Protease removal

Toxicants, which are proteins, may not be detoxified by the means so far described. They may however be degraded by proteases, removed by antibodies if antigenic, and recognized as foreign. Also those toxicants that rely on disulfide bonds for structure and activity can be inactivated by the enzyme thioredoxin, which reduces these bonds to sulfydryl. 

If an artificial enzyme can solve an important biological problem that cannot be treated with natural enzymes, it may be regarded as an artificial dream enzyme. For example, artificial proteases removing protein aggregates allegedly are responsible for Alzheimer s, Parkinson s, or mad cow disease, are among the artificial dream enzymes. 

The apphcation of a protease treatment to silk was introduced as an alternative to the degumming process using alkaline soap solutions (Freddi et al., 2003 Gulrajani et al., 2000). Alkaline soap has a deleterious effect on the silk, resulting in a harsh feel to the material. Enzyme degumming with protease removes the sericin without damaging the fibre. Results of enzyme treatment have shown that the fibre is stronger than that obtained by traditional alkaline soap treatments. 

Enzymatic Gravimetric Methods for TDF, SDF, and IDF. These methods use an a-amylase and protease to remove starch and reduce protein. They differ from each other in the conditions for gelatinization of starch. Elimination of detergent permits recovery of soluble fiber, which is not possible with the detergent methods. 

Urea Enzymatic Dialysis Method. This method (16) uses 8 M urea [57-13-6] to gelatinize and facUitate removal of starch and promote extraction of the soluble fiber at mild (50°C) temperatures. EoUowing digestion with heat-stable a-amylase and protease, IDE is isolated by filtration or I DE is obtained after ethanol precipitation. Values for I DE are comparable to those obtained by the methods described eadier, and this method is less time-consuming than are the two AO AC-approved methods. Corrections for protein are required as in the AO AC methods. 

Proteia and starch stains are removed by proteases and amylases, respectively. Fats and oils are generally difficult to remove at low wash temperatures by conventional detergents. By usiag Upases, it is possible to improve the removal of fats/oils of animal and vegetable origin even at temperatures where the fatty material is ia a soUd form. Particulate soils can be difficult to remove, especially if the particle sise is small. Removal of particulate soil from cotton fabric can be improved by use of a ceUulase which removes cellulose fibrils from the surface of the yam. 

Protein residues, eg, soft-boiled egg yolk, are difficult stains to handle. If the stains are not totally denatured, proteases can decompose them. There are commercial proteases with a high temperature optimum (60°C) that can remove most protein soils in a dishwasher (63). Patents on the use of Upases in ADDs have claimed that Upases can reduce the formation of spots and films on glasses (62,64—66) however, no commercial appUcation of Upases in ADDs has been implemented. 

Cured hides must be properly soaked to obtain satisfactory rehydration and removal of unwanted material. InterfibnUar proteins should be degraded in order to increase water uptake. Bacterial proteases and pancreatic proteases are normally preferred, and are compatible with most tannery chemicals used in soaking, ie, most surfactants and preservatives containing sodium chlorite. 

In the first publication describing the preparative use of an enzymatic reaction in ionic liquids, Erbeldinger et al. reported the use of the protease thermolysin for the synthesis of the dipeptide Z-aspartame (Entry 6) [34]. The reaction rates were comparable to those found in conventional organic solvents such as ethyl acetate. Additionally, the enzyme stability was increased in the ionic liquid. The ionic liquid was recycled several times after the removal of non-converted substrates by extraction with water and product precipitation. Recycling of the enzyme has not been reported. It should be noted, however, that according to the log P concept described in the previous section, ethyl acetate - with a value of 0.68 - may interfere with the pro-... 

Bacterial protease B. subtilis Desizing fibres, spot remover... 

The Rieske protein in mitochondrial bci complexes is assembled when the protein is incorporated into the complex. The Rieske protein is encoded in the nucleus and synthesized in the cytosol with a mitochondrial targeting presequence, which is required to direct the apoprotein to the mitochondrial matrix. The C-terminus is then targeted back to the outside of the inner mitochondrial membrane where the Rieske cluster is assembled. In addition, the presequence is removed and the protein is processed to its mature size after the protein is inserted into the bci complex. In mammals, the presequence is cleaved in a single step by the core proteins 1 and 2, which are related to the general mitochondrial matrix processing protease (MPP) a and (3 subunits the bovine heart presequence is retained as a 8.0 kDa subunit of the complex (42, 107). In Saccharomyces cerevis-iae, processing occurs in two steps Initially, the yeast MPP removes 22 amino acid residues to convert the precursor to the intermediate form, and then the mitochondrial intermediate protease (MIP) removes 8 residues after the intermediate form is in the bci complex (47). Cleavage by MIP is independent of the assembly of the Rieske cluster Conversion of the intermediate to the mature form was observed in a yeast mutant that did not assemble any Rieske cluster (35). However, in most mutants where the assembly of the Rieske cluster is prevented, the amount of Rieske protein is drastically reduced, most likely because of instability (35, 44). 

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