kappa-Carrageenan as a new smart macroaffinity ligand for the purification of pullulanase.
kappa-Carrageenan is a polysaccharide from red seaweed which gets precipitated by K+ ions and dissolves again in water. This smart, K(+)-responsive polymer was found to selectively bind pullulanase activity from Bacillus acidopullulyticus. Gel filtration on Sephadex G-200 showed the formation of the polymer-pullulanase complex at the pre-precipitation stage. On the other hand, phospholipase D, an enzyme which did not co-precipitate with kappa-carrageenan, did not form any complex with the polymer. Thus, K+ ions could be used to selectively precipitate the pullulanase activity. Then, 92% enzyme activity could be eluted with 1 M maltose solution. The single step protocol resulted in 50-fold purification, with a single band on sodium dodecylsulfate-polyacrylamide gel electrophoresis.
Roy
Ipsita
I
Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India.
Gupta
Munishwar N
MN
D000345:Affinity Labels; D002351:Carrageenan / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006026:Glycoside Hydrolases / Q000302:isolation & purification*; D008024:Ligands
D000345:Affinity Labels; D008024:Ligands; D002351:Carrageenan; D006026:Glycoside Hydrolases; C100181:pullulanase