PDF 
Views 
Share
Interaction of Di-cyclohexyl Phthalate (DCHP) with Catalase and Its Effect on the Function and Structure of Catalase: An In Vitro and In Silico Study
CC BY 4.0 · Indian J Med Paediatr Oncol 2024; 45(S 01): S1-S16
DOI: DOI: 10.1055/s-0044-1788224
*Corresponding author: (e-mail: mylalitkumar@gmail.com).
Abstract
Background: Phthalates are lipophilic diesters, which can get readily metabolized into their respective monoesters and introduce toxicity in living organisms. The main objective of this study was to determine the inhibitory effect of DCHP on rat liver catalase (CAT) activity in vitro and in silico.
Materials and Methods: The Wistar rat liver was excised and the liver homogenate (10% w/v) was prepared in 0.25 M sucrose solution under ice-cold conditions (4°C). The homogenate was centrifuged at 9,000 rpm for 30 minutes at 4°C. The postnuclear supernatants were stored in aliquots at −20°C and used for further experiments.
Results: The protein content was determined by using the Lowry method. CAT exhibited optimum activity at pH 7.4 and temperature 37°C. The K m, K cat, and V max values for CAT were calculated from Lineweaver–Burk’s double reciprocal plot obtained after assaying the enzyme activity in the absence and presence of the DCHP. The docking study shows the maximum binding efficiency with ALA7 and GLN10, these amino acids are present in other than the catalytic active site of the enzyme.
Conclusion: This in vitro and in silico study indicated significant alterations in the activity of CAT due to DCHP. The results showed a negative modulatory effect of DCHP on CAT activity. The results of this study may be useful in the management of toxicity induced by phthalates.
Publication History
Article published online:
08 July 2024
© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India
*Corresponding author: (e-mail: mylalitkumar@gmail.com).
Abstract
Background: Phthalates are lipophilic diesters, which can get readily metabolized into their respective monoesters and introduce toxicity in living organisms. The main objective of this study was to determine the inhibitory effect of DCHP on rat liver catalase (CAT) activity in vitro and in silico.
Materials and Methods: The Wistar rat liver was excised and the liver homogenate (10% w/v) was prepared in 0.25 M sucrose solution under ice-cold conditions (4°C). The homogenate was centrifuged at 9,000 rpm for 30 minutes at 4°C. The postnuclear supernatants were stored in aliquots at −20°C and used for further experiments.
Results: The protein content was determined by using the Lowry method. CAT exhibited optimum activity at pH 7.4 and temperature 37°C. The K m, K cat, and V max values for CAT were calculated from Lineweaver–Burk’s double reciprocal plot obtained after assaying the enzyme activity in the absence and presence of the DCHP. The docking study shows the maximum binding efficiency with ALA7 and GLN10, these amino acids are present in other than the catalytic active site of the enzyme.
Conclusion: This in vitro and in silico study indicated significant alterations in the activity of CAT due to DCHP. The results showed a negative modulatory effect of DCHP on CAT activity. The results of this study may be useful in the management of toxicity induced by phthalates.
No conflict of interest has been declared by the author(s).
Publication History
Article published online:
08 July 2024
© 2024. The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution License, permitting unrestricted use, distribution, and reproduction so long as the original work is properly cited. (https://creativecommons.org/licenses/by/4.0/)
Thieme Medical and Scientific Publishers Pvt. Ltd.
A-12, 2nd Floor, Sector 2, Noida-201301 UP, India