EurekaMag.com logo
+ Site Statistics
References:
53,869,633
Abstracts:
29,686,251
+ Search Articles
+ Subscribe to Site Feeds
EurekaMag Most Shared ContentMost Shared
EurekaMag PDF Full Text ContentPDF Full Text
+ PDF Full Text
Request PDF Full TextRequest PDF Full Text
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter
Follow on LinkedInFollow on LinkedIn

+ Translate

Irreversible thermoinactivation of alpha-chymotrypsin in buffer and water miscible organic solvent: Comparison with a reverse micellar system



Irreversible thermoinactivation of alpha-chymotrypsin in buffer and water miscible organic solvent: Comparison with a reverse micellar system



Journal of Molecular Catalysis B Enzymatic 7(1-4): 191-205, Sept 15



The mechanism of irreversible thermoinactivation of alpha-chymotrypsin in buffer medium, dimethylformamide, dimethyl-sulfoxide/buffer and reverse micelles was studied. Experiments for the enzyme thermoinactivation in the different systems were accompanied by studies on protein structure alterations. The formation of free SH groups was followed during the stability tests. This formation is due to the cleavage of the enzyme primary structure which leads to small peptides. These were identified by SDS-PAGE, in the case of the buffer medium, and also by HPLC in the three other systems. An increase of SH groups with the residence time of the enzyme in the buffer and organic solvent/buffer system was observed. This was corroborated by the disappearance of the electrophoresis bands of the protein and also by HPLC analysis. In the latter technique, the peaks corresponding to alpha-chymotrypsin disappeared with the concomitant appearance of small peaks in the chromatogram. For the enzyme encapsulatedin reverse micelles, the formation of free SH groups was not detected and the HPLC analysis revealed that the protein peak stayed intact during the residence time in this system. The thermoinactivation of chemically modified alpha-chymotrypsin by the introduction of dianhydride pyromellitic on the Lys residues of the protein was also studied in some of the systems. The results showed that the chemical modification stabilized the enzyme when the system used was buffer or organic solvent mixed with buffer being this stabilization partially due to a slower process of cysteine bond cleavage.

(PDF emailed within 1 workday: $29.90)

Accession: 010885760

Download citation: RISBibTeXText



Related references

Kinetic analysis of deactivation of immobilized alpha-chymotrypsin by water-miscible organic solvent in kyotorphin synthesis. Biotechnology and Bioengineering 65(2): 170-175, 1999

Kinetic analysis of deactivation of immobilized a-chymotrypsin by water-miscible organic solvent in kyotorphin synthesis. Biotechnology and Bioengineering 65(2): 0-5, 1999

Influence of water miscible organic solvents on alpha-chymotrypsin in solution and immobilized on Eupergit CM. Biotechnology Letters 28(12): 929-935, 2006

Control of water content by reverse micellar solutions for peroxidase catalysis in a water-immiscible organic solvent. Journal of Bioscience & Bioengineering 95(4): 425-427, 2003

Control of water content by reverse micellar solutions for peroxidase catalysis in a water-immiscible organic solvent. Journal of Bioscience and Bioengineering 95(4): 425-427, 2005

Denaturing agents (urea, acrylamide) protect enzymes against irreversible thermoinactivation: a study with native and immobilized alpha-chymotrypsin and trypsin. Febs Letters 51(1): 152-155, 1975

Synthesis of L-tyrosine glyceryl ester catalyzed by alpha-chymotrypsin in water-miscible organic solvents: A possible sun-tan accelerator product. Biotechnology Letters 15(12): 1223-1228, 1993

Micellar solubilization of bio polymers in organic solvents 5. activity and conformation of alpha chymotrypsin in iso octane bis 2 ethylhexyl sodium sulfo succinate reverse micelles. Journal of the American Chemical Society 103(14): 4239-4244, 1981

Biocatalysis of chlorophyllase from the alga Phaeodactylum tricornutum in a water/miscible-organic-solvent system. Biotechnology & Applied Biochemistry 18(3): 285-298, 1993

Phase diagrams of a CTAB/organic solvent/buffer system applied to extraction of enzymes by reverse micelles. Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences 862(1-2): 58-63, 2007

Thermostability of alpha-chymotrypsin in water/organic solvent systems. Biotechnology Letters 14(11): 1041-1044, 1992

A reverse micellar mesophase of face-centered cubic Fm3m symmetry in phosphatidylcholine/water/organic solvent ternary systems. Langmuir 29(51): 15805-15812, 2014

Liquid-liquid extraction of alpha -lactalbumin using reverse micellar organic solvent. BioFactors 22(1/4): 347-351, 2004

Liquid-liquid extraction of alpha-lactalbumin using reverse micellar organic solvent. Biofactors 22(1-4): 347-351, 2005

The kinetics of the alpha-chymotrypsin-catalyzed hydrolysis of p-nitrophenyl acetate in organic solvent-water mixtures. Journal of Biological Chemistry 241(21): 4825-4834, 1966