## 3 Results
### Influence of Gemini Surfactants on Enzymatic Profile of Mycelia  
In the first stage of the analysis, we studied the effects of the gemini surfactants on the enzymatic profile of _A. brasiliensis_ ATCC 16404. The results are shown in Table 1.  
The lipolytic enzymes alkaline phosphatase (2), esterase (3), esterase lipase (4), acid phosphatase (11), naphthol-AS-BI-phosphohydrolase (12), and the amyloidic enzymes \(\alpha\)-galactosidase (13), \begin{table}
\begin{tabular}{c c c c c c c c c} \hline \multirow{2}{*}{**Type of Sample**} & \multirow{2}{*}{**Biocide Concentration**} & \multicolumn{6}{c}{**Symbol of Enzyme in API-ZYM Test**} \\ \cline{3-10}  & & **2** & **3** & **4** & **11** & **12** & **13** & **16** & **18** \\ \hline K & 0 & & & & & & & & \\ \hline \multirow{3}{*}{C5} & \% MIC & & & & & & & & \\  & \% MIC & & & & & & & \\  & 2 MIC & & & & & & & \\ \hline \multirow{3}{*}{C6} & \% MIC & & & & & & & & \\  & 2 MIC & & & & & & & \\ \hline \multirow{3}{*}{A5} & \% MIC & & & & & & & & \\  & 2 MIC & & & & & & & \\ \hline \multirow{3}{*}{A6} & \% MIC & & & & & & & & \\  & 2 MIC & & & & & & & \\ \hline \end{tabular}  
2–18 refer to the enzyme numbers in the API -ZYM test: 2—alkaline phosphatase, 3—esterase (C4), 4—esterase lipase (C8), 11—acid phosphatase, 12—naphthol-AS-BI-phosphohydrolase, 13—\(\alpha\)-galactosidase, 16—\(\alpha\)-glucosidase and 18—N-acetyl-\(\beta\)-glucosaminidase, respectively. Concentration of hydrolyzed substrate: light grey—5 nM, grey—10 nM, orange—20 nM, yellow—30 nM, green—\(\geq\)40 nM.  
The lipolytic enzymes alkaline phosphatase (2), esterase (3), esterase lipase (4), acid phosphatase (11), naphthol-AS-BI-phosphohydrolase (12), and the amyloidic enzymes \(\alpha\)-galactosidase (13),  
\end{table} 
Table 1: Effect of cationic C6 or C5 and neutral A5 or A6 surfactants at 1% MIC, MIC, and 2 MIC on the enzymatic profile of _Aspergillus brasiliensis_ mycelia. \(\alpha\)-glucosidaseidase (16), and N-acetyl-\(\beta\)-glucosaminidase (18) were detected in the control sample. When surfactants were added to the cultures, several enzymes were suppressed. The amylases activity observed in control sample was almost completely inhibited in the samples with cationic and neutral gemini surfactants. The exceptions were \(\alpha\)-galactosidaseosidase (13) and N-acetyl-\(\beta\)-glucosidaseaminidase (18), for which negligible activity (5 nmol) was observed for cationic gemini surfactant C5 across the entire concentration range, and for C6 at 1% MIC and MIC. During detailed analysis of the influence of the gemini surfactants on the overall levels of enzymatic activity of _A. brasiliensis_ mycelia, we observed interesting results in the cases of acid phosphatase (11) and naphthol-AS-BI-phosphohydrolase (12). The highest activity of both lipases, exceeding 40 nmol, was noted in the control sample. As shown in Table 1, cationic gemini surfactants C5 and C6 had no influence on the levels of lipolytic activity by either enzyme. At 1% MIC, MIC, and 2 MIC (0.155 mM, 0.31 mM, and 0.62 mM, respectively), these biocides released the same amounts of substrate as the control sample. An exception was compound C5 at 2 MIC, for which the concentration of naphthol-AS-BI-phosphohydrolase was only 30 nM. A relationship was observed between the concentrations of non-ionic gemini surfactants A5 and A6 at 1% MIC, MIC, and 2 MIC (15 mM, 30 mM, and 60 mM, respectively) and the activities of acid phosphatase (11) and naphthol-AS-BI-phosphohydrolase (12). As the surfactant concentration increased, the amounts of released substrate decreased: from 40 nM to 5 nM for acid phosphatase (11) and from 30 nM to 5 nM for naphthol-AS-BI-phosphohydrolase (12) for both biocides, A5 and A6. None of the samples treated with cationic or neutral gemini surfactants showed activity for the lipolytic enzymes alkaline phosphatase (2), esterase (3), or esterase lipase (4). Based on the effect of these compounds on the metabolism of _A.