From the journal RSC Chemical Biology Peer review history
Investigation of the biological functions of heparan sulfate using a chemoenzymatic synthetic approach
Round 1
Manuscript submitted on 10 lis 2020![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-letter.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
12-Dec-2020
Dear Prof Liu:
Manuscript ID: CB-REV-11-2020-000199
TITLE: Investigation of the biological functions of heparan sulfate using a chemoenzymatic synthetic approach
Thank you for your submission to RSC Chemical Biology, published by the Royal Society of Chemistry. I sent your manuscript to reviewers and I have now received their reports which are copied below.
I have carefully evaluated your manuscript and the reviewers’ reports, and the reports indicate that major revisions are necessary.
Please submit a revised manuscript which addresses all of the reviewers’ comments. Further peer review of your revised manuscript may be needed. When you submit your revised manuscript please include a point by point response to the reviewers’ comments and highlight the changes you have made. Full details of the files you need to submit are listed at the end of this email.
Please submit your revised manuscript as soon as possible using this link:
*** PLEASE NOTE: This is a two-step process. After clicking on the link, you will be directed to a webpage to confirm. ***
https://mc.manuscriptcentral.com/rsccb?link_removed
(This link goes straight to your account, without the need to log on to the system. For your account security you should not share this link with others.)
Alternatively, you can login to your account (https://mc.manuscriptcentral.com/rsccb) where you will need your case-sensitive USER ID and password.
You should submit your revised manuscript as soon as possible; please note you will receive a series of automatic reminders. If your revisions will take a significant length of time, please contact me. If I do not hear from you, I may withdraw your manuscript from consideration and you will have to resubmit. Any resubmission will receive a new submission date.
Supporting our community through Covid-19
While our publishing services are running as usual, we also know that this is a very challenging time for everyone, for many different reasons. If any aspect of the publishing process is worrying you – for example you think you may struggle to meet a pre-determined deadline – please let us know, and we will work out an answer together.
RSC Chemical Biology strongly encourages authors of research articles to include an ‘Author contributions’ section in their manuscript, for publication in the final article. This should appear immediately above the ‘Conflict of interest’ and ‘Acknowledgement’ sections. I strongly recommend you use CRediT (the Contributor Roles Taxonomy from CASRAI, https://casrai.org/credit/) for standardised contribution descriptions. All authors should have agreed to their individual contributions ahead of submission and these should accurately reflect contributions to the work. Please refer to our general author guidelines http://www.rsc.org/journals-books-databases/journal-authors-reviewers/author-responsibilities/ for more information.
The Royal Society of Chemistry requires all submitting authors to provide their ORCID iD when they submit a revised manuscript. This is quick and easy to do as part of the revised manuscript submission process. We will publish this information with the article, and you may choose to have your ORCID record updated automatically with details of the publication.
Please also encourage your co-authors to sign up for their own ORCID account and associate it with their account on our manuscript submission system. For further information see: https://www.rsc.org/journals-books-databases/journal-authors-reviewers/processes-policies/#attribution-id
Please note: to support increased transparency, RSC Chemical Biology offers authors the option of transparent peer review. If authors choose this option, the reviewers’ comments, authors’ response and editor’s decision letter for all versions of the manuscript are published alongside the article. Reviewers remain anonymous unless they choose to sign their report. We will ask you to confirm whether you would like to take up this option at the revision stages.
I look forward to receiving your revised manuscript.
Yours sincerely,
Dr Gonçalo Bernardes
Associate Editor, RSC Chemical Biology
************
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-comments.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
Reviewer 1
The manuscript summarizes the heparan sulfate (HS) and heparin oligosaccharides synthesized recently by chemoenzymatic strategies and their applications in functional studies. General background information about heparin, HS, and related proteoglycans is also provided. While the contents are highly important and are well suited for the journal, the general organization of the manuscript and the writing need substantial improvement before it can be accepted for publication.
