Compositions and methods for enzymatic detachment of...

Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase

Reexamination Certificate

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C435S041000, C424S094600, C536S023200

Reexamination Certificate

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10538902

ABSTRACT:
Isolated nucleic acid sequences and amino acid sequences for soluble, β-N-acetylglucosaminidase or active fragments or variants thereof which promote detachment of bacterial cells from a biofilm are provided. An isolated mutant bacteria which forms biofilm colonies which tightly adhere to surface but which are unable to release cells into the medium or spread over the surface is also provided. In additions, methods are described for modulating detachment of bacterial cells from biofilm by mutating soluble,β-N-acetylglucosamimidase or altering its expression or activity are also provided. Also provided are compositions, methods and devices for preventing, inhibiting and treating bacterial infections.

REFERENCES:
patent: WO 98/50512 (1998-11-01), None
Clarke et al., Cloning and Expression of the β-N-Acetylglucosaminidase Gene fromStreptococcus pneumoniae(1995) J. Biol. Chem., vol. 270(15), pp. 8805-8814.
Graham et al., Molecular Cloning of the cDNA Which Encodes β-N-Acetylhexosaminidase A fromDictyostelium discoideum(1988) J. Biol. Chem, vol. 263(32), pp. 16823-16829.
Kaplan et al., “Detachment ofActinobacillus actinomycetemcomitansBiofilm Cells by an Endogenous β-Hexosaminidase Activity”, J. Bacteriology(2003) 185(16):4693-4698.
Somerville et al., “Sequence analysis of the β-N-acetylglucosaminidase gene ofVibrio vulnificus: Evidence for a common evolutionary origin of hexosaminidases”, Proc. Natl. Acad. Sci. USA(1993) 90:6751-6755.
Kaplan et al., 2001, “Structural and genetic analyses of O polysaccharide fromActinobacillus actinomycetemcomitansserotype f,”Infect. Immun. 69: 5375-5384.
Kaplan et al., 2002, “Biofilm dispersal ofNeisseria subflavaand other phylogenetically diverse oral bacteria,”Appl. Environ. Microbiol. 68(10): 4943-4950.
Kaplan et al., Feb. 2003, “Biofilm growth and detachment ofActinobacillus actinomycelecomitans”, J. Bacteriol., 185(4):1399-1404.

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