PONE-D-21-34850Degradation of Chondroitin Sulfate A by a PUL-like operon in Tannerella forsythiaPLOS ONE

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“PN, BG, RE, and AD were supported by studentships provided by Wilfrid Laurier University. This work was supported by a National Science and Engineering Research Council of Canada Discovery Development Grants (RPGIN-2014-05018 and DDG-2020-00007). Additional support was provided by GlycoNet Grant AM-21. We are grateful for the support of the Canadian Light Source and Cornell High Energy Synchrotron Source where diffraction data were collected.”

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Reviewer #2: Yes

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Reviewer #1: Yes

Reviewer #2: Yes

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Reviewer #2: Yes

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Reviewer #2: Yes

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Reviewer #1: The aim of the paper by Nguyen et al. was to identify a new polysaccharide utilization locus (PUL) like operon from Tannerella forsythia one of the bacterial triad causing advanced periodontitis. Cristallization data are reported from three proteins encoded by this new PUL-like operon: Bfo2290, Bfo2291 and Bfo2294. Digestion assays using chondroitin sulfate A demonstrate that Bfo2291 and Bfo2290 are a chondroitin sulfate A lyase and a ΔDi-4S sulfatase, respectively. Data from this paper further expands the database of PULs involved in periodontal disease.

The experiments are described in sufficient details and the approaches used to support the results are up to date.

SPECIFIC COMMENTS:

The results provided are convincing; there are some inaccuracies that should be checked.

Materials and Methods session.

Page 9. Line 14 and page 10 line 2. Please check the FPLC equipment, ATKA or AKTA?

Page 10. line 13. Please check the 2-aminoacridone concentration used to label sugars.

Reviewer #2: Authors here present that chondroitin sulfate A may be digested by the enzymes Bfo2290 and Bfo2291 produced by Tanerella forsythia that belong to the PUL-like operon Bfo2285-Bfo2295. The manuscript is well written and the data very interesting. Conclusions support the results. There are some minor comments to concern:

1. In the Introduction, it is better to take away the words "possible" and "frequently" in the phrases "GAGs are a possible carbohydrate component of proteoglycans..." and "GAGs are frequently composed of repeating disaccharide subunits..." as GAGs do compose the glycan part of proteoglycans (this is not a possibility) and always contain repeating disaccharides, not frequently.

2. In Materials and Methods, in the description of FACE analysis at the 3rd lane, please cite the enzymes that you used and the units required for the digestion of Chondroitin sulfate A.

3. In Results, in FACE analysis figure 3, lane A presents the chondroitin sulfate A. There are three bands presented here. In the next lanes, authors support that Bfo2291 digests this GAG. Could you please show in the figure of the bands what kind of GAGs are likely expected to be? Are they long chains, disaccharides or oligo- mono-saccharides? It is not clear the kind of poly-, oligo- or mono-mers or, alternatively, the molecular weight of the resulted bands.

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Reviewer #1: No

Reviewer #2: No

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Degradation of chondroitin sulfate A by a PUL-like operon in Tannerella forsythia

PONE-D-21-34850R1

Dear Dr. Suits,

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Kind regards,

Nikos K Karamanos, Ph.D.

Academic Editor

PLOS ONE

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