Some general suggestions and specific comments are provided below:
1. The general information about HSPG, endothelial glycocalyx, HS, HS functions can be combined in one section.
2. The organization of the manuscript can be improved. It is hard to follow in the current version.
3. Conclusions can be improved to capture the key points in the review.
4. Numerous grammatic and typographic errors need to be corrected. Some are listed below:
5. Page 2, Summary, line 4, recommend to change “a mixture of polysaccharides” to “polysaccharide mixtures”; the last 5th line, recommend to change “to use” to “in using”; the last sentence seems redundant.
6. Page 3, the first paragraph, related references should be cited; line 8, it should be “serves as a receptor for herpes simplex virus” instead of “serves a receptor for herpes simplex virus receptor”; line 14, “potential therapeutic potential” has redundancy which should be corrected.
7. Page 4, line 3, “its” should be replaced by “it”; line 4, misses an “an” before “anti-inflammatory”; the last second sentence in the second paragraph, it should be “…the enhanced diffuse alveolar damage by heparinase from…”; the first sentence of the last paragraph, recommend to change “HS” to “HS preparations”; change “and form HSPGs” to “as HSPGs”; the last sentence, “syndecan 1-4” should be “syndecans 1-4”.
8. Page 5, line 3, it should be “a number of”; line 6, it should be “enzymes”
9. Page 6, the last 5th line, it is meaningless to refer to “5B” in the original report and should be removed.
10. Page 8, line 5, it should be “demonstrated in synthesizing”; “HS oligosaccharides” is shown twice; line 14, it is important to list the specific mutant that is being described.
11. Fig 2B, recommend to use “PmHS2” instead of “pmHS2”. It is also recommended to assign compound numbers to individual structures so that the compounds can be referred by their numbers in the text (Page 10).
12. Page 22, lines 4-5, “a wide audience of biologist” does not seem to be a proper description here.
13. Figure 5, the positions of asterisks seems to have shifted too much to the top left direction.
14. Figure 4 needs to be referred in the text.
15. Figure numbers in the text do not match to those in the figures. For example, Figure 4 is not referred in the text and Fig 6A and Fig 6C in the text seem to be Figure 5.
Reviewer 2
This manuscript by Liu et al. provides insights into the biological roles of heparan sulfate (HS) oligosaccharides that can be accessed in well-defined form using chemoenzymatic synthetic strategies. It is an interesting review article highlighting the role of HS as a medicinally relevant biomolecule in diverse biological contexts, while also emphasizing important aspects relating to its structural determination and quantitation.
However, important structural studies on chemoenzymatically synthesize HS/heparin derivatives are missing and should be added. Indeed, methods for analyzing heparin/HS sequence and conformation are important for understanding biological activity and structure-activity relationships. For instance, JACS 2008, 130, 12998–13007 by Linhardt and coworkers.
Before acceptance at RSC Chem Biol, the authors should address a number of points that should lead to a considerably improved manuscript:
The structure of the paper is not very well organized and the sections/subsections and their respective titles should be better defined for clarity. This should in turn increase the focus of the review.
The paper will also benefit from inclusion of additional examples of the actual chemoenzymatic synthesis of the HS derivatives, which would be appreciated by the chemical biology community of the journal.
Finally, a thorough revision/proof-reading of the text is needed to correct a number of language usage issues before acceptance.
Copied below are some further minor points and corrections that should be addressed, including addition of some references:
---
Structure/Organization of sections not totally clear by ambiguous use of italics/bold
Summary –
The availability of structurally defined HS oligosaccharides is critical in (...)
It also serves as a receptor for herpes simplex virus entry, allowing
Overview of HS – this section would be better named HS structure/metabolism
HS metabolism is carried out in lysosomes involving a number...
HS interactions with chemokines
(…) in nature, which lends to tight binding* (!)
Chemoenzymatic synthesis of HS – should be not italics.
Page 7:
Structural descriptions (first three/four sentences) would be better introduced early, in a previous introductory section.
