Refolding of thermally denatured cholesterol oxidases by magnetic nanoparticles

Shubhrima Ghosh; Razi Ahmad; S K Khare

doi:10.1016/j.ijbiomac.2019.07.103

Refolding of thermally denatured cholesterol oxidases by magnetic nanoparticles

Int J Biol Macromol. 2019 Oct 1:138:958-965. doi: 10.1016/j.ijbiomac.2019.07.103. Epub 2019 Jul 17.

Authors

Shubhrima Ghosh¹, Razi Ahmad¹, S K Khare²

Affiliations

¹ Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India.
² Enzyme and Microbial Biochemistry Laboratory, Department of Chemistry, Indian Institute of Technology, Delhi, Hauz Khas, New Delhi 110016, India. Electronic address: skkhare@chemistry.iitd.ac.in.

PMID: 31325504
DOI: 10.1016/j.ijbiomac.2019.07.103

Abstract

Proteins are prone to unfolding and subsequent denaturation by changes in temperature, pH and other harsh conditions. Nanoparticles act as artificial 'chaperones' due to favourable orientation of the proteins on their scaffold which prevents aggregation and reconfigures denatured proteins into their native functional state. In the present study, thermal denaturation of Cholesterol oxidases from Pseudomonas aeruginosa PseA, Rhodococcus erythropolis MTCC 3951 and Streptomyces sp. were studied at temperatures 50-70 °C. Further, these thermally denatured proteins were refolded using functionalized Magnetic Iron (II, III) oxide nanoparticles which was confirmed using DLS, Zeta Potential Measurements, fluorescence and CD spectroscopy. The refolded proteins were found to regain their secondary structure and activity to a great extent.

Keywords: Cholesterol oxidase; Nanoparticles; Refolding.

MeSH terms

Cholesterol Oxidase / chemistry*
Enzyme Activation
Ferric Compounds / chemistry
Magnetite Nanoparticles / chemistry*
Particle Size
Protein Denaturation*
Protein Refolding*
Spectrum Analysis
Temperature
Thermodynamics

Substances

Ferric Compounds
Magnetite Nanoparticles
ferric oxide
Cholesterol Oxidase