Review Reports - Expanding the Horizons of <i>Saccharomyces cerevisiae</i>: Nutrition, Oenology, and Bioethanol Production

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The content of this review is comprehensive, and the research content can also arouse the interest of researchers. Some of the problems are as follows:

LINES 309-313 Can supplement and summarize the relevant applications at this stage by illustrating the above issues with examples? What innovative end synergistic microbial reactions have been employed, such as non-high-temperature or short-time reactions? What is the future trend? If applied, what are the associated costs, and how much do they save compared to traditional technologies?

In FIGURE. 1- FIGURE. 3, there are some words and chemical formulas that are not clear. Could you please further enrich and improve these diagrams according to the content of the review?

The form of the article needs to be proofread again, such as:

LINE 138 [32] is missing a period.

LINE 146 Is a new segment and does it require a space before it?

LINE 200 Where are the references placed?

LINE 439 The last sentence is missing a period.

LINE 489 Is reference 2014 missing representation?

LINE 559 Is Fig.1C correct?

Author Response

Comment 1:

Response 1:

I appreciate the reviewer’s insightful comments. In response, I have supplemented and summarized the relevant applications with additional examples and context. The revised text now includes examples of innovative microbial reactions, such as enzymatic hydrolysis at ambient temperatures and the synergistic co-cultivation of microbes, which reduce processing times and energy consumption. Future trends, including the use of multi-enzyme cocktails and advancements in metabolic engineering, have been highlighted to illustrate the potential for further innovation. Additionally, I have addressed the economic implications by providing cost comparisons with traditional technologies, demonstrating significant cost savings. Lines 293-318

Comment 2:

In FIGURE. 1- FIGURE. 3, there are some words and chemical formulas that are not clear. Could you please further enrich and improve these diagrams according to the content of the review?

Response 2:

All figures are revised

Comment 3:

The form of the article needs to be proofread again:

Response 3:

Thank you for pointing out these issues. The manuscript has been proofread and updated accordingly, examples as follows:

LINE 138: A period has been added after the reference. Revised Line 136

LINE 146: A space has been inserted before the new segment. Revised Line 144

LINE 200: The references have been properly placed and cited in Table 1.

LINE 439: The last sentence has been corrected to include a period. Revised Line 470

LINE 489: The missing reference (2014) has been properly added. Revised Line 520

LINE 559: deleted.

Reviewer 2 Report

Comments and Suggestions for Authors

In this review manuscript, the author systematically summarizes the recent achievements of Saccharomyces cerevisiae in the fields of nutrition, oenology, and bioethanol production. Specifically, the manuscript focuses on the introduction of specialty S. cerevisiae, highlighting cutting-edge methodologies for their isolation and optimization. Overall, this review offers a comprehensive and novel perspective on S. cerevisiae, providing a valuable and accessible introduction for readers interested in this research field. However, a few points in the manuscript required attention and revision. I suggest that the paper be considered for publication in Sustainability after addressing the following concerns and suggestions.

1. Figure clarity and readability: The Figure in this manuscript contains substantial information, making it comprehensive. However, the schematic diagram is somewhat unclear due to low resolution, and the font size is too small. I suggest that the authors enlarge both the diagram and text in the revised manuscript for improved readability.

2. Inconsistent formatting: In the main text, some words are inconsistently formatted. For example, the S. cerevisiae should be italicized, but it is not in several instances, including lines 15, 17, 21, 122, 199-200, 777, and 1228. Additionally, other terms should also follow proper formatting, such as “Yarrowia and Pichia” in line 127, and “Zymomonas mobilis” in line 1164. The authors are advised to carefully review and correct similar issues throughout the manuscript before it is accepted.

3. Abbreviation: It is recommended to introduce the abbreviation “ROS” for Reactive Oxygen Species in line 458 to ensure consistency and clarity.

4. Lack of necessary citations: The first two paragraphs of the second section, “2. Why S. cerevisiae” (lines 95-113), lack sufficient citations to support the statements. I suggest the authors add some references to better substantiate the content.

Author Response

Comment 1:

Figure clarity and readability: The Figure in this manuscript contains substantial information, making it comprehensive. However, the schematic diagram is somewhat unclear due to low resolution, and the font size is too small. I suggest that the authors enlarge both the diagram and text in the revised manuscript for improved readability.

Response

I appreciate the reviewer’s feedback regarding the clarity and readability of the figures. In response, I have revised all three figures in the manuscript to improve their resolution and enhance readability. Specifically:

Diagram clarity: The schematic diagrams have been enhanced to ensure high resolution.

Font size: The text within the figures has been enlarged to improve legibility.

Overall layout: Adjustments have been made to the figure layouts to make them more visually appealing and easier to interpret.