HS oligosaccharides are very difficult to synthesize using chemical methods (in plural better).
Long HS chains are necessarily required for investigating* (not investing) the biological functions.
The chemoenzymatic method (replace with approach/strategy” better).
Page 8:
The N-sulfated backbone is subjected to the enzymatic modifications by: “epi”(?)
Preparation of synthetic heparin
Page 9:
Batches of contaminated heparin entered to the marketplaces
The chemoenzymatic method approach is becoming a promising method…
Page 10: not sure about the suitability of including glycantherapeutics.com website on the text.
Design of anti-inflammatory HS to treat liver diseases
Sterile inflammation
Page 11
ECM acronym – spell out
Synthetic HS oligosaccharides protect against acute liver injury caused by – this is the actual live disease part.
Using anticoagulant HS oligosaccharides to protect liver damage against ischemica-reperfusion.
Page 15
Synthetic HS oligosaccharides decreased liver I/R (spell out acronym) injury in mice.
This hypothesis was approved proved using several…
A 3-O-sulfate group on the glucosamine sugar unit, the rarest sulfation in HS, is linked to biological functions including anticoagulant activity, binding of FGF receptors [cite Revuelta et al. Chem Commun, 2018, 54, 13455-13458, in the context of 3-O-sulfated GlcNAc synthetic HS oligosaccharide mimics, which might have also been further explored in the review]...
Quantitative analysis of HS using synthetic disaccharide and tetrasaccharide standards
Structural analysis of HS
Page 16
The molecular probe, or called sensor, – which one?
The method has been demonstrated the ability…
To obtain the structural information of HS, NMR can be used.
Suggestion to add NMR structural/conformational studies of chemoenzymatically synthesized isotopically-labeled HS/heparin-derivatives N as less-complex heparin analogues that can also be used as novel reagents for studying the interaction of heparin with proteins, [citing JACS 2008, 130, 12998–13007, Solution Structures of Chemoenzymatically Synthesized Heparin and Its Precursors].
Quantitative disaccharide analysis of HS – Page 18
The molecular probe, or called sensor, – which one?
The method has been demonstrated the ability
Conclusions
Page 22
Compared (d missing) with other…
Refs to add:
Acc Chem Res 2020 Linhardt and coworkers – Chemoenzymatic Synthesis of Glycosaminoglycans.
JACS 2008, 130, 12998–13007 – Solution Structures of Chemoenzymatically SynthesizedHeparin and Its Precursors.
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-response.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
January 15, 2021
Dr. Goncalo Bernades
Associate Editor
RCS Chemical Biology
Re: resubmission of manuscript CB-REV-11-2020-000199
Dear Dr. Bernades,
We are now resubmitting a revised manuscript entitled “Investigation of the biological functions of heparan sulfate using a chemoenzymatic synthetic approach” to be published as a review article in RCS Chemical Biology. We are thankful for reviewers’ constructive critique and suggestions. In the revised manuscript, you will find that we have made substantial changes to improve the clarity as suggested by reviewers. The changes include shortening the manuscript and rearranging the order of the sections. Conclusion section has been rewritten. A paragraph describing the impact from the surrounding saccharide sequences on the conformation of IdoA and IdoA2S residues is also included as suggested by Reviewer #2. The changes are highlighted in yellow. We also submitted a clean version of the manuscript.
Reviewer #1
1. “While the contents are highly important and are well suited for the journal, the general organization of the manuscript and the writing need substantial improvement before it can be accepted for publication.”
The revised manuscript has gone through substantial revision.
2. “The general information about HSPG, endothelial glycocalyx, HS, HS functions can be combined in one section.”
We have combined the sections as suggested.
3. “The organization of the manuscript can be improved. It is hard to follow in the current version.”
We have reorganized and shortened the presentation.
4. “Conclusions can be improved to capture the key points in the review.”
This section has been rewritten.
5. “Numerous grammatic and typographic errors need to be corrected. Some are listed below:..”
