Alpha-chymotrypsin

Birktoft J J and D M Blow 1972. The structure of Crystalline Alpha-Chymotrypsin V. The Atomic Structure of Tosyl-Alpha-Qiymotrypsin at 2 Angstroms Resolution. Journal of Molecular Biology 68 187-240. 

A pepsin hydrolysate of flounder fish protein isolate has been used as the substrate (40% w/v) for plastein synthesis, using either pepsin at pH 5 or alpha chymotrypsin at pH 7, with an enzyme—substrate ratio of 1 100 w/v at 37°C for 24 h (151). The plastein yields for pepsin and alpha chymotrypsin after precipitation with ethanol were 46 and 40.5%, respectively. 

By adding 1-alkanols to AOT-based w/o microemulsions, some proteins (ribonucle-ase, lysozyme, alpha-chymotrypsin, pepsin, bovine serum albumin, and catalase) are readily expelled, while the major part of the surfactant remained in solution [171]. 

These high performance size exclusion separations of alpha-chymotrypsin SI and myosin light chains compare favorably with those achieved by ion exchange chromatography but require only a fraction of the time to accomplish. Furthermore, the very short retention times allow for separation of these labile proteins at room temperature, whereas operation at 0-40 C would otherwise be mandatory to avoid the loss of enzymatic activity. 

The effects of various enzymes on the activity of HPLC fractions that inhibited 3H-PCP binding were investigated. As shown in table 1, pronase (0.5 pg/ml), carboxypeptidase A (0.1 unit/ml), and trypsin (3.0 g/ml ) markedly decreased the potency of 10 n units of PCP-like activity. No significant change in activity was. seen when fractions were incubated with alpha-chymotrypsin. 

PK Banerjee, GL Amidon. Physicochemical Property modification strategies based on enzyme substrate specificities II Alpha-chymotrypsin hydrolysis of aspirin derivatives. J Pharm Sci 70 1304, 1981. 

E.V. Kudryashova, A.K. Gladilin, A.V. Vakurov, F. Heitz, A.V. Levashov, and V.V. Mozhaev, Enzyme-poly electrolyte complexes in water-ethanol mixtures negatively charged groups artificially introduced into alpha-chymotrypsin provide additional activation and stabilization effects. Biotechnol. Bioeng. 55, 267-277 (1997). 

Sigler, P. B. (1970). lodination of a single tyrosine in crystals of alpha-chymotrypsin. Biochem. 9,3609. 

T.C. Liang, R.H. Abeles, Complex of alpha-chymotrypsin and N-acetyl-L-leucyl-L-phenylalanyl trifluoromethyl ketone Structural studies with NMR spectroscopy. Biochemistry 26 (1987) 7603-7608. 

A. Tulinsky, R.A. Belvins, Structure of a tetrahedral transition state complex of alpha-chymotrypsin dimer at 1.8-A resolution, J. Biol. Chem. 262(16) (1987) 7737-7743. 

Blocher, M., Walde, R, and Dunn, I. J. (1999). Modeling of enzymatic reactions in vesicles the case of alpha-chymotrypsin. / Biotechnol Bioeng., 62, 36 3. 

Fadnavis, N. W. and Luisi, P. L. (1989). Immobilized enzymes in reverse micelles studies with gel-entrapped Trypsin and alpha-Chymotrypsin in AOT reverse micelles. Biotechnol. Bioeng., 33, 1277-82. 

Berezin, I. V., A. V. Levashov, and K. Martinek, On the modes of interaction between competitive inhibitors and the alpha-chymotrypsin active centre , FEBS Lett., 7,20-22 (1970). 

D. N. Reinhoudt, 1998, Large acceleration of alpha-chymotrypsin-catalyzed dipeptide formation by 18-crown-6 in organic solvents, Biotechnol. Bioeng. 59, 553-556. 

Suenaga, H., et al. 1995. Strong inhibitory effect of sugar biphenylboronic acid complexes on the hydrolytic activity of alpha-chymotrypsin. J Chem Soc 13 1733. 

Li, Y., Z. Shao, and A.K. Mitra. 1992. Dissociation of insulin oligomers by bile salt micelles and its effect on alpha-chymotrypsin-mediated proteolytic degradation. Pharm Res 7 864. 

Kozlova, N., et al. 1999. Catalytic properties and conformation of hydrophobized alpha-chymotrypsin incorporated into a bilayer lipid membrane. FEBS Lett 461 141. 

Park, H. S., Lin, Q., Hamilton, A. D., Protein surface recognition by synthetic receptors A route to novel submicromolar inhibitors for alpha-chymotrypsin. J. Am. Chem. Soc. 1999, 121, 8-13. 

