Extraction and preliminary characterizaion of a mycobacteriolytic preparation (stazyme) from aStaphylococcus strain: mycobactericidal activity and its use in rapid extraction of mycobacterial DNA

Stazyme, a mycobacteriolytic preparation from aStaphylococcusstrain, is able to break the permeability barrier in multiple drug resistantMycobacterium avium

“…Stazyme was produced from a coagulase‐negative Staphylococcus isolate designated strain Clavelis and deposited at the Institut Pasteur, Paris in the Staphylococcus Culture Collection under reference numbers 89‐325, 89‐326 and 89‐327 [7]. The new batch of stazyme was prepared essentially as reported earlier using a modified Luria‐Bertani (LB) medium in which the Bacto tryptone (1%, w/v) was replaced by 2% (w/v) casamino acids [7]. Briefly, organisms were cultured in 1 l of medium for 48 h at 37°C with agitation, the culture filtrate was centrifuged at 10 000× g for 30 min, followed by filtration through a 0.2 μm pore size filter, and concentrated with 20 M polyethylene glycol.…”

“…Briefly, organisms were cultured in 1 l of medium for 48 h at 37°C with agitation, the culture filtrate was centrifuged at 10 000× g for 30 min, followed by filtration through a 0.2 μm pore size filter, and concentrated with 20 M polyethylene glycol. The filtrate proteins were precipitated in saturated ammonium sulfate solution, washed, dialyzed in 50 mM Tris (pH 8), lyophilized, and stocked refrigerated as reported earlier [7]. The stocks from 3–4 batches were reconstituted, pooled to give a total protein content of about 1 mg ml −1 , gel filtered on Sephadex G‐75 (Pharmacia, St. Quentin, France) in 50 mM Tris, pH 8 in a column with a bed volume of 20 ml.…”

“…The stocks from 3–4 batches were reconstituted, pooled to give a total protein content of about 1 mg ml −1 , gel filtered on Sephadex G‐75 (Pharmacia, St. Quentin, France) in 50 mM Tris, pH 8 in a column with a bed volume of 20 ml. Successive 1 ml fractions were collected, and assayed for their protein content and lytic activity against M. smegmatis ATCC 607 as described earlier [7]. Parallel controls included lytic activity of proteins extracted from the same quantity of uninoculated bacterial growth medium treated likewise (which was devoid of any lytic activity), and a previous 1993 batch of stazyme prepared using LB medium [7].…”

“…Native proteins were analyzed on 10% (w/v) acrylamide gel by loading 25 μl of enzyme preparation [7]. Gels were run for 2 h at 90 V, and stained with Coomassie blue.…”

Isolation and identification of a staphylococal strain with an anti-mycobacterial activity and study of it’s mode of action

“…To our knowledge, there's only one isolated staphylococcal strain, reported to have an antimycobacterial activity, but unfortunately, the active molecule wasn't identified (Clavel-Sérès et al, 1993). However, it was reported that several species belonging to the Staphylococcus genus are known for their capacity to produce various antibacterial substances (Beaudet et al, 1982;Frenette et al;1984).…”