Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The article is very interesting as it presents the most relevant aspects that must be considered in the hyaluronic acid (HA) production, specifically when produced in a biorefinery and present a review about different processes being specified the type of bioreactor, growing conditions, C and N source, supplements and additives, the molecular weight and HA production. Are described too the economic aspects and mentioned several studies about environmental sustainability of HA production in both cases.

The article is well described and detailed and is an asset to all those researchers, students, businesspeople, public and private sector personnel who are implementing studies about the HA.

The conclusions are very well detailed, described and succinct as they present the most important aspects that must be considered in each topic. However, I consider that two paragraphs (one for each point) should be included with the most relevant aspects of point 5 and point 6 because these are not mentioned in the conclusions. Taking all these criteria into consideration, I consider that the article should be accepted with minor revisions.

I recommend the following changes based on text:

Line 24: please replace “GBAER” by “Environmental Biotechnology and Renewable Energies Group (GBAER, for its acronym in Spanish)”.

Lines 32 and 168: please replace “C and N” by “Carbon (C) and Nitrogen (N)”.

Line 45: please delete the word “management”.

Line 70: please replace “high-value value-added” by “high-value-added”.

Line 85: please replace “ethanol, biodiesel, methane, and hydrogen” by “as ethanol, biodiesel, methane, hydrogen, synthesis gas, eletricity/heat and bio-oil”.

Line 91: please replace “(methane) biogas” by “biogas (methane)”.

Lines 94 and 95: please replace “t CO₂/TJ” by “t CO₂ TJ^-1”. It is necessary to uniformize all the units.

Line 96: please replace “value added” by “value-added”.

Line 134: please replace “Phys-chem” by “physicochemical”.

Line 135: please replace “bio energies” by “bioenergies”.

Line 143: please replace “ISO standards” by “International Organization Standardization (ISO)”.

Line 149: please replace “Is” by “is”.

Lines 173, 364, 374 and 736: please replace “AH” by “HA”.

Line 191, 195 and 530: please replace “various” by “several”.

Line 214: please replace “y” by “and”.

Line 231: please replace “electricity, organic acids” by “electricity, heat, organic acids”.

Line 245: please replace “52] In” by “52]. In”.

Line 248: please replace “GBAER” by “Environmental Biotechnology and Renewable Energies Group type...”. You should never start a sentence with acronyms.

Lines 250, 254, 255, 257, 272 and 282: please replace “Principle” by “principle”.

Line 255: please replace “footprint, (iv) Principle” by “footprint and (iv) Principle”.

Line 277: Why is there a parenthesis in this sentence?.

Line 281: please replace “25].” by “25]).”.

Line 295: please replace “kg/CO₂e” by “kg CO₂e^-1”.

Line 316: please replace “hyaluronic acid (HA stage)” by “HA (HA stage)”.

Figure 1: in the “Stage H” the hydrogen stream exits through the top and in the “Stage M” the biogas stream exits through the top too. In all cases, the streams with gases exits through the top.

Line 325: please replace “HMEZS” by “HMZS”.

Line 357: please replace “MRI” by “Magnetic resonance imaging (MRI)”.

Line 415: please replace “Peptone” by “peptone”.

Line 426: please replace “(ii)” by “(iii)”.

Line 434: please replace “5 °C” by “5°C”.

Line 469: please replace “MgSO₄, K₂HPO₄, KH₂PO₄, and K₂SO₄.” by “magnesium sulphate (MgSO₄), dipotassium phosphate (K₂HPO₄), monopotassium phosphate (KH₂PO₄) and potassium sulphate (K₂SO₄)”.

Line 471: please replace “CaCl₂, FeSO₄, (NH₄)₂SO₄, MnCl₂, ZnSO₄, and CuSO₄” by “calcium chloride (CaCl₂), iron(II) sulphate (FeSO₄), ammonium sulphate ((NH₄)₂SO₄), manganese(II) chloride (MnCl₂), zinc sulphate (ZnSO₄), and copper(II) sulphate (CuSO₄)”.

Lines 471-474: please replace “as Alanine, Glycine, Leucine, Isoleucine, Valine, Methionine, Serine, Threonine, Proline, Asparagine, Arginine, Lysine, Ornithine, Glutamate, Aspartate, Phenylalanine, Tryptophan, Histidine, Cysteine, adenine, Guanine,…” by “alanine, glycine, leucine, isoleucine, valine, methionine, serine, threonine, proline, asparagine, arginine, lysine, ornithine, glutamate, aspartate, phenylalanine, tryptophan, histidine, cysteine, adenine, guanine, ...”.

Line 476: please replace “Beta-glycerophosphate” by “beta-glycerophosphate”.

Lines 497-501: I don´t understand the sentence between these lines.

Line 517, Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, line 720, Table 9 and lines 922, 923, 929, 932, 934 and 936: please replace “g/L” by “g L^-1”.

Line 571: please replace “High” by “high”.

Line 610: please replace “E. coli” by “E. coli”.

Line 618: please replace “26, 30,…” by “26°C, 30°C,…”.

Table 5: please replace “(NH₄)2SO₄” by “(NH₄)₂SO₄”.

Table 6: please replace “L/L” by “L L^-1”.

Table 7: please replace “v/v” by “v v^-1”.

Line 691: please replace “municipal solid waste” by “MSW” and “organic fraction of municipal solid waste” by “OFMSW”.

Line 758: please replace “billion)” by “billion).”.

Line 761: please replace “€ 1,500” by “1,500 €”.

Line 113: please replace “€ 4,000” by “4,000 €”.

Lines 772 and 773: please replace “€/g” by “€ g^-1”.

Line 782: please replace “€ 2.4” by “2.4 €”.

Line 784: please replace “€ 1.4” by “1.4 €”.

Line 799: please replace “$/kg” by “USD kg^-1”.

Lines 801 and 802: please replace “kg/year” by “kg year^-1”.

Line 803: please replace “abovementioned” by “above mentioned”.

Line 817: please replace “HA” by “hyaluronic acid”.

Lines 819-821: please replace “(Life Cycle Assessment Hyaluronic Acid Production, Environmental Impacts from Hyaluronic Acid Production)” by “(LCA HA production and environmental impacts from HA production)”.

Lines 831-832: please replace “International Organization for Standardization (ISO) standards” by “ISO standards”.

Lines 866-867: please replace “Precautionary Principle.” by “precautionary principle”.

Line 875: please replace “Hyaluronic Acid.” by “hyaluronic acid.”.

Table 9: please replace “ND” by “NA”.

Author Response

Response to Reviewers Document

 

 

 

Content

 

 

 

 

Section A.  Editors’ Decision and Comments………………………………………...2

 

Section B. Answers to Reviewers’ Comments………………………………………. 5

 

B1. Answers to Reviewer 1 Comments……………………………………………….5

 

B2. Answers to Reviewer 2 Comments………………………………………………23

 

References………………………………………………………………………………33

 

 

 

Section A.  Editors’ Decision and Comments

 

Below please find the text sent by Fermentation Editorial Office regarding the first evaluation of our MS

[Fermentation] Manuscript ID: fermentation-3013052 - Major Revisions

Fermentation Editorial Office <[email protected]>      16 de mayo de 2024, 8:47 a.m.

Responder a: Cathy Lu <[email protected]>, Fermentation Editorial Office <[email protected]> Para: Héctor Mario Poggi-Varaldo <[email protected]>

CC: Guadalupe Pérez-Morales <[email protected]>, Teresa Ponce-Noyola <[email protected]>, Abigail Pérez-Valdespino <[email protected]>, Everardo Curiel-Quesada <[email protected]>, Juvencio Galíndez-Mayer <[email protected]>, Nora Ruiz-Ordaz <[email protected]>, Perla Xochitl Sotelo-Navarro

<[email protected]>, Fermentation Editorial Office <[email protected]>, Cathy Lu <[email protected]> Dear Dr. Poggi-Varaldo,

Thank you again for your manuscript submission:

 

Manuscript ID: fermentation-3013052 Type of manuscript: Review

Title: A review of the production of Hyaluronic Acid in the context of its integration into GBAER-type biorefineries

Authors: Guadalupe Pérez-Morales, Héctor Mario Poggi-Varaldo *, Teresa Ponce-Noyola, Abigail Pérez-Valdespino, Everardo Curiel-Quesada, Juvencio Galíndez-Mayer, Nora Ruiz-Ordaz, Perla Xochitl Sotelo-Navarro

Received: 30 Apr 2024

E-mails: [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Industrial Fermentation https://www.mdpi.com/journal/fermentation/sections/Industrial_fermentation Microbial Fermentation of Organic Wastes for Production of Biofuels and Biochemicals 2.0 https://www.mdpi.com/journal/fermentation/special_issues/79SH3ZEMTE

 

Your manuscript has now been reviewed by experts in the field. Please find your manuscript with the referee reports at this link:

https://susy.mdpi.com/user/manuscripts/resubmit/37a40c4ce5bd266ebee717ffffc1e9ed

Please revise the manuscript according to the referees' comments and upload

the revised file within 10 days.