These errors are corrected.
6. “Page 2, summary, line 4, recommend to change “a mixture of polysaccharides” to “polysaccharide mixtures.”
The change has been made.
7. “Page 3, the first paragraph, related references should be cited; line 8, it should be “severs as a receptor for herpes simplex virus...”
Additional references have been added. The suggested change has been made.
8. “Page 4, line 3, “its should be replaced by “it”; line 4, missing an “an” before “anti-inflammatory”; the last second sentence in the second paragraph, it should be “…the enhanced diffuse alveolar damage by heparinase from..”; the first sentence of the last paragraph. Recommend to change “HS” to “HS preparation”; change “ad form HSPGs” to as “HSPGs”; the last sentence, “syndecan 1-4” should be syndecans 1-4”.”
Those changes are included.
9. “Page 5, line 3, it should be “a number of”; line 6, it should be “enzymes”.”
Change has been made as suggested
10. “Page 6, the last 5th line, it is meaningless to refer to 5B in the original report and should be removed.”
Removed
11. “Page 8, line 5, it should be “demonstrated in synthesizing; “HS oligosaccharides” is shown twice; line 14, it is important to list the specific mutant that is being described.”
The information is included.
12. “Fig 2B, recommend to use “PmHS2” instead of “pmHS2”. It is also recommended to assign compound numbers to individual structures so that the compounds can be referred by their numbers in the text (page 10).”
The change is made in the text and in Fig 2B. We also add compound numbers in Fig 2B and refer those in the text.
13. “Page 22, lines 4-5, “a wide audience of biologist” does not seem to be a proper description here.”
The statement has been removed.
14. “Fig 5, the positions of asterisks seems to have shifted too much to the top left direction.”
This is due to the conversion to PDF file. We have corrected the problem.
15. “Figure 4 needs to be referred in the text.”
It is referenced now.
16. “Figure numbers in the text do not match to those in the figures. For example, …”
The figure numbers are now properly presented.
Reviewer #2
17. “However, important structural studies on chemoenzymatically synthesize HS/heparin derivatives are missing and should be added. Indeed, methods for analyzing heparin/HS sequence and conformation are important for understanding biological activity and structure-activity relationship. For instance, JACS 2008, 130, 12998-13007 by Linhardt and coworkers.”
The reference is included in the revised manuscript. We added a paragraph describing the synthesis of polysaccharides using the chemoenzymatic method. Because the current manuscript is focused on the synthesis of homogeneous oligosaccharides, it is somewhat outside the scope of current review. We encourage readers to find the related information in an article published elsewhere [1].
18. “ The structure of the paper is not very well organized and the sections/subsections and their respective titles should be better defined for clarity. This should in turn increase the focus of the review.”
The revised manuscript has been reorganized, and the clarity should be improved.
19. “The paper will also benefit from inclusion of additional examples of the actual chemoenzymatic synthesis of the HS derivatives, which would be appreciated by the chemical biology community of the journal.”
We agree with Reviewer 2’s comment. However, the actual examples for the chemoenzymatic synthesis of HS have been described in great details in a review article published previously [2]. The emphasis of this article is to show the use of the synthetic method for biological activity related studies.
20. “A thorough revision/proof-reading of the text is needed to correct a number of language usage issues before acceptance.”
We have carefully edited the revised manuscript.
21-24. “The availability of structurally defined ..”; “Overview of HS- this section …”; “HS interactions with chemokines …:; and “Chemoenzymatic synthesis …”
Same comments are made by Reviewer #1, and are corrected as described above.
25. “Page 7. Structural descriptions (first three/four sentences) would be better introduced early, in a previous introductory section. HS oligosaccharides are very difficult to synthesize using chemical methods (in plural better). Long HS are necessarily required for investigating (not investing) the biological functions. The chemoenzymatic method (replace with approach/strategy” better).”
Those changes have been made in the revised manuscript.
26. “Page 8: The N-sulfated backbone is subjected to the enzymatic modifications by: “epi”(?).”
