title: Probing the interaction induced conformation transitions in acid phosphatase with cobalt ferrite nanoparticles: Relation to inhibition and bio-activity of Chlorella vulgaris acid phosphatase.
abstract: The present study explored the interaction and kinetics of cobalt ferrite nanoparticles (NPs) with acid phosphatase (ACP) by utilizing diverse range of spectroscopic techniques. The results corroborate, the CoFe2O4 NPs cause fluorescence quenching in ACP by static quenching mechanism. The negative values of van't Hoff thermodynamic expressions (ΔH=-0.3293Jmol(-1)K(-1) and ΔG=-3.960kJmol(-1)K(-1)) corroborate the spontaneity and exothermic nature of static quenching. The positive value of ΔS (13.2893Jmol(-1)K(-1)) corroborate that major contributors of higher and stronger binding affinity among CoFe2O4 NPs with ACP were electrostatic. In addition, FTIR, UV-CD, UV-vis spectroscopy and three dimensional fluorescence (3D) techniques confirmed that CoFe2O4 NPs binding induces microenvironment perturbations leading to secondary and tertiary conformation changes in ACP to a great extent. Furthermore, synchronous fluorescence spectroscopy (SFS) affirmed the comparatively significant changes in microenvironment around tryptophan (Trp) residue by CoFe2O4 NPs. The effect of CoFe2O4 NPs on the activation kinetics of ACP was further examined in Chlorella vulgaris. Apparent Michaelis constant (Km) values of 0.57 and 26.5mM with activation energy values of 0.538 and 3.428kJmol(-1) were determined without and with 200μM CoFe2O4 NPs. Apparent Vmax value of -7Umml(-1) corroborate that enzyme active sites were completely captured by the NPs leaving no space for the substrate. The results confirmed that CoFe2O4 NPs ceased the activity by unfolding of ACP enzyme. This suggests CoFe2O4 NPs perturbed the enzyme activity by transitions in conformation and hence the metabolic activity of ACP. This study provides the pavement for novel and simple approach of using sensitive biomarkers for sensing NPs in environment.
authors.affiliation: College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, China.
authors.forename: Farooq
authors.initials: F
authors.lastname: Ahmad
authors.affiliation: College of Atmospheric Science, Lanzhou University, Lanzhou, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Zhou
authors.affiliation: College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, China.
authors.forename: Hongzhou
authors.initials: H
authors.lastname: Yao
authors.affiliation: College of Chemical Engineering, Zhejiang University of Technology, Hangzhou, China; Research Center of Analysis and Measurement, Zhejiang University of Technology, Hangzhou, China. Electronic address: yingzhou@zjut.edu.cn.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Institute of Environmental Science, College of Environment, Zhejiang University of Technology, Hangzhou 310032, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Xu
mesh_terms: D000135:Acid Phosphatase / Q000378:metabolism*; D048408:Chlorella vulgaris / Q000378:metabolism; D003035:Cobalt / Q000378:metabolism; D005290:Ferric Compounds / Q000378:metabolism; D007700:Kinetics; D053758:Nanoparticles / Q000737:chemistry
chemical_list: D005290:Ferric Compounds; C569492:cobalt ferrite; D003035:Cobalt; D000135:Acid Phosphatase
keywords: Acid phosphatase; Algae; Environment; Interaction; Kinetics; Nanotoxicity