 

Please use the version of your manuscript found at the above link for your revisions.

 

(I)      Please check that all references are relevant to the contents of the manuscript.

 

Our answer to Editor: We have checked all references for relevance to the article

 

(II)     Any revisions to the manuscript should be highlighted, such that any changes can be easily reviewed by editors and reviewers.

 

Our answer to Editor: We highlighted all the revisions in the RMS with red characters and yellow background

 

(III)    Please provide a cover letter to explain, point by point, the details

of the revisions to the manuscript and your responses to the referees’ comments.

 

Our answer to Editor: We are providing two documents: A cover letter that explains the modifications made to the RMS and describes other issues, and a Response-to-Reveiwers document that addresses, point by point, the details of the revisions to the RMS as well as our answers to Reviewers’ comments

 

(IV)   If the reviewer(s) recommended references, please critically analyze them to ensure that their inclusion would enhance your manuscript. If you believe these references are unnecessary, you should not include them.

 

Our answer to Editor: Reviewers did not recommend references. However, we have conducted approx. three literature searches  on certain subjects to write selected responses to Reviewers. Therefore, some new references will appear in the RMS. All of them are relevant to the content of the RMS.

 

(V)    If you found it impossible to address certain comments in the review reports, please include an explanation in your appeal.

 

Our answer to Editor: We feel that we could address all the Comments of the Reviewers. In fact, we agree with the remarks made by Reviewers. We feel that we did not have to defend a statement of ours or to deny a requested modification.

 

(VI)   The revised version will be sent to the editors and reviewers.

 

Our answer to Editor: We are sending to Editors and Reviewers the RMS, the Response-to-Reviewers document, and the Conveying Letter Revised.

 

(VII) If one of the referees has suggested that your manuscript should undergo extensive English revisions, please address this issue during revision. We propose that you use one of the editing services listed at https://www.mdpi.com/authors/english or have your manuscript checked by a colleague fluent in English writing.

 

Our answer to Editor: Only Reviewer 2 suggested minor revision of English. We subjected the RMS to two rounds of Grammarly revisions, and one round of revision by Trinka. Also, Dr Juvencio Galíndez-Mayer, one of our co-authors and very fluent in English, corrected the RMS for English usage.

Grammarly score was 99%

 

(VIII) We would like to draw your attention to the status of this invitation “Publish Author Biography on the webpage of the paper” -

https://susy.mdpi.com/user/manuscript/author_biography/37a40c4ce5bd266ebee717ffffc1e9ed.

If you decide to publish your biography, please remember to fill in it before your paper is accepted.

 

Our answer to Editor: We have sent our biographical capsule to Dr C. Lu for her uploading to MDPI webpage.

 

Please do not hesitate to contact us if you have any questions regarding the revision of your manuscript or if you need more time. We look forward to hearing from you soon.

 

Kind regards, Ms. Lucille Liu

E-Mail: [email protected]

 

 

 

Section B. Answers to Reviewers’ Comments

 

 

B1. Answers to Reviewer 1 Comments

 

 

Notation

BNBP bionanobioparticle

BRF               biorefinery

GBAER         Spanish abbreviation for the Environmental Biotechnology and

Renewable Energies Group

GT                  genetically transformed

HA                  hyaluronic acid

LL                   line or lines

MS                  manuscript

PP                  page or pages

RMS               revised manuscript

 

 

Reviewer 1 wrote:

“Comments and Suggestions for Authors

The article is very interesting as it presents the most relevant aspects that must be considered in the hyaluronic acid (HA) production, specifically when produced in a biorefinery and present a review about different processes being specified the type of bioreactor, growing conditions, C and N source, supplements and additives, the molecular weight and HA production. Are described too the economic aspects and mentioned several studies about environmental sustainability of HA production in both cases.

The article is well described and detailed and is an asset to all those researchers, students, businesspeople, public and private sector personnel who are implementing studies about the HA.

The conclusions are very well detailed, described and succinct as they present the most important aspects that must be considered in each topic. However, I consider that two paragraphs (one for each point) should be included with the most relevant aspects of point 5 and point 6 because these are not mentioned in the conclusions. Taking all these criteria into consideration, I consider that the article should be accepted with minor revisions.

          I recommend the following changes based on text:”

 

Answer:    we thank Reviewer 1 for his/her proactive summary of our article and its potential impact if published. Please see the detailed answers to Comments of Reviewer 1 below, one by one.

 

Comment 1

“Line 24: please replace “GBAER” by “Environmental Biotechnology and Renewable Energies Group (GBAER, for its acronym in Spanish)”.

 

Answer: We  appreciate your remark. However, the use of the abbreviation  “GBAER” in this line is to refer to the name assigned to this type of refineries, so in the text we place in parentheses the explanation of the concept to the type of biorefineries that we deal with in this article (biorefineries that use municipal solid waste as raw material, not the particular name of the Group).

Additionally, at the suggestion of another reviewer, the abstract had to be modified to 200 words to comply with the journal's guidelines. therefore, the word "GBAER" was kept so as not to exceed the required size of the abstract.

 

Before it read:

“Biorefineries (BRF) that process the organic fraction of municipal solid waste (OFMSW) and generate bioproducts and bioenergies have attracted attention because they can simultaneously address energy and environmental problems/needs. Obtaining high-value-added products (VAP) plays a determining role in achieving economic sustainability. Hyaluronic acid (HA) is a VAP that could be integrated into modern biorefinery systems. The objective of this article was to conduct a critical review of the microbial production of HA (MPHA) and its production profile for its integration into a GBAER-type BRF (a type of BRF based on organic wastes) and to identify the environmental and economic sustainability aspects of the modified BRF that would confirm it as a sustainable option. Two types of pathogenic Streptococci that naturally produce AH, exhibited the highest HA production: S. equi ssp. equi and S. zooepidemicus. The first has produced up to 12.0 gL^-1, while the second yielded 6.6 gL^-1. Interestingly, these bacteria reached a production of up to 29.4 gL^-1 when subjected to genetic transformation (GT). A strain of Corynebacterium glutamicum was also identified with a maximum HA production of 71.4 gL^-1. Indeed, this strain is not a naturally HA-producing bacterium, but it achieved that high production after GT. We found that the MPHA under organic wastes as sources of C and N is scarce. Only Streptococcus zooepidemicus without GT showed a moderate HA production of 6.7 gL^-1 when starch was used as an alternative C source and yeast extract as a conventional source of N. When an alternative source of N was supplemented (i.e., cheese whey or fermented corn liquor) or when alternative sources of C and N were used simultaneously, HA production was lower (only 4 and 3.54 gL^-1, respectively.) In addition, our review identified other knowledge gaps that must be addressed regarding aspects of process scale-up, HA industrial production, economic aspects, and environmental sustainability of the MPHA. The integration of MPHA into GBAER-type BRF using saccharified liquors as a source of C is promising, but at the same time, it constitutes a technical challenge. The studies of this specific integration and the necessary advances in other R&D areas identified above will allow for evaluating the environmental and economic sustainability of the completely modified GBAER-BRF and other systems that want to integrate MPHA into their background processes.”

 

Now it reads (in the revamped Abastract, modified according to Reviewer 1 Comment 55):

 

“Biorefineries (BRF) that process the organic fraction of municipal solid waste and generate bioproducts and bioenergies have attracted attention because they can simultaneously address energy and environmental problems/needs. The objective of this article was to critically review the microbial production of hyaluronic acid (MPHA) and its production profile for its integration into a GBAER-type BRF (a type of BRF based on organic wastes) and to identify the environmental and economic sustainability aspects of the modified BRF that would confirm it as a sustainable option. It was found that the MPHA by selected strains of pathogenic Sterptococci wer moderate to high, although the trend to work with genetically transformed (GT) (innocuous) bacteria is gaining momentum. For instance, A GT strain of Corynebacterium glutamicum reached a maximum HA production of 71.4 gL^-1. We found that the MPHA reports that use organic wastes as sources of carbon (C) and nitrogen (N) is scarce. When alternative sources of C and N were used simultaneously, HA production by S. zooepidemicus was lower than that with conventional sources. We identified several knowledge gaps that must be addressed regarding aspects of process scale-up, HA industrial production, economic feasibility and sustainability, and environmental sustainability of the MPHA.”

 

Comment 2 


“Lines 32 and 168: please replace “C and N” by “Carbon (C) and Nitrogen (N)”.”

 

Answer: We thank Reviewer 1 for his/her comment. Attending to the suggestions, the names of chemical elements were added.

 

Before it read:

“MPHA under organic wastes as sources of C and N is scarce. Only Streptococcus zooepidemicus”

 

Now it reads:

“….as sources of carbon (C) and nitrogen (N) is scarce…”

 

Comment 3

 “Line 45: please delete the word “management”.”

