Peer Review History

Original SubmissionJuly 6, 2019
Decision Letter - Pradeep Kumar, Editor

PONE-D-19-19023

Production, purification and evaluation of biodegrading potential of PHB depolymerase of Stenotrophomonas sp. RZS 7

PLOS ONE

Dear Dr. Sayyed,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

We would appreciate receiving your revised manuscript by 17.07.2019. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter.

To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

Please include the following items when submitting your revised manuscript:

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'.
  • A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'.
  • An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'.

Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

We look forward to receiving your revised manuscript.

Kind regards,

Pradeep Kumar

Academic Editor

PLOS ONE

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Additional Editor Comments (if provided):

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #1: Partly

Reviewer #2: Yes

**********

2. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #1: No

Reviewer #2: Yes

**********

3. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #1: Yes

Reviewer #2: Yes

**********

4. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #1: No

Reviewer #2: Yes

**********

5. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #1: the author failed to explain concentration of am.sulphate used for Isolation of protein. doing SDS PAGE is outdated and not enough for protein work. technically 2D PAGE and MALDI is required. Results are not statistically expressed(SD values, Vlaue, Pvalue are missing).

Reviewer #2: Dear Editor and authors:

Best regards, I appreciated the research idea where it concerning by the biodegradation of certain polymeric waste by microbial enzyme. Since, there is no perfect work, so I have some comments on it:

1. Biodegrading word in the title should be "biodegradation".

2. The 9 lines in the beginning of the abstract should be transfer to the introduction section.

3. All the paragraphs have not included the start space.

4. The materials and methods section included lot of expressing mistakes which I corrected them in my comments in the attached file.

5. In the results; Tables did not express the results and data mentioned in the text as follow:

• Table 1 not express the results of Ammonium salt precipitation method as mentioned in the text, but it expressed the results of Octyl sepharose column method. Also, what is these values contained between brackets in the table?

• Data in Table 2 are differed from data mentioned in the text for all purification methods.

6. The phrase" The dialyzed precipitate of Stenotrophomonas sp. RZS 7 obtained after ammonium salt precipitation when applied onto an Octyl-Sepharose CL-4B column……" was mentioned under title of Column chromatography ; this really confusing where, are the three purification methods separated from each others? Or they are complementary?

7. According to materials and methods: Crotonic acid was produced as a result of degradation of PHB film so there is a reverse relation between the amount and concentrations of them which is differed with results in table 3 which recorded decreasing in conc. of crotonic acid by degradation of PHB film!!!!!! Logically It must be increase. This point needs clarification for the reader.

8. Fig 1 and Fig 2 are the same!!!!!! Insert fig 2 which reflect the data mentioned in the text.

9. Correct titles in table 4 where all are before degradation, while it should be before and after.

10. Put A, B,C on photos of Fig 9

**********

6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #1: No

Reviewer #2: Yes: Samah El-Debaiky

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step.

Attachments
Attachment
Submitted filename: PONE-D-19-19023_reviewer.doc
Revision 1

Reviewer #1:

Que. The author failed to explain concentration of am.sulphate used for Isolation of protein. doing SDS PAGE is outdated and not enough for protein work. technically 2D PAGE and MALDI is required. Results are not statistically expressed (SD values, Vlaue, Pvalue are missing).

Ans : Concentration of ammonium per sulphate is now included in SDS-PAGE under Materials and Methods part. SDS-PAGE is still used as one of the basic methods for determining the molecular weight. Data in results is statistically analyzed and the values in Tables are the average of triplicates. Values were taken to be statistically significant at P ≤ 0.05, and values of standard deviation (SD) are already given in the parenthesis.

Reviewer #2:

1. Biodegrading word in the title should be "biodegradation".

Ans : Revised in the title

2. The 9 lines in the beginning of the abstract should be transfer to the introduction section.

Ans : Sentence from line 3 to 5 is now shifted to introduction. Rest of the text is more relevant in Abstract and hence not shifted

3. All the paragraphs have not included the start space.

Ans : Now every paragraph has given 1Tab start spacing

4. The materials and methods section included lot of expressing mistakes which I corrected them in my comments in the attached file.

Ans : All the typos and mistakes in Materials and Methods have been rectified.

5. In the results; Tables did not express the results and data mentioned in the text as follow:

• Table 1 not express the results of Ammonium salt precipitation method as mentioned in the text, but it expressed the results of Octyl sepharose column method. Also, what is these values contained between brackets in the table?

Ans : Tables are now placed at proper place and Table Numbers have been revised. The values in the brackets are Standard Deviation (SD) value.

