Aspergillus oryzae protease

In all the reported examples, the enzyme selectivity was affected by the solvent used, but the stereochemical preference remained the same. However, in some specific cases it was found that it was also possible to invert the hydrolases enantioselectivity. The first report was again from iQibanov s group, which described the transesterification of the model compound (13) with n-propanol. As shown in Table 1.6, the enantiopreference of an Aspergillus oryzae protease shifted from the (l)- to the (D)-enantiomer by moving from acetonitrile to CCI4 [30]. Similar observations on the inversion of enantioselectivity by switching from one solvent to another were later reported by other authors [31]. 

Following Mosher s report, several publications appeared showing the preparation of Mosher s acid. One example is the chemoenzymatic preparation of Mosher s acid using Aspergillus oryzae protease (Scheme 1-5)24 ... 

Aspergillus oryzae protease Scheme 1-5. Chemoenzymatic preparation of Mosher s acid. 

For the preparation of the (S)-enantiomer an enzymatic racemic resolution process using Aspergillus oryzae protease was developed by Sepracor ... 

Protease from Aspergillus saitoi Protease from Bacillus subtilis Protease from Aspergillus oryzae Protease from Streptomyces griseus... 

The use of subtilisin (a protease which is widely used in detergent formulations) as a biocatalyst for the stereospecific hydrolysis of esters is well established [271-273]. Along the same lines, a protease derived from Aspergillus oryzae, which has hitherto mainly been used for cheese processing, has been shown to be particularly useful for the resolution of sterically hindered substrates such as a,a,a-trisubstituted carboxylates [274] (Scheme 2.42). While traditionaT proteases such as subtilisin were plagued by slow reaction rates and low selectivities, the a-tri-fluoromethyl mandelic ester (which constitutes a precursor of a widely used chiral derivatization agent, Mosher s acid [275]) was successfully resolved by Aspergillus oryzae protease [276]. 

Scheme 2.42 Resolutimi of bulky esters by subtilisin and Aspergillus oryzae protease...

Hatori, M., OhM, K., Hirano, S., Yang, X.-P., Kuboki, H., Abe, C., 2008. Effects of a casein hydrolysate prepared from Aspergillus oryzae protease on adjuvant arthritis in rats. Bioscience, Biotechnology, and Biochemistry 72, 1983—1991. 

In the early 1970s, Puski carried out a systematic investigation on functional soy protein hydrolysates. Puski hydrolyzed soy protein isolate with Aspergillus oryzae protease at pH 7 for 3 hours with different levels of enzyme/... 

Aspergillus oryzae protease Small-scale spray drier 160.0 75.0... 

Enz5mies Proteases Amylases Cellulases Various Bacilli, e.g. Bacillus licheniformis Bacillus subtilis, Aspergillus oryzae Trichoderma viride, Penicillium pinophilum... 

Polyethylene glycol) Protease (Aspergillus oryzae) Pyridine 8 35... 

Tatsumi, H., Murakami, S., Tsuji, R. E., Ishida, Y., Murakami, K., Masaki, A., Kawabe, H., Arimura, H., Nakano, E., and Motai, H. (1991). Cloning and expression in yeast of a cDNA clone encoding Aspergillus oryzae neutral protease II, a unique metalloprotease. Mol. Gen. Genet., 228, 97-103. 

Another Example, an Alkaline Protease from Aspergillus oryzae... 

On the basis of sequence alignment, subtilisin-type proteases can be subdivided into class I and class II.42 Subtilisins, thermitase and others, none of which has a disulfide bond, belong to class I, and ten proteases including aqualysin I and proteinase K to class II. An alkaline protease from Aspergillus oryzae, which has no cysteine residue, belongs to class II. The sequence identity between aqualysin I and the alkaline protease is 44%.49 ... 

After the introduction of pronase E, other more or less nonsubstrate-specific proteolytic enzymes have been applied to assist Se speciation. Most of them were derived from DNA/RNA clean-up protocols. The new enzymes (subtilisin from Bacillus licheniformis, also named protease VIII, EC 3.4.21.14 proteinase K from Tritirachium album, EC 3.4.21.64 the crude Novo Nordisk product of Flavourzyme from Aspergillus oryzae) proved to be capable of extracting Se with varying yields and chromatographic recovery of Se species. It is important to highlight that the latter parameter also depends on the instrumentation available. In this regard, different recovery values for the same samples reported by independent research groups do not necessarily indicate successful or unsuccessful sample preparation. Similarly, extraction efficiency (defined as the ratio of extracted Se to total Se in the sample) cannot be used as such for comparison purposes because sample preparation may include some extra steps, for example, TCA precipitation or ultrafiltration, which may reduce this value even by 10-20 percent. 

Protease (Aspergillus oryzae var.) Produced by controlled fermentation using Aspergillus oryzae var. The purified enzyme occurs as an off white to tan, amorphous powder. Soluble in water (the solution is usually light yellow), but practically insoluble in alcohol, in chloroform, and in ether. Major active principle protease. Typical applications used in the chillproofing of beer, in the production of bakery products, in the tenderizing of meat, in the production of protein hydrolysates, and in the development of flavor in processed foods. 

Application and Principle This procedure is used to determine the proteolytic activity, expressed as hemoglobin units on the tyrosine basis (HUT), of preparations derived from Aspergillus oryzae var. and Aspergillus niger var., and it may be used to determine the activity of other proteases at pH 4.7. The test is based on the 30-min enzymatic hydrolysis of a hemoglobin substrate at pH 4.7 and 40°. Unhydrolyzed substrate is precipitated with trichloroacetic acid and removed by filtration. The quantity of solubilized hemoglobin in the filtrate is determined spectrophotometrically. 

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