 

Answer: Thank you for your comment; the word “management” was deleted.

 

Before it read:

“….municipal solid waste management----”

 

Now it reads:

“…municipal solid waste…”

 

Comment 4

“Line 70: please replace “high-value value-added” by “high-value-added”.

 

Answer: I appreciate your observation. Attending to the suggestions, the word was modified.

 

Before it read:

“….BRF can generate high-value value-added products….”

 

Now it reads:

“…BRF can generate high-value-added products….”

 

Comment 5

 “please replace “ethanol, biodiesel, methane, and hydrogen” by “as ethanol, biodiesel, methane, hydrogen, synthesis gas, electricity/heat and bio-oil”.”

 

Answer:

Thank you for your comment. We have implemented the suggested changes

 

Before it read:

“….range of biofuels such as ethanol, biodiesel, methane, and hydrogen….”

 

Now it reads: (LL84)

“…..range of biofuels such as ethanol, biodiesel, methane, hydrogen, synthesis gas, electricity /heat and bio-oil…..”

 

Comment 6

“Line 91: please replace “(methane) biogas” by “biogas (methane)”. “

 

Answer:

We are grateful to Reviewer 1 for his/her comment. We have modified the RMS.

 

Before it read:

“….In the case of (methane) biogas….”

 

Now it reads:

“……In the case of biogas (methane)….”

 

Comment 7

“Lines 94 and 95: please replace “t CO₂/TJ” by “t CO₂ TJ^-1”. It is necessary to uniformize all the units”

 

Answer: We have armonized the units throughout the RMS

 

Before it read:

“t CO₂/TJ”

 

Now it reads:

“ t CO₂ TJ^-1”

 

Comment 8

“Line 96: please replace “value added” by “value-added”.”

 

Answer: We have hyphenated the expression.

 

Before it read:

“…..A second advantage of BRF is that value added products (VAP)….”

 

Now it reads:

“…..A second advantage of BRF is that value-added products (VAP)…..”

 

Comment 9

“Line 134: please replace “Phys-chem” by “physicochemical”.”

 

Answer: we have modified the RMS accordingly.

 

Before it read:

“…..These BRF consist of a network of physical, Phys-chem, biological….”

 

Now it reads:(LL133)

“…..These BRF consist of a network of physical, physicochemical, biological….”

 

Comment 10

“Line 135: please replace “bio energies” by “bioenergies”.”

 

Answer: we appreciate the careful reading of the MS by Reviewer 1. We have replaced the suggested word.

 

Before it read:

“…..auxiliary processes that convert the OFMSW into bio energies and bioproducts….”

 

Now it reads:

“ ……auxiliary processes that convert the OFMSW into bioenergies and bioproducts……”

 

Comment 11

“Line 143: please replace “ISO standards” by “International Standards Organization (ISO)”.

 

Answer: We have replaced the expression

 

Before it read:

“aligned to ISO standards”

 

Now it reads:

“……aligned to International Standards Organization (ISO)…...”

 

Comment 12

“Line 149: please replace “Is” by “is”.”

 

Answer: I appreciate your observation. Attending to the suggestions, the word was replaced.

 

Before it read:

“….. It Is known that HA is a product….”

 

Now it reads:

“…..It is known that HA is a product….”

 

Comment 13

“Lines 173, 364, 374 and 736: please replace “AH” by “HA”.”

 

Answer: The changes were made in the lines 173, 364, 374 and 736:

 

Before it read:

“….AH….”

 

Now it reads:

“…..HA……”

 

Comment 14

“Line 191, 195 and 530: please replace “various” by “several”.”

 

Answer: The suggested changes were made in lines 191, 195 and 530:

 

Before it read:

“….various…..”

 

Now it reads:

 “….several…..”

 

Comment 15

Line 214: please replace “y” by “and”.

 

Answer: Thanks for the meticulous reading of our MS. The word “y” was replaced by “and”

 

Before it read:

“…..(astaxanthin, lutein, echinenone, chlorophyll y fucoxanthin)…..”

 

Now it reads:

“……(astaxanthin, lutein, echinenone, chlorophyll and fucoxanthin)……”

 

Comment 16

“Line 231: please replace “electricity, organic acids” by “electricity, heat, organic acids”.”

 

Answer: We thank Reviewer 1 for his/her comment. We have modified the RMS accordingly.

 

Before it read:

“…….ethanol, electricity, organic acids…..”

 

Now it reads:

“……ethanol, electricity, heat, organic acids……”

 

Comment 17

“Line 245: please replace “52] In” by “52]. In”.”

 

Answer: I appreciate your observation. To Attend it, Added missing punctuation. Additionally, because 11 new references were added, the numbering changed throughout the text.

 

Before it read:

“….[17, 19, 52] In this way…..”

 

Now it reads:

“…..[17, 19, 54]. In this way…..”

 

Comment 18

“Line 248: please replace “GBAER” by “Environmental Biotechnology and Renewable Energies Group type...”. You should never start a sentence with acronyms.”

 

Answer: We appreciate this remark.

 

Before it read:

“…..2.1 GBAER type biorefineries and the potential for integration of the microbial hyaluronic acid production…..”

 

Now it reads:  

“……2.1 Biorefineries of GBAER type and the potential for integration of the microbial hyaluronic acid production……”

 

Comment 19

“Lines 250, 254, 255, 257, 272 and 282: please replace “Principle” by “principle”.”

 

Answer: The suggested changes were made in lines 250, 254, 255, 257, 272 and 282:

 

Before it read:

“….Principle…..”

 

Now it reads: (LL 294-303)

“…….principle……”

 

Comment 20

“Line 255: please replace “footprint, (iv) Principle” by “footprint and (iv) Principle”.”

 

Answer: Suggested changes were implemented in the RMS

 

Before it read:

 “…..carbon footprint, (iv) principle of production……”

 

Now it reads:

“……..carbon footprint and (iv) principle of production…….”

 

Comment 21

“Line 277: Why is there a parenthesis in this sentence?.”

 

Answer: We appreciate your comment. We have rephrased the sentence.

 

Before it read:

“…..handling of waste (avoiding the discharge……”

 

Now it reads:

“…….handling of waste. This, in turn avoids the discharge…..”

 

Comment 22

“Line 281: please replace “25].” by “25]).”.”

 

Answer: Modification was made.

 

Before it read:

“……..(enzymes, bionanoparticles, succinic acid, [18, 25]…….”

 

Now it reads:

“………(enzymes, bionanoparticles, succinic acid, [18, 25])……..”

 

Comment 23

 

“Line 295: please replace “kg/CO₂e” by “kg CO₂e^-1”.”

 

Answer: We appreciate this remark. Please note that there was a mistake in the MS. The units should have read as kg CO₂ e without a backlash /

 

Before it read:

“….avoiding emissions of 128 kg/CO₂e for each ton of MSW and reducing eutrophication ….”

 

Now it reads:

“…..avoiding emissions of 128 kg CO₂e (ton MSW)^-1 and reducing eutrophication…..“

 

Comment 24

“Line 316: please replace “hyaluronic acid (HA stage)” by “HA (HA stage)”.”

 

Answer: Done.

 

Before it read:

“………hyaluronic acid (HA stage)………”

 

Now it reads:

“……HA  (HA stage)…..”

 

 

Comment 25

“Figure 1: in the “Stage H” the hydrogen stream exits through the top and in the “Stage M” the biogas stream exits through the top too. In all cases, the streams with gases exits through the top.”

 

Answer: Figure 1 was modified to indicate the outflow of gases in stages H and M

 

Before it read:

 

 

Now it reads:

 

 

Comment 26

“Line 325: please replace “HMEZS” by “HMZS”.”

 

Answer: Thank you for your comment. However, the correct name of the configuration is HMEZS. We modified Figure 1 to indicate the place where stage E is located (the organic acid extraction).

 

Before it read:

NA

 

Now it reads:

NA

 

 

Comment 27

“Line 357: please replace “MRI” by “Magnetic resonance imaging (MRI)”.”

 

Answer: Done

 

Before it read:

“…….a specific MRI contrast agent…..”

 

Now it reads: (LL 403)

“…..a specific Magnetic resonance imaging (MRI) contrast agent…….”

 

Comment 28

“Line 415: please replace “Peptone” by “peptone””

 

 

Answer: We have corrected the sentence

 

Before it read:

“Others growth factor can be Peptone and tryptone”

 

Now it reads:

“Other growth factors can be peptone and tryptone”

 

Comment 29

“Line 426: please replace “(ii)” by “(iii)”.”

 

Answer: We thank Reviewer 1 for his/her careful reading of our MS. The changes were made in line 426

 

Before it read:

“…..(ii) precipitation of the supernatant and (ii) centrifugation…..”

