Lipase from Yarrowia lipolytica: Prospects as an Industrial Biocatalyst for Biotechnological Applications

Round 1

Reviewer 1 Report

* Ensure to write all scientific name in italic format.

* I suggest making a special topic for lipase biosynthesis from extreme yeasts including thermotolerant, halotolerant, osmotolerant, etc.

I suggest revising this MS by native English speaker, moderate editing will improve the quality of language.

Author Response

Dear Editors,

First, we thank the reviewers for their crucial and constructive criticism, which significantly improved the scientific quality of our work. So, below are the answers to your comments and questions. For ease of reference, all changes have been marked in yellow (reviewer 1), in the revised version of the manuscript.

Evaluator #1:

 

Thank you very much for your comments and contributions, they helped to improve the scientific quality of our work. We accept all proposed revisions.

 

• Be sure to write all scientific names in italics.

 

Response: Thank you for your comments and contributions, your suggestion was accepted and we made the necessary adjustments to the text of the article in the scientific name in italics.

 

• I suggest making a special topic for lipase biosynthesis of extreme yeasts, including thermotolerant, halotolerant, osmotolerant, etc.

 

Response: Thank you for your comments and contributions, your suggestion was of great value for the context of our work and we have included topic 2: “Biosynthesis of extreme yeast lipases”.

 

• Comments on the quality of the English language, I suggest reviewing this MS by a native English speaker, moderate editing will improve the language quality.

 

Response: Thank you for your comments and contributions, we have made the corrections in the new version of the manuscript.

Author Response File: Author Response.pdf

Reviewer 2 Report

The manuscript has an interesting theme, but it is not clear what is the novelty. Between 2021 until now, there are 70 review papers in Scopus on the same subject. I think, if authors organize better the idea of this review, valuing the bibliometric tools, the manuscript will be better.

Author Response

Dear Editors,

First, we thank the reviewers for their crucial and constructive criticism, which significantly improved the scientific quality of our work. So, below are the answers to your comments and questions. For ease of reference, all changes have been marked in yellow (reviewer 1), green (reviewer 2), and blue (reviewer 3) in the revised version of the manuscript.

Evaluator #2:

 

Thank you very much for your comments and contributions, they helped to improve the scientific quality of our work. We accept all proposed revisions.

 

• The manuscript has an interesting theme, but it's not clear what's new. Between 2021 and now, there are 70 review articles in Scopus on the same subject. I think that if the authors better organize the idea of this review, valuing bibliometric tools, the manuscript will be better.

 

Response: Thank you for your comment and contribution, your suggestion was accepted and we highlight the structure of our work in terms of bibliometric research and we highlight in the first paragraph of topic 4 what will be the differential of this work in relation to the review articles present in literature related to the subject.

 

Author Response File: Author Response.pdf

Reviewer 3 Report

This is an interesting review dedicated to industrial applications of the lipases from yeasts Yarrowia lipolytica. The paper outlines the methods for preparation of immobilized lipases, physical and chemical modifications of lipases and bioreactor designs used in enzymatic reactions with lipases from Y. lipolytica.

The list of the used literature covers 260 sources.

The text of the article is of high professional level, well illustrated but has a number of drawbacks:

 

1. The structure of the article requires correction. Where is section 1?

Abstract

Introduction

2. Applications of the Lipases from Yarrowia lipolytica

2.1. Use of Lipases from Yarrowia lipolytica

2.2. Applications of the lipases from Yarrowia lipolytica - The name of the subsection 2.2.repeats the name of the section 2. 

3. Preparation of immobilized biocatalysts of Lipases from Yarrowia lipolytica

3.1. Lipases from Yarrowia lipolytica immobilization

3.1.1. Immobilization of Lipases from Yarrowia lipolytica on hydrophobic supports

3.1.2. Use of heterobifunctional supports to prevent enzyme release.

3.1.3. Modulation of Lipases from Yarrowia lipolytica Properties via immobilization on

different supports

3.2. Physical or chemical modification of Yarrowia lipolytica lipases to modulate enzymatic

properties

3.3. Co-immobilization of Lipases from Yarrowia lipolytica and other enzymes

4. Bioreactor designs for uses of Lipases from Yarrowia lipolytica

5. Future trends

6. Patents with lipase Yarrowia lipolytica  

7. Conclusions

 

Subsections 2.1 and 2.2. have almost synonymous names – "Use…" and "Applications…" and do not reflect the content of the text. For example, in subsection2.1 we read:

"Another critical application of Y. Lipolytica yeast is in the treatment of oily wastewater or oily wastewaters, which have high values of chemical oxygen demand (COD)."

