of eukaryotic origin, but the similarity between these proteins is confined mainly to the N-terminal halves.

Full Text

The Full Text of this article is available as a PDF (204.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Christensen J. E., Lin D. L., Palva A., Steele J. L. Sequence analysis, distribution and expression of an aminopeptidase N-encoding gene from Lactobacillus helveticus CNRZ32. Gene. 1995 Mar 21;155(1):89–93. doi: 10.1016/0378-1119(94)00924-h. [DOI] [PubMed] [Google Scholar]
  3. Delley M., Mollet B., Hottinger H. DNA Probe for Lactobacillus delbrueckii. Appl Environ Microbiol. 1990 Jun;56(6):1967–1970. doi: 10.1128/aem.56.6.1967-1970.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Foucaud C., Kunji E. R., Hagting A., Richard J., Konings W. N., Desmazeaud M., Poolman B. Specificity of peptide transport systems in Lactococcus lactis: evidence for a third system which transports hydrophobic di- and tripeptides. J Bacteriol. 1995 Aug;177(16):4652–4657. doi: 10.1128/jb.177.16.4652-4657.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Gilbert C., Atlan D., Blanc B., Portailer R., Germond J. E., Lapierre L., Mollet B. A new cell surface proteinase: sequencing and analysis of the prtB gene from Lactobacillus delbruekii subsp. bulgaricus. J Bacteriol. 1996 Jun;178(11):3059–3065. doi: 10.1128/jb.178.11.3059-3065.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hagting A., Kunji E. R., Leenhouts K. J., Poolman B., Konings W. N. The di- and tripeptide transport protein of Lactococcus lactis. A new type of bacterial peptide transporter. J Biol Chem. 1994 Apr 15;269(15):11391–11399. [PubMed] [Google Scholar]
  7. Higgins C. F. ABC transporters: from microorganisms to man. Annu Rev Cell Biol. 1992;8:67–113. doi: 10.1146/annurev.cb.08.110192.000435. [DOI] [PubMed] [Google Scholar]
  8. Kunji E. R., Mierau I., Hagting A., Poolman B., Konings W. N. The proteolytic systems of lactic acid bacteria. Antonie Van Leeuwenhoek. 1996 Oct;70(2-4):187–221. doi: 10.1007/BF00395933. [DOI] [PubMed] [Google Scholar]
  9. Kunji E. R., Smid E. J., Plapp R., Poolman B., Konings W. N. Di-tripeptides and oligopeptides are taken up via distinct transport mechanisms in Lactococcus lactis. J Bacteriol. 1993 Apr;175(7):2052–2059. doi: 10.1128/jb.175.7.2052-2059.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Mathiopoulos C., Mueller J. P., Slack F. J., Murphy C. G., Patankar S., Bukusoglu G., Sonenshein A. L. A Bacillus subtilis dipeptide transport system expressed early during sporulation. Mol Microbiol. 1991 Aug;5(8):1903–1913. doi: 10.1111/j.1365-2958.1991.tb00814.x. [DOI] [PubMed] [Google Scholar]
  12. Olson E. R., Dunyak D. S., Jurss L. M., Poorman R. A. Identification and characterization of dppA, an Escherichia coli gene encoding a periplasmic dipeptide transport protein. J Bacteriol. 1991 Jan;173(1):234–244. doi: 10.1128/jb.173.1.234-244.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Poolman B., Kunji E. R., Hagting A., Juillard V., Konings W. N. The proteolytic pathway of Lactococcus lactis. Soc Appl Bacteriol Symp Ser. 1995;24:65S–75S. [PubMed] [Google Scholar]
  14. Sasaki M., Bosman B. W., Tan P. S. Comparison of proteolytic activities in various lactobacilli. J Dairy Res. 1995 Nov;62(4):601–610. doi: 10.1017/s0022029900031332. [DOI] [PubMed] [Google Scholar]
  15. Steiner H. Y., Naider F., Becker J. M. The PTR family: a new group of peptide transporters. Mol Microbiol. 1995 Jun;16(5):825–834. doi: 10.1111/j.1365-2958.1995.tb02310.x. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES