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Gewählte Publikation:

Nguyen, HM; Mathiesen, G; Stelzer, EM; Pham, ML; Kuczkowska, K; Mackenzie, A; Agger, JW; Eijsink, VGH; Yamabhai, M; Peterbauer, CK; Haltrich, D; Nguyen, TH.
(2016): Display of a beta-mannanase and a chitosanase on the cell surface of Lactobacillus plantarum towards the development of whole-cell biocatalysts
MICROB CELL FACT. 2016; 15: FullText FullText_BOKU

Abstract:
Background: Lactobacillus plantarum is considered as a potential cell factory because of its GRAS (generally recognized as safe) status and long history of use in food applications. Its possible applications include in situ delivery of proteins to a host, based on its ability to persist at mucosal surfaces of the human intestine, and the production of food-related enzymes. By displaying different enzymes on the surface of L. plantarum cells these could be used as whole-cell biocatalysts for the production of oligosaccharides. In this present study, we aimed to express and display a mannanase and a chitosanase on the cell surface of L. plantarum. Results: ManB, a mannanase from Bacillus licheniformis DSM13, and CsnA, a chitosanase from Bacillus subtilis ATCC 23857 were fused to different anchoring motifs of L. plantarum for covalent attachment to the cell surface, either via an N-terminal lipoprotein anchor (Lp_1261) or a C-terminal cell wall anchor (Lp_2578), and the resulting fusion proteins were expressed in L. plantarum WCFS1. The localization of the recombinant proteins on the bacterial cell surface was confirmed by flow cytometry and immunofluorescence microscopy. The highest mannanase and chitosanase activities obtained for displaying L. plantarum cells were 890 U and 1360 U g dry cell weight, respectively. In reactions with chitosan and galactomannans, L. plantarum CsnA- and ManB-displaying cells produced chito-and manno-oligosaccharides, respectively, as analyzed by high performance anion exchange chromatography (HPAEC) and mass spectrometry (MS). Surface-displayed ManB is able to break down galactomannan (LBG) into smaller manno-oligosaccharides, which can support growth of L. plantarum. Conclusion: This study shows that mannanolytic and chitinolytic enzymes can be anchored to the cell surface of L. plantarum in active forms. L. plantarum chitosanase-and mannanase-displaying cells should be of interest for the production of potentially xxxprebioticxxx oligosaccharides. This approach, with the enzyme of interest being displayed on the cell surface of a food-grade organism, may also be applied in production processes relevant for food industry.
Autor*innen der BOKU Wien:
Haltrich Dietmar
Nguyen Hoang Minh
Nguyen Thu Ha
Peterbauer Clemens Karl
Stelzer Elena Maria
BOKU Gendermonitor:

Find related publications in this database (using NML MeSH Indexing)
Biocatalysis;Cell Surface Display Techniques*;Cloning, Molecular;Food Industry/methods;Glycoside Hydrolases/genetics;Glycoside Hydrolases/metabolism*;Humans;Lactobacillus plantarum/enzymology*;Lactobacillus plantarum/genetics*;Lactobacillus plantarum/growth & development;Lactobacillus plantarum/metabolism;Lipoproteins/metabolism;Mannans/metabolism;Oligosaccharides/metabolism;Prebiotics;Recombinant Proteins/metabolism;beta-Mannosidase/genetics;beta-Mannosidase/metabolism*;

Find related publications in this database (Keywords)
Cell-surface display
Whole-cell biocatalyst
Lactobacillus plantarum
Mannanase
Chitosanase
Lipoprotein anchor
Cell wall anchor


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