“epi” is referred to “C5-epimerase”.
27. “Page 9: Batches of contaminated heparin entered to the marketplaces. The chemoenzymatic method approach is becoming a promising method… Page 10: not sure about the suitability of including glycantherapeutics.com website on the text.”
The website has been deleted from the text.
28. “Design of anti-inflammatory HS to treat liver diseases Sterile inflammation Page 11 ECM acronym-spell out Synthetic HS oligosaccharides protect against acute liver injury caused by-this is the actual live disease part.”
The full name of ECM is given.
29. “Using anticoagulant HS oligosaccharides to protect liver damage against ischemia reperfusion. Page 15 Synthetic HS oligosaccharides decreased liver I/R (spell out acronym) injury in mice. This hypothesis was approved proved using several…”.
The changes have been made.
30. “A 3-O-sulfate group on the glucosamine sugar unit, the rarest sulfation in HS, is linked to biological functions including anticoagulant activity, binding of FGF receptors [cite Revuelta et al. Chem Commun, 2018, 54, 13455-13458, in the context of 3-O-sulfated GlcNAc synthetic HS oligosaccharide mimics, which might have also been further explored in the review]…”
Thanks for Reviewer #2 pointing out the reference. It appears that the paper referred by the reviewer describes the synthesis of a disaccharide that has the glycosidic linkage that does not exist in authentic in HS. We feel that review a derivative that is not an authentic HS disaccharide and is outside the scope of current review.
31. “Quantitative analysis of HS using synthetic disaccharides and tetrasaccharide standards structural analysis of HS Page 16 the molecular probe, or called sensor-which one? The method has been demonstrated the ability … To obtain the structural information of HS, NMR can be used.”
The changes have been incorporated.
32. “Suggestion to add NMR structural/conformational studies of chemoenzymatically synthesized isotopically-labeled HS/heparin-derivatives N as less-complex heparin analogues that can also be used as novel reagents for studying the interaction of heparin with proteins, [citing JACS 2008, 130, 12998-13007, solution structures of chemoenzymatically synthesized heparin and its precursors].”
We include a paragraph reviewing the use of NMR to analyze the conformation of IdoA2S and IdoA residues under different saccharide sequences [3]. Currently, synthesis of site-specific isotopically labeled oligosaccharides is possible; however, these oligosaccharides have not been used to study the interactions with proteins.
The paper referred by Reviewer #2 has been cited in the revised manuscript.
34. “Quantitative disaccharide analysis of HS-Page 18 The molecular probe or called sensor, -which one?”
This question has been answered under 31.
35. “Refs to add: Acc Chem Res 2020 Linhardt and coworkers-Chemoenzymatic synthesis of Glycosaminoglycans. JACS 2008, 130, 12998-13007-Solution structures of chemoenzymatically synthesized heparin and its precursors.”
Both references are included in the revised manuscript.
We hope that you will find that the revised manuscript has adequately addressed reviewers’ questions. We look froward to hearing the formal acceptance of our manuscript from you.
Sincerely,
Jian Liu, PhD
John A and Deborah S. McNeill, Jr. Distinguished Professor
Eshelman School of Pharmacy
University of North Carolina
Email: jian_liu@unc.edu
Reference
1. Zhang X, Lin L, Huang H, and Linhardt RJ. Chemoenzymatic Synthesis of Glycosaminoglycans. Acc Chem Res. 2020;53:335-46.
2. Liu J, and Linhardt RJ. Chemoenzymatic synthesis of heparan sulfate and heparin. Nat Prod Rep. 2014;31:1676-85.
3. Hsieh P-H, Thieker DF, Guerrini M, Woods RJ, and Liu J. Uncovering the relationship between sulfation patterns and conformation of iduronic acid in heparan sulfate. Sci Rep. 2016;6:29602.
