Application of the effects of ionic strength reducing agents in the purification and crystallization of chitinase A.
The effects of ionic strength reducing agents may find a large number of applications. Based on these effects, we have redesigned the purification scheme of Chitinase A (ChiA) from Serratia marcescen. This scheme led to reproducibly crystallizable enzyme in both salting-in and salting-out conditions, which are presented here. Herein, we demonstrate some experimental applications of the ionic strength reducing agents theory and, in parallel, provide further evidence of the theory's correctness. Finally, we report a new crystal form produced recently in salting-in crystallization experiments. This form may allow the co-crystallization of ChiA mutants with longer substrates.
D000645:Ammonium Sulfate; D011232:Chemical Precipitation; D002688:Chitinases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D002852:Chromatography, Ion Exchange; D003451:Cryoprotective Agents; D003460:Crystallization / Q000379:methods; D009154:Mutation; D009994:Osmolar Concentration; D011092:Polyethylene Glycols; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D012706:Serratia marcescens / Q000201:enzymology / Q000235:genetics
D003451:Cryoprotective Agents; D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D002688:Chitinases; D000645:Ammonium Sulfate