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Journal of Bacteriology, October 1999, p. 6347-6353, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Functional Analysis of Glycosyltransferase Genes
from Lactococcus lactis and Other Gram-Positive Cocci:
Complementation, Expression, and Diversity
Richard
van
Kranenburg,*
Harmjan R.
Vos,
Iris I.
van Swam,
Michiel
Kleerebezem, and
Willem M.
de Vos
Microbial Ingredients Section, NIZO Food
Research, Ede, The Netherlands
Received 10 February 1999/Accepted 23 March 1999
Sixteen exopolysaccharide (EPS)-producing Lactococcus
lactis strains were analyzed for the chemical compositions of
their EPSs and the locations, sequences, and organization of the
eps genes involved in EPS biosynthesis. This allowed the
grouping of these strains into three major groups, representatives of
which were studied in detail. Previously, we have characterized the eps gene cluster of strain NIZO B40 (group I) and
determined the function of three of its glycosyltransferase (GTF)
genes. Fragments of the eps gene clusters of strains NIZO
B35 (group II) and NIZO B891 (group III) were cloned, and these encoded
the NIZO B35 priming galactosyltransferase, the NIZO B891 priming
glucosyltransferase, and the NIZO B891 galactosyltransferase involved
in the second step of repeating-unit synthesis. The NIZO B40 priming
glucosyltransferase gene epsD was replaced with an
erythromycin resistance gene, and this resulted in loss of EPS
production. This epsD deletion was complemented with
priming GTF genes from gram-positive organisms with known function and
substrate specificity. Although no EPS production was found with
priming galactosyltransferase genes from L. lactis or
Streptococcus thermophilus, complementation with priming
glucosyltransferase genes involved in L. lactis EPS and
Streptococcus pneumoniae capsule biosynthesis could
completely restore or even increase EPS production in L. lactis.
*
Corresponding author. Mailing address: Microbial
Ingredients Section, NIZO Food Research, Kernhemseweg 2, 6718 ZB Ede,
The Netherlands. Phone: 31-318-659511. Fax: 31-318-650400. E-mail: kranenbu{at}nizo.nl.
Journal of Bacteriology, October 1999, p. 6347-6353, Vol. 181, No. 20
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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