But Applications is a topic of subsection 2.2. Section 2 is unduly wordy and repetitive.  For example, in both subsections the information about the use of Y. lipolytica to clean wastewaters of oil mills is given (the fourth paragraph of the submission 2.1. and the first paragraph of the subsection 2.2). Such repetitions make it difficult to perceive the text. I think that section 2 should be written more concisely and consistently, especially considering that the Table 1 already shows concisely and consistently the applications of Y. lipolytica. The sections 3-7 are clearly written.

 

2. The paragraph "From the database…" (p. 3) discusses the variety of articles dedicated to Y. lipolytica lipases. I also looked in PubMed the last reviews on this topic and found a recent review Nascimento et al. A Temporal Evolution Perspective of Lipase Production by Yarrowia lipolytica in Solid-State Fermentation. Processes 2022, 10, 381. https://doi.org/10.3390/pr10020381 I would like to read in this paragraph, what are the differences and novelty of your review in comparison with Nascimento et al, 2022.

3. There are many too long incomprehensible phrases in this review, such as:

·        This makes the valuable enzyme in applications that require the hydrolysis of fats with these types of fatty acids being used in the production of cleaning products, such as detergents and products that act in cleaning surfaces, making the removal more accessible, after the action of this enzyme breaking down oils and fats present in the surface area.

·        In a recent study, the combination of the characteristics present in Y. Lipolytica in a fungal lipase for the treatment of oily wastewater was carried out to improve the performance of the lipase activity of Yarrowia lipolytica using the technique of functional domain of locculation of Saccharomyces cerevisiae, in which a significant decrease in the removal of chemical oxygen demand (COD) by 91.8% was observed, proving its efectiveness in the treatment of oily wastewater [39].

·        Because Y. Lipolytica  is capable of consuming a range of carbon sources, such as hydrocarbons, fats, and diferent types of oils, its biotechnological applications are increasingly being studied [68] and it is due to the ability of the yeast Y. Lipolytica to produce diferent types of lipase that can be used as biocatalysts in the production of biodiesel, in addition to accumulating a signiicant amount of lipids, that this application in the production of biodiesel becomes efective.  

Perhaps some phrases should be simplified.

4. "This paper aims to present and discuss the advances obtained by the study of lipase Yarrowia lipolytica (Figure 1) and the provisions of the future of its industrial applications".-  It seems to me that this is not a suitable place to quote a figure depicting a lipase structure. It would be better to insert it below in text explaining what is lipase.

5. Figure 2. (a). The picture does not look entirely correct. Perhaps this is a transformation bug to the PDF format.

 

Minor comments

Figure 2. (a) and Figure 9. It would be more correct to write the names of countries with the title letter.

Y. Lipolytica  - the species epithet "lipolytica" should be written with the lowercase letter, even in the case of a heading, where the rest of the words are written with the title letter. Check carefully the entire text, including the list of literature.

Yarrowia  lipolytica, Y.  lipolytica - There is no need to write completely the genus name "Yarrowia" with repeated use. The genus name should be written in full upon its first use in a paper. Now Yarrowia lipolytica and Y.  lipolytica are used chaotic in the article.

Figure 4. Representation of the step-by-step procedure of the preparation of immobilized biocatalysts of Lipases from Yarrowia lipolytica. - There is no need to write the word "Lipase" with a title letter.

P. 5. Saccharomyces cerevisiae needs to be written in italics.

 

[25] , [26] - incorrectly   [25, 26] - correctly.   In the text, reference numbers should be placed in square brackets [ ], and placed before the punctuation; for example [1], [1–3] or [1,3].

research [24]. It is an aerobic…  Put a point at the end of the sentence.

I made some minor corrections and comments in the manuscript (see pdf file).