• Data in Table 2 are differed from data mentioned in the text for all purification methods.

Ans : Tables are now placed at proper place and Table Numbers have been revised. The values in the brackets are Standard Deviation (SD) value.

6. The phrase" The dialyzed precipitate of Stenotrophomonas sp. RZS 7 obtained after ammonium salt precipitation when applied onto an Octyl-Sepharose CL-4B column……" was mentioned under title of Column chromatography ; this really confusing where, are the three purification methods separated from each others? Or they are complementary?

Ans : All three methods were separate. Dialyzed precipitate of ammonium sulphate method was used for column chromatography. Octyl-Sepharose is a resin used in CL-4B column under column chromatography

7. According to materials and methods: Crotonic acid was produced as a result of degradation of PHB film so there is a reverse relation between the amount and concentrations of them which is differed with results in table 3 which recorded decreasing in conc. of crotonic acid by degradation of PHB film!!!!!! Logically It must be increase. This point needs clarification for the reader.

Ans : Crotonic acid is not the end product of PHB degradation. It is used as standard for spectrophotometric estimation of PHB. As per Law and Slepecky method [22] when PHB is heated with concentrated sulphuric acid it gets converted into crotonic acid. Clarification is now given at the footnote of Table 3

8. Fig 1 and Fig 2 are the same!!!!!! Insert fig 2 which reflect the data mentioned in the text.

Ans : Fig 2 is now replaced with another figure

9. Correct titles in table 4 where all are before degradation, while it should be before and after.

Ans : Corrected

10. Put A, B,C on photos of Fig 9

Ans : Revised

Attachments
Attachment
Submitted filename: PONE-D-19-19023 Response to Reviewers.doc
Decision Letter - Pradeep Kumar, Editor

PONE-D-19-19023R1

Production, purification and evaluation of biodegradation potential of PHB depolymerase of Stenotrophomonas sp. RZS 7

PLOS ONE

Dear Sayyed,

Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process.

Reviewers suggested few minor changes in the manuscript. Kindly go through it and do the necessary changes accordigly. 

We would appreciate receiving your revised manuscript by 05.11.2019. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter.

To enhance the reproducibility of your results, we recommend that if applicable you deposit your laboratory protocols in protocols.io, where a protocol can be assigned its own identifier (DOI) such that it can be cited independently in the future. For instructions see: http://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols

Please include the following items when submitting your revised manuscript:

  • A rebuttal letter that responds to each point raised by the academic editor and reviewer(s). This letter should be uploaded as separate file and labeled 'Response to Reviewers'.
  • A marked-up copy of your manuscript that highlights changes made to the original version. This file should be uploaded as separate file and labeled 'Revised Manuscript with Track Changes'.
  • An unmarked version of your revised paper without tracked changes. This file should be uploaded as separate file and labeled 'Manuscript'.

Please note while forming your response, if your article is accepted, you may have the opportunity to make the peer review history publicly available. The record will include editor decision letters (with reviews) and your responses to reviewer comments. If eligible, we will contact you to opt in or out.

We look forward to receiving your revised manuscript.

Kind regards,

Pradeep Kumar

Academic Editor

PLOS ONE

[Note: HTML markup is below. Please do not edit.]

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: Dear editor and authors,

Have a good day, my regards to all of you. The authors stand on all the comments except 4 points: 1) line 3 in abstract contains double dots. 2)under title PHB depolymerase assay in methods: correct "a 10,000 rpm" to "at 10,000 rpm". 3) under each table: change "Figures in the parenthesis " to " Values in the parenthesis ". 4) Equation in page 9: change "= X100" to "X100"

Reviewer #3: The submitted paper was well-written, clearly followed an interesting field of research to update the mind of respected academic readers about one of the most important natural enzymes that deal with environmental issues. Generally, authors have done their best to provide high-quality data for both journal and reviewers. The highlighted points withing this paper has enough worth to publish with PLOS ONE journal. Meanwhile, I highly recommend the respected authors to add the figure of their protein purification steps gel to their paper. Due to having critical and functional roles among bacterial species, it is quite quintessential the authors discuss the possible structure of this protein by considering other closest species to highlight valued points for further studies. Also, figure 1 has not enough quality to put it within the MS body text. It is better the authors try to annex it to their paper as supplementary record. Figures 5-7 also have a low quality. Please change their resolution option through an image editor software.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes

Reviewer #3: No

[NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.]

While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step.

Revision 2

PONE-D-19-19023R1

Production, purification and evaluation of biodegradation potential of PHB depolymerase of Stenotrophomonas sp. RZS 7

Reviewer #2:

The authors stand on all the comments except 4 points:

1) line 3 in abstract contains double dots.

