Microbiology
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Microbiology 150 (2004), 621-630; DOI  10.1099/mic.0.26671-0
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van Hijum, S. A. F. T.
Right arrow Articles by Dijkhuizen, L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van Hijum, S. A. F. T.
Right arrow Articles by Dijkhuizen, L.
Agricola
Right arrow Articles by van Hijum, S. A. F. T.
Right arrow Articles by Dijkhuizen, L.
Microbiology 150 (2004), 621-630; DOI  10.1099/mic.0.26671-0
© 2004 Society for General Microbiology

Biochemical and molecular characterization of a levansucrase from Lactobacillus reuteri

S. A. F. T. van Hijum1,2, E. Szalowska1,2, M. J. E. C. van der Maarel1,3 and L. Dijkhuizen1,2

1 Centre for Carbohydrate Bioengineering, TNO-RUG, University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
2 Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, PO Box 14, 9750 AA Haren, The Netherlands
3 Innovative Ingredients and Products Department, TNO Nutrition and Food Research, Rouaanstraat 27, 9723 CC Groningen, The Netherlands

Correspondence
L. Dijkhuizen
L.Dijkhuizen{at}biol.rug.nl

Lactobacillus reuteri strain 121 employs a fructosyltransferase (FTF) to synthesize a fructose polymer [a fructan of the levan type, with {beta}(2->6) linkages] from sucrose or raffinose. Purification of this FTF (a levansucrase), and identification of peptide amino acid sequences, allowed isolation of the first Lactobacillus levansucrase gene (lev), encoding a protein (Lev) consisting of 804 amino acids. Lev showed highest similarity with an inulosucrase of L. reuteri 121 [Inu; producing an inulin polymer with {beta}(2->1)-linked fructosyl units] and with FTFs from streptococci. Expression of lev in Escherichia coli resulted in an active FTF (Lev{Delta}773His) that produced the same levan polymer [with only 2–3 % {beta}(2->1->6) branching points] as L. reuteri 121 cells grown on raffinose. The low degree of branching of the L. reuteri levan is very different from bacterial levans known up to now, such as that of Streptococcus salivarius, having up to 30 % branches. Although Lev is unusual in showing a higher hydrolysis than transferase activity, significant amounts of levan polymer are produced both in vivo and in vitro. Lev is strongly dependent on Ca2+ ions for activity. Unique properties of L. reuteri Lev together with Inu are: (i) the presence of a C-terminal cell-wall-anchoring motif causing similar expression problems in Escherichia coli, (ii) a relatively high optimum temperature for activity for FTF enzymes, and (iii) at 50 °C, kinetics that are best described by the Hill equation.

Abbreviations: FTF, fructosyltransferase; HPSEC-MALLS, high-performance size-exclusion chromatography multi-angle laser light scattering

The GenBank accession number for the L. reuteri 121 gene encoding levansucrase (lev) and its flanking regions is AF465251.




This article has been cited by other articles:


Home page
 Appl. Environ. Microbiol.Home page
M. A. Anwar, S. Kralj, M. J. E. C. van der Maarel, and L. Dijkhuizen
The Probiotic Lactobacillus johnsonii NCC 533 Produces High-Molecular-Mass Inulin from Sucrose by Using an Inulosucrase Enzyme
Appl. Envir. Microbiol., June 1, 2008; 74(11): 3426 - 3433.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
L. K. Ozimek, S. Kralj, M. J. E. C. van der Maarel, and L. Dijkhuizen
The levansucrase and inulosucrase enzymes of Lactobacillus reuteri 121 catalyse processive and non-processive transglycosylation reactions.
Microbiology, April 1, 2006; 152(Pt 4): 1187 - 1196.
[Abstract] [Full Text] [PDF]

Home page
 Microbiol. Mol. Biol. Rev.Home page
S. A. F. T. van Hijum, S. Kralj, L. K. Ozimek, L. Dijkhuizen, and I. G. H. van Geel-Schutten
Structure-Function Relationships of Glucansucrase and Fructansucrase Enzymes from Lactic Acid Bacteria
Microbiol. Mol. Biol. Rev., March 1, 2006; 70(1): 157 - 176.
[Abstract] [Full Text] [PDF]

Home page
 Appl. Environ. Microbiol.Home page
M. Malten, R. Biedendieck, M. Gamer, A.-C. Drews, S. Stammen, K. Buchholz, L. Dijkhuizen, and D. Jahn
A Bacillus megaterium Plasmid System for the Production, Export, and One-Step Purification of Affinity-Tagged Heterologous Levansucrase from Growth Medium
Appl. Envir. Microbiol., February 1, 2006; 72(2): 1677 - 1679.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
J. G. Arrieta, M. Sotolongo, C. Menendez, D. Alfonso, L. E. Trujillo, M. Soto, R. Ramirez, and L. Hernandez
A Type II Protein Secretory Pathway Required for Levansucrase Secretion by Gluconacetobacter diazotrophicus
J. Bacteriol., August 1, 2004; 186(15): 5031 - 5039.
[Abstract] [Full Text] [PDF]

Home page
 MicrobiologyHome page
S. Kralj, G. H. van Geel-Schutten, M. J. E. C. van der Maarel, and L. Dijkhuizen
Biochemical and molecular characterization of Lactobacillus reuteri 121 reuteransucrase
Microbiology, July 1, 2004; 150(7): 2099 - 2112.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
INT J SYST EVOL MICROBIOL MICROBIOLOGY J GEN VIROL
J MED MICROBIOL ALL SGM JOURNALS
Copyright © 2004 Society for General Microbiology.