 

Now it reads:

“…….(ii) precipitation of the supernatant, and (iii) centrifugation…..”

 

Comment 30

“Line 434: please replace “5 °C” by “5°C”.”

 

Answer: Done.

 

Before it read:

“….. is dissolved in ethanol at 5 °C….”

 

Now it reads:

“..….is dissolved in ethanol at  5°C….”

 

Comment 31

“Line 469: please replace “MgSO₄, K₂HPO₄, KH₂PO₄, and K₂SO₄.” by “magnesium sulphate (MgSO₄), dipotassium phosphate (K₂HPO₄), monopotassium phosphate (KH₂PO₄) and potassium sulphate (K₂SO₄)”.”

 

Answer: Changes were implemented.

 

Before it read:

“……. such as MgSO₄, K₂HPO₄, KH₂PO₄, and K₂SO_{4 …….}”

 

Now it reads:(LL 516-521)

“……. such as magnesium sulphate (MgSO₄), dipotassium phosphate (K₂HPO₄), monopotassium phosphate (KH₂PO₄) and potassium sulphate (K₂SO₄)……”

 

Comment 32

“Line 471: please replace “CaCl₂, FeSO₄, (NH₄)₂SO₄, MnCl₂, ZnSO₄, and CuSO₄” by “calcium chloride (CaCl₂), iron(II) sulphate (FeSO₄), ammonium sulphate ((NH₄)₂SO₄), manganese(II) chloride (MnCl₂), zinc sulphate (ZnSO₄), and copper(II) sulphate (CuSO₄)”.

 

Answer: We appreciate the meticulous reading of our MS by Reviewer 1..

 

Before it read:

“……trace metals such as CaCl₂, FeSO₄, (NH₄)₂SO₄, MnCl₂, ZnSO₄, and CuSO₄……”

 

Now it reads:

“…….trace metals such as calcium chloride (CaCl₂), iron(II) sulphate (FeSO₄), ammonium sulphate ((NH₄)₂SO₄), manganese(II) chloride (MnCl₂), zinc sulphate (ZnSO₄), and copper(II) sulphate (CuSO₄)……..”

 

Comment 33

“Lines 471-474: please replace “as Alanine, Glycine, Leucine, Isoleucine, Valine, Methionine, Serine, Threonine, Proline, Asparagine, Arginine, Lysine, Ornithine, Glutamate, Aspartate, Phenylalanine, Tryptophan, Histidine, Cysteine, adenine, Guanine,…” by “alanine, glycine, leucine, isoleucine, valine, methionine, serine, threonine, proline, asparagine, arginine, lysine, ornithine, glutamate, aspartate, phenylalanine, tryptophan, histidine, cysteine, adenine, guanine, ...”.

 

Answer: Done

 

Before it read:

“….amino acids such as Alanine, Glycine, Leucine, Isoleucine, Valine, Methionine, Serine, Threonine, Proline, Asparagine, Arginine, Lysine, Ornithine, Glutamate, Aspartate, Phenylalanine, Tryptophan, Histidine, Cysteine, adenine, Guanine, uracil vitamins like…..”

 

Now it reads:

“….amino acids such as alanine, glycine, leucine, isoleucine, valine, methionine, serine, threonine, proline, asparagine, arginine, lysine, ornithine, glutamate, aspartate, phenylalanine, tryptophan, histidine, cysteine, adenine, guanine….”

 

Comment 34

“Line 476: please replace “Beta-glycerophosphate” by “beta-glycerophosphate”.”

 

Answer: Change made.

 

Before it read:

“…… and Beta-glycerophosphate…..”

 

Now it reads:

“……and beta-glycerophosphate……”

 

Comment 35

“Lines 497-501: I don´t understand the sentence between these lines.”

 

Answer 35: We appreciate very much this critical observation. The following corrections were made:

 

-Line 497, which is part of the footer of Table 2, was placed in the corresponding place on the line 527,

 

-Line 498 was supplemented with the information that was mistakenly deleted.

 

Before it read:

 

“Trypto-casein soy broth.

Like the production of HA using Streptococcus zooepidemicus without GT, most

Among vitamin and other additives, glutamine, glutamate and oxalic acid addition have been reported [108], as well as pyruvate, N-acetyl glucosamine, phosphatidylcholine [37].”

 

Now it reads:

“Like the production of HA using Streptococcus zooepidemicus without GT, most fermentations were batch-type. They used low-capacity bioreactors (0.2 to 5 L), except in the study [108], which was the only one to report the production of HA on a pilot scale (70 L). The operation conditions were established at 37°C, pH 7, agitation 150-600 rpm. When aeration was applied, the values were in the range of  0.5 to 2 vvm.

The sources of C and N in the culture media were generally glucose and yeast extract, respectively. However, it was found that peptone and casein hydrolysates were used as supplementary sources of N. Mineral supplements, included MgSO₄ and K₂HPO₄, and trace metals such as CaCl₂, FeSO₄, MnSO₄, ZnCl₂, and CuSO₄, were used as is the case of the study reported by Lu et al.,[119].

Among vitamins and other additives, glutamine, glutamate and oxalic acid addition have been reported [120], as well as pyruvate, N-acetyl glucosamine and phosphatidylcholine [39].”

 

Comment 36

“Line 517, Table 1, Table 2, Table 3, Table 4, Table 5, Table 6, Table 7, Table 8, line 720, Table 9 and lines 922, 923, 929, 932, 934 and 936: please replace “g/L” by “g L^-1”.”

 

Answer: Thank you very much for these comments. We have harmonized the units throughout the RMS.

 

Before it read:

“….g/L….”

 

Now it reads:

“…….g L^-1 ……“

 

Comment 37

“Line 571: please replace “High” by “high”.”

 

Answer: Done.

 

Before it read:

“. ……High MW…..”

 

Now it reads:

“….high MW……”

 

Comment 38

“Line 610: please replace “E. coli” by “E. coli”.”

 

Answer: Done.

 

Before it read:

“……E. coli can produce low and high MW…..”

 

Now it reads:

“…….E. coli can produce low and high MW…….”

 

Comment 39

“Line 618: please replace “26, 30,…” by “26°C, 30°C,…”.”

 

Answer: The RMS was modified accordingly.

 

Before it read:

“……fungal strain: 26, 30, and 37°C…..”

 

Now it reads:

“…….fungal strain: 26°C, 30°C, and 37°C…..”

 

Comment 40

“Table 5: please replace “(NH₄)2SO₄” by “(NH₄)₂SO₄”.”

 

Answer: Done

 

Before it read:

“…..(NH₄)2SO₄ …….“

 

Now it reads:

“……. (NH₄)₂SO₄……. “

 

Comment 41

“Table 6: please replace “L/L” by “L L^-1”.”

 

Answer: Done.

 

Before it read:

“…….L/L……”

 

Now it reads:

“…….L L^-1……. “

 

Comment 42

“Table 7: please replace “v/v” by “v v^-1”.”

 

Answer: Done.

 

Before it read:

“……v/v…..”

 

Now it reads:

“….. v v^-1…… “

 

Comment 43

“Line 691: please replace “municipal solid waste” by “MSW” and “organic fraction of municipal solid waste” by “OFMSW”.”

 

Answer: We appreciate your comment; the change was made in line 691

 

Before it read:

“…….waste from municipal solid waste or the organic fraction of municipal solid waste……”

 

Now it reads:

“…..waste from MSW or the OFMSW…..”

 

Comment 44

“Line 758: please replace “billion)” by “billion)….”

 

Answer: Done

 

Before it read:

“….. USD 16.8 billion)…..”

 

 

Now it reads:

“……USD 16.8 billion)……”

 

 

Comment 45

“Line 761: please replace “€ 1,500” by “1,500 €”.”

 

Answer: Change made. The units were harmonized throughout the RMS.

 

Before it read:

“……it varies between € 1,500…..”

 

Now it reads:

“…….it varies between 1,500 €……”

 

Comment 46

“Line 762: please replace “€ 4,000” by “4,000 €”.”

 

Answer: Done. The units were harmonized throughout the RMS.

 

Before it read:

“……and € 4,000 per kg…..”

 

Now it read:

“…….and 4,000 € per kg……”

 

Comment 47

“Lines 772 and 773: please replace “€/g” by “€ g^-1”.”

 

Answer: Done. The units were harmonized throughout the RMS.

 

Before it read:

“….€/g…..”

 

Now it reads: (LL 845-846)

“…..€ g^-1….. “

 

Comment 48

Line 782: please replace “€ 2.4” by “2.4 €”.

 

Answer: Done. Please see our Answer in Comments 45 and 46

 

 

Before it read:

“….€ 2.4….”

 

Now it reads:

“….2.4 €…….”

 

Comment 49

“Line 784: please replace “€ 1.4” by “1.4 €”.”

 

Answer: Done.

 

Before it read:

“……€ 1.4…..”