In spite of aforementioned faults it should be recommended for print after major revision of section 2.

Waste processing and bioremediation are urgent problems that lipase of Y. lipolitica helps to fight. This is the main reason why every attempt to integrate knowledge in this area must be warmly welcomed.

Author Response

Dear Editors,

First, we thank the reviewers for their crucial and constructive criticism, which significantly improved the scientific quality of our work. So, below are the answers to your comments and questions. For ease of reference, all changes have been marked in yellow (reviewer 1), green (reviewer 2), and blue (reviewer 3) in the revised version of the manuscript.

 

Reviewer #3:

 

Thank you very much for your comments and contributions, they helped to improve the scientific quality of our work. We accept all proposed revisions.

 

• The structure of the article requires correction. Where is section 1?

Abstract

Introduction

2. Applications of Lipases from Yarrowia lipolytica

2.1. Use of lipases from Yarrowia lipolytica

2.2. Applications of lipases from Yarrowia lipolytica - The name of subsection 2.2. repeats the name of section 2.

3. Preparation of immobilized lipase biocatalysts from Yarrowia lipolytica

3.1. Lipases from immobilization of Yarrowia lipolytica

3.1.1. Immobilization of Lipases from Yarrowia lipolytica on hydrophobic supports

3.1.2. Use of heterobifunctional scaffolds to prevent enzyme release.

3.1.3. Modulation of Yarrowia lipolytica lipases Properties via immobilization on different supports

3.2. Physical or chemical modification of lipases from Yarrowia lipolytica to modulate enzymes.

properties

3.3. Co-immobilization of Yarrowia lipolytica Lipases and other enzymes

4. Bioreactor projects for the use of lipases from Yarrowia lipolytica
5. Future trends
6. Patents with Yarrowia lipolytica lipase
7. Conclusions

 

To respond:  Thank you for your comments and contributions, we consider your suggestions of great value, we have included one more topic, but we kept the introduction as topic 1 , due to the journal rule that indicates that the introduction must be topic 1 and the conclusion numbered as the last topic , but about structuring topic 2 into just one topic, which is now topic 4 , filtering the information in your text to avoid unnecessary repetitions in our work.

 

• Subsections 2.1 and 2.2. have almost synonymous names – "Use…" and "Applications…" and do not reflect the content of the text. For example, in subsection 2.1 we read: "Another critical application of Y. Lipolytica yeast is in the treatment of oily wastewater or oily wastewater, which have high chemical oxygen demand (COD) values." But Applications is a topic of subsection 2.2. Section 2 is unduly wordy and repetitive. For example, in both subsections information is provided on the use of Y. lipolytica to clean wastewater from oil mills (the fourth paragraph of submission 2.1. and the first paragraph of subsection 2.2). These repetitions make it difficult to understand the text. I think section 2 should be written more concisely and consistently, especially considering that Table 1 already concisely and consistently shows the applications of Y. lipolytica.

 

Response: Thank you for your comments and contributions, we consider your suggestions of great value and have chosen to structure topic 2 into just one topic, which is now topic 4 “Applications of the Lipases from Y. lipolytica”, and filter the information in your text to avoid unnecessary repetitions, applying it in our work.

 

• The paragraph "From the database..." (p. 3) discusses the variety of papers devoted to Y lipases . lipolytica. I also searched PubMed for the latest reviews on the subject and found a recent review by Nascimento et al. A Perspective on the Time Evolution of Lipase Production by Yarrowia lipolytica in Solid State Fermentation. Processes 2022, 10, 381. https://doi.org/10.3390/pr10020381 I would like to read in this paragraph, what are the differences and novelties of your review compared to Nascimento et al, 2022.

 

 

Response: Thank you for your comments and contributions, your suggestion was accepted and added to the text after reformulating the bibliographical research and structure of this article as mutually requested by reviewer 2, we highlight in this paragraph what will be the differential of this work in relation to the review articles present in the literature related to the topic, including that by Nascimento et al, 2022.

 

• There are too many incomprehensible phrases in this review, such as:
• This makes the enzyme valuable in applications that require the hydrolysis of fats with these types of fatty acids being used in the production of cleaning products such as detergents and products that work to clean surfaces, making removal more accessible, after the action of this enzyme breaks down oils and fats present on the surface area.

