Papain Hydrolysates of Lupin Proteins with Antioxidant, Antimicrobial, and Acetylcholinesterase Inhibitory Activities
, Dasha Mihaylova 2,*, Hafize Fidan 3, Margarita Terziyska 4 and Atanas Pavlov 1,5Round 1
Reviewer 1 Report
Abstract:
The second statment line 16 is not relevant as it still connotes nutritional value.
This is because health is also obtained as a result of good nutrition.
In line 25, the sentence should start with "To the best of our knowledge..." and
not "For the best....."
Introduction
Line 39-41. It is better to reword the sentence as follows
"Bioactive peptides are believed to have properties such as......."
Line 43-44
Please replace "Sensitive" with "important"
Line 44. The statment "We can consider that meat is no longer a sufficient
and acceptable 44 supplier that’s why in recent years a lot of alternatives emerge."
must be reworded as its meaning is very clear.
Figures must be improved, error bars are not showing in some cases. There are situations where there is a comma after Sample. Please be consistent.
Conclusion does not clearly reflect the topic and abstract as the Anticholinesterase
activity has been clearly left out.
Author Response
Please find the answers to your comments and questions in the attached file.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
This work shows the potential bioactivity of hydrolysates obtained from lupin under different processing conditions.
There are some major issues in the experimental design, statistical analysis, and discussion section that should be address to make this work suitable for publication. Furthermore, it is not clear why authors did not include some important parts in the analysis and discussion section.
General comments:
Highlight the novelty of this work, since several publications on protein hydrolysates from lupin have been already published.
English should be revised, sometimes it is confusing, especially in L32-33, L51, and the results and discussion section.
The effect of the hydrolysis conditions was not analyzed or discussed, this was an essential part of the study.
Abstract:
L25-27: These lines are repeated
L27: Authors cannot state this with the analysis they performed in this work
Introduction:
Introduction is poor, in this section authors should have included the state-of-the-art on the role of proteins as bioactive compounds, described previous publications, included the importance of lupin proteins or the advantages of using papain and measuring AChE. A few works have been cited although literature is plenty of publications addressing this topic. Some of this information was described in the discussion section instead of introduction.
Methods
L79: include more detailed sample preparation. Why alkaline extraction was carried out? What is the advantage over acidic extraction?
L89: What was the weight of lupin proteins dissolved in PBS?
L92: delete “-“ before 6, it is confusing. Do not use abbreviations for respectively.
L102: indicate the concentration range used for BSA
L121: indicate the solvent
Results
This section is poor also. More in-depth discussion is needed in this section. Information is just described and not discussed.
Authors should identify the peptides corresponding to each fraction or at least those with the highest bioactivity.
L245: Please describe and discuss these fractions in the main text
L285-313: This information should be at the beginning of section 3.2
L359-385: Just one citation to support all this information.
L394: It should say DH-AChE
The effect of E:S ratio and incubation for hydrolysis was not analyzed, this was an essential part of the study. A two-way ANOVE should be performed for these results.
Table 1:
Define E:S
The experimental design depicted in the table seems incorrect.
Figure 1 and 2: include the statistical analysis
Why ABTS is presented in another chart?
Author Response
Please find the answers to your comments and questions in the attached file.
Author Response File: Author Response.pdf
Reviewer 3 Report
Herein, authors have extracted papain hydrolysates from lupin flour and analyzed it for its antioxidant properties. While the work carried out by the authors is good and well presented, some additional experiments are required to confirm the structure of the extracted compound.
1) English language and grammar require minor revisions throughout the article.
2) Is the DDPH test performed in the dark?
3) Please provide IR, GPC, and 2D NMR of extracted compound and feedstock for structural analysis.
4) Compare these results with previously reported papain hydroxylates.
Author Response
Please find the answers to your comments and questions in the attached file.
Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
I have no comment
Author Response
Thank you very much for your comment.
Reviewer 2 Report
The manuscript was substantially improved, only minor revisions should be made.
Results:
Table 2 - the letters obtained by Tukey analysis were not included in this table. Please include them.
L424 — correct ANOWA for ANOVA
In the case of AChE inhibition, shouldn't it be better to conduct Tukey for Two-way ANOVA since there is a significant effect of the interaction?
Conclusions: this section still needs some improvement. Statements are too general, for example in line 475-476 authors declare “our findings are of significant importance “. An explanation of this relevance should be included.
Supplementary material:
Please be sure to have the same format in all tables.
Author Response
The authors would like once again to show their appreciation for the reviewer’s helpful, attentive, and thorough review. We accept his\her very detailed suggestions and improvements for the manuscript.
Below, the reviewer can find a point-by-point answer to all comments:
The manuscript was substantially improved, only minor revisions should be made.
Answer: Thank you very much for your evaluation
Results:
Table 2 - the letters obtained by Tukey analysis were not included in this table. Please include them.
Answer: Appropriate addition has been made.
L424 — correct ANOWA for ANOVA
Answer: Appropriate correction has been made.
In the case of AChE inhibition, shouldn't it be better to conduct Tukey for Two-way ANOVA since there is a significant effect of the interaction?
Answer: Appropriate tests and addition have been made.
Conclusions: this section still needs some improvement. Statements are too general, for example in line 475-476 authors declare “our findings are of significant importance “. An explanation of this relevance should be included.
Answer: Appropriate edition and addition have been made.
Supplementary material:
Please be sure to have the same format in all tables
Answer: Appropriate formatting has been made.
