Stimulation of Heme-Dependent Catalase Enhanced the Cytoprotective Effect of Lactobacillus plantarum against Oxidative Stress
Round 1
Reviewer 1 Report
Dear; Editor
Thank you very much for giving me the opportunity to read this distinguished research work.
The scientific paper is very, very distinctive, and the effort expended in presenting the valuable results is very clear in the way the manuscript is written, also presenting the results and interpreting them in a specific scientific way that helps the reader to complete the work in an enjoyable way.
Best Regards
Author Response
We appreciate the positive comments from the reviewer.
Reviewer 2 Report
1- Page 2 line 72: added method of identification of bacterial isolate.
2- Page 2 line 81: washed three times by ......................???
3- Page 2 line 91: A unit enzyme was defined as the protein amount ... change to A unit enzyme was defined as the enzyme amount .....
4- Page 3 line 107: why incubated accuratelt for 10 min only????
5-Page 4 line 154: plase added A unit of CAT, A unit of GSH-px, A unit of SOD.
Author Response
Reviewer #2:
Thanks for your positive comments. We have made corresponding changes according to your good suggestions. We hope that the changes in the manuscript can enable our manuscript to win your satisfaction.
Point 1: added method of identification of bacterial isolate
Response: Thanks for your good advice. We have added the method of identification of bacterial isolate as follows:
Briefly, 1 g of yogurt sample was added to 10 mL sterile phosphate-buffered saline (PBS) and the mixture was serially diluted in PBS, followed by surface spreading on de Man, Rogosa and Sharpe (MRS) agar and cultivation at 37℃ for 48h. Single colonies were picked from the MRS agar and incubated in MRS broth statically at 37℃ for 48h. Then, cells were harvested from the above cultures and the genomic DNA were extracted using as templates for PCR amplification of 16S rRNA gene with primers 27F (5’- AGAGTTTGATCMTGGCTCAG-3’) and 1492R (5’- GGTTACCTTGTTACGACTT-3’). The 16S rRNA gene of CGMCC6888 showed 99% identities to those of Lb. plantarum by sequence alignment.
Point 2: washed three times by ......................???
Response: Thanks for your reminding, we have added more in 2.2. We have added “washed three times by sterile saline solution, and resuspended with sterile saline solution to 108 CFU / mL” at line 81.
Point 3: A unit enzyme was defined as the protein amount ... change to A unit enzyme was defined as the enzyme amount .....
Response: Thanks for your suggestion. We have changed “A unit enzyme was defined as the protein amount that breaks down 1 μM of H2O2 at 37℃” to “A unit enzyme was defined as the enzyme amount that breaks down 1 μM of H2O2 at 37℃ per minute”.
Point 4: why incubated accuratelt for 10 min only????
Response: Thank you for your question. In weak basic conditions, pyrogallol could be spontaneously oxidized to superoxide anion. Under the monitor of 325nm, the content of superoxide anion quickly increases at the initial period of the reaction, and reaches max and gradually reduces. During 10 min of the reaction, the content of superoxide anion is increasing linearly, thus ensuring the detection accuracy of the superoxide anion scavenging activity.
Point 5: please added A unit of CAT, A unit of GSH-px, A unit of SOD.
Response: Thanks for your suggestion. According to the comment of the reviewer 2, we have added A unit of CAT, A unit of GSH-px, A unit of SOD. Briefly, A unit CAT was defined as the enzyme amount that breaks down 1 μM of H2O2 at 37℃ per minute. A unit CAT was defined as above. A unit GSH-px was defined as the enzyme amount that catalyzes 1 nmol GSH oxidation per minute. A unit SOD was defined as the enzyme amount inhibits the reaction rate of xanthine oxidase by 50% per minute.
Author Response File: 
Author Response.docx
Reviewer 3 Report
Page No 1 Line 8 : Lb. plantarum expansion to : Lactobacillus plantarum in first time
Commonly change entire manuscript: Lb. plantarum change to L. plantarum
Page No 2 Line 56 Lactobacillus changes to Italics
Page No 6 Line 209 As per the table 2, folding of radical values is not matched with the text. How to calculate the O2 radical values
In overall manuscript, check the values of fold increase in various parameters
Author Response
Please see the attachment
Author Response File: 
Author Response.pdf