In a recent study, the combination of the characteristics present in Y. lipolytica in a fungal lipase for the treatment of oily effluents was carried out to improve the performance of the lipase activity of Yarrowia lipolytica using the technique of functional domain of loculation of Saccharomyces cerevisiae , in the which a significant decrease in chemical oxygen demand (COD) removal was observed by 91.8%, proving its effectiveness in the treatment of oily wastewater [39].

• Because Y. Lipolytica is able to consume a variety of carbon sources, such as hydrocarbons, fats and different types of oils, its biotechnological applications are being increasingly studied [68] and it is due to the ability of yeast Y. Lipolytica to produce different types of lipases that can be used as biocatalysts in the production of biodiesel, in addition to accumulating a significant amount of lipids, that this application in the production of biodiesel becomes effective.

Perhaps some sentences should be simplified.

 

Response: Thanks for your comments and contributions, we've taken your suggestions and restructured and simplified these sentences in the text.

• "This work aims to present and discuss the advances obtained by the study of the lipase Yarrowia lipolytica (Figure 1) and the predictions of the future of its industrial applications".- It seems to me that this is not a suitable place to quote a figure representing a lipase structure. It would be better to insert it below in the text explaining what lipase is.

 

 

Response: Thank you for your comments and contributions, we took your suggestion and repositioned it (Figure 1) in the text below after explaining what lipase is.

 

• Figure 2. (a). The image doesn't look quite right. Maybe this is a transformation bug for PDF format.

 

 Response: Thank you for your comments and contributions, we understand your suggestion, but as a result of further guidance aimed at improving the use of bibliometric research in our research, Figure 2. (a ) has been deleted.

 

• Figure 2. (a) and Figure 9. It would be more correct to write the country names with the title letter.

 

Response: Thank you for your comments and contributions, we understand your suggestion, but as a result of further guidance aimed at improving the use of bibliometric research in our research, Figure 2.(a ) and Figure 9 have been deleted.

 

• Lipolytica - the epithet of the species "lipolytica" must be written in lower case, even in the header, where the rest of the words are written in the title letter. Carefully check all text, including the list of literature.

 

Response: Thank you for your comments and contributions, the first letter “l” of the species name “Y. lipolytica” has been changed to lowercase throughout the text.

 

• Yarrowia lipolytica, Y. lipolytica - No need to completely write the genus name "Yarrowia" with repeated use. The genus name must be spelled out the first time it is used in an article. Now Yarrowia lipolytica and Y. lipolytica are used chaotic in the article.

 

Response: Thank you for your comments and contributions, following your suggestion the text has been replaced from “ Yarrowia lipolytica” to “Y. lipolytica”, keeping it complete in the context of the presentation of the researched data according to Tables 4 and 5, and in their discussions of the same, since it is necessary to present the searched words with all the characters correctly to corroborate the legitimacy of the data, in the same way that the term was kept in the references because they are titles of academic works.

 

• Figure 4. Step-by-step representation of the preparation of immobilized lipase biocatalysts from Yarrowia lipolytica. - There is no need to write the word "Lipase" with the title letter.

Response: Thank you for your comments and contributions, your suggestion has been corrected in the text, see Figure 7.

 

• 5. Saccharomyces cerevisiae should be written in italics.

 

Response: Thank you for your comments and contributions, your suggestion has been corrected in the text.

 

• [25], [26] - incorrectly [25, 26] - correctly. In the text, reference numbers must be enclosed in square brackets [] and placed before punctuation; for example [1], [1–3] or [1,3].

Response: Thank you for your comments and contributions, your suggestion was very important to improve the structure of our work, corrections were made to the text.

 

• research [24]. It's an aerobic… Put a period at the end of the sentence.

 

Response: Thank you for your comments and contributions, punctuation correction has been corrected in the text.

 

 

 

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The authors improved the quality of this manuscript, and it can be accepted. However, I just ask the authors the review the term lipolytica which in some cases is written lipolitica

Reviewer 3 Report

The authors have greatly improved their manuscript, changing the structure of the article, adding additional information and correcting minor shortcomings.

I think, the manuscript can be accepted in the present form.