Bacillus subtilis, proteases from

Bacillus subtilis, proteases from, 28 327, 328 Bacillus thermoproteolyticus, thermolysin from, 28 326... 

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

Mukesh Kurtrar, D. J., Rajarr, R., Lawrence, L., Priyadarshini, S., Sandhiya, C., Kalaichelvan, P. T. Destairring and dehairing capabUities of partially pitrified Bacillus subtilis protease from optimized fermentation medirrm. Asian J Exp Biol Sci 2012,3,613-620. 

Excellent yields were achieved in the selective acylation at 0-3 of 6-0-acelyl-l,5-anhydro-2-deoxy-D-arafe/no-hex-l-enitol (25) by lipase mediated acyltransfer from several vinylesters. As shown in Scheme 7, the starting material (25) could be recovered in 80% yield from two of the products (26) by enzymatic hydrolysis. Reports have been published on the lipase mediated selective synthesis of 2-functionalised 3-monoesters (27) of methyl 5-0-decyl-a-D-arabinofuranoside, on the regioselective, lipase-catalysed acylation of methyl a- and -D-arabino- and -xylo-pyranoside, on the regioselective acylation and deacylation of 2 -deoxynucleoside derivatives by use of a Pseudomonas fluorescens lipase and a Bacillus subtilis protease, respectively, and on the regioselective acylation of castanospermin with a variety of enzymes in pyridine. 

Alkaline protease from Bacillus subtilis DY, pH 8, 37°, 80-85% yield. Methyl esters are cleaved similarly. 

Isolation of a-amylase and protease from Bacillus subtilis fermentation broth [20]... 

N-Acetvlneuraminic Acid Aldolase. A new procedure has also been developed for the synthesis of 9-0-acetyl-N-acetylneuraminic acid using the aldolase catalyzed reaction methodology. This compound is an unusual sialic acid found in a number of tumor cells and influenza virus C glycoproteins (4 ). The aldol acceptor, 6-0-acetyl-D-mannosamine was prepared in 70% isolated yield from isopropenyl acetate and N-acetyl-D-mannosamine catalyzed by protease N from Bacillus subtilis (from Amano). The 6-0-acetyl hexose was previously prepared by a complicated chemical procedure (42.) The target molecule was obtained in 90% yield via the condensation of the 6-0-acetyl sugar and pyruvate catalyzed by NANA aldolase (Figure 6). With similar procedures applied to KDO, 2-deoxy-NANA and 2-deoxy-2-fluoro-NANA were prepared from NANA. 

More recently, the chemo-enzymatic synthesis of inulin-containing hydrogels was reported [54]. The key point was the solubility of inulin [a mixture of oligomers and polymers containing 2-60 (or more) 5-2,1 linked D-fructose molecules having a glucose unit as the initial residue] in dimethylformamide (DMF), a fact that allowed its esterification by action of a protease from Bacillus subtilis. 

It is commonly agreed that enzymes are inactive in nearly anhydrous dimethyl sulfoxide (DMSO) [55] and that such inactivity might be a direct result of protein solubilization in the organic milieu, which causes deleterious changes in the proteins secondary and tertiary structures [56]. However, different authors have recently reported that some proteases, namely thermolysin (from Bacillus thermo-proteolyticus) and Proleather (from Bacillus subtilis), were still active in pure DMSO, despite the fact that proteins were indeed solubilized. The former enzyme cata-... 

Irvine, D. M., Puhan, Z. and Gruetzner, V. 1969. Protease complex from a mutated strain of Bacillus subtilis as a milk coagulant for cheese manufacture. J. Dairy Sci. 52, 889-889. 

Thermolysin belongs to a class of proteases (called neutral proteases) which are distinct from the serine proteases, sulfhydryl proteases, metal-loexopeptidases, and acid proteases. Neutral proteases A and B from Bacillus subtilis resemble thermolysin in molecular weight, substrate specificity, amino acid content, and metal ion dependence. Since physiological substrates are most likely proteins, it is difficult to design simple experiments that can be interpreted in terms of substrate specificity and relative velocities. Therefore, studies of substrate specificity and other kinetic parameters must be carried out on di- and tripeptides so that details of the mechanism of catalysis can be obtained and interpreted simply. 

The zinc ion in a neutral protease from Bacillus subtilis has been exchanged with other metal ions (139—141). The Co(II) enzyme is reported to be active (140). 

Bovine lung or mucous QAE Sephadex A50 Proteases of Bacillus subtilis Proteolytic enzymes from pig pancreas... 

In the brewing industry, there is a development toward substitution of malt with unmalted barley and amylase, by use of glu-canase and protease of microbial origin. The neutral protease from Bacillus amyloliquefaciens and the thermostable neutral protease Bacillus subtilis var. thermoproteolyticus have been used by brewers successfully to hydrolyze barley proteins into amino acids and peptides. 

