Thermodynamic study of β- N-acetylhexosaminidase enzyme heterogeneity in human seminal plasma

Abstract

Background It has been suggested that the activity of β- N-acetylhexosaminidase (Hex) in seminal plasma may be used as a biochemical marker of azoospermia. The purpose of our study was to evaluate this hypothesis using a thermodynamic procedure developed to determine total Hex activity and that of its isoenzymes in this biological fluid. Methods Using the substrate 3,3′-dichlorophenolsulphoftaleinil N-acetyl-β- d-glucosaminide, a highly significant difference ( p<0.001) is found between the activation energy of Hex A (41.5 kJ/mol) and of Hex B (72.3 kJ/mol), making it possible to determine the activity of these isoenzymes from the apparent activation energy of the total Hex in seminal plasma. Results A significant difference between the normozoospermic and azoospermic groups was only found for Hex A isoenzyme activity ( p<0.05), although with considerable overlapping between the values of both groups. Significant partial correlations were found for the total Hex, Hex A and Hex B activities with the immobile spermatozoa count ( p<0.01) and for total Hex and Hex B with the dead spermatoza count ( p<0.05). In turn, Hex A had a significant partial correlation with the live spermatozoa count ( p<0.05); however, Hex activity in seminal plasma of acromosomal origin appears to be of little importance in quantitative terms. Conclusions It was not possible to confirm that total Hex activity in seminal plasma, or even of its isoenzymes Hex A and Hex B, is a suitable biochemical marker of azoospermia (efficiency≤67%). The thermodynamic procedure described may be a useful alternative for the study of the Hex enzyme heterogeneity in spermatozoa.