Round 2
Revised manuscript submitted on 15 led 2021![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-letter.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
10-Feb-2021
Dear Dr Liu:
Manuscript ID: CB-REV-11-2020-000199.R1
TITLE: Investigation of the biological functions of heparan sulfate using a chemoenzymatic synthetic approach
Thank you for your submission to RSC Chemical Biology, published by the Royal Society of Chemistry. I sent your manuscript to reviewers and I have now received their reports which are copied below.
After careful evaluation of your manuscript and the reviewers’ reports, I will be pleased to accept your manuscript for publication after revisions.
Please revise your manuscript to fully address the reviewers’ comments. When you submit your revised manuscript please include a point by point response to the reviewers’ comments and highlight the changes you have made. Full details of the files you need to submit are listed at the end of this email.
Please submit your revised manuscript as soon as possible using this link :
*** PLEASE NOTE: This is a two-step process. After clicking on the link, you will be directed to a webpage to confirm. ***
https://mc.manuscriptcentral.com/rsccb?link_removed
(This link goes straight to your account, without the need to log in to the system. For your account security you should not share this link with others.)
Alternatively, you can login to your account (https://mc.manuscriptcentral.com/rsccb) where you will need your case-sensitive USER ID and password.
You should submit your revised manuscript as soon as possible; please note you will receive a series of automatic reminders. If your revisions will take a significant length of time, please contact me. If I do not hear from you, I may withdraw your manuscript from consideration and you will have to resubmit. Any resubmission will receive a new submission date.
Supporting our community through Covid-19
While our publishing services are running as usual, we also know that this is a very challenging time for everyone, for many different reasons. If any aspect of the publishing process is worrying you – for example you think you may struggle to meet a pre-determined deadline – please let us know, and we will work out an answer together.
RSC Chemical Biology strongly encourages authors of research articles to include an ‘Author contributions’ section in their manuscript, for publication in the final article. This should appear immediately above the ‘Conflict of interest’ and ‘Acknowledgement’ sections. I strongly recommend you use CRediT (the Contributor Roles Taxonomy from CASRAI, https://casrai.org/credit/) for standardised contribution descriptions. All authors should have agreed to their individual contributions ahead of submission and these should accurately reflect contributions to the work. Please refer to our general author guidelines http://www.rsc.org/journals-books-databases/journal-authors-reviewers/author-responsibilities/ for more information.
The Royal Society of Chemistry requires all submitting authors to provide their ORCID iD when they submit a revised manuscript. This is quick and easy to do as part of the revised manuscript submission process. We will publish this information with the article, and you may choose to have your ORCID record updated automatically with details of the publication.
Please also encourage your co-authors to sign up for their own ORCID account and associate it with their account on our manuscript submission system. For further information see: https://www.rsc.org/journals-books-databases/journal-authors-reviewers/processes-policies/#attribution-id
Please note: to support increased transparency, RSC Chemical Biology offers authors the option of transparent peer review. If authors choose this option, the reviewers’ comments, authors’ response and editor’s decision letter for all versions of the manuscript are published alongside the article. Reviewers remain anonymous unless they choose to sign their report. We will ask you to confirm whether you would like to take up this option at the revision stages.
I look forward to receiving your revised manuscript.
Yours sincerely,
Dr Gonçalo Bernardes
Associate Editor, RSC Chemical Biology
************
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-comments.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
Reviewer 1
The revision is much improved compared to the previous manuscript. Several additional changes need to be made before the manuscript can be accepted for publication.
1. Page 3, the last sentence, recommend to change “based on the success in the synthesis” to “on the synthesis”.
2. Page 4, the first paragraph, the last sentence, the “it” before “serves” should be deleted. The second paragraph, the fifth sentence, it should be “bacterial” not “bacteria”. The second last sentence, recommend to change “In this study” to “In that study”
3. Page 5, the first sentence, “a complex mixture” should be changed to “complex mixtures”.
4. Page 7, “Chemoenzymatic synthesis of HS” section, line 4, it should be “…chemoenzymatic syntheses of HS are…”
5. Page 8, the last third line, the sentence “One limitation for the …” is not clear. Please revise.
6. Page 11, the last paragraph, the third line, it should be “plays a critical role…”
7. Page 37 and 38, the alpha linkages in Figures need to be in the axial position, not somewhere between axial and equatorial.
