Purification of catalase from human placenta.
The therapeutic use of an antioxidant complex containing superoxide dismutase and catalase has been proposed for the treatment of several diseases in which reactive oxygen species have an important role. Although superoxide dismutase for human use is commercially available, methods for the production of catalase for human use have not been described. An industrial process was developed for the purification of catalase for human use as a by-product of albumin production from human placenta, comprising two parts: (1) albumin and catalase co-purification steps, including blood extraction from ground placentas, precipitation of haemoglobin with ethanol/chloroform, concentration/diafiltration by tangential filtration and anionic chromatography, by which non-adsorbed catalase was separated from albumin; and (2) catalase purification steps after albumin separation, including a second anionic chromatography step and dye-affinity chromatography. This method provided a final recovery of 27% (70-100% in each step) with 670-fold purification of catalase (85% pure) and a specific activity of 49000 units/mg, which is higher than that of commercially available human catalase. This process permits the co-purification of catalase and albumin and can easily be scaled up.
Maimoni Gonçalves
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Centro de Biotecnologia, Instituto Butantan, Av. Vital Brasil 1500, CEP 05503 900, São Paulo, Brazil.
Cezar de Cerqueira Leite
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L
Raw
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Cabrera-Crespo
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J
D002374:Catalase / Q000302:isolation & purification*; D011232:Chemical Precipitation; D002725:Chloroform / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange; D000431:Ethanol / Q000737:chemistry; D005260:Female; D005374:Filtration / Q000379:methods; D006801:Humans; D010920:Placenta / Q000098:blood supply / Q000201:enzymology*; D011247:Pregnancy; D012709:Serum Albumin / Q000302:isolation & purification
D012709:Serum Albumin; D000431:Ethanol; D002725:Chloroform; D002374:Catalase