The use of enzymes to remove stains comprising proteins and/or carbohydrates, in combination with various detergents, is well known in the art of detergent formulations. Such enzyme formulations are designed to remove various types of stains from soft surfaces such as cloth and hard surfaces such as porcelain and metal. Thus, for example, proteases such as trypsin, pancreatin, papain and bromelain have reportedly been used in detergent formulations to remove proteinaceous stains with variable degrees of success. Specific glycosidases such as cellulase, lysozyme, amylase and glucanase, on the other hand, have been formulated with various detergents for removal of certain carbohydrate stains. Other detergent formulations have combined proteases and glycosidases for stain treatment.
Some of the glycosidases used in detergent formulations, e.g., .beta.-amylase, .alpha.-galactosidase and .beta.-galactosidase, are exoglycosidases which cleave one or more terminal residues from an oligosaccharide or polysaccharide. Other glycosidases, e.g. cellulase and .alpha.-amylase are endoglycosidases which are reactive with specific internal linkages within an oligo- or polysaccharide substrate. Such endoglycosidases are referred to herein as Type I endoglycosidases. Although formulations of detergent with one or more proteases and/or glycosidases (including Type I endoglycosidases) have greatly improved stain removal, many stains, e.g. blood, fecal material and body soil stains, often leave a residual stain after treatment.
In the art of contact lens cleaning, similar enzyme/detergent formulations have been used to clean and sterilize hard and soft contact lenses. In many cases, these formulations have been used to degrade the biofilm which forms on the surface of contact lenses and which is used by various ophthalmic pathogens such as Pseudomonas aeruginosa and Staphylococcus epidermidis to adhere to such lens. See, e.g., Duran, J. A., et al. (1987), Arch. Ophthalmol, 105 106-109; Stern, G. A., et al. (1987), Ophthalmology, 94, 115-119 (which reports the treatment of mucin coated contact lenses with various enzymes such as pancreatin, papain, trypsin and neuraminidase to inhibit Pseudomonas adherence); and Slucher, M. M., et al. (1987), Arch. Ophthalmol, 105, 110-115.
The use of biofilms for microbial adhesion is not limited to contact lenses. Thus, Streptococcus mutans reportedly uses extracellular polysaccharides to adhere to tooth enamel. EPO Publication No. 0195672 reports the use of .alpha.-1,3 glucanase or .alpha.-1,6 glucanase to cleave the extracellular polysaccharides used by Streptococcus mutans to adhere to tooth enamel.
The effect of certain enzymes on cells adhered to glass surfaces has also been reported by Danielsson, A., et al. (1977), Botanica Marina, 20, 13-17. As reported therein, Pseudomonas species isolated from sea water was adhered to glass slides. Thereafter, the slides were treated with either pronase, trypsin, .alpha.-amylase (a Type I endoglycosidase), or lysozyme (also a Type I endoglycosidase). In this report, treatment with the proteolytic enzymes pronase and trypsin resulted in the release of a portion of the population of adhered bacteria, whereas the cell degradative enzyme lysozyme showed diminished activity compared to the proteolytic enzymes. The .alpha.-amylase reportedly had no effect at all. In addition to the attachment of microorganisms to contact lenses, tooth enamel and glass surfaces, many other surfaces are subject to microbial attachment. See, e.g., Marrie, T. J., et al. (1984), J. Clin. Microbiology, 19, 991-914 (bacterial attachment to cardiac pacemaker leads and powerpacks); Freimer, N. B., et al. (1978), Acta. Path. Microbiol. Scand. Sectb., 86, 53-57 (binding of microorganisms to macrophages); and Mirelman, et al. (1982), Tokai J. Exp. Clin. Med., 7, 77-183 (microbial adherence to mammalian mucosal surfaces). Various mechanisms have been proposed to describe the adhesion of microorganisms, such as bacteria, to non-biological solid surfaces. See, e.g. Fletcher, M. (1987), Microbiological Sciences, 4, 133-136, and Duddridge, J. E., et al. (1983), Factors Affecting the Adhesion of Bacteria to Surfaces in Microbial Corrosion, Delco Printing Co., Ltd., pp. 28-35. Although these references discuss microbial adherence to various surfaces and the factors which may be involved in such attachment, they do not discuss the control of microorganism growth on such surfaces or their removal therefrom.