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-response.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
February 18, 2021
Dr. Goncalo Bernades
Associate Editor
RCS Chemical Biology
Re: resubmission of manuscript CB-REV-11-2020-000199
Dear Dr. Bernades,
We are now resubmitting a revised manuscript entitled “Investigation of the biological functions of heparan sulfate using a chemoenzymatic synthetic approach” to be published as a review article in RCS Chemical Biology. We are pleased to learn that our manuscript is nearly acceptable. We are thankful for reviewer’ positive comment of our revised manuscript. In this revised manuscript, we have made changes as the referee # 1 suggested. The changes are highlighted in yellow. We also submitted a clean version of the manuscript. We look forward to hearing formal acceptance to this manuscript from you.
Referee # 1
1. Page 3, the last sentence, recommend to change “based on the success in the synthesis” to “on the synthesis”.
This change has been made.
2. Page 4, the first paragraph, the last sentence, the “it” before “serves” should be deleted. The second paragraph, the fifth sentence, it should be “bacterial” not “bacteria”. The second last sentence, recommend to change “In this study” to “In that study”
Those changes have been made and included in the revised manuscript.
3. Page 5, the first sentence, “a complex mixture” should be changed to “complex mixtures”.
Change has been made.
4. Page 7, “Chemoenzymatic synthesis of HS” section, line 4, it should be “…chemoenzymatic syntheses of HS are…”
Change “synthesis” to “syntheses”
5. Page 8, the last third line, the sentence “One limitation for the …” is not clear. Please revise.
This sentence has been rewritten.
6. Page 11, the last paragraph, the third line, it should be “plays a critical role…”
The error is corrected.
7. Page 37 and 38, the alpha linkages in Figures need to be in the axial position, not somewhere between axial and equatorial.
We redrew the chemical structures in Fig 3 and Fig 4.
Sincerely,
Jian Liu, PhD
John A. and Deborah S. McNeill, Jr. Distinguished Professor
Eshelman School of Pharmacy
University of North Carolina
Email: jian_liu@unc.edu
Round 3
Revised manuscript submitted on 19 úno 2021![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/icon-letter.png)
![](https://www.rsc-cdn.org/pubs-core/2022.0.146/content/NewImages/chevron-down.png)
19-Feb-2021
Dear Prof Liu:
Manuscript ID: CB-REV-11-2020-000199.R2
TITLE: Investigation of the biological functions of heparan sulfate using a chemoenzymatic synthetic approach
Thank you for submitting your revised manuscript to RSC Chemical Biology. After considering the changes you have made, I am pleased to accept your manuscript for publication in its current form. I have copied any final comments from the reviewer(s) below.
You will shortly receive a separate email from us requesting you to submit a licence to publish for your article, so that we can proceed with publication of your manuscript.
You can highlight your article and the work of your group on the back cover of RSC Chemical Biology, if you are interested in this opportunity please contact me for more information.
Discover more Royal Society of Chemistry author services and benefits here:
https://www.rsc.org/journals-books-databases/about-journals/benefits-of-publishing-with-us/
Thank you for publishing with RSC Chemical Biology, a journal published by the Royal Society of Chemistry – connecting the world of science to advance chemical knowledge for a better future.
With best wishes,
Dr Gonçalo Bernardes
Associate Editor, RSC Chemical Biology
Transparent peer review
To support increased transparency, we offer authors the option to publish the peer review history alongside their article. Reviewers are anonymous unless they choose to sign their report.
We are currently unable to show comments or responses that were provided as attachments. If the peer review history indicates that attachments are available, or if you find there is review content missing, you can request the full review record from our Publishing customer services team at RSC1@rsc.org.