Comment 2:

Inconsistent formatting: In the main text, some words are inconsistently formatted. For example, the S. cerevisiae should be italicized, but it is not in several instances, including lines 15, 17, 21, 122, 199-200, 777, and 1228. Additionally, other terms should also follow proper formatting, such as “Yarrowia and Pichia” in line 127, and “Zymomonas mobilis” in line 1164. The authors are advised to carefully review and correct similar issues throughout the manuscript before it is accepted.

Respond 2:

Thank you for the detailed feedback. I have carefully reviewed the manuscript and made the necessary corrections to address the inconsistencies in formatting. A thorough review was conducted to ensure all terms follow standard scientific formatting conventions. Examples Lines 15, 17, 21, 122, 126, etc.

Comment 3:

Abbreviation: It is recommended to introduce the abbreviation “ROS” for Reactive Oxygen Species in line 458 to ensure consistency and clarity.

Response 3:

ROS added Line 489

Comment 4:

Lack of necessary citations: The first two paragraphs of the second section, “2. Why S. cerevisiae” (lines 95-113), lack sufficient citations to support the statements. I suggest the authors add some references to better substantiate the content.

Response 4:

Revised and all the necessary references were added. Lines 95-113

Reviewer 3 Report

Comments and Suggestions for Authors

The manuscript entitled “Expanding the horizonts of Saccharomyces cerevisiae: nutrition, oenology, and bioethanol production” presented a bibliographic revision of S. cerevisiae studies focused on nutrition, wine making and bioethanol production. In my opinion, only probiotic use of yeasts section suffers from a lack of depth. Overall, this review is interesting and valuable; the work seems to be well structured and written. However, the manuscript needs some minor changes:

L 66-70: I suggest to move this paragraph to section 6.
L 82: At this point, maybe could be necessary to explain the differences between “specialty” and “conventional” S. cerevisiae strains, instead of at lines 146 to 150.
L 108: The author could check the entire manuscript to insert a space between the numeric values from the unit: temperature data and numerical values, v.g. “2ºC”, the correct form is “2 ºC”.
L 122, 127: Please use italic letters: “non-Saccharomyces”.
L 125: “Its extensive applications across industries, ranging from bread and beer to wine and bioethanol,” This sentence should be deleted, because of it is repetitive.

6. L 256: please, to change: “Figure 2” with “(Figure 2)”.

L 318: “(OSAM)” this acronym it is not necessary.
L 320: the author should select only one “stile” to insert the units throughout the manuscript, please choose between: “mg/g” or “mg g⁻¹”.
L 338: The author should explain the origin of the OTA found in the yeast-based products, and that it is adsorbed by yeasts during the fermentation of cereals that may be contaminated to a greater or lesser extent.
L 384: please, change “(Regodón Mateos et al., 2006b)” with "[217]”.
L 408: please, amend this sentence as “ALE is also being explored…”.
L 439: please, insert a full stop after “yields”.
L 489, 548: introduce brackets at the reference cites “Pereira et al. (2014)”; “Pim et al. (1998)”.
L 559: Where is the Figure 1C in the text?
L 678, 880, 1051, 1106: please, change “Adaptive laboratory evolution” with its acronym.
L 890: insert the correct units at “8º alcohol”.
L 899: please, change “(Oliveira et al. 2014)” with “[xx]”. This study is not included in the references list.
L 984: please, cite the references as “[xx]” according to author´s instructions.
L 1012-1013: please, complete this sentence as “…with a 50% theoretical ethanol yield…” and “…achieving theoretical ethanol yields above 70%”.
L 1061-1063: please, delete this duplicate sentence.
L 1155: delete “hydroxymethylfurfural”.
L 1164, 1205, 1228: please, use italic letters in all genes.

23. In the References chapter, many references cited are not according to author’s instructions. For instance, L 1285, 1309, 1372, 1379, 1415, 1442, 1459, 1478, 1487… Please, the author has to carefully verify line by line, the format of references should be standardized according to the requirement of SUSTAINABILITY journal.

Author Response

Response

Thank you for pointing this out. I have expanded the section on the probiotic use of yeasts to provide greater depth and detail. The revised section now includes a more comprehensive discussion of Saccharomyces cerevisiae's health benefits, such as its role in glycemic control, immune modulation, and antioxidant properties. Additionally, specific mechanisms like gut flora restoration, pathogen inhibition, and biofilm formation have been highlighted, alongside its applications in managing metabolic disorders and supporting immune function. For further context, references to recent reviews that explore these aspects in depth have been incorporated to ensure the section is robust and informative. Lines 342-356

Comment 1:

L 66-70: I suggest to move this paragraph to section 6.