Strop, P., Cechova, D., and Tomasek, V. (1993). Model study of hydrophobic interaction of alpha- and beta-trypsin and alpha-chymotrypsin. J. Chromatogr. 259, 255-268. 

Castro MJ, Anderson S. Alanine point-mutations in die reactive region of bovine pancreatic trypsin inhibitor effects on the kinetics and thermodynamics of binding to beta-trypsin and alpha-chymotrypsin. Biochemistry 1996 35 11435-11446. 

Alpha-chymotrypsin is an enzyme which hydrolyses peptide bonds (preferentially those formed by aromatic amino acyl residues and a variety of esters and amides forming an acylated intermediate). This property has been exploited for spin labelling, and by reacting the enzyme with a nitroxide substrate analogue. A, X, it is possible to record, at low pH, the ESR spectrum of the acyl enzyme [138]. This showed strong... 

Packer AJ, Fraioli AJ, Epstein DL. The effect of timolol and acetazolamide on transient intraocular pressure elevation following cataract extraction with alpha-chymotrypsin. Ophthalmology 1981 88(3) 239-43. 

Beck, B., Glen, R.C. and Clark, T. (1996). The Inhibition of Alpha Chymotrypsin Predicted Using Theoretically Derived Molecular Properties. J.MoLGraphics, 14,130. 

Hasegawa, K., Yokoo, N., Watanabe, K., Hirata, M., Miyashita, Y. and Sasaki, S. (1996b). Multivariate Free-Wilson Analysis of Alpha Chymotrypsin Inhibitors Using Pis. Chemom. Intel Lab.Syst.,33, 63-69. 

The approach of the substrate towards the active site of SP was studied with electrostatic field potentials (ESP) by Lamotte-Brasseur et al. (1990). They examined the force exerted by the active sites of the serine proteinases alpha-chymotrypsin and subtilisin on an approaching substrate. About 20 residues from each of these enzymes were employed to construct electrostatic potential maps, using point charges derived from CNDO calculations (Pople and Beveridge, 1970) with e =l.Net charges were obtained from a mulliken analysis... 

Molecular mechanics minimization and molecular dynamics were chosen to examine the possible conformations for the two acyl enzymes and conclusions were drawn from the time evolution of the two systems. The starting point was a crystal structure of phenylethaneboronic acid bound to alpha-chymotrypsin. QUANTA/CHARMM (Brooks et al, 1983) was employed for the calculations. Ninety-five water molecules from the X-ray structure were included. Distance monitoring and the creation ofH-bonds were the main criteria for differentiating between the two molecules. Both acyl enzymes have their ketone carbonyls H-bonded to Gly-216 NH. Both start with their ester carbonyl in the oxyanion hole (H-bonded to Ser-195 and to Gly-193). The R-acyl enzyme looses both of these hydrogen bonds during the simulation. Attack of water on the R-species should, thus, be less frequently successful. Values for differences in energy were not used because of a small... 

Haynes, C.A., Tarnura, K., Korfer, H. R., Blanch, H. W., and Prausnitz, J.M. Thermodynamic properties of aqueous alpha-chymotrypsin solutions from membrane osmometry measurements. Journal of Physical Chemistry, 1992, 96, No. 2, p. 905-912. 

Other results of extraction of proteins using different trialkylammonium salts differ somewhat from this tendency. Jolivalt (36) studied the solubilization of a-chymotrypsin in the Aliquat 336 system using isotridecanol as a cosurfactant. Alpha-chymotrypsin was most significantly extracted for pH above its isoelectric point but as the pH decreased below the pi, the extraction yield decreased very slowly and became negligible at 4 pH units below the pi. This means that a-chymotrypsin can be extracted in significant proportion by a cationic surfactant even when positively charged overall. 

Eckstein M, Sesing M, KraglU, AdlercreutzP (2002) Allow water activity alpha-chymotrypsin is more active in an ionic liquid than in non-ionic organic solvents. Biotechnol Lett 24 867-872... 

Laszlo JA, Compton DL (2001) Alpha-chymotrypsin catalysis in imidazolium-based ionic liquids. Biotechnol Bioeng 75 181-186... 

Alkaline phosphatase Alpha chymotrypsin Alanine aminotransferase Adenosine phosphate Acousto optic modulator Aspartate aminotransferase Adenosine triphosphate... 

Barros, R.J., Wehtje, E., and Adlercreutz, P. (2001) Modeling the performance of immobilized alpha-chymotrypsin catalyzed peptide synthesis in acetonitrile medium. 

Salam, S.M.A., Kagawa, K., and Kawashiro, K. (2006) alpha-Chymotrypsin-catalyzed peptide synthesis in frozen aqueous solution using N-protected amino acid carbamoylmethyl esters as acyl donors. Tetrahedron Asymmetry, 17 (1), 22-29. 

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