By Deniz Ekinci
This booklet includes an outline targeting the study zone of protein purification, enzymology, supplementations, antioxidants, biotransformation, gene supply, signaling, legislation and association. specific emphasis is dedicated to either theoretical and experimental points.
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Figure 7 presents the results of the autolysin assay for the whole cell extracts. 96 Awc – LiCl on whole cells Dwc – SDS on whole cells Table 1. presents the results of the levels of protein recovery using these different extraction methods. Fig. 7. Autolysin activity for three dilutions of protein (1/10, 1/5 and 1) extracted from cell debris with LiCl (lanes 1-3), from whole cells with SDS (lanes 4-6) and from whole cells with LiCl (lanes 7-9). We noticed that when used on whole cells, SDS showed the same autolysin extraction pattern (3 bands) as when used on cell debris (Fig.
1967), the treatment of the cell wall by sodium dodecyl sulfate (SDS) would resolve the problem of the insolubility of the autolysins of lactic bacteria during purification. , 1989), its presence may affect the tertiary structure of proteins because it is one of the most strongly denaturing detergents. It is also one of the most difficult to remove with dialysis. , 1997) to be measured. Thus, we propose that the use of other non ionic (Triton X-100) or zwitterion (CHAPS) non-denaturing detergents will have the advantage of resolving the problem of the insolubility of autolysins whilst protecting their structure.
The detergent CHAPS is the only compound capable of being dialysed that does not compromise the activity of the autolysin (Hjelmeland & Chrambach, 1984), however, it must not be complexed. All these results bring us back to our starting point, the difficulty in purifying this autolysin. 5-CHAPS complex, or insoluble compound-CHAPS complex. In the first case, we have the situation where the insoluble complex is present in The Problems Associated with Enzyme Purification 37 the actual extraction conditions used.