 

Now it reads:

“….1.4 €……”

 

Comment 50

“Line 799: please replace “$/kg” by “USD kg^-1”.”

 

Answer: Done. We have harmonized this unit throughout the RMS.

 

Before it read:

“…..$/kg…..”

 

Now it reads:

“…..USD kg^-1….. “

 

Comment 51

“Lines 801 and 802: please replace “kg/year” by “kg year^-1”.”

 

Answer: Done.

 

Before it read:

“……kg/year….”

 

Now it reads:

“…..kg year^-1……. “

 

Comment 52

“Line 803: please replace “abovementioned” by “above mentioned”.”

 

Answer: The change was made.

 

Before it read:

“……results of the abovementioned studies…..”

 

Now it reads:

“ ….results of the above-mentioned studies….”

 

Comment 53

“Line 817: please replace “HA” by “hyaluronic acid”.”

 

Answer: Done

 

Before it read:

“…..Environmental sustainability studies of HA production……”

 

Now it reads:

“…..Environmental sustainability studies of hyaluronic acid production….”

 

Comment 54 


“Lines 819-821: please replace “(Life Cycle Assessment Hyaluronic Acid Production, Environmental Impacts from Hyaluronic Acid Production)” by “(LCA HA production and environmental impacts from HA production)”.”

 

Answer: Done.

 

Before it read:

“(Life Cycle Assessment Hyaluronic Acid Production, Environmental Impacts from Hyaluronic Acid Production).”

 

Now it reads:

“……(LCA HA production and environmental impacts from HA production)……”

 

Comment 55

“Lines 831-832: please replace “International Organization for Standardization (ISO) standards” by “ISO standards”.

 

Answer: I appreciate your observation.  To Attend it, the sentence was modified

 

Before it read:

“The LCA was standardized under the International Organization for Standardization (ISO) standards. Since 2006, Since 2006, this methodology has been established”

 

Now it reads: (LL915)

“….The LCA was standardized under ISO standards since 2006, this methodology has been established….”

                                                                                                                    

Comment 56

“Lines 866-867: please replace “Precautionary Principle.” by “precautionary principle”.”

 

Answer: We appreciate very much the careful reading of the MS by Reviewer 1.

 

Before it read:

“….the Precautionary Principle….”

 

Now it reads:

“…..the precautionary principle…..”

 

Comment 57

“Line 875: please replace “Hyaluronic Acid.” by “hyaluronic acid.”.”

 

Answer: Done.

 

Before it read:

“ …production of Hyaluronic Acid…..”

 

Now it reads:

“…. production of hyaluronic acid…...”

 

Comment 58

“Table 9: please replace “ND” by “NA”.”

 

Answer:

We have modified the Table.

 

Before it read:

” …ND….”

 

Now it reads:

“ ….NA....”

Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

Dear Authors! I included all my comments and questions in the attached file.

Comments for author File: Comments.pdf

Comments on the Quality of English Language

Language correction is necessary.

Author Response

Response to Reviewers Document

 

 

 

Content

 

 

 

 

Section A.  Editors’ Decision and Comments………………………………………...2

 

Section B. Answers to Reviewers’ Comments………………………………………. 5

 

B1. Answers to Reviewer 1 Comments……………………………………………….5

 

B2. Answers to Reviewer 2 Comments………………………………………………23

 

References………………………………………………………………………………33

 

 

 

Section A.  Editors’ Decision and Comments

 

Below please find the text sent by Fermentation Editorial Office regarding the first evaluation of our MS

[Fermentation] Manuscript ID: fermentation-3013052 - Major Revisions

Fermentation Editorial Office <[email protected]>      16 de mayo de 2024, 8:47 a.m.

Responder a: Cathy Lu <[email protected]>, Fermentation Editorial Office <[email protected]> Para: Héctor Mario Poggi-Varaldo <[email protected]>

CC: Guadalupe Pérez-Morales <[email protected]>, Teresa Ponce-Noyola <[email protected]>, Abigail Pérez-Valdespino <[email protected]>, Everardo Curiel-Quesada <[email protected]>, Juvencio Galíndez-Mayer <[email protected]>, Nora Ruiz-Ordaz <[email protected]>, Perla Xochitl Sotelo-Navarro

<[email protected]>, Fermentation Editorial Office <[email protected]>, Cathy Lu <[email protected]> Dear Dr. Poggi-Varaldo,

Thank you again for your manuscript submission:

 

Manuscript ID: fermentation-3013052 Type of manuscript: Review

Title: A review of the production of Hyaluronic Acid in the context of its integration into GBAER-type biorefineries

Authors: Guadalupe Pérez-Morales, Héctor Mario Poggi-Varaldo *, Teresa Ponce-Noyola, Abigail Pérez-Valdespino, Everardo Curiel-Quesada, Juvencio Galíndez-Mayer, Nora Ruiz-Ordaz, Perla Xochitl Sotelo-Navarro

Received: 30 Apr 2024

E-mails: [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected], [email protected]

Industrial Fermentation https://www.mdpi.com/journal/fermentation/sections/Industrial_fermentation Microbial Fermentation of Organic Wastes for Production of Biofuels and Biochemicals 2.0 https://www.mdpi.com/journal/fermentation/special_issues/79SH3ZEMTE

 

Your manuscript has now been reviewed by experts in the field. Please find your manuscript with the referee reports at this link:

https://susy.mdpi.com/user/manuscripts/resubmit/37a40c4ce5bd266ebee717ffffc1e9ed

Please revise the manuscript according to the referees' comments and upload

the revised file within 10 days.

 

Please use the version of your manuscript found at the above link for your revisions.

 

(I)      Please check that all references are relevant to the contents of the manuscript.

 

Our answer to Editor: We have checked all references for relevance to the article

 

(II)     Any revisions to the manuscript should be highlighted, such that any changes can be easily reviewed by editors and reviewers.

 

Our answer to Editor: We highlighted all the revisions in the RMS with red characters and yellow background

 

(III)    Please provide a cover letter to explain, point by point, the details

of the revisions to the manuscript and your responses to the referees’ comments.

 

Our answer to Editor: We are providing two documents: A cover letter that explains the modifications made to the RMS and describes other issues, and a Response-to-Reveiwers document that addresses, point by point, the details of the revisions to the RMS as well as our answers to Reviewers’ comments

 

(IV)   If the reviewer(s) recommended references, please critically analyze them to ensure that their inclusion would enhance your manuscript. If you believe these references are unnecessary, you should not include them.

 

Our answer to Editor: Reviewers did not recommend references. However, we have conducted approx. three literature searches  on certain subjects to write selected responses to Reviewers. Therefore, some new references will appear in the RMS. All of them are relevant to the content of the RMS.

 

(V)    If you found it impossible to address certain comments in the review reports, please include an explanation in your appeal.

 

Our answer to Editor: We feel that we could address all the Comments of the Reviewers. In fact, we agree with the remarks made by Reviewers. We feel that we did not have to defend a statement of ours or to deny a requested modification.

 

(VI)   The revised version will be sent to the editors and reviewers.

 

Our answer to Editor: We are sending to Editors and Reviewers the RMS, the Response-to-Reviewers document, and the Conveying Letter Revised.

 

(VII) If one of the referees has suggested that your manuscript should undergo extensive English revisions, please address this issue during revision. We propose that you use one of the editing services listed at https://www.mdpi.com/authors/english or have your manuscript checked by a colleague fluent in English writing.

 

Our answer to Editor: Only Reviewer 2 suggested minor revision of English. We subjected the RMS to two rounds of Grammarly revisions, and one round of revision by Trinka. Also, Dr Juvencio Galíndez-Mayer, one of our co-authors and very fluent in English, corrected the RMS for English usage.

Grammarly score was 99%

 

(VIII) We would like to draw your attention to the status of this invitation “Publish Author Biography on the webpage of the paper” -

https://susy.mdpi.com/user/manuscript/author_biography/37a40c4ce5bd266ebee717ffffc1e9ed.

If you decide to publish your biography, please remember to fill in it before your paper is accepted.

 

Our answer to Editor: We have sent our biographical capsule to Dr C. Lu for her uploading to MDPI webpage.

 

Please do not hesitate to contact us if you have any questions regarding the revision of your manuscript or if you need more time. We look forward to hearing from you soon.

 

Kind regards, Ms. Lucille Liu

E-Mail: [email protected]

 

 

 

B2. Answers  to Reviewer 2 Comments

 

Notation

BNBP bionanobioparticle

BRF               biorefinery

GBAER         Spanish abbreviation for the Environmental Biotechnology and

Renewable Energies Group

GT                  genetically transformed

HA                  hyaluronic acid

LL                   line or lines

MS                  manuscript

PP                  page or pages

RMS               revised manuscript

 

 

Comment 59. “Comment 1. The abstract is too long - it has 378 words instead of 200. It is also inappropriate to the content of the entire article. Please refer to the journal's guidelines: "The abstract should be an objective representation of the article and it must not contain results that are not presented and substantiated in the main text and should not exaggerate the main conclusions." The abstract also requires language correction.”

