Alginate as a macroaffinity ligand and an additive for enhanced activity and thermostability of lipases.
Lipases are being employed increasingly for the synthesis of drug intermediates and pharmaceutically important molecules as well as for the resolution of racemic mixtures for obtaining physiologically active enantiomers. Alginate was used as a macroaffinity ligand to purify lipases from Chromobacterium viscosum, porcine pancreas and wheatgerm by employing the technique of affinity precipitation. In all the cases, the purified preparation showed a single band on SDS/PAGE. The process gave adequate yields of 87, 75 and 62% in the case of Chromobacterium, porcine and wheatgerm lipases respectively. Alginate was also found to activate the enzymes; this effect was most dramatic in the case of wheatgerm lipase, for which a 4-5-fold activity increase was observed. Furthermore, alginate was also found to protect the enzyme against thermoinactivation.
Sharma
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Chemistry Department, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India.
Gupta
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MN
D000464:Alginates / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D001709:Biotechnology / Q000379:methods; D011232:Chemical Precipitation; D002861:Chromobacterium / Q000737:chemistry / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D004795:Enzyme Stability; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D008024:Ligands; D008049:Lipase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D008322:Mammals; D014908:Triticum / Q000201:enzymology
D000464:Alginates; D006603:Hexuronic Acids; D008024:Ligands; D020723:Glucuronic Acid; D008049:Lipase