Galactosidase, purification

B. Assay with Galactosylceramide P-Galactosidase Purification of Activator Proteins... 

Affinity chromatography of human liver acid /3-D-galactosidase Purification of /J-D-galactosidase from rat mammary gland... 

Purification of human-liver P-D-galactosidases Purification of human placental p-D-2-acetamido-2-deoxyhexosi-dases A and B... 

The authors wish to thank Maijo Searle-van Leeuwen for purification of the P-galactosidase from Aspergillus niger and Novo Nordisk Ferment Ltd Dittingen (Switzerland) for assistance in the purification of the RG-hydrolase and Novo Nordisk A/S Bagsvaerd (Denmark) for the gift of the crude recombinant RG-lyase. 

Ferric ion was immobilized on a Chelating Sepharose Fast Flow column preparatory to the separation of seven enkephalin-related phosphopep-tides.17 Non-phosphorylated peptides flowed through the column, and the bound fraction contained the product. The capacity of the column was found to be 23 pmol/mL by frontal elution analysis. Cupric ion was immobilized on Chelating Superose for the isolation of bovine serum albumin.18 Cupric ion was immobilized on a Pharmacia HiTrap column for the separation of Protein C from prothrombin, a separation that was used to model the subsequent apparently successful separation of Factor IX from prothrombin Factor IX activity of the eluate was, however, not checked.19 Imidazole was used as the displacement agent to recover p-galactosidase from unclarified homogenates injected onto a nickel-loaded IMAC column.20 Pretreatment with nucleases and cleaning in place between injections were required procedures. A sixfold purification factor was observed. 

Since then it has been established that emulsin is a mixture of enzymes comprising in addition to /3-glucosidase28 the enzymes a-galactosidase, a-mannosidase, /3-glucuronidase and /3-(N-acetyl)-glucosaminidase. Concerning the process of purification of the Rohferment and the mode of differentiation of the constituent enzymes of emulsin, the most informa-... 

Dissolve the enzyme to a concentration of 1 - 2 mg/ml in Soln. A (if it is delivered in another buffer, dialyze against Soln. A). Mix 1 mg -galactosidase in Soln. A with 0.25 mg SMCC activated antibody (concentration about 1 mg/ml for activation see Protocol 4.1.3). Shake at RT for 2 h, dialyze or desalt on a Sephadex G-25 column against Soln. B and concentrate to about 2 mg/ml. Mix 9 vol. of the concentrated conjugate with 1 vol. Soln. C and store without further purification at 4 °C. 

Chinen, 1., T. Nakamura, and N. Fukuda. Purification and properties of alpha-galactosidase from immature stalks of Saccharum officinarum (sugar cane). J Biochem (Tokyo) 1981 90(5) 1453-1461. 

Anema, P. J. 1964. Purification and properties of 3-galactosidase of Bacillus subtilis. Biochim. Biophys. Acta 89, 495-502. 

Goncalves, J. A. and Castillo, F. J. 1982. Partial purification and characterization of (3-D-galactosidase from Kluyveromyces marxianus. J. Dairy Sci. 65, 2088-2094. 

Itoh, T., Ohashi, M., Toba, T. and Adachi, S. 1980. Purification and properties of 0-galactosidase from Lactobacillus bulgaricus. Milchwissensckaft 35, 593-597. 

During the purification of /3-galactosidase, a student found that 1 ml of a 1 10,000 dilution of crude extract gave A420 readings as follows ... 

Day 1 Partial Purification of Antibodies (IgG) from Normal Rabbit Serum and Anti-j8-Galactosidase Rabbit Serum... 

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