Modified simplified method for isolation of lysostaphin from the culture filtrate of Staphylococcus staphylolyticus.
The present paper reports a modified method for isolation of lysostaphin--a bacteriolytic agent with specific affinity for staphylococcal cell wall. The proposed purification scheme includes three steps. The first procedure is ultrafiltration through a membrane filter giving a yield of 75.6%. The result of ultrafiltration is a concentrated, 10-times purified preparation of lysostaphin with specific activity 0.62 U/mg which can be used for digestion of S. aureus cells. Further step, performed by ion-exchange chromatography on DEAE-cellulose, yields a 60-times purified preparation containing a mixture of enzyme components of lysostaphin. The yield of this step is 47.2%, the preparation contains 3.54 U/mg protein. Using gel filtration on Sephadex G-50 a component with hexosaminidase activity was separated from the endopeptidase component on the basis of molar mass difference. A 270-times purified preparation of lysostaphin-endopeptidase with minimum of contaminating substances was obtained in this step. The yield of gel filtration was 22.1%, specific activity increased up to 16.3 U/mg protein.
Márová
I
I
1st Department of Dermatovenerology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
Dadák
V
V
D000595:Amino Acid Sequence; D002240:Carbohydrate Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D005374:Filtration; D008248:Lysostaphin / Q000302:isolation & purification*; D008828:Microbiological Techniques; D008969:Molecular Sequence Data; D013210:Staphylococcus / Q000737:chemistry* / Q000254:growth & development
D003470:Culture Media; D008248:Lysostaphin