 

Answer

We appreciate the careful reading of our MS by Reviewer 2, as well as the quote on the Abstract characteristics. We agree with Reviewer 2 and consequently, we have shortened the Abstract and improved the English writing (please see also our Answer to Comment 71.4.)

 

Before it read

“… Biorefineries (BRF) that process the organic fraction of municipal solid waste (OFMSW) and generate bioproducts and bioenergies have attracted attention because they can simultaneously address energy and environmental problems/needs. Obtaining high-value-added products (VAP) plays a determining role in achieving economic sustainability. Hyaluronic acid (HA) is a VAP that could be integrated into modern biorefinery systems. The objective of this article was to conduct a critical review of the microbial production of HA (MPHA) and its production profile for its integration into a GBAER-type BRF (a type of BRF based on organic wastes) and to identify the environmental and economic sustainability aspects of the modified BRF that would confirm it as a sustainable option. Two types of pathogenic Streptococci that naturally produce AH, exhibited the highest HA production: S. equi ssp. equi and S. zooepidemicus. The first has pro-duced up to 12.0 gL-1, while the second yielded 6.6 gL-1. Interestingly, these bacteria reached a production of up to 29.4 gL-1 when subjected to genetic transformation (GT). A strain of Coryne-bacterium glutamicum was also identified with a maximum HA production of 71.4 gL-1. Indeed, this strain is not a naturally HA-producing bacterium, but it achieved that high production after GT. We found that the MPHA under organic wastes as sources of C and N is scarce. Only Streptococcus zooepidemicus without GT showed a moderate HA production of 6.7 gL-1 when starch was used as an alternative C source and yeast extract as a conventional source of N. When an alterna-tive source of N was supplemented (i.e., cheese whey or fermented corn liquor) or when alterna-tive sources of C and N were used simultaneously, HA production was lower (only 4 and 3.54 gL-1, respectively.) In addition, our review identified other knowledge gaps that must be ad-dressed regarding aspects of process scale-up, HA industrial production, economic aspects, and environmental sustainability of the MPHA. The integration of MPHA into GBAER-type BRF us-ing saccharified liquors as a source of C is promising, but at the same time, it constitutes a tech-nical challenge. The studies of this specific integration and the necessary advances in other R&D areas identified above will allow for evaluating the environmental and economic sustainability of the completely modified GBAER-BRF and other systems that want to integrate MPHA into their background processes.…”

 

Now it reads

“…Biorefineries (BRF) that process the organic fraction of municipal solid waste and generate bioproducts and bioenergies have attracted attention because they can simultaneously address energy and environmental problems/needs. The objective of this article was to critically review the microbial production of hyaluronic acid (MPHA) and its production profile for its integration into a GBAER-type BRF (a type of BRF based on organic wastes) and to identify the environmental and economic sustainability aspects of the modified BRF that would confirm it as a sustainable option. It was found that the MPHA by selected strains of pathogenic Sterptococci was moderate to high, although the trend to work with genetically transformed (GT) (innocuous) bacteria is gaining momentum. For instance, a GT strain of Corynebacterium glutamicum reached a maximum HA production of 71.4 gL-1. MPHA reports that use organic wastes as carbon (C) and nitrogen (N) sources are scarce. When alternative sources of C and N were used simultaneously, HA production by S. zooepidemicus was lower than that with conventional sources. We identified several knowledge gaps that must be addressed regarding aspects of process scale-up, HA industrial production, economic feasibility and sustainability, and environmental sustainability of the MPHA.….”

 

Comment 60. PP 4 LL 143 & ff “Comment 2. BRF is still to be determined because information on Mexican markets and the economic benefits is scarce, and it is a topic in which research and development still need to be improved” - and what about the perspective of global markets?”

 

Answer

We thank Reviewer 2 for the remark. It will allow to expand the issue on BRF markets and improve the RMS. information about the perspective of global market  of BRF was added after line 145 and three new references were used to support the statements.

 

Before it read

“… BRF is still to be determined because information on Mexican markets and the economic benefits is scarce, and it is a topic in which research and development still need to be improved …”

 

Now it reads

“…….BRF is still to be determined because information on Mexican markets and the economic benefits is scarce, and it is a topic in which research and development still need to be improved. Although positive economic results are expected to be obtained, as shown by BRF that process food waste and produce biodiesel and biogas, in countries such as the USA and India [27]. Ladakis et al. [21], found that BRF that process MSW from Greek regions and generate VAP suchs lipids, proteins, and succinic acid, showed positive economic results. Jacob-Lopes et al. [28] conducted a market review for several VAPs (for instance, bioactive compounds used as food additives) produced for macroalgae-based BRFs. They reported that these products had a consolidated market share in USA, Japan, and Brazil, and presented positive economic benefits.”

 

 

Comment 61. “Comment 3. What are bionanobioparticles? The authors use this name for, presumably carrier, in subsection 2.1 and in Figure 1.”

 

Answer

We thank Reviewer 2 for this question. Its answer will allow to clarify the issue in the RMS since the bionanobioparticles constitute one of the bioproducts of the GBAER-type BRF.

 

Before it read

“… This core BRF was expanded with a solvent extraction of organic acids and low-molecular-weight solvents from a fraction of the fermented solids stream generated in the H stage and with a final stage of bionanobioparticle production (NN) for the environ-mental industry. The NN stage used the saccharified liquors from the S stage. The expanded BRF was coined HMEZS-NN [18, 64]. …”

 

Now it reads  

“…….This core BRF was expanded with a solvent extraction of organic acids and low-molecular-weight solvents from a fraction of the fermented solids stream generated in the H stage and with a final stage of bionanobioparticle production (NN) for the environmental industry Bionanobioparticles (BNBP) are nanodecorated bioparticles (BP) with iron-material nanoparticles (magnetite, mixed oxides, etc.) [66]. The bioparticles, in turn, are biological catalysts sampled from anaerobic fluidized bed reactors [67,68] treating saccharified liquors (the latter produced in the BRF [69]). The BP consist of granular activated carbon (mesh 30) colonized by an anaerobic methanogenic consortium [68]. This consortium is immobilized on the carrier and typically forms a biofilm, thus leading to BP. With adequate acclimation, BP can degrade chlorinated organic compounds and surfactants [67,68,60-72]. The nano decoration is carried out using the biological reduction power (bioreduction) of the biomass immobilized in the BP, using a mixture of a FeCl₃ solution and a degradable source of carbon (such as saccharified liquors from the BRF) in an ancillary methanogenic bioreactor [66,69]. In this way iron-material nanoparticles are formed on the surface of the bioparticles leading to BNBP. The latter typically show enhanced degradation of organo-chlorinated compounds in polluted waters [66].

The NN stage used the saccharified liquors from the S stage. The expanded BRF was coined HMEZS-NN…...”

 

Comment 62. “Comment 4. Lines 354-357: It is necessary to indicate the appropriate citation.”

 

Answer

We made an effort to follow Reviewer 2’s advice by indicating the appropriate citation in this passage.  Please note that this Comment is similar to Comments 64,67,68,70.

 

Before it read

“… Currently, HA has a potential focus in cancer treatment as a drug carrier and in the design of nanoparticles or liposomes encapsulated with or conjugated to therapeutic agents, as well as its use to detect CD44 hyaladherins (CD44 cluster of differentiation), which is used in the diagnosis of specific tumors. Other uses are as a specific MRI contrast agent for cancer diagnosis and treatment [66, 72, 73].

 

Now it reads

“ Currently, HA has a potential focus in cancer treatment as a drug carrier and in the design of nanoparticles or liposomes encapsulated with or conjugated to therapeutic agents [75,81]. It is also used to detect CD44 hyaladherins (CD44 cluster of differentiation), which is used in the diagnosis of specific tumors [82]. Other uses are as a specific Magnetic resonance imaging (MRI) contrast agent for cancer diagnosis and treatment [75,82]…..”

 

Comment 63. “Comment 5- Line 364: “...AH..” – misspelling”

 

Answer

We appreciate the careful reading of our MS by Reviewer 2. We have replaced AH by HA throughout the text of the RMS. Please note that this Comment is similar to the Comments 65 and 66

 

Before it read

“… Regarding food applications, the use of AH as an edible film or coating that improves food products' quality and shelf life has been reported [75, 76]. …”

 

Now it reads

“…….Regarding food applications, the use of HA as an edible film or coating that improves food products' quality and shelf life has been reported [75, 76……..”

 

 

Comment 64. “Comment 6. Lines 372-374, 374-376: It is necessary to indicate the appropriate citations.”

