Enzymatically Hydrolysed Common Buckwheat (Fagopyrum esculentum M.) as a Fermentable Source of Oligosaccharides and Sugars

Round 1

Reviewer 1 Report

Dear authors,

I have reviewed your manuscript and, although I found the idea interesting, the work fails to present the results and discuss them properly. In addition, I found the dataset to be very primary, which does not support some sentences, especially, the results found in the antimicrobial evaluation. The discussion was often speculative. Therefore, I do not recommend its publishing in this journal. Below, you can see details about my review. 

At abstract the authors cite that Common buckwheat (Fagopyrum esculentum M.) is highly rich in starches, but in the introduction they say otherwise (see line 36).

Methodology 

How the fermantation process was monitored? pH initial and pH final? acidity?

3.4. Static In Vitro Digestion 124 The static in vitro digestion (IVD) of enzymatically hydrolysed hydrophilic extracts 125 of F. esculentum was carried out in three different stages as described by Minekus et al. 126 and Bradford et al. [24]. Modifications to the procedure, as described by Streimikyte et al. 127 in a study conducted in 2020, were incorporated [25]. The digestion procedure was per- 128 formed twice. Where are the results of this stage of the work discussed?

Results: The released rutin structure observed in Figure 2b is 174 comparable to SEM images of rutin presented by Abdelcader et al. in 2018 [27].... This sentence is speculative.

Where is the figure 4?

Table 1. Potencial of antimicrobial properties of common buckwheat fermented with Tibetan kefir grains and birch sap. This must show values instead of negative and positive. 

These results demonstrate promising potential pharmaceutical applications for the afore- 246 mentioned extracts, especially for use against E. Coli due to the wide range of foodborne 247 infections and urinary illnesses it causes [37].  Why????? 

Conclusion.... leading to increased compound release and higher bioavailability... Where it is demonstrated in the manuscript?

 

 

 

Author Response

The authors are grateful for the Reviewers report, which impactfully improves the manuscript. Below we answered your comments.

Point 1: At abstract the authors cite that Common buckwheat (Fagopyrum esculentum M.) is highly rich in starches, but in the introduction they say otherwise (see line 36).

Answer 1: Thank you for your comment. We agree it may be confusing with the abstract and introduction part. However, common buckwheat is a great source of fibers synonymously: non-starch polysaccharides. We changed this confusion to more appropriate. (Line 36)

Point 2: Methodology 

How the fermantation process was monitored? pH initial and pH final? acidity?

3.4. Static In Vitro Digestion 124 The static in vitro digestion (IVD) of enzymatically hydrolysed hydrophilic extracts 125 of F. esculentum was carried out in three different stages as described by Minekus et al. 126 and Bradford et al. [24]. Modifications to the procedure, as described by Streimikyte et al. 127 in a study conducted in 2020, were incorporated [25]. The digestion procedure was per- 128 formed twice. Where are the results of this stage of the work discussed?

Answer 2: Thank you for your insight. We created figure 1 as a graphical overview of the experimental design, which includes pH values and where we presented the results after static in vitro digestion. Hopefully, it will be clearer.

Point 3: Results: The released rutin structure observed in Figure 2b is 174 comparable to SEM images of rutin presented by Abdelcader et al. in 2018 [27].... This sentence is speculative.

Answer 3: Thank you for your comment. We agree; perhaps we concluded it was too straightforward and had to leave just a discussion of images and released particles. We deleted this sentence.

Point 4: Where is the figure 4?

Answer 4: Thank you, we corrected the mistake.

Point 5: Table 1. Potencial of antimicrobial properties of common buckwheat fermented with Tibetan kefir grains and birch sap. This must show values instead of negative and positive. 

Answer 5: Thank you. We corrected it and changed it to “inhibition zone, mm “ (Table 1)

Point 6: These results demonstrate promising potential pharmaceutical applications for the afore- 246 mentioned extracts, especially for use against E. Coli due to the wide range of foodborne 247 infections and urinary illnesses it causes [37].  Why????? 

