Comparative static and shaking culture of metabolite derived from methyl red degradation by Lysinibacillus fusiformis strain W1B6

This paper reports on the comparative characteristics and properties of the metabolites derived from methyl red (MR) decolorization by Lysinibacillus fusiformis strain W1B6 under static and shaking conditions. A batch culture system was used to investigate the effect of aeration on azoreductase activity in the biodegradation process, transformation of colour removal and the metabolite products. Biodegradation analysis was monitored using Fourier transform infrared spectroscopy and high-performance liquid chromatography while metabolites were determined using gas chromatography–mass spectroscopy. Phytotoxicity and anti-microbial tests were also conducted to detect the toxicity of metabolites. The results showed that this strain grew more rapidly under shaking conditions while azoreductase activity increased more rapidly under static conditions. Despite that, no significant difference in the decolorization was observed under both static and shaking conditions with up to 96% and 93.6% decolorization achieved, respectively, within 4 h of incubation. MR was degraded into two fragmented compounds, i.e. 2-aminobenzoic acid and N,N-dimethyl-1.4-benzenediamine. The concentration of 2-amino benzoic acid was higher under static conditions resulting the biotransformation of 2-amino benzoic acid into methyl anthranilate more rapidly under static conditions. Other metabolites were also detected as intermediate biotransformation products and by-products. Less or no toxic effect was found in the metabolite degradation products under both culture conditions.


Recommendation? Accept with minor revision (please list in comments)
Comments to the Author(s) 1-All the results are total data based. No dye decolorization photographs and Toxicity results are shown. 2-Some sentences are not clear"A 100 ppm metabolite extracted and untreated MR was used for the test." and grammatical errors are also there. 3-Results and conclusion is not too strong.

Review form: Reviewer 2
Is the manuscript scientifically sound in its present form? Yes

Do you have any ethical concerns with this paper? No
Have you any concerns about statistical analyses in this paper? No

Recommendation?
Accept with minor revision (please list in comments)

Comments to the Author(s)
This manuscript reports the results on a study of methyl red dye decolorisation by Lysinibacillus fusiformis bacteria. The study reports interesting, though quite specific results. This manuscript can be published after minor revisions noted: -please provide more details on how the Lysinibacillus fusiformis species was identified. -add scale bars to Fig 1 and

Comments to the Author(s)
This manuscript is poorly written to be considered suitable for publication. Many sentences have not sense, they are not written in intelligible English. It is very difficult to follow and understand the manuscript. Therefore, I do not recommend its publication.

24-Apr-2019
Dear Dr SIMARANI: Title: Comparative static (anoxic) and shaking culture of metabolite derived from Methyl Red degradation by Lysinibacillus fusiformis strain W1B6 Manuscript ID: RSOS-190152 Thank you for your submission to Royal Society Open Science. The chemistry content of Royal Society Open Science is published in collaboration with the Royal Society of Chemistry.
The editor assigned to your manuscript has now received comments from reviewers. We would like you to revise your paper in accordance with the referee and Subject Editor suggestions which can be found below (not including confidential reports to the Editor). Please note this decision does not guarantee eventual acceptance.
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Once again, thank you for submitting your manuscript to Royal Society Open Science and I look forward to receiving your revision. If you have any questions at all, please do not hesitate to get in touch. ********************************************** RSC Associate Editor: Comments to the Author: The work reported in this manuscript could have some interest to our readership, but the presentation is currently below our standard. I recommend the authors revise the manuscript in order to address the concerns raised by reviewers 1 and 2. They must also completely revise the manuscript with regard to grammar, readability, and communicating their message. Seeking assistance in the latter is highly recommended.

Reviewer: 2
Comments to the Author(s) This manuscript reports the results on a study of methyl red dye decolorisation by Lysinibacillus fusiformis bacteria. The study reports interesting, though quite specific results. This manuscript can be published after minor revisions noted: -please provide more details on how the Lysinibacillus fusiformis species was identified. -add scale bars to Fig 1 and check magnification (100[ is not correct) -revise Fig.4 to make it clearly visible -the toxicity study models (plants and bacteria), why did you use them, not human cell cultures, which would be better for this study. Please provide some rationale on this Reviewer: 3 Comments to the Author(s) This manuscript is poorly written to be considered suitable for publication. Many sentences have not sense, they are not written in intelligible English. It is very difficult to follow and understand the manuscript. Therefore, I do not recommend its publication.

Recommendation?
Accept as is It is a pleasure to accept your manuscript in its current form for publication in Royal Society Open Science. The chemistry content of Royal Society Open Science is published in collaboration with the Royal Society of Chemistry.
The comments of the reviewer(s) who reviewed your manuscript are included at the end of this email. I apologise this has taken longer than usual. The work reported in this manuscript could have some interest to our readership, but the presentation is currently below our standard. I recommend the authors revise the manuscript in order to address the concerns raised by reviewers 1 and 2. They must also completely revise the manuscript with regard to grammar, readability, and communicating their message. Seeking assistance in the latter is highly recommended.

Reviewer: 1
Comments to the Author(s) 1-All the results are total data based. No dye decolorization photographs (DONE. It has done by Figure 4) and Toxicity results are shown (DONE).
The toxicity results are available in Table 2 and 3 and also in this link: https://datadryad.org/review?doi=doi:10.5061/dryad.2cb0g0h 2-Some sentences are not clear"A 100 ppm metabolite extracted and untreated MR was used for the test."(DONE It has already revised in main document and Table 1 and 2) and grammatical errors are also there (DONE, it has been revised).

Reviewer: 2
Comments to the Author(s) This manuscript reports the results on a study of methyl red dye decolorisation by Lysinibacillus fusiformis bacteria. The study reports interesting, though quite specific results. This manuscript can be published after minor revisions noted: -please provide more details on how the Lysinibacillus fusiformis species was identified. It has already been written on sub chapter 3.2 that isolation and identification of the strain was according to Reference no 13. (DONE) -add scale bars to Fig 1 and check magnification (100[ is not correct). It has been revised to 1000 magnification with addition the scale bars (DONE)