## 3 Results and Discussion
### Protease and Cellulase Activity and Total Protein Content  
Following the determination of protease activity, it was noticed that _T. atroviride_ did not present significant protease activity after 4 h of incubation, having less than 1 \(\upmu\)M Tyrosine. After 5 days of incubation, there was a relevant increase in protease activity in both non-irradiated and laser-irradiated strains. Statistically significant higher protease activity was observed for the 60 s irradiated sample (104.3 \(\pm\) 9.6 \(\upmu\)M) compared to the non-irradiated sample (74.9 \(\pm\) 3.2 \(\upmu\)M) and 300 s irradiated sample (69.7 \(\pm\) 2.6 \(\upmu\)M), 39% and 50% higher, respectively (Figure 2a). The protease activity increased further after 12 days of incubation, and there was no significant difference between the non-irradiated and irradiated samples at this time. A noticeable decrease could be observed in all samples after 15 days of incubation, the lowest value being for 3 \(\times\) 300 s irradiated samples (33.8 \(\pm\) 4.2 \(\upmu\)M, 56% less than the non-irradiated samples). There was no significant difference between the control and the 60 s irradiated sample. The cellulase activity of _T. atroviride_ was less than 50 \(\upmu\)M glucose after 4 h of incubation. After 5 days of incubation from the first irradiation, there was a similar increase in cellulosolytic enzyme synthesis in both non-irradiated and irradiated samples (Figure 2b). There was no significant difference between the irradiated and non-irradiated samples. After 12 days of incubation and two rounds of irradiation, there was a significant increase in cellulase production only in the non-irradiated sample (1027.4 \(\pm\) 22.4 \(\upmu\)M) (Figure 2b). The irradiated samples, both 60 s and 300 s, maintained the same cellulase activity after 12 days as after 5 days, and this activity was around 50% less than the non-irradiated sample. After 15 days of incubation and three irradiations, there was an increase in the cellulase activity for the samples irradiated for 60 s (851.2 \(\pm\) 54.2 \(\upmu\)M) and 300 s (803.2 \(\pm\) 31.2 \(\upmu\)M) in comparison to the corresponding cellulase activity measured after 12 days of incubation (540 \(\pm\) 51.4 and 487.2 \(\pm\) 41.3 \(\upmu\)M, respectively), i.e., the cellulase activity increased by 58% and 65%, respectively, from the 12 th day to the 15 th day of incubation. The cellulase activity for the non-irradiated sample (730.2 \(\pm\) 27.9 \(\upmu\)M) was 29% lower after 15 days than after 12 days of incubation (Figure 1b). The cellulase activity became higher by 17% and 10%  
Figure 2: Protease and cellulase production: (**a**) protease activity of _T. atroviride_ as a response to being exposed for different time intervals (60 and 300 s) to 0.271 \(\upmu\)moles/s 400 nm laser irradiation; (**b**) cellulase activity of _T. atroviride_ as a response to being exposed for different time intervals to 0.271 \(\upmu\)moles/s 400 nm laser irradiation; Tyr = tyrosine; The columns with the same letter do not differ significantly. Bars are \(\pm\) error bars, \(\alpha\) = 0.05, \(n\) = 3.  
in the 60 s and 300 s irradiated samples, respectively, compared with the non-irradiated samples on the 15 th day of incubation and after three rounds of irradiation. These data indicate that the first effect of irradiation with blue-light laser is the stimulation of protease production, accompanied by a stagnation of cellulase production, followed by an increase in cellulase production. Cellulase synthesis seems to be delayed due to laser irradiation. Too much of an irradiation dose can have an inhibitory effect on protease production. We quantified the total protein from the culture using the Bradford and Biuret methods. Biuret gave 10\(\times\) higher protein concentration compared to Bradford. It is known that both methods can be affected by interference from other compounds such as polyphenols and flavonoids.