Abstract
Plastics are ubiquitous in the aquatic environment and their degradation of fragments down to the nanoscale level have raised concerns given their ability to pervade cells. The accumulation of nanoparticles could lead to molecular crowding which can alter the normal functioning of enzymes. The purpose of this study was to examine the influence of polystyrene nanoparticles (NPs) on the fractal kinetics of the lactate dehydrogenase reaction: pyruvate + NADH ↔ lactate + NAD(+). The influence of NPs on LDH activity was examined first in vitro to highlight specific effects and secondly in mussels exposed to NPs in vivo for 24h at 15 °C. The reaction rates of LDH were determined with increasing concentrations of pyruvate to reach saturation at circa 1 mM pyruvate. The addition of F-actin, a known binding template for LDH, revealed a characteristic change in reaction rates associated with fractal organization. The addition of 50 and 100 nm transparent NPs also produced these changes. The fractal dimension was determined and revealed that both F-actin and NPs reduced the fractal dimension of the LDH reaction. The addition of viscosity sensor probe in the reaction media revealed viscosity waves during the reaction at low substrate concentrations thought to be associated to synchronized switching between the relaxed and tensed states of LDH. The amplitude and the frequency of viscosity waves were increased by both NPs and F-actin which were associated with increased reaction rates. In mussels exposed to NPs, the isolation of digestive gland subcellular fraction revealed that LDH activity was significantly influenced by the fractal dimension of the LDH reaction where a loss of affinity (high fractal K(M)) was detected in mussels exposed to the high concentrations of NPs. It is concluded that polystyrene NPs could change the biophysical properties of the cytoplasm such as the fractal organization of the intracellular environment during the LDH reaction.