PONE-D-24-37344Characterization of Bacillus licheniformis IBRL-CHS2 Lipase: A novel subfamily 1.4 enzyme.PLOS ONE

Dear Dr. Abdul Rahim,

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.

==============================

ACADEMIC EDITOR: Both reviewers have commented about a major revision are needed before the acceptation of this MS therefore, I invite the authors to revise carefully the MS following closely all recommendations given by the reviewers.

==============================

Please submit your revised manuscript by Nov 01 2024 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. When you're 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.

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). You should upload this letter as a separate file labeled 'Response to Reviewers'.
• A marked-up copy of your manuscript that highlights changes made to the original version. You should upload this as a separate file labeled 'Revised Manuscript with Track Changes'.
• An unmarked version of your revised paper without tracked changes. You should upload this as a separate file labeled 'Manuscript'.

If you would like to make changes to your financial disclosure, please include your updated statement in your cover letter. Guidelines for resubmitting your figure files are available below the reviewer comments at the end of this letter.

If applicable, we recommend that you deposit your laboratory protocols in protocols.io to enhance the reproducibility of your results. Protocols.io assigns your protocol its own identifier (DOI) so that it can be cited independently in the future. For instructions see: https://journals.plos.org/plosone/s/submission-guidelines#loc-laboratory-protocols. Additionally, PLOS ONE offers an option for publishing peer-reviewed Lab Protocol articles, which describe protocols hosted on protocols.io. Read more information on sharing protocols at https://plos.org/protocols?utm_medium=editorial-email&utm_source=authorletters&utm_campaign=protocols.

We look forward to receiving your revised manuscript.

Kind regards,

Estibaliz Sansinenea

Academic Editor

PLOS ONE

Journal requirements:

When submitting your revision, we need you to address these additional requirements.

1. Please ensure that your manuscript meets PLOS ONE's style requirements, including those for file naming. The PLOS ONE style templates can be found at

https://journals.plos.org/plosone/s/file?id=wjVg/PLOSOne_formatting_sample_main_body.pdf and

https://journals.plos.org/plosone/s/file?id=ba62/PLOSOne_formatting_sample_title_authors_affiliations.pdf

2. We suggest you thoroughly copyedit your manuscript for language usage, spelling, and grammar. If you do not know anyone who can help you do this, you may wish to consider employing a professional scientific editing service.

The American Journal Experts (AJE) (https://www.aje.com/) is one such service that has extensive experience helping authors meet PLOS guidelines and can provide language editing, translation, manuscript formatting, and figure formatting to ensure your manuscript meets our submission guidelines. Please note that having the manuscript copyedited by AJE or any other editing services does not guarantee selection for peer review or acceptance for publication.

Upon resubmission, please provide the following:

The name of the colleague or the details of the professional service that edited your manuscript

A copy of your manuscript showing your changes by either highlighting them or using track changes (uploaded as a *supporting information* file)

A clean copy of the edited manuscript (uploaded as the new *manuscript* file).

3. We note that your Data Availability Statement is currently as follows: [All relevant data are within the manuscript and its Supporting Information files.]

Please confirm at this time whether or not your submission contains all raw data required to replicate the results of your study. Authors must share the “minimal data set” for their submission. PLOS defines the minimal data set to consist of the data required to replicate all study findings reported in the article, as well as related metadata and methods (https://journals.plos.org/plosone/s/data-availability#loc-minimal-data-set-definition).

For example, authors should submit the following data:

- The values behind the means, standard deviations and other measures reported;

- The values used to build graphs;

- The points extracted from images for analysis.

Authors do not need to submit their entire data set if only a portion of the data was used in the reported study.

If your submission does not contain these data, please either upload them as Supporting Information files or deposit them to a stable, public repository and provide us with the relevant URLs, DOIs, or accession numbers. For a list of recommended repositories, please see https://journals.plos.org/plosone/s/recommended-repositories.

If there are ethical or legal restrictions on sharing a de-identified data set, please explain them in detail (e.g., data contain potentially sensitive information, data are owned by a third-party organization, etc.) and who has imposed them (e.g., an ethics committee). Please also provide contact information for a data access committee, ethics committee, or other institutional body to which data requests may be sent. If data are owned by a third party, please indicate how others may request data access.

4. PLOS requires an ORCID iD for the corresponding author in Editorial Manager on papers submitted after December 6th, 2016. Please ensure that you have an ORCID iD and that it is validated in Editorial Manager. To do this, go to ‘Update my Information’ (in the upper left-hand corner of the main menu), and click on the Fetch/Validate link next to the ORCID field. This will take you to the ORCID site and allow you to create a new iD or authenticate a pre-existing iD in Editorial Manager.

5. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels.  

In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions.

6. Please include a separate caption for each figure in your manuscript.

Additional Editor Comments:

Both reviewers have commented about a major revision are needed before the acceptation of this MS therefore, I invite the authors to revise carefully the MS following closely all recommendations given by the reviewers.

