Optimization, Probiotic Characteristics, and Rheological Properties of Exopolysaccharides from Lactiplantibacillus plantarum MC5
Round 1
Reviewer 1 Report
Dear authors,
Your manuscript is interesting and may be attractive for its use for commercial purposes. I consider it valuable that the manuscript contains a detailed procedure of the methods used with formulas. But some information must be completed:
Introduction
It would be appropriate to include information on the reclassification of the genus Lactobacillus in the introduction.
3.1 Materials
What methods were used to identify and confirm the strain used?
Author Response
Response to Reviewer 1 Comments
Dear Editor,
We appreciate you and the reviewers for your precious time in reviewing our paper and providing valuable comments. It was your valuable and insightful comments that led to possible improvements in the current version. The authors have carefully considered the comments and tried our best to address every one of them. We hope the manuscript after careful revisions meet your high standards. The authors welcome further constructive comments if any. Below we provide the point-by-point responses to the reviewer’s comments. All modifications in the manuscript are in red, using the tracking function.
Point 1: It would be appropriate to include information on the reclassification of the genus Lactobacillus in the introduction.
Response 1: Please we have added the information on the reclassification of the genus Lactobacillus plantarum MC5 in the introduction. Please check lines 68-69.
Point 2: What methods were used to identify and confirm the strain used in 3.1 Materials?
Response 2: Please strain MC5 was first identified as Lactobacillus plantarum by 16S rRNA sequencing technology and was later identified as Lactobacillus plantarum by whole genome technology. The phylogenetic tree of strain MC5 is shown below. Please check 3.1 Materials and Figure 1.
Author Response File: 
Author Response.pdf
Reviewer 2 Report
1. The title does not reflect the content of the text, in the title Subcritical water-assisted extraction is treated as a highlight however in the abstract is mentioned that the best treatment was a combination with hypochlorite digestion. It is necessary to align the title to the relevance of the results, especially because in the text it is mentioned that chemical agents such as hypochlorite are a danger to the environment and health (I totally agree), however, their best result is obtained by using an amount of hypochlorite that although it is low, it still has the same effect. It is recommended to contextualize and align the text with the title, reflecting in a clear and concise manner the contribution of the work in context with the results obtained.
2. The document indicates that the strain used in the work was isolated and specifically specifies that it is L. plantarum MC5, but does not show any evidence of macro or microscopic morphological characterization or molecular identification. In order to be sure which strain, it is, it is necessary to present molecular identification. Since subcritical water assisted extraction (and its impact) is, in my opinion, the most relevant contribution of the work, it would be very important to explain it in more detail using a diagram or scheme that allows understanding the process and knowing the reactor so that the process can be reproducible.
3. It is necessary to indicate which exopolysaccharide is being produced or, failing that, once you are sure by molecular identification of the identity of the microorganism, you can inquire about the possible identities of the EPS produced.
4. It is necessary to update the references as most of them (60%) are prior to 2015.
Author Response
Response to Reviewer 2 Comments
Dear Editor,
We appreciate you and the reviewers for your precious time in reviewing our paper and providing valuable comments. It was your valuable and insightful comments that led to possible improvements in the current version. The authors have carefully considered the comments and tried our best to address every one of them. We hope the manuscript after careful revisions meet your high standards. The authors welcome further constructive comments if any. Below we provide the point-by-point responses to the reviewer’s comments. All modifications in the manuscript are in red, using the tracking function. Please see the attachment.
Point 1: The title does not reflect the content of the text, in the title Subcritical water-assisted extraction is treated as a highlight however in the abstract is mentioned that the best treatment was a combination with hypochlorite digestion. It is necessary to align the title to the relevance of the results, especially because in the text it is mentioned that chemical agents such as hypochlorite are a danger to the environment and health (I totally agree), however, their best result is obtained by using an amount of hypochlorite that although it is low, it still has the same effect. It is recommended to contextualize and align the text with the title, reflecting in a clear and concise manner the contribution of the work in context with the results obtained.
Response 1: Thank you for your valuable advice. But this paper does not cover the content Subcritical water-assisted extraction and hypochlorite digestion. The main content of this paper is to optimize the exopolysaccharide production conditions of Lactobacillus plantarum MC5, and further study the probiotic and rheological properties of exopolysaccharides. Therefore, we have not modified the title of the article for the time being.
Point 2: The document indicates that the strain used in the work was isolated and specifically specifies that it is L. plantarum MC5, but does not show any evidence of macro or microscopic morphological characterization or molecular identification. In order to be sure which strain, it is, it is necessary to present molecular identification. Since subcritical water assisted extraction (and its impact) is, in my opinion, the most relevant contribution of the work, it would be very important to explain it in more detail using a diagram or scheme that allows understanding the process and knowing the reactor so that the process can be reproducible.
Response 2: We agree with you. We have added the results of 16S rRNA identification of strain MC5 to the manuscript. Please check Figure 1and 2.1 section.
