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Applied and Environmental Microbiology, June 2002, p. 2917-2923, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.2917-2923.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Lactobacillus bulgaricus Proteinase Expressed in Lactococcus lactis Is a Powerful Carrier for Cell Wall-Associated and Secreted Bovine ß-Lactoglobulin Fusion Proteins

Eric Bernasconi,1 Jacques-Edouard Germond,2 Michèle Delley,2 Rodolphe Fritsché,2 and Blaise Corthésy1*

R & D Laboratory of the Division of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois, CH-1000 Lausanne 11,1 Nestec Ltd., Nestlé Research Center, CH-1000 Lausanne 26, Switzerland2

Received 27 November 2001/ Accepted 1 April 2002

Lactic acid bacteria have a good potential as agents for the delivery of heterologous proteins to the gastrointestinal mucosa and thus for the reequilibration of inappropriate immune responses to food antigens. Bovine ß-lactoglobulin (BLG) is considered a major allergen in cow's milk allergy. We have designed recombinant Lactococcus lactis expressing either full-length BLG or BLG-derived octapeptide T6 (IDALNENK) as fusions with Lactobacillus bulgaricus extracellular proteinase (PrtB). In addition to constructs encoding full-length PrtB for the targeting of heterologous proteins to the cell surface, we generated vectors aiming at the release into the medium of truncated PrtB derivatives lacking 100 (PrtB{partial}, PrtB{partial}-BLG, and PrtB{partial}-T6) or 807 (PrtB{Delta}) C-terminal amino acids. Expression of recombinant products was confirmed using either anti-PrtB, anti-BLG, or anti-peptide T6 antiserum. All forms of the full-length and truncated recombinant products were efficiently translocated, irrespective of the presence of eucaryotic BLG sequences in the fusion proteins. L. lactis expressing PrtB{partial}-BLG yielded up to 170 µg per 109 CFU in the culture supernatant and 9 µg per 109 CFU at the bacterial cell surface within 14 h. Therefore, protein fusions relying on the use of PrtB gene products are adequate for concomitant cell surface display and secretion by recombinant L. lactis and thus may ensure maximal bioavailability of the eucaryotic antigen in the gut-associated lymphoid tissue.

* Corresponding author. Mailing address: R & D Laboratory, Division of Immunology and Allergy, Hôpital Orthopédique, Avenue Pierre Decker 4, CH-1005 Lausanne, Switzerland. Phone: 41 21 314 07 83. Fax: 41 21 314 07 71. E-mail: blaise.corthesy{at}chuv.hospvd.ch.

Applied and Environmental Microbiology, June 2002, p. 2917-2923, Vol. 68, No. 6
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.6.2917-2923.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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  • Chatel, J.-M., Nouaille, S., Adel-Patient, K., Le Loir, Y., Boe, H., Gruss, A., Wal, J.-M., Langella, P. (2003). Characterization of a Lactococcus lactis Strain That Secretes a Major Epitope of Bovine Beta-Lactoglobulin and Evaluation of Its Immunogenicity in Mice. Appl. Environ. Microbiol. 69: 6620-6627 [Abstract] [Full Text]  
  • Germond, J.-E., Delley, M., Gilbert, C., Atlan, D. (2003). Determination of the Domain of the Lactobacillus delbrueckii subsp. bulgaricus Cell Surface Proteinase PrtB Involved in Attachment to the Cell Wall after Heterologous Expression of the prtB Gene in Lactococcus lactis. Appl. Environ. Microbiol. 69: 3377-3384 [Abstract] [Full Text]  


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