Myoglobin purification

Mixture of proteins (Myoglobin, Ovalbumin, Lysozyme and Chymotrypsinogen) Purification Hydrophobic Interaction [52]... 

A separation of myoglobin and lysozyme has been presented by Nicoud [44]. This purification was performed on SMB containing 8 columns using ACA 54 (Biosepra, France) as support. Very pure extracts (>98%) and raffinates (>98%) were obtained from a 50-50 mixture. An internal profile is given in Fig. 7. Another recently presented example is the separation of cyclosporine A from cyclic oligopeptide and other impurities in the reversed phase mode or by adsorption on silica gel presented by Schulte et al. [10]. 

Prior to the purification of heme oxygenase in sufficient quantities to carry out in vitro studies, the coupled oxidation of myoglobin was... 

Meat discoloration studies typically involve a maximum of 5 days, with discoloration analy-sis being performed every day or on alternate days. The actual experimental time involved in the objective assessment of discoloration is not extensive and depends on the number of samples being analyzed. Colorimetric measurements with hand-held colorimeters are very rapid (three measurements per meat surface in < 1 min). Spectral scans of meat surfaces require 1 to 2 min. Extraction and analysis of ground meat products has the added step of homogenization and filtration prior to spectrophotometry, but relative to many laboratory procedures, this is relatively quick. Isolation and purification of preparative amounts of myoglobin requires only 2 to 3 days once appropriate preparations are made. Finally, metmyoglobin can be reduced to oxymyoglobin in 15 to 20 min. 

Three analytical procedures are usually applied to the purification of myoglobin from crude meat extracts. 

Angiotensin-I, methionine enkephalin, substance-P, bovine trypsin, horse cytochrome-C, horse myoglobin, bovine insulin and egg-white lysozyme were purchased from Sigma Chemical Co. (St. Louis, MO) and used without further purification. 

Dimyristoylphosphatidylethanolamine (DMPE), biotinyl-N-hydroxysuccinimide ester (BNHS), triethylamine, myoglobin, bovine serum albumin (BSA), lysozyme, guanidinium chloride, dimethylamino-cinnamaldehyde, acrylamide, ammonium persulfate, sodium dodecyl sulfate (SDS), and molybdenum blue reagent were all obtained from Sigma Chemicals and used without further purification as received. The compounds N, N, N, N -tetramethylethylenediamine (TEMED), N, N -methylene-bis-acrylamide, and Coomassie Brilliant Blue R-250 were from BioRad Laboratories. Avidin was obtained from Vector Laboratories with a quoted activity of 14 units/mg. All solvents were from Fisher Scientific. Water was passed through a Barnstead Nanopure system. 

Applications. The number of reported uses of ion exchange dextrans is increasing rapidly. Examples are the use of DEAE-Sephadex for the separation of mucopolysaccharides by stepwise elution with NaCl [18] in dilute HCl, soluble RNA [19], and the separation of myoglobin and haemoglobin [20]. Cation exchange dextrans have been used for the separation of Q- and /3-globulin [21], alkaloids [21], and for the purification of enzymes [22]. 

In general, in any separation and purification procedure, the target molecule (a protein in this case) must be able to be identified by some physical or chemical property. This might be the red color of myoglobin or a specific enzyme assay. Separation of molecules is carried out by exploiting physical or chemical differences between proteins in a mixture. 

Bovine serum aprotinin (Aprotinin), bovine serum albumin (BSA), bovine serum immunoglobulin G (IgG), bovine serum thyroglo-bulin (Thyroglobulin), and horse heart myoglobin (Myoglobin) were purchased from Sigma Co. Ltd. (Osaka, Japan) and used without further purification. AH other chemicals were commercially available and used as received. 

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