Proteases alcalase

In 1958, the microbial alkaline protease Alcalase (Novo Industries) was produced by fermentation of a strain of Bacillus licheniformis. It had high StabiHty and activity at pH 8—10, was marketed in 1961, and was incorporated into Bio 40. However, it was not until the successful marketing of the presoaking agent Biotex in 1963 that detergent manufacturers saw the tme possibiHties of enzymes. 

A practical enzymatic procedure using alcalase as biocatalyst has been developed for the synthesis of hydrophilic peptides.Alcalase is an industrial alkaline protease from Bacillus licheniformis produced by Novozymes that has been used as a detergent and for silk degumming. The major enzyme component of alcalase is the serine protease subtilisin Carlsberg, which is one of the fully characterized bacterial proteases. Alcalase has better stability and activity in polar organic solvents, such as alcohols, acetonitrile, dimethylformamide, etc., than other proteases. In addition, alcalase has wide specificity and both l- and o-amino acids that are accepted as nucleophiles at the p-1 subsite. Therefore, alcalase is a suitable biocatalyst to catalyse peptide bond formation in organic solvents under kinetic control without any racemization of the amino acids (Scheme 5.1). 

Chen, S.-T., Chen, S.-Y. and Wang, K.-T., Kinetically controlled peptide bond formation in anhydrous alcohol catalyzed by the industrial protease alcalase. J. Org. Chem., 1992, 57, 6960. 

Aldred N, Phang IY, Conlan SL et al. (2008) The effects of a serine protease, Alcalase (R), on the adhesives of barnacle cyprids (Balanus amphitrite). Biofouling 24 97-107... 

An industrial process has been developed for production of isoelectric soluble soy protein hydrolysate with no bitterness and a bland taste (13). The raw material may be acid washed soy white flakes, soy protein concentrate or soy protein isolate. The raw material is hydrolyzed by the alkaline protease ALCALASE to a specified degree of hydrolysis using the pH-stat at pH 8.0... 

Effect of bleaching agents Chlorinated oxidizing agents carmot be used in enzymatic HDDs, even in the prewash cycle. The residual chlorine level in the washing bath is enough to quickly destroy the enzyme activity. For example, the activity of Novo s protease Alcalase disappears after 5 min at 38°C (100 F) at about 2 ppm available chlorine [10]. 

Several new detergent enzymes have emerged on the market (Table 1). Truly alkaline proteases, introduced in 1974 and 1982, were fermented on strains of Bacillus lentus firmus. These enzymes have a pH optimum between 9 and 11, and have taken important market shares from Alcalase. 

ACE inhibitory peptides have been separated from the skin of skate by Lee et ah (2011). The purified peptides showed IC50 values of 95 and 148 M, respectively, for both peptides isolated from the skin of skate. Further, Lineweaver-Burk plots indicated that the peptides act as noncompetitive inhibitor against ACE. Recently, many inhibitory peptides against ACE are reported (Table 15.2) as natural alternative biofunctional peptides that are safer than that of the existing artificial ACE inhibitory compounds in the market that show some side effects. Various peptides have been isolated from seafood by-products from the fisheries industry such as backbones from tuna (Lee et al., 2010). Tuna backbone has hydrolyzed using various proteases such as alcalase, a-chymotrypsin, neutrase, papain, pepsin, and trypsin to obtain an antioxidant peptide (Je et ah, 2007). 

Recently the effects of alcalase (another protease from subtilisin family), bovine pancreatic chymotrypsin, and papain (from papaya) have been evaluated on the desquamatory process.45 Alcalase (or Optimase) is an alkaline serine proteases derived from Bacillus licheniformis with... 

Alaska pollack skin Alcalase + protease + Gly-Phe-Lys 2.65 Byun and Kim, 2001... 

The enzymes used were Alcalase(R) 0.6 L, a liquid, food-grade preparation of subtilisin Carlsberg, and Neutrase(R) 0.5 L, a liquid, food-grade preparation of a B. subtilis neutral protease. Both enzymes are commercially available from Novo Industri A/S (7 ). All other reagents were analytical grade laboratory chemicals. ... 

Enzyme-catalyzed hydrolysis, exploiting the esterase activity of proteases such as trypsin and chymotrypsint ° l or carboxypeptidase has opened alternative routes to the deprotection of several peptide methyl, ethyl, and ferf-butyl esters. In fact, methyl, ethyl, and benzyl esters are successfully hydrolyzed from protected peptides using the alkaline protease from Bacillus subtilis or alcalase from Bacillus licheniformis which accepts... 

GSE is potentially less costly and more stable in comparison to V8 protease. The commercial detergent Alcalase is a rich source of GSE, from which it can be purified. GSE has the same Pi specificity as the protease from the V8 strain of Staphylococcus aurens (V8 protease, endoproteinase Glu-C).t ... 

Protease-catalyzed kinetic resolution of N-protected hydrophobic amino acids in SCCO2 was described by Chen et al. (88) as a method for the production of unnatural amino acids. With the alcalase protease from Bacillus licheniformus, carbobenzyloxy-protected racemic amino acid esters... 

Modification of Ultrafiltered versus Acid Precipitated Soy Protein. When the retentate obtained from the ultrafiltration of soybean extract is subjected to an enzymatic hydrolysis as described earlier (2) for acid precipitated protein, a hydrolysis curve (DH versus time) may be drawn. A comparison of such hydrolysis curves is shown in Fig. 2 for acid precipitated soy protein isolate and ultrafiltered soy protein isolate. The curves are drawn on the basis of the same hydrolysis parameters. The enzyme used is the microbial alkaline protease subtilisin Carlsberg (ALCALASE ). 

The question of accumulated, inert matter. The data of la-cobucci et al. (18) indicate that even considerable amounts of accumulated inert matter (half of total solids content in the reactor) did not disturb the steady state. How the case will be in the present system, where Alcalase is used instead of P.duponti protease has not yet been investigated, but as it will be shown in the following it seems plausible that a similar result could be obtained. 

In the course of our development work it turned out that Optimase M 440 was no longer available [39]. Since it was known from another project that the liquid enzyme formulations exhibit a less pronounced emulsifying behavior (cf. [37]), the different commercial bulk preparations, now Alcalases and Solvay Proteases, were re-evaluated. All yielded very good results with respect to chemical purity and enantiomeric excess (>99% each). However, Solvay Protease L 660 (the successor of Optimase L 660) was clearly the most active enzyme, thus requiring the smallest amount of enzyme for the reaction. The clearly higher price of this enzyme [40] should be of no consequence because of the expected simpler work-up and, hence, lower manpower costs. 

As Subtilisin Carlsberg was, and still is, one of the cheapest enzymes available on the market, no further proteases were tested. Four Subtilisin Carlsberg preparations from Solvay Enzymes [9] and Novo Nordisk [10] were tested successfully Solvay-Protease M 440 (solid form) and L 660 (liquid form) as well as the Novo enzymes Alcalase 2.0 T (solid) and Alcalase 2.5 L (liquid). 

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