Application and Principle This procedure is used to determine protease activity, expressed as PC units, of preparations derived from Bacillus subtilis var. and Bacillus licheniformis var. The assay is based on a 30-min proteolytic hydrolysis of casein at 37° and pH 7.0. Unhydrolyzed casein is removed by filtration, and the solubilized casein is determined spectro-photometrically. 

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... 

An enzyme which generally displays a high esterase/protease ratio, such as the alkaline protease from Bacillus subtilis DY, selectively removes methyl, ethyl, and benzyl esters from a variety of Trt-, Z-, and Boc-protected di- and tripeptides and a pentapeptide at pH 8 and 37 °C (Scheme 14).P9]... 

Superior to lipases with regard to activity at higher solvent polarity are other hydrolases such as proteases. For example, subtilisin from Bacillus subtilis can be used to acetylate sucrose in dimethylformamide to the 1-mono-acetylated derivative [13], whereas the chemical acetylation reaction regioselectively leads to the 6-mono-acetylated product [ 14]. Thus, the two procedures nicely complement each other. 

There is also preliminary evidence for a protein secretion apparatus in Bacillus subtilis. Caulfield et al. (1984, 1985) have studied the S complex, a particle consisting of four proteins that appears to be involved in protein secretion. The complex is present on ribosomes as a small particle, essentially a third ribosomal subunit its proteins can be cross-linked to the 50 S ribosomal subunit. In addition, a 64-kDa protein present in the S complex is protected from added protease in the presence of both ribosomes and membrane, but not by either alone. The S complex does not appear to cause an arrest of translation (Caulfield et al., 1984). The S complex aggregates to form a clathrin-like structure when it is removed from ribosomes (Caulfield et al., 1985). The authors have proposed that such a structure might serve to form a cage around a nascent secretory polypeptide, isolating it from the cytoplasm until it reaches the membrane. Subsequent to membrane binding, three of the proteins of the S complex dissociate and the 64-kDa protein remains associated. This latter protein may then play a role in secretion. 

Subtilisin (from Bacillus subtilis) [9014-01-1] Mr 27,000 (sedimentation equilibrinm) [EC 3.4.21.62], This alkaline protease is purified 211-fold by affinity chromatography using 4-(4-aminophenylazo)phenylarsonic acid complex to activated CH-Sepharose 4B. It is inhibited by 2-phenylethane boronic acid, PMSF, 3,4-dichloroisocoumarin, acetone and benzamide. [Chandraskaren Dhar Anal Biochem 150 141 1985, Schomburg Schomburg Springer Handbook of Enzymes 2nd Edn vol 7 p 286 2002.]... 

Figure 18-9. C-terminal deprotection of peptide esters by the alkaline protease from Bacillus subtilis DY and alcalase.

Gallagher et al. [138] investigated the future application of two enzymes, bromelain and a bacillus protease (Bacillus subtilis) in the production of peptides from casein in point of view of the functional properties of the products. Bromelain action resulted in a hydrolysate with a great number of high-molecular-mass peptides this may have improved the functional properties of a food product. The bacillus protease seemed to be more suitable for producing bitter peptides for future research and/or for future food. 

The transesterifications of divinyl adipate with glucose in DMF catalyzed by alkaline protease from Bacillus subtilis at various water contents were examined. The enzymatic reaction by the Bacillus protease was carried out in the presence of more than 2% water and maximum reaction rate was observed at a water content of 20%. It seems that the protease activity in DMF can be recovered by the addition of water (Figure 14 B). 

Three other plant enzymes, papain, bromelain, and to a lesser extent ficin, have found acceptance in the food industry as proteases. Papain is derived from the latex of the fruit, leaves, and trunk of Carica papaya, and bromelain from the fruit and stems of pineapple plants. These enzymes are used to prevent the hazing of beer when chilled (Chill-Proofing) by modifying the protein. Other applications for these plant proteases are in meat tenderizers and digestive aids. Ficin from the latex of Ficus carica is used to a much lower extent, perhaps because of its marked action on native protein and difficult handling. Proteases from Aspergillus Jlavus-oryzae, and to a lesser extent from Bacillus subtilis, have been used to replace and supplement these plant proteases in all applications, but papain continues to have the widest acceptance. 

Processes carried out at a pH of 7-9 represent the most efficient use of this type of enzyme. The first industrial process involving this type of protease was in the bating of hides to remove the debris from the skin after liming, and to impart softness and air exchange to the finished leather. Although pancreatic bates are still in general use proteases derived from Aspergillus flavus-oryzae and Bacillus subtilis now supple ment them. 

Pancreatic preparations have been widely used as digestive aids, because they contain proteases, amylase and lipase. They have been prescribed for patients who have pancreatic disorders or after removal of the pancreas. The various activities present in the pancreatic preparations can be duplicated by in vitro methods from blends of microbial enzymes derived from Bacillus subtilis, Aspergillus flavus-oryzae and Aspergillus niger. Cellulase derived from Aspergillus niger is often added to the microbial preparation. The pancreatic preparations still hold the major share of the market, but this could be a useful application for the right combination of microbial enzymes. 

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