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(Received for publication, April 21, 1997)
From the Department of Bacteriology, Institute of Infectious
Diseases and Immunology, School of Veterinary Medicine, University of
Utrecht, 3508 TD Utrecht, The Netherlands.
Bacteria belonging to the species
Streptococcus pneumoniae vary in their capsule. Presently,
90 capsular serotypes are known, all possessing their own specific
polysaccharide structure. Little is known about the biosynthesis of
these capsular polysaccharides. The cps locus of S. pneumoniae serotype 14 was cloned. So far, 7 open reading frames
have been sequenced, cps14B to cps14H. The gene
products are similar to proteins involved in bacterial polysaccharide biosynthesis, both of Gram-negative and -positive micro-organisms. Gene-specific mutants were created for cps14D to
cps14H by insertional mutagenesis. All mutants no longer
agglutinated with a monoclonal antibody against type 14 capsule
polysaccharides. The biosynthetic function of cps14E and
cps14G was determined by analysis of the intermediates in
the synthesis of the oligosaccharide subunit, formed in membrane
preparations of the wild-type and mutant strains and in membrane
preparations of Escherichia coli expressing the pneumococcal glycosyltransferases. The enzyme encoded by
cps14E is a glucosyl-1-phosphate transferase that links
glucose to a lipid carrier, the first step in the biosynthesis of the
type 14 repeating unit. The gene product of cps14G encodes
a 
-1,4-galactosyltransferase, the enzyme responsible for the second
step in the subunit synthesis, the transfer of galactose to
lipid-linked glucose.
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