Proteins crystallins

Proteins. Crystalline lysozyme, bovine serum albumin, ribonucle-ase,and pepsin were obtained from the Armour Laboratories and crystalline /3-lactoglobulin was prepared from skimmed milk. a-Lactalbu-min was obtained from W. G. Gordon of this laboratory. Crystalline ovalbumin and human serum albumin were obtained from Nutritional Biochemical Corp. and purified pigskin gelatin from the Eastman Kodak Co. 

The half-lives of proteins range over several orders of magnitude (Table 23.1). Ornithine decarboxylase, at approximately 11 minutes, has one of the shortest half-lives of any mammalian protein. This enzyme participates in the synthesis of polyamines, which are cellular cations essential for growth and differentiation. The life of hemoglobin, on the other hand, is limited only by the life of the red blood cell, and the lens protein, crystallin, by the life of the organism. 

Bacillus thuringiensis Protein crystalline inclusions within sporulating cells... 

In ENDOR studies, maximum spectroscopic information is obtained, if the paramagnetic compounds are diluted into diamagnetic single crystal hosts. However, it is often not possible to find suitable host compounds or to grow sufficiently large crystals. This is particularly true with biological systems, e.g. copper or iron proteins. Crystalline powders or frozen solution samples, on the other hand, can normally be prepared more easily. For such randomly oriented compounds, Rist and Hyde have shown that... 

EXAMPLE 25.1 The phase diagrcim for the eye lens protein crystallin in water. Figure 25.4 shows the phase cUagram for mixtures of water and the lens protein crystallin. 

Trichloroethanoic acid, CCI3COOH. A crystalline solid which rapidly absorbs water vapour m.p. 58°C, b.p. 196-5" C. Manufactured by the action of chlorine on ethanoic acid at 160°C in the presence of red phosphorus, sulphur or iodine. It is decomposed into chloroform and carbon dioxide by boiling water. It is a much stronger acid than either the mono- or the dichloro-acids and has been used to extract alkaloids and ascorbic acid from plant and animal tissues. It is a precipitant for proteins and may be used to test for the presence of albumin in urine. The sodium salt is used as a selective weedkiller. 

Urease is one of the enzymes which have been obtained in the crystalline state. This has been done by stirring jack bean meal with 30°o aqueous acetone, filtering and allowing the filtrate to remain at o for several hours. The urease which crystallises out is separated by centrifuging and is then recrystallised. Like crystalline pepsin and trypsin, it is a protein. 

At very low concentrations of water, or in foods held below the free2ing point of water, physical conditions may be such that the available water may not be free to react. Under these conditions, the water may be physically immobi1i2ed as a glassy or plastic material or it may be bound to proteins (qv) and carbohydrates (qv). The water may diffuse with difficulty and thus may inhibit the diffusion of solutes. Changes in the stmcture of carbohydrates and proteins from amorphous to crystalline forms, or the reverse, that result from water migration or diffusion, may take place only very slowly. 

When water activity is low, foods behave more like mbbery polymers than crystalline stmctures having defined domains of carbohydrates, Hpids, or proteins. Water may be trapped in these mbbery stmctures and be more or less active than predicted from equiUbrium measurements. As foods change temperature the mobiUty of the water may change. A plot of chemical activity vs temperature yields a curve having distinct discontinuities indicating phase... 

In Parenteral and Enteral Nutrition. Amino acid transfusion has been widely used since early times to maintain basic nitrogen metaboHsm when proteinaceous food caimot be eaten. It was very difficult to prepare a pyrogen-free transfusion from protein hydrolysates. Since the advances in L-amino acid production, the crystalline L-amino acids have been used and the problem of pyrogen in transfusion has been solved. The formulation of amino acid transfusion has been extensively investigated, and a solution or mixture in which the ratio between essential and nonessential amino acid is 1 1, has been widespread clinically. Special amino acid mixtures (eg, branched chain amino acids-enriched solution) have been developed for the treatment of several diseases (93). 

Flavin mononucleotide was first isolated from the yellow en2yme in yeast by Warburg and Christian in 1932 (4). The yellow en2yme was spHt into the protein and the yellow prosthetic group (coen2yme) by dialysis under acidic conditions. Flavin mononucleotide was isolated as its crystalline calcium salt and shown to be riboflavin-5Lphosphate its stmeture was confirmed by chemical synthesis by Kuhn and Rudy (94). It is commercially available as the monosodium salt dihydrate [6184-17 /, with a water solubiUty of more than 200 times that of riboflavin. It has wide appHcation in multivitamin and B-complex solutions, where it does not require the solubili2ers needed for riboflavin. 

Purification. Enzyme purity, expressed in terms of the percent active enzyme protein of total protein, is primarily achieved by the strain selection and fermentation method. In some cases, however, removal of nonactive protein by purification is necessary. The key purification method is selective precipitation of the product or impurities by addition of salt, eg, sodium sulfate, or solvent, eg, ethanol or acetone by heat denaturation or by isoelectric precipitation, ie, pH adjustments. Methods have been introduced to produce crystalline enzyme preparations (24). 

Tetrodotoxin is one of the most powerful non-protein neurotoxins known. It occurs in the liver and ovaries of the Japanese puffer fish, Sphoerides rubripes and S. phyreus, and its lethal effects have been known for centuries, although it was isolated in crystalline form... 

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