# Effect of a Conical Cellulose Structure on Horseradish Peroxidase Biomacromolecules
## 4 Discussion  
The results of our study indicate that the incubation of the HRP enzyme solution near the apex of the cellulose conical structure for 10 min does not affect the HRP's properties. In contrast, a longer incubation time (40 min) results in a change in the degree of the enzyme oligomerization on the mica surface. This indicates an effect of the incubation time on the properties of the enzyme. At the same time, no effect on the enzymatic activity of HRP is revealed.  
The change in the degree of HRP oligomerization is likely to be caused by a change in the topology of the background electromagnetic field near the conical structure. Indeed, a theoretical investigation performed by [PERSON] et al. showed that a square pyramid can concentrate an electromagnetic field in the area of the pyramid's base [10]. A cone, in turn, also pertains to pyramid-like figures [43]. Namely, a cone represents a pyramid with a circular base [43]. The latter can be considered a polygon with an infinite number of sides [44]. Accordingly, the cone is also expected to interact with the background electromagnetic field, thus altering its topography, since such an effect has been observed for a square pyramid [11, 13]. It has been shown that electric and electromagnetic fields of even low intensity can substantially alter the ratio between ortho- and para-isomers of water [45, 46, 47]. [PERSON] et al. demonstrated that the hydration shell of a biological macromolecule predominantly contains para-H\({}_{2}\)O [48]. In our experiments, the background electromagnetic field could well be concentrated by the cellulose cone, thus affecting the hydration of the enzyme globule by altering the ratio between ortho- and para-H\({}_{2}\)O in this shell. Additionally, the adsorption properties of the enzyme were changed at the expense of altered enzyme hydration [49, 50]. This is how we explain the effect observed in our experiments.  
The adsorption of macromolecules from their solution onto a solid substrate is determined by the interaction between the substrate (in our case, mica) surface and the surface of a hydrated enzyme globule, as well as by the interaction of the enzyme macromolecules with adjacent macromolecules in the solution, and with the solvent molecules [51]. This is why a change in enzyme hydration leads to a change in its oligomeric state on mica. Accordingly, there is a change in the contribution of aggregates of various orders to the relative distribution of AFM-visualized objects.  
The results of our work can be of use in the development of biosensors intended for highly sensitive analysis, since a change in the hydration of biomacromolecules can result in a change in the functional properties [49, 50]. The results obtained should also be considered in direct measurements using biosensors with conical constructive elements in order to protect the highly sensitive biosensors from external electromagnetic interference by means of anechoic chambers with conical elements. Since HRP is widely used as a model enzyme in studies of external impacts on enzymes determining food quality, our data can also be useful in the development of food-processing methods.