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Heterogeneity of S-layer proteins from aggregating and non-aggregating Lactobacillus kefir strains

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Abstract

Since the presence of S-layer protein conditioned the autoaggregation capacity of some strains of Lactobacillus kefir, S-layer proteins from aggregating and non-aggregating L. kefir strains were characterized by immunochemical reactivity, MALDI-TOF spectrometry and glycosylation analysis. Two anti-S-layer monoclonal antibodies (Mab5F8 and Mab1F8) were produced; in an indirect enzyme-linked immunosorbent assay Mab1F8 recognized S-layer proteins from all L. kefir tested while Mab5F8 recognized only S-layer proteins from aggregating strains. Periodic Acid-Schiff staining of proteins after polyacrylamide gel electrophoresis under denaturing conditions revealed that all L. kefir S-layer proteins tested were glycosylated. Growth of bacteria in the presence of the N-glycosylation inhibitor tunicamycin suggested the presence of glycosydic chains O-linked to the protein backbone. MALDI-TOF peptide map fingerprint for S-layer proteins from 12 L. kefir strains showed very similar patterns for the aggregating strains, different from those for the non-aggregating ones. No positive match with other protein spectra in MSDB Database was found. Our results revealed a high heterogeneity among S-layer proteins from different L. kefir strains but also suggested a correlation between the structure of these S-layer glycoproteins and the aggregation properties of whole bacterial cells.

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Acknowledgments

This work was supported by the Agencia Nacional de Investigaciones Científicas y Tecnológicas (ANPCyT), Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Comisión de Investigaciones Científicas de la Provincia de Buenos Aires (CIC PBA) and, Facultad de Ciencias Exactas, Universidad Nacional de La Plata (UNLP). The IBB-UAB is a member of ProteoRed, funded by Genoma Spain and follows the quality criteria set up by ProteoRed standards. Authors are grateful to Diana Velasco for bibliographic assistance.

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Authors and Affiliations

  1. Centro de Investigación y Desarrollo en Criotecnología de Alimentos (CIDCA), CCT La Plata, CONICET, 47 y 116 (s/n), La Plata, 1900, Argentina

    Pablo Mobili, Analía G. Abraham & Graciela L. De Antoni

  2. Cátedra de Microbiología, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina

    María de los Ángeles Serradell & Graciela L. De Antoni

  3. Institut de Biotecnología y Biomedicina (IBB), de la Universitat Autònoma de Barcelona, Barcelona, Spain

    Sebastián A. Trejo & Francesc X. Avilés Puigvert

  4. Área Bioquímica y Control de Alimentos, Facultad de Ciencias Exactas, Universidad Nacional e La Plata, La Plata, Argentina

    Analía G. Abraham

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  1. Pablo Mobili
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  2. María de los Ángeles Serradell
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  3. Sebastián A. Trejo
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  4. Francesc X. Avilés Puigvert
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  5. Analía G. Abraham
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  6. Graciela L. De Antoni
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Correspondence to Analía G. Abraham.

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Mobili, P., de los Ángeles Serradell, M., Trejo, S.A. et al. Heterogeneity of S-layer proteins from aggregating and non-aggregating Lactobacillus kefir strains. Antonie van Leeuwenhoek 95, 363–372 (2009). https://doi.org/10.1007/s10482-009-9322-y

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  • DOI: https://doi.org/10.1007/s10482-009-9322-y

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