Type II endoglycosidases, as used herein, are a category of endoglycosidases which are capable of cleaving specific internal glycosidic linkages found in glycoproteins. These endoglycosidases cleave all or part of the carbohydrate moiety from a glycoprotein depending on the location of the reactive glycosidic linkage in the glycoprotein. Examples include endo-.beta.-N-acetylglucosaminidases (Endo-D, Endo-H, Endo-L, Endo-CI, Endo-CII, Endo-F-Gal type and Endo-F), endo-.alpha.-N-acetylgalactosaminidase and endo-.beta.-N-galactosidases. See, e.g. Tarentino, A. L., et al. (1985), Biochem, 24, 4665-4671; Arakawa, M., et al. (1974), J. Biochem., 76, 307-317; Plummer, T. H., et al. (1984), J. Biochem, 259, 10700-10704; Tarentino, A. L., et al. (1975), Biochem. and Biophys. Res. Comm., 67, 455-462; and Trimble, R. B., et al. (1984), Anal. Biochem., 141, 515-522; and "Glycoprotein and Proteoglycan Techniques" (1985) by J. G. Beeley, Chapter 6, pp. 153-300, Elsevier, Amsterdam, N.Y., Oxford. In addition to having a specificity for the internal glycosidic linkages of glycoproteins, at least one endoglycosidase (endo-.beta.-N-acetylglucosaminidase H) has also demonstrated a specificity which produces the cleavage of lipid-linked oligosaccharides (Chalifour, R. J., et al. (1983), Archives of Biochem. and Biophys., 229, 386-394) and reportedly di-N-acetylchitobiose linkages in oligosaccharides and glycoproteins (Tarention, A. L., et al. (1974), J. Biol. Chem., 249, 811-817).
Such Type II endoglycosidases, in general, have been used primarily for analytical purposes, e.g. the determination of protein or carbohydrate sequence and/or the structure and function of specific glycoproteins. See, e.g. Hsieh, P., et al. (1982), J. Biolchem., 258, 2555-2561, and Geyar, R., et al. (1984), Eur. J. Biochem., 143, 531-539. In a recent report, a Type II endoglycosidase was reportedly used to analyze a glycoprotein antigen from Leishmania mexicana amazonensis. Chin Shen Chang, et al. (1986), Mol. Biochem. Parasitol 18, 197-210. This glycoprotein antigen was first immunologically bound to immunobeads. After reacting the immunologically bound glycoprotein with analytical amounts of Endo-H, the immunobeads were washed and boiled in buffer containing 1% SDS in preparation for polyacrylamide gel electrophoresis. This analysis revealed a decrease in molecular weight attributed to the cleavage of carbohydrate from the immunologically bound glycoprotein antigen.
Type II endoglycosidases, however, have not been used to remove substances, including glycoproteins and glycolipids, from surfaces of substances such as fabric, contact lenses, metals, ceramics, cells, tissue and the like. Nor have they been used to control microorganism growth in suspension or on such surfaces.
Glycosidases have been used in combination with other enzymes for removal of various materials. .beta.-glycosidases are described as carbohydrate-metabolizing enzymes in Anderson, et al. (1964), Biochem. J., 90, 30. Neuraminidase (N-acetylneuraminiate glycohydrolase) inhibitors are viewed as possible anti-viral, antibacterial agents in Khorlin, et al. (1979), FEBS Letters, 8, 17; and Haskell, et al. (1970), J. Med. Chem,, 13, 48. Dextranase is described as catalyzing hydrolysis of bacterial polysaccharide, dextran (.alpha.-1,6-glucan), to isomaltose residues in Chalet, et al. (1970), Appl. Microbiol., 20, 421. Lysozyme (muramidase) is described as hydrolyzing glycosidic linkages in the mucopolysaccharide cell wall structure of a variety of microbes in Chipman, et al. (1969), Science, 165, 454 and Montague (1964), Biochem. Biophys. Acta., 86, 588. Lastly, inhibition of lysozyme by D-glucosamine derivatives is described in Neuberger, et al. (1967), Nature, 215, 524.
Type II endoglycosidases such as endo-.beta.-N-acetylglucosaminidase H, D, F and/or PNGase F have not, however, previously been combined with antimicrobial agents to form antimicrobial compositions.
The references discussed above are provided solely for their disclosure prior to the filing date of the instant case, and nothing herein is to be construed as an admission that such references are prior art or that the inventors are not entitled to antedate such disclosure by virtue of prior invention or priority based on earlier filed applications.