A Novel Salt-Tolerant L-Glutaminase: Efficient Functional Expression, Computer-Aided Design, and Application

Round 1

Reviewer 1 Report

In general, the theme approached by the authors in this study is very interesting and topical. the research of fermentation is a subject that deserves more investigation because of its interest for the sustainable human development. While this manuscript is a well sectioned paper with a good written manner, following concerns need to be addressed.

 The introduction can be improved;

 The authors should review the way they cite figures in the body of the text;

 The authors can improve the way they present the methods section, there are many sentences in methods section should be added to discussion section.

 The authors can improve the way they present the discussion.

 My comments are listed below:

 Figure 1: lanes of figure B and C should be numbered and specify these lanes in the figure caption.

Figure 2: authors should preform statistics analysis.

Figure 4: authors should preform statistics analysis

 Figure 5: authors should preform statistics analysis.

 Figure 6: authors should preform statistics analysis.

 In conclusion section: authors should specify this section for their obtained results and for the future uses not for background.

 References should be updated.

 

Author Response

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Author Response File: Author Response.docx

Reviewer 2 Report

In this paper, a novel l-glutaminase with high salt tolerance was expressed and characterized. The expression level of the enzyme was improved by improving mRNA stability, and the thermal stability of the enzyme was improved by site directed mutagenesis. The excellent characteristics of this L-glutaminase indicates that it has potential value in industry. The study is interesting and suitable for publish on “fermentation”.

1. The data for the effect of thermal stability at optimal temperatures (50-52°C )on LreuglsA activity is unclear.

2. It is better to check the thermal stability of l-glutaminase in the presence of 20% NaCl.

3. For the section of 3.3, comparing the change of specific activity of enzyme is more reasonable than comparing the change of total activity. The specific activity directly shows the effect of mutation on enzyme, while the total activity will be affected by different expression levels. Although the author calculates the average value, the change between different batches may be relatively large.

4. I suggest to give a table to compare the reported l-glutaminase with this study to highlight the benefit and advantages of this novel L-glutaminase.

Author Response

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Author Response File: Author Response.docx

Reviewer 3 Report

In this work, authors reported a novel L-glutaminase 10 (LreuglsA) with eminent salt-tolerance which  achieved more than 70% of the activity with 30% NaCl. High  salt-tolerance is very important for industry biocatalysis. However, molecular mechanism for the high salt-tolerance should be given fully from both model and experiment research.

In addition, English should be checked thoroughly to verify the correctness, such as P1L32-33

 

Author Response

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Author Response File: Author Response.docx

Round 2

Reviewer 1 Report

The authors responded to all comments and I now recommend accepting this manuscript for publication.

Reviewer 3 Report

The ms can be accepted and published in the present form because the questions arose are all issued.