Point 3: It is necessary to indicate which exopolysaccharide is being produced or, failing that, once you are sure by molecular identification of the identity of the microorganism, you can inquire about the possible identities of the EPS produced.
Response 3: Thank you for your careful reading of our manuscript. Because the main content of this study was to optimize the conditions for the production of extracellular polysaccharide of Lactobacillus plantarum MC5 and to determine the probiotic properties (anti-digestive activity, simulated gastroenteric fluid tolerance, antioxidant activity) and rheological properties of the polysaccharide, the content of the composition and structure of extracellular polysaccharide was not included in the paper considering the overall structure and hierarchy of the paper. The next step for our team is to determine the structure of the exopolysaccharides produced by strain MC5 and write a paper focusing on the composition, structure, and molecules of exopolysaccharides produced by Lactobacillus plantarum MC5.
Point 4: It is necessary to update the references as most of them (60%) are prior to 2015.
Response 4: Thank you for pointing this out. We have updated the references before 2015. Please check the references in the manuscript.
Reviewer 3 Report
This manuscript titled " Optimization, probiotic characteristics, and rheological properties of exopolysaccharides from Lactobacillus plantarum MC5" is appropriate for the journal. There is originality and relevance in the research. Authors should make minor formatting corrections throughout the manuscript. Comments are specified in the document.
Comments for author File: Comments.pdf
Author Response
Response to Reviewer 3 Comments
Dear Editor,
We appreciate you and the reviewers for your precious time in reviewing our paper and providing valuable comments. It was your valuable and insightful comments that led to possible improvements in the current version. The authors have carefully considered the comments and tried our best to address every one of them. We hope the manuscript after careful revisions meet your high standards. The authors welcome further constructive comments if any. Below we provide the point-by-point responses to the reviewer’s comments. All modifications in the manuscript are in red, using the tracking function. Please see the attachment.
Cmments and Suggestions for Authors: This manuscript titled "Optimization, probiotic characteristics, and rheological properties of exopolysaccharides from Lactobacillus plantarum MC5" is appropriate for the journal. There is originality and relevance in the research. Authors should make minor formatting corrections throughout the manuscript. Comments are specified in the document.
Point 1: Lactobacillus plantarum MC5. In particular, the authors must attend, and homogenize to a single format, the one indicated in the authors' guide.
Response 1: Thank you for pointing this out. We have homogenized “Lactobacillus plantarum MC5” to “L. plantarum MC5” According to the author's guide. Please check them in the manuscript.
Point 2: Column main text (EPS production), omit on other charts to avoid text clutter in Figure 3.
Response 2: We agree with you. We have omitted the EPS production from the other charts in Figure 3. Please check Figure 3.
Point 3: Requires improving the presentation of the graphics, particularly in the format of letters.
Response 3: Please we have changed the format of letters in Figure 3. Please check Figure 3.
Point 4: Note of Table 3 moves to table feet.
Response 4: Thank you for your valuable advice. We have moved the note of Table 3 to the table feet. Please check lines 158-159.
Point 5: Uniform format of units throughout the manuscript.
Response 5: Please we have unified the units in the manuscript. Please check the manuscript.
Point 6: Correct wording, redundant, add bibliographic citation in line 176.
Response 6: Please we have corrected this sentence and added bibliographic citation. Please check lines 176-177.
Point 7: Place (a) and (b) off the graph in Figure 4.
Response 7: Please we have placed (a) and (b) off the graph in Figure 4. Please check Figure 4.
Point 8: Change Nacl to NaCl in Figure 4.
Response 8: Please we have changed Nacl to NaCl in Figure 4. Please check Figure 4.
Point 9: Place (a), (b), (c), and (d) off the graph in Figure 6.
Response 9: Please we have placed (a), (b), (c), and (d) off the graph in Figure 6. Please check Figure 6.
Point 10: ”in vitro “ in italics.
Response 10: Please we have changed ”in vitro “ to ”in vitro “ in the manuscript. Please check the manuscript.
Point 11: Standardize abbreviations throughout the document in the 3.2 section.
Response 11: Please we have normalized abbreviations throughout the document in the 3.2 section and added an abbreviations list at line 473. Please check them in the 3.2 section and line 473.
Point 12: ph ? in the 3.4.1 section.
Response 12: Please the pH is 6.4 in the 3.4.1. Please check it in the 3.4.1 section.
Point 13: Change 500mL to 500 mL in line 366.
Response 13: Please we have changed 500mL to 500 mL in line 366. Please check line 366.
Point 14: Change 2 mol/L to 2 Mol/L in line 367.
Response 14: Please we have changed 2 mol/L to 2 Mol/L in line 367. Please check line 367.
Point 15: correct units 0.1 mM in line 376.
Response 15: Please we have changed 0.1 mM to 0.1 mMol/L in line 376. Please check line 376.