Answer 6: Thank you for your answer, we changed this discussion part (line 249)

Point 7: Conclusion.... leading to increased compound release and higher bioavailability... Where it is demonstrated in the manuscript?

Answer 7: Thank you for your comment, we agree with your opinion and changed this part

Sincerely

Reviewer 2 Report

 

Review on Enzymatically hydrolyzed common buckwheat as a fermentable source of oligosaccharides

 

Page 1 Line 30: has high potential in….

Check for typos and singular plural mistakes in the manuscript.

 

Very interesting work. I would have liked to see more incorporation of existing literature

Author Response

The authors are grateful for the Reviewers report, which improves the manuscript. Below we answered your comments.

 

Point 1: Page 1 Line 30: has high potential in….

Check for typos and singular plural mistakes in the manuscript. 

Very interesting work. I would have liked to see more incorporation of existing literature

Answer 1: Thank you for your comment. We incorporated more literature and checked the mistakes.

Sincerely,

Round 2

Reviewer 1 Report

Dear authors,

Even though the manuscript had improved after the review, item 3.4 (in vitro digestion) has not yet been discussed in the work. It is not clear how and why the digestion was performed.  The dataset should be extended to determine the phytochemical profile and antioxidant capacity of extracts before and after the processes. Also, item numbering throughout the manuscript should be checked. Thus, I don't recommend your manuscript for publishing. 

Author Response

Reviewer comment: Even though the manuscript had improved after the review, item 3.4 (in vitro digestion) has not yet been discussed in the work. It is not clear how and why the digestion was performed.  The dataset should be extended to determine the phytochemical profile and antioxidant capacity of extracts before and after the processes. Also, item numbering throughout the manuscript should be checked. Thus, I don't recommend your manuscript for publishing. 

Answer to the reviewer 2

Dear Reviewer,

Thank you for Your comments. We added a static in vitro digestion procedure to the materials and methods part (Line 129-150).

The results are presented in 3.2. High-Pressure Liquid Chromatography Size Exclusion for Molecular Mass Distribution (HPLC-SEC) section (Line 213). Figure 3a,c, and d chromatographs represent the molecular mass distribution of polysaccharides, oligosaccharides, disaccharides, and monosaccharides of F. esculentum extracts after enzymatic hydrolysis using different enzymes cocktails before the digestion (IVD). Figure 3b,d, e represent extracts' molecular mass distribution using various enzyme cocktails after IVD. 

The authors agree that the dataset should be extended. However, this is communication - a short report of findings. We wanted to share our findings which connect with authors new published article:

Streimikyte, P.; Kailiuviene, J.; Mazoniene, E.; Puzeryte, V.; Urbonaviciene, D.; Balciunaitiene, A.; Liapman, T.D.; Laureckas, Z.; Viskelis, P.; Viskelis, J. The Biochemical Alteration of Enzymatically Hydrolysed and Spontaneously Fermented Oat Flour and Its Impact on Pathogenic Bacteria. Foods 2022, 11, 2055. https://doi.org/10.3390/foods11142055

Moreover, the phytochemical findings of F.esculentum extracts can be observed in the article that authors published in Plants:

Balčiūnaitienė, A.; Štreimikytė, P.; Puzerytė, V.; Viškelis, J.; Štreimikytė-Mockeliūnė, Ž.; Maželienė, Ž.; Sakalauskienė, V.; Viškelis, P. Antimicrobial Activities against Opportunistic Pathogenic Bacteria Using Green Synthesized Silver Nanoparticles in Plant and Lichen Enzyme-Assisted Extracts. Plants 2022, 11, 1833. https://doi.org/10.3390/plants11141833

However, in the future, we plan to make extensive metabolomics of hydrolysed and fermented extracts. 

Thank you,

Sincerely,

Round 3

Reviewer 1 Report

None