[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: Yes

Reviewer #2: Partly

**********

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

Reviewer #1: No

Reviewer #2: No

**********

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: No

**********

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: Yes

Reviewer #2: No

**********

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: What is the orginality of the study. Please mention in detailed.

The species name in the enzyme abbreviation should be removed.

Enzyme assay should be detailed.

Statistical analysis should be added.

Reviewer #2: The topic of the manuscript ID: PONE-D-24-37344 is appropriately suited for publication in the PLOS ONE Journal. Some precision, data, and explanation should be also added to improve quality and to facilitate the manuscript’s understanding. The study used classic methods to achieve the results. The latter were discussed well in light of current knowledge and are of relevance to future biotechnological and industrial applications. However, general and specific points need deep attention. Accordingly, in its current form the manuscript needs extensive revisions which I think might be worth considering.

The reviewer has other specific comments/suggestions (authors’ line counts):

- The title should be improved to be more attractive and concise.

- The first sentence in the Abstract should be removed or moved to the introduction section since there are no provided results.

- The acronyms “DMSO”, “SDS”, “PAGE”, “PET”, “OD”, and, “IPTG”, etc, should be spelt out. The abbreviations used in the paper should be firstly spelt out and should be double-checked.

- L25: Please correctly provide the expression unit of the Vmax where “+1” in “min” and “mg” are in superscript.

- Keywords: Please remove “LipAB.licheniformis” and “MLipAB.licheniformis (Mature lipase)”.

- Introduction section should be enhanced and needs revision to properly address the focus of the study. The Authors are invited to provide updated information about the classification lipases from LIPABASE: a database for 'true' lipase family enzymes available at http://www.lipabase-pfba-tun.org

- Introduction section needs enhancement to properly address the focus of the study. Next, since bacterial lipases have been described also in the patent literature it would have been worth to look at such literature as well and not only into the scientific literature.

- The authors are invited to provide the novelty of their work compared to the same one reported elsewhere by Reddy et al., (2016) Trop Life Sci Res. 27(supp1): 145–150. doi: 10.21315/tlsr2016.27.3.20 entitled “Cloning and Expression of a Subfamily 1.4 Lipase from Bacillus licheniformis IBRL-CHS2”!

- In the whole text: Please correct as: “min” instead of “mins” or “minutes”. Next, please correct as: “h” instead of “hours”, “hrs”, or “hour”.

- L145: The word “Stability” is missing in the sub-title. Next, L151: please add “H” after “Tris-”.

- In the whole text, please remove or change the personal words “we” by another expression.

- In Section 2: Please be sure to consistently list manufacturer locations (in this case, by consistently including the country).

- L193-L196: The authors said that “The control reaction, which did not contain any metal ions, was also prepared. The lipase activity in the presence of each metal ion was compared to the control reaction to calculate the relative activity (%).”: To be convinced, the author should mention that the control for the metal cations MUST be measured after dialyzed against used buffer and/or measured in the presence of strong metallic ions chelators (for example 10 mM EDTA or 1 mM EGTA).

- The are some errors in the presentation of citations e.g., “L212: …described in [28].”.

- L232 and elsewhere: The name of restriction enzymes should not be in italic only the restriction enzymes sites (Do not use italics for the first three letters and close up the entire name, e.g., AccI, HaeII. Removal of italics is a change made by IUPAC in 2003.).

- Table 1: How many times the experiments are conducted? Please give the standard error (±SD) values.

- L319 and L582: Please correctly provide the name of “MLipAB.licheniformis's”.

- L501: Please correct as “strain ARM”.

- There are some mistakes in the reference section. Are 123 references necessary? Please remove the oldest references. Please re-check and consult the updated guides to authors, which is available on this journal’s web site, to make them fully comply with the requested style and format of the journal and the name of microorganisms and Latin words should be written in italic some pagination are missing (Ref no.: 76) Please provide correct abbreviation of journal title in accordance with Index Medicus URL: (http://www.bg.ump.edu.pl/czasopisma/medicus.php?lang=eng) and Web of Science: (https://images.webofknowledge.com/images/help/WOS/M_abrvjt.html).

- Finally, some spelling and grammar mistakes should be corrected, and please ask a native speaker to carefully revise the manuscript.

**********

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: 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.]

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 PLOS at figures@plos.org. Please note that Supporting Information files do not need this step.

Characterization of a novel subfamily 1.4 Lipase from Bacillus licheniformis IBRL-CHS2: Cloning and expression optimization

PONE-D-24-37344R1

Dear Dr. Abdul Rahim,

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

Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.

An invoice will be generated when your article is formally accepted. Please note, if your institution has a publishing partnership with PLOS and your article meets the relevant criteria, all or part of your publication costs will be covered. Please make sure your user information is up-to-date by logging into Editorial Manager at Editorial Manager® and clicking the ‘Update My Information' link at the top of the page. If you have any questions relating to publication charges, 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 help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- 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.

Kind regards,

Estibaliz Sansinenea

Academic Editor

PLOS ONE

Additional Editor Comments (optional):

The authors have done all changes suggested by both reviewers according to their comments improving the Ms, therefore, the MS can be accepted in the current form.

Reviewers' comments: