Casein, amino acid incorporation

Limiting essential amino acids covalently attached to proteins by using activated amino acid derivatives can improve the nutritional quality and change the functional properties of proteins. The best chemical methods for incorporating amino acids into water-soluble proteins involve using car-bodiimides, N-hydroxysuccinimide esters of acylated amino acids, or N-carboxy-a-amino acid anhydrides. The last two methods can give up to 75% incorporation of the amount of amino acid derivative used. With the anhydride method, as many as 50 residues of methionine have been linked to the 12 lysine residues of casein. The newly formed peptide and isopeptide bonds are hydrolyzed readily by intestinal aminopeptidase, making the added amino acids and the lysine from the protein available nutritionally. 

Figure 3 Covalent incorporation of L-leucine n-dodecyl ester into succi-nylated asl casein by modification with papain and the resulting formation of a 20,000-dalton macropeptide having an amphiphilic structure. The position of hydrophilic amino acid residues on the protein molecule are indicated with vertical bars. (From Ref. 3. Reprinted by permission.)...

Functional properties of some enzymatically modified and EPM-treated products of milk proteins [136] were determined as follows. An enzymatically prehydrolyzed commercial milk protein concentrate (SR) without further hydrolysis, and casein hydrolyzed by alcalase, a-chymotrypsin, and papain, respectively, were used as substrates in the EPM reaction. The concentration of the hydrolysates was 20% w/ v in the EPM reactions. A methionine methyl ester hydrochloride/ substrate ratio of 1 5 was used for incorporating this amino acid. After incubation, the products with methionine incorporation were simultaneously dialyzed for 2 days through a cellophane membrane against distilled water. The nondialyzable fractions and the EPM products without amino acid enrichment were freeze-dried. Covalent methionine incorporation in the EPM products with amino acid enrichment was verified by exopeptidase hydrolysis of the protein chains. The functional properties of the different EPM products are summarized in Table 1. An important functional property of proteins and/or peptide mixtures is their emulsifying behavior. This is highly influenced by the molecular structure, the position and ratio of hydrophobic-hydrophilic amino acids. Emulsion activity was found to be low (34.0) for casein, and the values determined for enzyme hydrolyzed and modified products were in general even lower. The papain hydrolysate, sample H3, showed here a different behavior as well this was the one of the sample series that had the highest EAI value (43.0). The emulsion stability of the enzymatically modified products displayed tendencies quite opposite to the values of emul-... 

Amino acids are absorbed by the mammary gland in quantities sufficient to account for the protein synthesised within it. Considerable interconversion of amino acids occurs before synthesis, and certain amino acids are important as sources of others. Thus, ornithine, which does not appear in milk protein, is absorbed and retained in large quantities by the mammary gland and has been shown to be a precursor of proline, glutamate and aspartate. Synthesis of the carbohydrate moieties of the proteins takes place in the mammary gland, as does phosphorylation of serine and threonine before their incorporation into the caseins. 

In order to produce the amphoteric protein-based surfactant, the incorporation of lipophilic amino acid ester was attempted using the one-step method of plastein reaction with papain at pH 9. In a system containing succinylated ttsi-casein as a protein substrate and luecine n-dodecyl ester as a lipophile, the peptide bond between Phe and Tyr of casein was first hydrolyzed, and this is followed by the incorporation of luecine n-dodecyl ester at the same position, forming a new C-terminus [34]. The structure of the macropeptide with respect to the distribution of hydrophilic amino acid residues is shown in Fig. 4 [29,34]. Amphiphilic structure consisting of hydrophilic protein portion and lipophilic luecine n-dodecyl ester was clearly demonstrated. 

In the presence of all amino acids in the medium bacterial growth is facilitated, since amino acids are incorporated into proteins in a readily available form. Preparation methods of casein hydrolysate were found to have important effects on propionibacterial growth. Mixtures of amino acids produced by alkaline, acid and trypsin hydrolysis were tested (Zodrow et al., 1963a) and the best result was achieved with the trypsin hydrolysate. It was suggested that in this case tryptophane is preserved and some peptides are formed which have specific stimulatory effects on the bacterial growth. Nitrate inhibits the deamination of amino acids in cheese by propionic acid bacteria (Peltola and Antila, 1953). 

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