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Applied and Environmental Microbiology, September 2002, p. 4283-4291, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4283-4291.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Molecular Characterization of a Novel Glucosyltransferase from Lactobacillus reuteri Strain 121 Synthesizing a Unique, Highly Branched Glucan with {alpha}-(1->4) and {alpha}-(1->6) Glucosidic Bonds

S. Kralj,1,2 G. H. van Geel-Schutten,1,3 H. Rahaoui,4 R. J. Leer,3 E. J. Faber,5 M. J. E. C. van der Maarel,1,6 and L. Dijkhuizen1,2*

Centre for Carbohydrate Bioengineering, TNORUG,1 Department of Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute, University of Groningen, Haren,2 Department of Applied Microbiology and Gene Technology,3 Department of Food Microbiology TNO Nutrition and Food Research, Zeist,4 Bijvoet Center, Department of Bio-Organic Chemistry, Utrecht University, Utrecht,5 Carbohydrate Technology Department, TNO Nutrition and Food Research, Groningen, The Netherlands6

Received 24 January 2002/ Accepted 8 June 2002

Lactobacillus reuteri strain 121 produces a unique, highly branched, soluble glucan in which the majority of the linkages are of the {alpha}-(1->4) glucosidic type. The glucan also contains {alpha}-(1->6)-linked glucosyl units and 4,6-disubstituted {alpha}-glucosyl units at the branching points. Using degenerate primers, based on the amino acid sequences of conserved regions from known glucosyltransferase (gtf) genes from lactic acid bacteria, the L. reuteri strain 121 glucosyltransferase gene (gtfA) was isolated. The gtfA open reading frame (ORF) was 5,343 bp, and it encodes a protein of 1,781 amino acids with a deduced Mr of 198,637. The deduced amino acid sequence of GTFA revealed clear similarities with other glucosyltransferases. GTFA has a relatively large variable N-terminal domain (702 amino acids) with five unique repeats and a relatively short C-terminal domain (267 amino acids). The gtfA gene was expressed in Escherichia coli, yielding an active GTFA enzyme. With respect to binding type and size distribution, the recombinant GTFA enzyme and the L. reuteri strain 121 culture supernatants synthesized identical glucan polymers. Furthermore, the deduced amino acid sequence of the gtfA ORF and the N-terminal amino acid sequence of the glucosyltransferase isolated from culture supernatants of L. reuteri strain 121 were the same. GTFA is thus responsible for the synthesis of the unique glucan polymer in L. reuteri strain 121. This is the first report on the molecular characterization of a glucosyltransferase from a Lactobacillus strain.

* Corresponding author. Mailing address: Department of Microbiology, University of Groningen, P.O. Box 14, 9750 AA Haren, The Netherlands. Phone: 31.50.3632150. Fax: 31.50.3632154. E-mail: L.Dijkhuizen{at}biol.rug.nl.

Applied and Environmental Microbiology, September 2002, p. 4283-4291, Vol. 68, No. 9
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.9.4283-4291.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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