 

Answer

We made an effort to follow Reviewer 2’s advice by indicating the appropriate citations in the indicated text and two new references were added. Please note that this Comment is similar to Comments 62,67,68,70.

 

Before it read

“… The MW of the first type of HA is greater than 1MDa; it exhibits anti-inflammatory activity and is used for recovery and repair of joint tissue and cartilage degeneration, respectively, and in plastic and cosmetic surgeries..…”

 

“….The MW of the second type of AH is between 0.01-1 MDa; the human body better absorbs this, has pro-inflammatory activity, and promotes tissue remodeling in wound healing

 

Now it reads

“ ….The MW of the first type of HA is greater than 1MDa; it exhibits anti-inflammatory activity and is used for recovery and repair of joint tissue and cartilage degeneration, respectively, and in plastic and cosmetic surgeries [32, 90]…. “

 

“….The MW of the second type of HA is between 0.01-1 MDa; the human body better absorbs this, has pro-inflammatory activity, and promotes tissue remodeling in wound healing [75,91]. The MW of the third type of HA is less than 0.01 MDa and has several food, health, and medical applications [32]….”

 

Comment 65. “Comment 7. Line 374: “...AH..” – misspelling” ”

 

Answer

Please note that this Comment is similar to Comments 63 and 66. We have replaced AH by HA throughout the text of the RMS

 

 

Before it read

“… respectively, and in plastic and cosmetic surgeries. The MW of the second type of AH is …”

 

Now it reads

“…….respectively, and in plastic and cosmetic surgeries. The MW of the second type of HA is ……..”

 

 

Comment 66. “Comment 8. Figure 3 - “...AH..” – misspelling.”

 

Answer

Please note that this Comment is similar to Comments 63 and 65. We have replaced AH by HA throughout the text of the RMS as well as in Figure 3 and tables.

 

Before it read

“                                    

 

                                                                                                                                      “

 

Now it reads

“

 

                                                                                                                                      ”

 

Comment 67. “Comment 9. Lines 412-414: It is necessary to indicate the appropriate citations.”

 

Answer

We are very grateful to Reviewer 2 for the careful reading of our MS. We made an effort to follow Reviewer 2’s advice by indicating the appropriate citations in the indicated text. Please note that this Comment is similar to Comments 62, 64, 68, 70

 

Before it read

“… The C sources can be glucose, fructose, and maltose; the N sources are typically am-monium sulfate and yeast extract. Thiamine, riboflavin, and pyridoxine can be added as vitamins, and sodium, potassium, and calcium can be added as minerals. Others growth factor can be Peptone and tryptone [36, 85].…”

 

Now it reads

“ The C sources can be glucose, fructose, and maltose [38,96-98]; the N sources are typically ammonium sulfate and yeast extract [36, 95]. Thiamine, riboflavin, and pyridoxine can be added as vitamins [99,100], and sodium, potassium, and calcium can be added as minerals. Other growth factors can be peptone and tryptone [103,104].

 

Comment 68. “Comment 10. Lines 489-491: It is necessary to indicate the appropriate citations.”

 

Answer

We thank Reviewer 2 for the careful reading of our MS. We made an effort to follow Reviewer 2’s advice by indicating the appropriate citations in the indicated text. Please note that this Comment is similar to Comments 62, 64, 67, 70.

 

Before it read

“….This production is two times higher than the highest production reported with Streptococcus zooepidemicus in fed-batch culture, although this value was achieved in low-capacity bioreactors...…”

 

Now it reads

“This production is two times higher than the highest production reported with Streptococcus zooepidemicus in fed-batch culture [117], although this value, in Streptococcus equi, was achieved in low-capacity bioreactors”

 

Comment 69. “Comment 11. Lines 497-501: This fragment is incomprehensible. Please edit it. In addition, the abbreviation GT must be entered in the text in advance.”

 

Answer

We agree with Reviewer 2. We also thank very much the careful reading of the MS by Reviewer 2.

          There was a big mistake in this section of the article: two whole paragraphs from our original version were erased.

Fortunately, our original Word draft file had the complete sentences we were able to rescue the missing passage.

 We have re-written the paragraph and the presentation of the abbreviation GT in the RMS.

 

Before it read

“….

Trypto-casein soy broth.

Like the production of HA using Streptococcus zooepidemicus without GT, most

Among vitamin and other additives, glutamine, glutamate and oxalic acid addition have been reported [108], as well as pyruvate, N-acetyl glucosamine, phosphatidylcholine [37].…”

 

Now it reads

“…….Trypto-casein soy broth

Like the production of HA using Streptococcus zooepidemicus without GT, most fermentations were batch-type. They used low-capacity bioreactors (0.2 to 5 L), except in a study [108], which was the only one to report the production of HA on a pilot scale (70 L). The operation conditions were established at 37°C, pH 7, agitation 150-600 rpm. When aeration was applied, the values were in the range of 0.5 to 2 vvm

The sources of C and N in the culture media were generally glucose and yeast extract, respectively. However, peptone and casein hydrolysate were used as a supplementary sources of N. Mineral supplements, included MgSO₄ and K₂HPO₄, and trace metals such as CaCl₂, FeSO₄, MnSO₄, ZnCl₂, and CuSO₄ as is the case of the study repoted by Lu et al.,[119].

 Among vitamin and other additives, glutamine, glutamate and oxalic acid addition have been reported [120], as well as pyruvate, N-acetyl glucosamine, phosphatidylcholine [39].……..”

 

Comment 70. “Comment 12. Lines 686-689: It is necessary to indicate the appropriate citations.”

 

Answer

We thank Reviewer 2 for his/her indication. We made an effort to follow Reviewer 2’s advice by indicating the appropriate citations in the indicated text. Please note that this Comment is similar to Comments 62, 64, 67, and 68

 

Before it read

“… Differences were found in the HA's MW. In studies where only the alternative source of C is used, the HA has a low MW (typically 0.001 to 0.98 MDa), while it has a high MW (1.8 to 3.8 MDa) when using only the alternative source of N or both alternative sources of C and N.…”

 

Now it reads

“…..Differences were found in the HA's MW. In studies where only the alternative source of C is used, the HA has a low MW, typically 0.001 to 0.98 MDa [136-138], while it has a high MW (1.8 to 3.8 MDa) when using only the alternative source of N or both alternative sources of C and N [140,141,143].----.”

 

 

Comment 71. “Comment 13. The topic of the review work is very interesting. The material collected for the preparation of this work is significant, but the presentation of the literature review on the microbiological production of hyaluronic acid is chaotic. As a reader, in many places I had the impression that I was reading a collection of information that was in no way connected into a coherent whole. The manuscript requires significant re-editing and tidying up. In addition, language correction is necessary. It is also worth noting that most of the literature used by the authors (over 70 out of 144 items) comes from the last 5 years.”

 

Answer

This Comment is very long and deals with several issues. As a matter of order, we have subdivided the Comment 71 in Sub Comments, as follows

 

Comment 71.1 “The topic of the review work is very interesting.”

Answer

We appreciate the positive remark of Reviewer 2.

 

Before it read

NA

 

Now it reads

NA

 

 

Comment 71.2 “…The material collected for the preparation of this work is significant,…”

 

Answer

Again, we are grateful for Reviewer’s 2 positive feedback.

 

Before it read

NA

 

Now it reads

NA

 

Comment 71.3 “….but the presentation of the literature review on the microbiological production of hyaluronic acid is chaotic. As a reader, in many places I had the impression that I was reading a collection of information that was in no way connected into a coherent whole. The manuscript requires significant re-editing and tidying up.”

 

Answer

We agree with Reviewer 2. Regarding the comments of Reviewer 2, we made a major revision. We have examined the MS looking for opportunities for improving logic, organization, style, and the English of the RMS:

 

(i) content order improvement in the RMS, please compare the old and new Index of the review to check for the improvements and reorganization of the RMS

 

(ii) justification and explanation of the scope of sections and selected sub-sections as well as their interrelations and the relationships with the main objective of the MS (review the MPHA) as well as the secondary goal of examining the opportunities for integrating the MPHA to waste-based BRF.

Furthermore, each section was provided with a new justification and statement of the scope of the section. Also, section content was re-organized in subsections, including a Concluding remarks subsection at end of each section.

 

(iii) revamping in general the Dewey’s numeration system. An example follows: the section Conclusion and Perspective had the number 5, instead it should read 7. And so on.

 

(iv) more simple (direct) writing, whenever possible.

 

(v) awkward syntax detection and re-writing.

In particular, it is worth mentioning a big mistake due to missing text around the lines LL 497-501 (Comment 69 in our numbering, Comment 11 with Reviewer’s numbering), that contributed to the “chaotic” feeling of Reviewer 2 and any potential reader. This mistake was corrected by the addition of two full paragraphs included in our original draft.

 

(vi) the Abstract was completely re-written and its size is now 196 words.

