Surface-Layer Lattices as Patterning Element for Multimeric Extremozymes

A promising new approach for the production of biocatalysts comprises the use of surface‐layer (S‐layer) lattices that present functional multimeric enzymes on their surface, thereby guaranteeing most accurate spatial distribution and orientation, as well as maximal effectiveness and stability of th...

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Bibliographic Details
Published in:Small (Weinheim an der Bergstrasse, Germany) Germany), 2013-11, Vol.9 (22), p.3887-3894
Main Authors: Ferner-Ortner-Bleckmann, Judith, Gelbmann, Nicola, Tesarz, Manfred, Egelseer, Eva M., Sleytr, Uwe B.
Format: Article
Language:English
Subjects:
Aldose-Ketose Isomerases - genetics
Aldose-Ketose Isomerases - metabolism
Bacteria
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Bacteriology
biocatalysts
Catalysis
Crystallization
Enzymes
extremozymes
gene technology
Membrane Glycoproteins - genetics
Membrane Glycoproteins - metabolism
Nanotechnology
protein structures
Proteins
Recombinant Fusion Proteins - genetics
Recombinant Fusion Proteins - metabolism
self-assembly
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Summary:A promising new approach for the production of biocatalysts comprises the use of surface‐layer (S‐layer) lattices that present functional multimeric enzymes on their surface, thereby guaranteeing most accurate spatial distribution and orientation, as well as maximal effectiveness and stability of these enzymes. For proof of concept, a tetrameric and a trimeric extremozyme are chosen for the construction of S‐layer/extremozyme fusion proteins. By using a flexible peptide linker, either one monomer of the tetrameric xylose isomerase XylA from the thermophilic Thermoanaerobacterium strain JW/SL‐YS 489 or, in another approach, one monomer of the trimeric carbonic anhydrase from the methanogenic archaeon Methanosarcina thermophila are genetically linked to one monomer of the S‐layer protein SbpA of Lysinibacillus sphaericus CCM 2177. After isolation and purification, the self‐assembly properties of both S‐layer fusion proteins as well as the specific activity of the fused enzymes are confirmed, thus indicating that the S‐layer protein moiety does not influence the nature of the multimeric enzymes and vice versa. By recrystallization of the S‐layer/extremozyme fusion proteins on solid supports, the active enzyme multimers are exposed on the surface of the square S‐layer lattice with 13.1 nm spacing. Novel self‐assembling biocatalysts based on a bacterial surface‐layer (S‐layer) protein and multimeric extremozymes are successfully produced. A tetrameric or a trimeric enzyme is genetically fused to an S‐layer protein through a flexible peptide linker. By recrystallization of the S‐layer/extremozyme fusion proteins on solid supports, active enzyme multimers are exposed in an accurate spatial distribution on the surface of the square S‐layer lattice.
ISSN:1613-6810
1613-6829
DOI:10.1002/smll.201201014