Pepsin purification

The resuspended and formulated Fraction II precipitate normally contains some aggregated IgG and trace substances that can cause hypotensive reactions in patients, such as the enzyme prekail ikrein activator (186). These features restrict this type of product to intramuscular adininistration. Further processing is required if products suitable for intravenous adininistration are required. Processes used for this purpose include treatment at pH 4 with the enzyme pepsin [9001-75-6] being added if necessary (131,184), or further purification by ion-exchange chromatography (44). These and other methods have been fiiUy reviewed (45,185,187,188). Intravenous immunoglobulin products are usually suppHed in the freeze-dried state but a product stable in the solution state is also available (189). 

In 1833 an amylase from germinating barley was recovered and called diastase (1). Like malt itself, this product converted gelatinized starch into sugars, primarily maltose. Shordy thereafter, BerzeHus proclaimed the existence of non-living catalysts, and Schwaim (2) reported on his observation and purification of pepsin. 

Prabakaran, S., Tepp, W. and DasGupta, B.R., Botulinum neurotoxin types B and E purification, limited proteolysis by endoproteinase Glu-C and pepsin, and comparison of their identified cleaved sites relative to the three-dimensional structure of t q)e A neurotoxin, Toxicon, 39, 1515-1531, 2001. 

Kinekawa, Y.-I. and Kitabatake, N. 1996. Purification of (3-lactoglobulin from whey protein concentrate by pepsin treatment. J. Dairy Sci 79, 350-358. 

When the ultimate objective is to produce bioactive peptides for particular purposes, such as antioxidative or antihypertensive activities, the purification of target peptides from protein hydrolysate can be carried out using UF membranes with or without chromatographic techniques. Jun et al (2004) reported successful preparation of protein hydrolysates from yellowfin sole frame by first using extracted mackerel intestine crude enzyme at pH 10.0 and 50 °C, followed by treatment with pepsin at pH 2.0 and 37 °C. The resultant hydrolysate was further fractionated through five different UF membranes with... 

VIP is a 28-amino-acid residue peptide amide isolated in 1970 by Said and Mutt in the course of the purification of secretin from porcine duodenum (47). It was characterized 2 years later as a highly basic peptide related to gut peptides. It inhibits acid and pepsin secretion, acts as a neurotransmitter in peripheral autonomic nervous system, and increases secretion of water and electrolytes from pancreas and gut. 

Sephadex G-15 was purchased from Pharmacia Biotech urea was purchased from Fisher Scientific D2O and pepsin were purchased from Sigma Chemical Co. All chemicals were used without further purification. IDeuterated urea/D20 solution was prepared by repeated lyophilization. 

We owe to Nencki and Siber, to Samanoski and to Pekel-haring a series of careful anal3 s of pepsin. These have alwa3 s been made on purified products possessing a fairly high enzymic power. The methods of purification used were very different and the pepsins obtained were standardized according to their activity. Nevertheless, it is very curious... 

Brier, S., Maria, G., Carginale, V, et al. (200T) Purification and characterization of pepsins Al and A2 from the Antarctic rock cod Trematomus bemacchii. FEBS Journal, 274 (23), 6152-6166. 

The purification of the human growth hormone permitted Li to determine the N- and C-terminal amino acids, the amino acid composition, and the amino acid sequence of human growth hormone. The hormone was purified from a 1,000 hypophysis and hydrolyzed with trypsin, chymotrypsin, and pepsin. A molecule of phenylalanine occupies the N- and C-terminal positions. Human growth hormone is formed from 188 amino acids that are connected into a meandering chain, the segments of which are bridged by sulfhydryl bonds at two points [7] one joins cysteine residues 53-164, the other residues 181-189. 

Purification of pepsin glycopeptides from human immunoglobulin IgA myeloma glycoprotein Purification of porcine haptoglobin Purification of the serum prohormone of angiotensin Purification of the aa-trehalase from Saccharomyces cerevisiae Separation of different populations of membrane vesicles from thymocytes... 

Purification of a cyclic nucleotide phosphodiesterase Affinity chromatography of carbohydrate-specific immunoglobulins Purification of chicken pepsin... 

Purification of the anti-(blood-group N) lectin from Vicia graminea seeds Separation of pepsin and chymosin Demonstration of the heterogeneity of rat a-foetoprotein Comparison with kinetic and equililnium dialysis methods for analysis of the binding of nucleotides to staphylococcal nuclease... 

Purification of glycosulphatase Purification of horseradish peroxidase Purification of human-liver p-D-galactosidasei Purification of P-D-2-acetamido-2-deoxy-hexosidases A and B from human placenta Purification of pepsin glycopeptides from human immunoglobulin IgAj myeloma glycoprotein... 

From completely iodinated pepsin after alkaline hydrolysis and partial purification at least 82% of the total iodine was accounted for in a solution analyzing as diiodotyrosine (7). 53fo was actually crystallized. 

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