Ans: One extra dot is now removed

2) Under title PHB depolymerase assay in methods: correct "a 10,000 rpm" to "at 10,000 rpm".

Ans: Revised as “at 10,000 rpm”

3) Under each table: change "Figures in the parenthesis " to " Values in the parenthesis ".

Ans : "Figures in the parenthesis is now revised as " Values in the parenthesis " under all tables.

4) Equation in page 9: change "= X100" to "X100"

Ans : Revised as X100

Reviewer #3:

The submitted paper was well-written, clearly followed an interesting field of research to update the mind of respected academic readers about one of the most important natural enzymes that deal with environmental issues. Generally, authors have done their best to provide high-quality data for both journal and reviewers. The highlighted points withing this paper has enough worth to publish with PLOS ONE journal. Meanwhile, I highly recommend the respected authors to add the figure of their protein purification steps gel to their paper. Due to having critical and functional roles among bacterial species, it is quite quintessential the authors discuss the possible structure of this protein by considering other closest species to highlight valued points for further studies.

Ans : This figure has already been published, placing here it will be a duplication

Also, figure 1 has not enough quality to put it within the MS body text. It is better the authors try to annex it to their paper as supplementary record.

Ans : Fig 1 is the FTIR spectra of PHB (Standard) and is required for comparison with Fig 2 FTIR spectra of PHB (Sample). SO it is retained in the paper.

Figures 5-7 also have a low quality. Please change their resolution option through image editor software.

Ans : Quality of Figure 5-7 have been improved.

Attachments
Attachment
Submitted filename: PONE-D-19-19023 Response to Reviewers.doc
Decision Letter - Pradeep Kumar, Editor

Production, purification and evaluation of biodegradation potential of PHB depolymerase of Stenotrophomonas sp. RZS 7

PONE-D-19-19023R2

Dear Dr. Sayyed,

We are pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it complies with all outstanding technical requirements.

Within one week, you will receive an e-mail containing information on the amendments required prior to publication. When all required modifications have been addressed, you will receive a formal acceptance letter and your manuscript will proceed to our production department and be scheduled for publication.

Shortly after the formal acceptance letter is sent, an invoice for payment will follow. To ensure an efficient production and billing process, please log into Editorial Manager at https://www.editorialmanager.com/pone/, click the "Update My Information" link at the top of the page, and update your user information. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.

If your institution or institutions have a press office, please notify them about your upcoming paper to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, you must inform our press team as soon as possible and no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org.

With kind regards,

Pradeep Kumar

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

Reviewers' comments:

Reviewer's Responses to Questions

Comments to the Author

1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation.

Reviewer #2: All comments have been addressed

Reviewer #3: All comments have been addressed

**********

2. Is the manuscript technically sound, and do the data support the conclusions?

The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented.

Reviewer #2: Yes

Reviewer #3: Yes

**********

3. Has the statistical analysis been performed appropriately and rigorously?

Reviewer #2: Yes

Reviewer #3: Yes

**********

4. Have the authors made all data underlying the findings in their manuscript fully available?

The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified.

Reviewer #2: Yes

Reviewer #3: Yes

**********

5. Is the manuscript presented in an intelligible fashion and written in standard English?

PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here.

Reviewer #2: Yes

Reviewer #3: Yes

**********

6. Review Comments to the Author

Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters)

Reviewer #2: (No Response)

Reviewer #3: All comments were addressed correctly as I expected. After revision process, the manuscript seems have a good structure and well revised. The quality of figures has improved, and English text right now sounds so fluent. To me, publishing this work as a valued article in PONE has no problem. The respected editorial team, however, will determine the destiny of this paper either for publishing or rejecting its content.

**********

7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files.

If you choose “no”, your identity will remain anonymous but your review may still be made public.

Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy.

Reviewer #2: Yes: Dr. Samah El-Debaiky

Reviewer #3: No

Formally Accepted
Acceptance Letter - Pradeep Kumar, Editor

PONE-D-19-19023R2

Production, purification and evaluation of biodegradation potential of PHB depolymerase of Stenotrophomonas sp. RZS 7

Dear Dr. Sayyed:

I am pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department.

If your institution or institutions have a press office, please notify them about your upcoming paper at this point, to enable them to help maximize its impact. If they will be preparing press materials for this manuscript, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org.

For any other questions or concerns, please email plosone@plos.org.

Thank you for submitting your work to PLOS ONE.

With kind regards,

PLOS ONE Editorial Office Staff

on behalf of

Dr. Pradeep Kumar

Academic Editor

PLOS ONE

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