Functional Analysis of Surface Layer (S-Layer) Glycoprotein Assembly (P15612-MOB)
Abstract
This project focusses on the complete and detailed description of the genes located in the S-layer glycan biosynthesis cluster of the thermophilic, Gram-positive bacterium Geobacillus stearothermophilus NRS 2004/3a. The work to be accomplished includes the characterization of all glycosyltransferases and other enzymes involved in the assembly of the S-layer glycoprotein glycan. In addition, for the first time, the interaction between the biosynthesis pathways of the S-layer protein of G. stearothermophilus NRS 2004/3a, which is encoded by the recently identified sgsE gene, and the S-layer glycan, which is mainly encoded by the S-layer biosynthesis cluster, will be investigated with respect to protein expression. These studies are required for potential applications of the S-layer glycoprotein of G. stearothermophilus NRS 2004/3a in the field of biomedicine and biotechnology. For this purpose, the S-layer protein glycosylation will be modified by the generation of specific gene knock-out mutants; furthermore, glycan chain microheterogeneity will be controlled at the molecular level. For the introduction of novel glycose residues into a mutant of this organism, bearing a truncated S-layer glycan, the heptose operon from Aneurinibacillus thermoaerophilus DSM 10155 has been chosen. The construction of an S-layer neoglycoprotein with defined, foreign glycan structures would point to the principal suitability of the bacterial system for the generation of recombinant, potentially medically relevant glycoproteins. In the future, the expression of several other important sugar receptor structures as S-layer neogycoproteins will be very likely. The outlined investigations constitute the foundations for the principal application potential of S-layer neoglycoproteins in biomedicine and nanotechnology. As bacteria have rather simple growth requirements and potential problems occurring with protein glycosylation can be excluded in a bacterial system, recombinant S-layer neoglycoproteins provide an attractive alternative to glycoproteins produced in eukaryotic cell lines.
keywords S-Layer (surface layer) Prokaryotic glycoprotein Geobacillus stearothermophilus S-Layer neoglycoprotein
Publikationen
Isolation of three new surface layer protein genes (slp) from Lactobacillus brevis ATCC 14869 and characterization of the change in their expression under aerated and anaerobic conditions.
Autoren: Jakava-Viljanen, M; Avall-Jääskeläinen, S; Messner, P; Sleytr, UB; Palva, A Jahr: 2002
Journal articles
Glycosylation of Bacterial Surface Layer (S-layer) Proteins
Autoren: Schäffer, C., Novotny, R., Pföstl, A., Sleytr, U.B., Messner, P. Jahr: 2003
Conference & Workshop proceedings, paper, abstract
Novel aspects of S-layer protein glycosilation
Autoren: Schäffer, C., Pföstl, A., Novotny, R., Giry-Laterriere, M., Steiner, K., Scheberl, A., Zayni, S., Sleytr, U.B., Messner, P. Jahr: 2003
Conference & Workshop proceedings, paper, abstract
Genetic organization of the first S-layer glycosylation cluster
Autoren: Novotny, R., Pföstl, A., Giry-Laterierre, M., Steiner, K., Zayni, S., Scheberl, A., Sleytr, U.B., Messner, P. Jahr: 2003
Conference & Workshop proceedings, paper, abstract
Crystalline Bacterial Cell Surface Layers (S-Layers)
Autoren: Sleytr, U.B., Messner, P. Jahr: 2003
Chapter in collected volumes
Prokaryotic glycoproteins: unexplored but important.
Autoren: Messner, P; Jahr: 2004
Journal articles
Surface-layer glycoproteins: an example for the diversity of bacterial glycosylation with promising impacts on nanobiotechnology.
Autoren: Schäffer, C; Messner, P Jahr: 2004
Journal articles
S-layer glycan-specific loci on the chromosome of Geobacillus stearothermophilus NRS 2004/3a and dTDP-L-rhamnose biosynthesis potential of G. stearothermophilus strains.
Autoren: Novotny, R; Schäffer, C; Strauss, J; Messner, P Jahr: 2004
Journal articles
Genetic organization of chromosomal S-layer glycan biosynthesis loci of Bacillaceae.
Autoren: Novotny, R; Pfoestl, A; Messner, P; Schäffer, C Jahr: 2004
Journal articles
Classification of isolates from locations in Austria and Yellowstone National Park as Geobacillus tepidamans sp. nov.
Autoren: Schäffer, C; Franck, WL; Scheberl, A; Kosma, P; McDermott, TR; Messner, P Jahr: 2004
Journal articles
Gene cloning, functional expression and secretion of the S-layer protein SgsE from Geobacillus stearothermophilus NRS 2004/3a in Lactococcus lactis.
Autoren: Novotny, R; Scheberl, A; Giry-Laterriere, M; Messner, P; Schäffer, C Jahr: 2005
Journal articles
The structure of secondary cell wall polymers: how Gram-positive bacteria stick their cell walls together.
Autoren: Schäffer, C; Messner, P Jahr: 2005
Journal articles
N-acetylmuramic acid as capping element of alpha-D-fucose-containing S-layer glycoprotein glycans from Geobacillus tepidamans GS5-97T.
Autoren: Kählig, H; Kolarich, D; Zayni, S; Scheberl, A; Kosma, P; Schäffer, C; Messner, P Jahr: 2005
Journal articles
The secondary cell wall polymer of Geobacillus tepidamans GS5-97T: structure of different glycoforms.
Autoren: Steindl, C; Schäffer, C; Smrecki, V; Messner, P; Müller, N Jahr: 2005
Journal articles
Glycosylation of surface layer (S-layer) proteins,
Autoren: Messner, P. Jahr: 2005
Conference & Workshop proceedings, paper, abstract
Glycosylation of surface layer proteins: mechanisms for glycosylation in bacteria
Autoren: Steiner, K., Novotny, R., Schäffer, C, Messner, P. Jahr: 2004
Conference & Workshop proceedings, paper, abstract
Genetic organization of chromosomal S-layer glycan biosynthesis loci of Bacillaceae
Autoren: Novotny, R., Steiner, K., Giry-Laterriere, M., Zayni, S., Scheberl, A., Messner, P., Schäffer, C. Jahr: 2004
Conference & Workshop proceedings, paper, abstract
Glycosylation of surface layer proteins: glycosylation mechanisms in bacteria
Autoren: Steiner, K., Novotny, R., Schäffer, C., Messner, P. Jahr: 2005
Chapter in collected volumes
Project staff
Paul Messner
Ao.Univ.Prof.i.R. Dipl.-Ing. Dr.nat.techn. Paul Messner
paul.messner@boku.ac.at
Project Leader
01.08.2002 - 31.07.2004
Christina Schäffer
Univ.Prof. Dipl.-Ing. Dr.nat.techn. Christina Schäffer
christina.schaeffer@boku.ac.at
Tel: +43 1 47654-80203
Project Staff
01.08.2002 - 31.07.2004
BOKU partners
External partners
University of South Carolina
Prof. Dr. Alvin Fox
partner