Staphylococcus aureus purification

Peters, D. Frank, R. Hengstenberg, W. Lactose-specific enzyme II of the phosphoenolpyruvate-dependent phosphotransferase system of Staphylococcus aureus. Purification of the histidine-tagged transmembrane component IICBLac and its hydrophilic IIB domain by metal-affinity chromatography, and functional characterization. Eur. J. Biochem., 228, 798-804 (1995)... 

Purification of Antibiotic 66-40 — Dissolve 28 g of crude Antibiotic 66-40 in 100 ml of distilled water and charge to an anion exchange adsorption column (Dowex 1 X2) in the hydroxyl form. Slurry 2,000 g of the resin in water in to a column 2yj in diameter and 36 "high. Elute the column with distilled water at a rate of about 23 ml/min collecting 100 ml fractions and monitor with a conductivity meter and by disc testing against Staphylococcus aureus. 

Analysis of table two shows that a staphylococcus aureus count of 1 million colony forming units per gram was killed off on plate within 5 to 15 minutes using very high levels of antimicrobials at a level only suitable for feet application whereas the Myavert C in the face mask achieved the same level of kill within three minutes, yet it is very mild and suitable for face and eye area application. Three minutes and longer application time for a product such as face is mask is common and this would achieve normal cleansing as well as microbiological purification of the face of the customer. 

Protein A, produced by Staphylococcus aureus, binds IgG. It is used extensively in antibody purification protocols Lectins are a group of proteins capable of binding carbohydrates. 

Bissett, D.L. Anderson, R.L. Lactose and D-galactose metabolism in Staphylococcus aureus. Ill Purification and properties of D-tagatose-6-phosphate kinase. J. Biol. Chem., 255, 8745-8749 (1980)... 

LJ Frank, D Wisniewski, GG Hammond, J Hermes, A Marcy, PM Cameron. High-yield expression, refolding, and purification of penicillin-binding protein 2a from methicillin-resistant Staphylococcus aureus strain 27R. Prot Expr Purif 6 671-678, 1995. 

S Roychoudhury, JE Dotzlaf, S Ghag, WK Yeh. Purification, properties, and kinetics of enzymatic acylation with (3-lactams of soluble penicillin-binding protein 2a a major factor in methicillin-resistant Staphylococcus aureus. J Biol Chem 269 12067-12073, 1994. 

CYE Wu, LC Blaszczak, MC Smith, PL Skatrud. Construction of a modified penicillin-binding protein 2a from methicillin-resistant Staphylococcus aureus and purification by immobilized metal affinity chromatography. J Bacteriol 176 1539-1541, 1994. 

CR Harrington, DM O Hara, PE Reynolds. Characterization of a monoclonal antibody and its use in the immunoaffinity purification of penicillin-binding protein 2 of methicillin-resistant Staphylococcus aureus. FEMS Microbiol Lett 53 143-147, 1989. 

Gaskin, D. K., Bohach, G. A., Schlievert, P. M., and Hovde, C. J. (1997). Purification of Staphylococcus aureus /3-toxin Comparison of three isoelectric focusing methods. Protein Express. Purif. 9, 76-82. 

Protein A present in the cell wall of Staphylococcus aureus is a marker probe for the Fc portion of IgG-1, -2 and -4. Thus, a one step purification of protein A from extracts of 5. aureus was achieved using IgG coupled to Sepharose 4B affinity media [146]. The preparations obtained were pure as judged by electrophoresis. Conversely, affinity... 

Lind I, Ahnert-Hilger G, Fuchs G et at. (1987) Purification of alpha-toxin from staphylococcus aureus and application to cell permeabilization. Anolyt. Biochem. 164 84-89. 

The benchmark for the purification of most IgG species is Protein A, a 42 kDa molecular weight protein derived from a strain of Staphylococcus aureus. The natural molecule is located on the outer membrane surface of Staphylococcus aureus, and is linked to the cell surface through its C-terminal region [139]. This allows the pathogen to bind the Fc part of the IgG and re-direct the anti-gen-binding domain. This prevents interaction of the Fc part of the antibody with effector cells, thereby circumventing the activation of the immune system. 

To increase the purity or to concentrate an antibody solution, it may be purified. Purification is done with a range of techniques applied to whole serum, supernatant, or ascites fluid. At the first level, the purified Ig will be separated from other serum proteins and will select all IgGs including the IgG of interest and other IgG molecules. These purification steps can be done by using ammonium sulfate to precipitate the Ig molecules or it can be done by binding antibodies to a Protein A and/or Protein G columns. Proteins A and G are produced by the bacteria. Staphylococcus aureus, and bind to different species and subclasses of antibodies by the Fc receptor. After the antibodies have attached, they are washed out by changing the buffer. 

In the course of screening extracts from Bolivian plants against Trypanosoma cruzi, Leishmania spp., Bacillus subtilis and Staphylococcus aureus, Dunalia brachyacantha (Griseb.) Sleumer was found to be active. The bioassay-guided purification of the leaf extract led to the isolation of two known acetoxywithanolides (136 and 137), which displayed antiparasitic and antimicrobial activity (Table 4) [25]. This constitutes the first report of antileishmanial and antitrypanosomial (Chagas disease) activities for steroidal lactones. 

PHB biobeads displaying the ZZ domain of Protein A from Staphylococcus aureus as the result of N-terminal fusion to PhaC were found to be suitable to purify IgG from serum samples and culture supernatant with high binding capacity and purification power [20, 68]. Other binding domains were successfully displayed such as scFv (single-chain variable fragment) or streptavidin and enabled application of the respective beads as affinity purification resin [69,70). 

Protein-A-agarose. This protein, derived from cell wall of Staphylococcus aureus binds with very high affinity to the region of the immunoglobulin G. It has been extensively used in antibody purification. 

Muraoka, T., T. Ando, and H. Okuda. 1982. Purification and Properties of a Novel Lipase from Staphylococcus Aureus 226. Journal of Biochemistry 92 (6) 1933-1939. 

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