Proteolytic enzymes, hydrolysis

Hydrolyzed Vegetable Protein. To modify functional properties, vegetable proteins such as those derived from soybean and other oil seeds can be hydrolyzed by acids or enzymes to yield hydrolyzed vegetable proteins (HVP). Hydrolysis of peptide bonds by acids or proteolytic enzymes yields lower molecular weight products useful as food flavorings. However, the protein functionaHties of these hydrolysates may be reduced over those of untreated protein. 

Proteins have been hydrolyzed by treatment with sulfuric acid, hydrochloric acid, barium hydroxide, proteolytic enzymes, and other hydrolytic reagents, but no condition has been found which avoids some destruction or incomplete liberation of tryptophan, cystine, and some other amino acids. The early work on this problem has been reviewed by Mitchell and Hamilton (194). The literature and their own excellent experiments on the hydrolysis problem in relation to the liberation and destruction of tryptophan have been presented recently by Spies and Chambers (269). 

Peptidases are enzymes that catalyse the hydrolysis of peptide bonds - the bonds between amino acids that are found in peptides and proteins. The terms protease , proteinase and proteolytic enzyme are synonymous, but strictly speaking can only be applied to peptidases that hydrolase bonds in proteins. Because there are many peptidases that act only on peptides, the term peptidase is recommended. Peptidases are included in subclass 3.4 of enzyme nomenclature [1,5]. 

Proteolytic degradation of a protein is characterized by hydrolysis of one or more peptide (amide) bonds in the protein backbone, generally resulting in loss of biological activity. Hydrolysis is usually promoted by the presence of trace quantities of proteolytic enzymes, but can also be caused by some chemical influences. 

The stomach secretes pepsinogens, which are inactive proteolytic enzymes, and protons - the high concentration of which initiates hydrolysis of the pepsinogens to form active pepsins, which then continue their own activation, via an autocatalytic, hydrolysis (Appendix 4.1). 

These proteolytic enzymes are all endopeptidases, which hydrolyse links in the middle of polypeptide chains. The products of the action of these proteolytic enzymes are a series of peptides of various sizes. These are degraded further by the action of several peptidases (exopeptidases) that remove terminal amino acids. Carboxypeptidases hydrolyse amino acids sequentially from the carboxyl end of peptides. They are secreted by the pancreas in proenzyme form and are each activated by the hydrolysis of one peptide bond, catalysed by trypsin. Aminopeptidases, which are secreted by the absorptive cells of the small intestine, hydrolyse amino acids sequentially from the amino end of peptides. In addition, dipeptidases, which are structurally associated with the glycocalyx of the entero-cytes, hydrolyse dipeptides into their component amino acids. 

Several proteolytic enzymes have been shown to enhance the solubility of fish protein concentrate (38). Product inhibition and self destruction of enzymes occurred, so that rates of hydrolysis decreased with time. 

Enzyme hydrolysis of peanut flour also altered the physical characteristics of baked cookies (60). With the exception of the bromelain hydrolysate, the use of peanut flour in cookies resulted in increased specific volume when compared to the 100% wheat flour control. Untreated peanut flour substitution reduced the diameter and increased the height of cookies however, treatment with proteolytic enzymes reversed the behavior. As evidenced by substantial increases in spread ratios, the diameter of cookies containing treated flours increased proportionately more than did the height. These data promote the feasibility of decreasing or increasing the spread of cookies through the addition of various amounts of untreated or enzyme-treated peanut flour. 

Enzymatic modification of proteins applicable to foods is reviewed by Whitaker ( ). Described briefly are present uses of proteolytic enzymes for modifying proteins through partial hydrolysis. Major emphasis is placed on those enzymes which bring about aggregation of proteins, cross-link formation, and side chain modification through post-translational changes in the polypeptide chain. 

Proteins can be modified by proteolytic enzymes with limited reduction in their nutritional bioavailability. Enzymatic hydrolysis of peptide bonds of proteins will reduce their molecular size, affect their structures, expose different regions of their molecules to the environment, and thereby alter their contribution to functionality, e.g. by increasing and decreasing the solubility and viscosity properties, respectively, of aqueous solutions. These changes can be controlled by carefully selecting proteolytic enzymes, maintaining proper treatment conditions, and monitoring the hydrolysis reactions. 

Let us now consider the retarding action of proteolytic enzymes on hydrolysis. Attention should be drawn to the fact that the carbonyl absorption band splits into two parts as a result of interaction of the cured KL-3 with proteolytic enzymes and kidney extract. Evidently it is associated with the specific interaction of the enzyme and urethane group in the polymer, the structure of which resembles the peptide group of a protein molecule. Owing to the specific action of the enzyme, this interaction does not accelerate the hydrolysis of urethane groups but even retards it owing to the shielding effect of the enzyme protein molecule. 

Biological amide hydrolysis, as in the hydrolysis of peptides and proteins, is catalyzed by the proteolytic enzymes. These reactions will be discussed in Chapter 25. 

The best way to show you t>ow the overlap method of peptide sequencing works is by a specific example. In this example, we will illustrate the use of the two most commonly used enzymes for selective peptide cleavage. One is trypsin, a proteolytic enzyme of the pancreas (MW 24,000) that selectively catalyzes the hydrolysis of the peptide bonds of basic amino acids, lysine and... 

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