 

(vii) the concept of bionanobioparticles was addressed and explained in the RMS

 

Consequently, several passages of the RMS were rewritten. We guess that up to 30% of the RMS is new or modified and both the logic and style are sounder now.

          Below please find a few examples of the RMS revamping; it is not an exhaustive list otherwise we should include here up to 30% of the manuscript.

 

Example 1

Before it read

 

“Content

1. Introduction
2. Biorefineries that process organic wastes

2.1 GBAER type biorefineries and the potential for integration of the microbial hyaluronic acid

3. Microbial production of hyaluronic acid
   • Native hyaluronic acid-producing bacteria
   • Recombinant bacteria that produce hyaluronic acid
   • Microbial production of hyaluronic acid using organic waste
4. Scale-up of hyaluronic acid production
5. Economic aspects of hyaluronic acid production
6. Environmental sustainability studies of HA production”

”

 

Now it reads

“Content

1. Introduction
2. Biorefineries that process organic wastes
   • General context
   • Biorefineries of GBAER type and the potential for integration of the microbial hyaluronic acid production
3. Microbial production of hyaluronic acid
   • Hyaluronic acid and its significance/importance
   • Native hyaluronic acid-producing bacteria
   • Recombinant bacteria that produce hyaluronic acid
   • Microbial production of hyaluronic acid using organic waste
4. Scale-up of hyaluronic acid production
   • Antecedents so far
   • Concluding remarks
5. Economic aspects of hyaluronic acid production
   • Hyaluronic market and price
   • Cost and economic feasibility of microbial hyaluronic production of hyaluronic acid
   • Concluding remarks
6. Environmental sustainability studies of AH production
7. Conclusion and perspective “

 

Example 2

Before it read

“

2. Biorefineries that process organic waste

The development of BRF has emerged as a sustainable alternative to oil refineries [40, 41] that, due to (i) the scarcity of oil resources

 

Now it reads

 “ 2. Biorefineries that process organic waste

This section devotes to the definition of BRF that use organic waste as feedstock, short descriptions of main BRF variants, the discussion of the Main and Auxiliary Principles that should ideally rule BRF design, as well as the development of a type of BRF by our Group. The motive of this section is that the Review intends, in the end, to foster/consider the integration of the microbial production of hyaluronic acid (MPHA) to organic waste-based BRF

 

2.1. General context

The development of BRF has emerged as a sustainable alternative to oil refineries [40, 41] that, due to (i)”

”

 

 

Comment 71.4 “In addition, language correction is necessary.”

 

Answer

We have corrected the RMS twice using Grammarly Premium. The score of the first round was 98%, whereas the Grammarly score of the article in the second round was 99%. We also subjected the RMS to another round of English revision, this time with Trinka software.

          Also, one of our co-authors very fluent in English (Dr J. Galíndez Mayer) revised the final RMS for English language correction.

          We feel that the combined English revision of the RMS gives a correct English language level.

 

Comment 71.5 “It is also worth noting that most of the literature used by the authors (over 70 out of 144 items) comes from the last 5 years.”

 

Answer

We thank Reviewer 2’s positive comment. We also feel that our review is an updated one.

 

Before it read

NA

 

Now it reads

NA

 

         

References

 

18. Sotelo-Navarro, P. X.; Poggi-Varaldo, H. M.; Chargoy-Amador, J. P.; Sojo-Benitez, A.; Pérez-Angón, M. A.; Sánchez-Pérez, R. Impactos Ambientales de Una Biorrefinería Tipo HMEZS-NN. Int. Contam. Ambient. 2022, 38 (Especial), 48–57.
19. Li, R. Techno-Economic and Environmental Characterization of Municipal Food Waste-to-Energy Biorefineries: Integrating Pathway with Compositional Dynamics. Energy 2024, 223 (December 2023). https://doi.org/10.1016/j.renene.2024.120038.
20. Jacob-Lopes, E.; Maroneze, M. M.; Deprá, M. C.; Sartori, R. B.; Dias, R. R.; Zepka, L. Q. Bioactive Food Compounds from Microalgae: An Innovative Framework on Industrial Biorefineries. Opin. Food Sci. 2019, 25, 1–7. https://doi.org/10.1016/j.cofs.2018.12.003.
21. Romero-Cedillo, L.; Poggi-Varaldo, H. M.; Santoyo-Salazar, J.; Escamilla-Alvarado, C.; Matsumoto-Kuwabara, Y.; Ponce-Noyola, M. T.; Bretón-Deval, L.; García-Rocha, M. Biological Synthesis of Iron Nanoparticles Using Hydrolysates from a Waste-Based Biorefinery. Sci. Pollut. Res. 2020, 27 (23), 28649–28669. https://doi.org/10.1007/s11356-020-08729-w.
22. Garibay-Orijel, C.; Hoyo-Vadillo, C.; Ponce-Noyola, T.; García-Mena, J.; Poggi-Varaldo, H. M. Impact of Long-Term Partial Aeration on the Removal of 2,4,6-Trichlorophenol in an Initially Methanogenic Fluidized Bed Bioreactor. Bioeng. 2006, 94 (5), 949–960. https://doi.org/10.1002/bit.20918.
23. Poggi-Varaldo, H. M.; Bárcenas-Torres, J. D.; Moreno-Medina, C. U.; García-Mena, J.; Garibay-Orijel, C.; Ríos-Leal, E.; Rinderknecht-Seijas, N. Influence of Discontinuing Feeding Degradable Cosubstrate on the Performance of a Fluidized Bed Bioreactor Treating a Mixture of Trichlorophenol and Phenol. Environ. Manage. 2012, 113, 527–537. https://doi.org/10.1016/j.jenvman.2012.03.015.
24. Poggi-Varaldo, H. M. Análisis de Ciclo de Vida de Una Biorrefinería Que Procesa La Fracción Orgánica de Residuos Urbanos y Produce Bioenergías, Ácidos Orgánicos, Enzimas, y Nanobiopartículas; Final Report, Diploma 2019, CADIS A.C, Ciudad de México, 2019. Limited edition. Available upon request.
25. Bretón‐Deval, L.; Rios‐Leal, E.; Poggi‐Varaldo, H. M.; Ponce‐Noyola, T. Biodegradability of Nonionic Surfactant Used in the Remediation of Groundwaters Polluted with PCE. Water Environ. Res. 2016, 88 (11), 2159–2168. https://doi.org/10.2175/106143016x14733681695564.
26. Breton-Deval, L.; Rossetti, S.; Ríos-Leal, E.; Matturro, B.; Poggi-Varaldo, H. M. Effect of Coupling Zero-Valent Iron Side Filters on the Performance of Bioreactors Fed with a High Concentration of Perchloroethylene. Environ. Eng. 2016, 142 (11), 986–994. https://doi.org/10.1061/(ASCE)EE.1943-7870.0001093.
27. Moreno-Medina, C. U.; Poggi-Varaldo, H. M.; Breton-Deval, L.; Rinderknecht-Seijas, N. Effect of Sudden Addition of PCE and Bioreactor Coupling to ZVI Filters on Performance of Fluidized Bed Bioreactors Operated in Simultaneous Electron Acceptor Modes. Sci. Pollut. Res. 2017, 24 (33), 25534–25549. https://doi.org/10.1007/s11356-016-7275-z.
28. Peters, M. S.; Timmerhaus, K. D.; West, R. E. Plant Desing and Economics for Chemical Engineers; 2003.
29. Stern, R.; Asari, A. A.; Sugahara, K. N. Hyaluronan Fragments : An Information-Rich System. J. Cell Biol. 2006, 85, 699–715. https://doi.org/10.1016/j.ejcb.2006.05.009.
30. Ke, C.; Wang, D.; Sun, Y.; Qiao, D.; Ye, H.; Zeng, X. Immunostimulatory and Antiangiogenic Activities of Low Molecular Weight Hyaluronic Acid. Food Chem. Toxicol. 2013, 58, 401–407. https://doi.org/10.1016/j.fct.2013.05.032.

 

 

 

 

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

Comments and Suggestions for Authors

Line 24: Sterptococci – misspelling.

Line 55: “…AH..” – misspelling

Author Response

We appreciate very much the careful reading of our RMS by Reviewer 2. We have corrected the mistakes in the RMS 2. The corrected words are in blue characters and yellow background in the RMS 2.

2^nd Revision

Reviewer 2

Line 24: Sterptococci – misspelling.

Line 55: “…AH..” – misspelling

          We appreciate very much the careful reading of the first RMS by Reviewer 2.

 

Third, the mistakes were corrected in the RMS2 with blue characters and yellow background in the RMS2, as follows

Comment 1: “Line 24: Sterptococci – misspelling.”

Now it reads: “…..by selected strains of pathogenic Streptococci was…”

Comment 2: “Line 55: “…AH..” – misspelling”

Now it reads: “…Environmental sustainability studies of AH HA production…”

Author Response File: Author Response.pdf