Response 1:

I have moved this paragraph to section 6 as suggested. Lines 817-820

Comment 2:

L 82: At this point, maybe could be necessary to explain the differences between “specialty” and “conventional” S. cerevisiae strains, instead of at lines 146 to 150.

Response 2:

The explanation of differences between “specialty” and “conventional” S. cerevisiae strains has been relocated from lines 146–150 to this section for better contextual clarity. Lines 81-84

Comment 3:

L 108: The author could check the entire manuscript to insert a space between the numeric values from the unit: temperature data and numerical values, v.g. “2ºC”, the correct form is “2 ºC”.

Response 3:

The manuscript has been thoroughly reviewed, and spaces have been added between numeric values and their units throughout (e.g., "2 ºC" instead of "2ºC"). Examples lines 108, 134, 135, etc.

Comment 4:

L 122, 127: Please use italic letters: “non-Saccharomyces”.

Response 4:

The term non-Saccharomyces has been italicized, as well as others where required. Examples Line 122, 126, 180, etc.

Comment 5:

L 125: “Its extensive applications across industries, ranging from bread and beer to wine and bioethanol,” This sentence should be deleted, because of it is repetitive.

Response 5:

The repetitive sentence regarding the extensive applications of S. cerevisiae has been deleted.

Comment 6:

L 256: please, to change: “Figure 2” with “(Figure 2)”.

Response 6:

Corrected Line 236

Comment 7:

L 318: “(OSAM)” this acronym it is not necessary.

Response 7:

Deleted

Comment 8:

L 320: the author should select only one “stile” to insert the units throughout the manuscript, please choose between: “mg/g” or “mg g⁻¹”.

Respond 8:

The unit style throughout the manuscript has been standardized to “mg/g” for consistency. Line 324.

Comment 9:

L 338: The author should explain the origin of the OTA found in the yeast-based products, and that it is adsorbed by yeasts during the fermentation of cereals that may be contaminated to a greater or lesser extent.

Response 9:

Added an explanation about the origin of OTA in yeast-based products. Lines 365-376.

Comment 10:

L 384: please, change “(Regodón Mateos et al., 2006b)” with "[217]”.

Response 10:

The citation has been updated as requested. Note that the citation numbering reflects adjustments made throughout the manuscript to align with structural and content changes.

Comment 11:

L 408: please, amend this sentence as “ALE is also being explored…”.

Response 11:

The sentence has been amended to “ALE is also being explored. Lines 446, 457, etc.

Comment 12:

L 439: please, insert a full stop after “yields”.

Response 12:

A full stop has been inserted after “yields”. Line 470

Comment 13:

L 489, 548: introduce brackets at the reference cites “Pereira et al. (2014)”; “Pim et al. (1998)”.

Response 13:

Brackets have been introduced for the reference citations. Lines 520, 578

Comment 14:

L 559: Where is the Figure 1C in the text?

Response 14:

Deleted

Comment 15:

L 678, 880, 1051, 1106: please, change “Adaptive laboratory evolution” with its acronym.

Response 15:

The term “Adaptive laboratory evolution” has been replaced with its acronym (ALE) consistently throughout these lines, as mentioned above response 11.

Comment 16:

L 890: insert the correct units at “8º alcohol”.

Response 16:

Revised. Lines 923-926.

Comment 17:

L 899: please, change “(Oliveira et al. 2014)” with “[xx]”. This study is not included in the references list.

Response 17:

Updated the citation style for this reference to “[xx]” and noted that this study needs to be added to the references list. Lines, 934, 1818

Comment 18:

L 984: please, cite the references as “[xx]” according to author´s instructions.

Response 18

Revised. Line 1019.

Comment 19:

L 1012-1013: please, complete this sentence as “…with a 50% theoretical ethanol yield…” and “…achieving theoretical ethanol yields above 70%”.

Response 19:

Completed the sentences to include “...with a 50% theoretical ethanol yield...” and “...achieving theoretical ethanol yields above 70%”. Lines 1047, 1048.

Comment 20:

L 1061-1063: please, delete this duplicate sentence.

Response 20:

Revised and deleted. Lines 1090, 1095.

Comment 21:

L 1155: delete “hydroxymethylfurfural”.

Response 21:

Removed the mention of “hydroxymethylfurfural” from the text.

Comment 22:

L 1164, 1205, 1228: please, use italic letters in all genes.

Response 22

All gene names have been italicized consistently. Example lines 513, 514, 535, etc.

Comment 23.

In the References chapter, many references cited are not according to author’s instructions. For instance, L 1285, 1309, 1372, 1379, 1415, 1442, 1459, 1478, 1487… Please, the author has to carefully verify line by line, the format of references should be standardized according to the requirement of SUSTAINABILITY journal.

Response 23:

The formatting of all references has been verified and standardized according to the requirements of the Sustainability journal. Lines from 1297 till end.