Cloning and Sequence Analysis of the X-prolyl-dipeptidyl-aminopeptidase Gene (pepX) from Lactobacillus Delbrückii Ssp. Lactis DSM7290

Overview

Journal Appl Microbiol Biotechnol

Specialties Biomedical Engineering
Biotechnology
Microbiology

Date 1993 Oct 1

PMID 7765315

Citations 11

Authors

J R Klein

M Imam

R PLAPP

Affiliations

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Abstract

Lactobacillus delbrückii ssp. lactis DSM7290 possesses an X-prolyl-dipeptidyl-aminopeptidase, designated PepX, which catalyses the hydrolytic removal of N-terminal dipeptidyl residues from peptides containing proline in the penultimate position. Using the specific substrate L-Ala-L-Pro-p-nitroanilide, PepX was purified by a four-step procedure including ammonium sulphate fractionation, hydrophobic interaction chromatography, ion exchange chromatography, and affinity chromatography. The N-terminus of the purified protein was sequenced. Screening of a gene library of chromosomal Lactobacillus delbrückii ssp. lactis DSM7290 DNA in the low-copy-number vector pLG339 resulted in the identification of the pepX gene in Escherichia coli using a specific plate assay with Gly-L-Pro-beta-naphthylamide as substrate. Nucleotide sequence analysis revealed an open reading frame of 2376 bp, coding for a protein of 792 amino acids with a molecular mass of 88449 Da.

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References

PLAPP R . Peptidases of Lactobacillus casei and L. plantarum. J Basic Microbiol. 1987; 27(3):123-30. DOI: 10.1002/jobm.3620270302. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

van de Guchte M, Kok J, Venema G . Gene expression in Lactococcus lactis. FEMS Microbiol Rev. 1992; 8(2):73-92. DOI: 10.1111/j.1574-6968.1992.tb04958.x. View

Yan T, Ho S, Hou C . Catalytic properties of X-prolyl dipeptidyl aminopeptidase from Lactococcus lactis subsp. cremoris nTR. Biosci Biotechnol Biochem. 1992; 56(5):704-7. DOI: 10.1271/bbb.56.704. View

KYTE J, Doolittle R . A simple method for displaying the hydropathic character of a protein. J Mol Biol. 1982; 157(1):105-32. DOI: 10.1016/0022-2836(82)90515-0. View

Meyer J, Jordi R . Purification and characterization of X-prolyl-dipeptidyl-aminopeptidase from Lactobacillus lactis and from Streptococcus thermophilus. J Dairy Sci. 1987; 70(4):738-45. DOI: 10.3168/jds.S0022-0302(87)80068-1. View

Khalid N, Marth E . Purification and Partial Characterization of a Prolyl-Dipeptidyl Aminopeptidase from Lactobacillus helveticus CNRZ 32. Appl Environ Microbiol. 1990; 56(2):381-8. PMC: 183349. DOI: 10.1128/aem.56.2.381-388.1990. View

Queen C, Korn L . A comprehensive sequence analysis program for the IBM personal computer. Nucleic Acids Res. 1984; 12(1 Pt 2):581-99. PMC: 321072. DOI: 10.1093/nar/12.1part2.581. View

Eckerskorn C, Mewes W, Goretzki H, Lottspeich F . A new siliconized-glass fiber as support for protein-chemical analysis of electroblotted proteins. Eur J Biochem. 1988; 176(3):509-19. DOI: 10.1111/j.1432-1033.1988.tb14308.x. View

10.

Stoker N, Fairweather N, Spratt B . Versatile low-copy-number plasmid vectors for cloning in Escherichia coli. Gene. 1982; 18(3):335-41. DOI: 10.1016/0378-1119(82)90172-x. View

11.

Dower W, Miller J, Ragsdale C . High efficiency transformation of E. coli by high voltage electroporation. Nucleic Acids Res. 1988; 16(13):6127-45. PMC: 336852. DOI: 10.1093/nar/16.13.6127. View

12.

Miller C, Mackinnon K . Peptidase mutants of Salmonella typhimurium. J Bacteriol. 1974; 120(1):355-63. PMC: 245770. DOI: 10.1128/jb.120.1.355-363.1974. View

13.

Harley C, Reynolds R . Analysis of E. coli promoter sequences. Nucleic Acids Res. 1987; 15(5):2343-61. PMC: 340638. DOI: 10.1093/nar/15.5.2343. View

14.

SANGER F, Nicklen S, Coulson A . DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977; 74(12):5463-7. PMC: 431765. DOI: 10.1073/pnas.74.12.5463. View

15.

Weiss N, Schillinger U, Kandler O . Lactobacillus lactis, Lactobacillus leichmannii and Lactobacillus bulgaricus, Subjective Synonyms of Lactobacillus delbrueckii, and Description of Lactobacillus delbrueckii subsp. lactis comb. nov. and Lactobacillus delbrueckii subsp. bulgaricus.... Syst Appl Microbiol. 2012; 4(4):552-7. DOI: 10.1016/S0723-2020(83)80012-5. View

16.

Atlan D, Laloi P, Portalier R . X-Prolyl-Dipeptidyl Aminopeptidase of Lactobacillus delbrueckii subsp. bulgaricus: Characterization of the Enzyme and Isolation of Deficient Mutants. Appl Environ Microbiol. 1990; 56(7):2174-9. PMC: 184579. DOI: 10.1128/aem.56.7.2174-2179.1990. View

17.

Birnboim H, DOLY J . A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979; 7(6):1513-23. PMC: 342324. DOI: 10.1093/nar/7.6.1513. View

18.

Moran Jr C, Lang N, LeGrice S, Lee G, Stephens M, Sonenshein A . Nucleotide sequences that signal the initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet. 1982; 186(3):339-46. DOI: 10.1007/BF00729452. View

19.

Casey M, Meyer J . Presence of X-prolyl-dipeptidyl-peptidase in lactic acid bacteria. J Dairy Sci. 1985; 68(12):3212-5. DOI: 10.3168/jds.S0022-0302(85)81229-7. View

20.

Vieira J, Messing J . Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985; 33(1):103-19. DOI: 10.1016/0378-1119(85)90120-9. View