&bgr;1,3-galactosyltransferase and DNA encoding the same

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical

Reexamination Certificate

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C435S193000, C435S252300, C435S252330, C435S320100, C536S023200

Reexamination Certificate

active

06828128

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a protein having a &bgr;1,3-galactosyltransferase activity, a DNA encoding the protein, a recombinant DNA containing the DNA, a transformant containing the recombinant DNA, a method for producing a protein having a &bgr;1,3-galactosyltransferase activity using the transformant, and a method for producing a galactose-containing carbohydrate using the transformant.
2. Background Art
Regarding &bgr;1,3-galactosyltransferase genes, the genes derived from higher animal (
J. Biol. Chem
., 273: 58 (1998),
J. Biol. Chem
., 273: 12770 (1998),
J. Biol. Chem
., 274: 12499 (1999)) have been obtained. However, since it is generally difficult to express the genes derived from &bgr;1,3-galactosyltransferase gene derived from higher-animal has not been expressed as an active protein in a microorganism such as
Escherichia coli
or the like.
On the other hand, in microorganisms, there is a report stating that a &bgr;1,3-galactosyltransferase gene was obtained from
Campylobacter jejuni
and the gene was expressed in
Escherichia coli
. However, although this enzyme has an activity of transferring galactose to N-acetylgalactosamine, there is no report about the activity of transferring galactose to N-acetylglucosamine (
J. Biol. Chem
., 275: 3896 (2000)).
Human milk abundantly contains galactose-containing carbohydrates, lacto-N-tetraose being one of the main components (
Acta Paediatr
., 82: 903 (1993),
J. Pediatr. Gastroenterol. Nutr
., 30: 181 (2000)). Since it is known that lacto-N-tetraose and lacto-N-neotetraose, which are the galactose-containing carbohydrates, are recognized by
Pseudomonas aeruginosa
(
Infect. Immun
., 59: 700 (1991)), the galactose-containing carbohydrates are considered to be strong candidates for safe antiinfection drugs which can prevent human body from infection with
Pseudomonas aeruginosa.
Regarding production of a galactose-containing carbohydrate such as lacto-N-tetraose or the like, both the methods of extraction from human milk and chemical synthesis are known but such methods have problems in terms of cost and productivity, so that its industrial production method has not yet been established.
SUMMARY OF THE INVENTION
An object of the present invention is to provide a protein having a &bgr;1,3-galactosyltransferase activity, and a DNA encoding the protein.
Another object of the present invention is to provide a method for producing a protein having a &bgr;1,3-galactosyltransferase activity using a transformant containing the DNA, and a method for producing a galactose-containing carbohydrate using the protein.
These and other objects have been attained by the present invention which relates to a
Streptococcus agalactiae
protein having a &bgr;1,3-galactosyltransferase activity, a DNA encoding the protein, a recombinant DNA comprising the DNA, a transformant containing the recombinant DNA, a method for producing a protein having a &bgr;1,3-galactosyltransferase activity using the transformant, and a method for producing a galactose-containing carbohydrate using the transformant.


REFERENCES:
Isshiki, et al., “Cloning, Expression, and Characterization of a Novel . . . ”, J. Biol. Chem., vol. 274, No. 18 (1999), pp. 12499-12507.
Amado, et al., “A Family of Human &bgr;3-Galactosyltransferases”, J. Biol. Chem., vol. 273, No. 21 (1998), pp. 12770-12778.
Hennet, et al., “Genomic Cloning and Expression of Three Murine UDP-galactose: . . . ”, J. Biol. Chem., vol. 273, No. 1 (1998), pp. 58-65.
Gilbert, et al., “Biosynthesis of Ganglioside Mimics . . . ”, J. Biol. Chem., vol. 275, No. 6 (2000), pp. 3896-3906.

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