## 4 Conclusions  
This study provides valuable information about the potential of papain-generated lupin protein hydrolysates to exert antimicrobial, antioxidant, and acetylcholinesterasecholinesterase inhibitory activities. Three E:S ratios were studied with three hydrolysis duration times. A total of nine different hydrolysates were generated. Both factors and combination influenced the DH and AchE inhibition (\(p<0.05\)). All the hydrolysates possessed antioxidant activity, acetylcholinesterasecholinesterase inhibitory activity, and inhibited the growth of _E. coli_ ATCC 8739. When tested with ABTS\({}^{\text{++}}\) scavenging assay, a thousand times higher AOA was measured for all the hydrolysates, compared to the other three techniques (DPPH, CUPRAC, FRAP). While antioxidant peptides from various sources have been more or less studied, research on protein acetylcholinesterasecholinesterase inhibitors is rather scarce. In this regard, our findings are of significant importance. We measured IC\({}_{50}\) from 155.56 to 199.63 mg/g protein between samples that, in our opinion, highlights the need for more detailed studies on that topic. Further studies of the peptide structure-activity relationship are needed.  
The following supporting information can be downloaded at: [[https://www.mdpi.com/article/10.3390/app122312370/s1](https://www.mdpi.com/article/10.3390/app122312370/s1)]([https://www.mdpi.com/article/10.3390/app122312370/s1](https://www.mdpi.com/article/10.3390/app122312370/s1)), Figure S1: The densitometric profile of low molecular weight standard markers (1.060-17.0 kDa); Figure S2: Molecular weight calibration curve (based on the densitometric profile from Figure S1); Figure S3: The densitometric profile of sample P1; Figure S6: The densitometric profile of sample P2; Figure S7: The densitometric profile of sample P3; Figure S8: The densitometric profile of sample P6; Figure S9: The densitometric profile of sample P7; Figure S10: The densitometric profile of sample P8; Figure S11: The densitometric profile of sample P9. Table S1. Two-way ANOVA of the impact of E:S ratio and enzyme hydrolysis duration-tion (EHD) on the DH. Table S2. Two-way ANOVA of the impact of E:S ratio and enzyme hydrolysis data-tion (EHD) on the AChE inhibitory activity. Table S3. Tukey HSD multiple comparisons on the AChE inh. between E:S ratio levels. Table S4. Tukey HSD multiple comparisons on the AChE inh. between EHD. Table S5. Two-way ANOVA of the impact of E:S ratio and enzyme hydrolysis duration (EHD) on AOA (FUPRAC). Table S6. Two-way ANOVA of the impact of E:S ratio and enzyme hydrolysis data-tion (EHD) on AOA (UCPRAC). Table S7. Two-way ANOVA of the impact of E:S ratio and enzyme hydrolysis data-tion (EHD) on AOA (DPPH). Table S8. Two-way ANOVA of the impact of E:S ratio and enzyme hydrolysis data-tion (EHD) on AOA (ABTS).  
Conceptualization, I.D.; methodology, I.D.; software, I.D. and M.T.; validation, I.D. and M.T.; formal analysis, A.G., I.D., D.M., and H.F.; resources, A.G. and I.D.; writing--original draft preparation, A.G., I.D., D.M., H.F., and M.T.; writing--review and editing, I.D. and D.M.; visualization, I.D. and D.M.; supervision, I.D. and A.P.; project administration, I.D.; funding acquisition, I.D. All authors have read and agreed to the published version of the manuscript.  
This research was funded by the Bulgarian Scientific Fund, grant No. KP-06-M36/2.  
Not applicable.  
The authors declare no conflict of interest.