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Reviewers' comments:
Reviewer #1: This manuscript describes hydrogen production by Rhodobacter
sphaeroides using the byproducts derived from pretreated lignocellulose. Using biomass such as lignocellulose and cellulose to produce sustainable energy is an interesting topic.It has been well-known that photosynthetic purple nonsulfur bacteria (Rhodobacter,Rhodopseudomonas and Rhodospirillum) have capacity of hydrogen production mediated by nitrogenase using volatile fatty acides in the absent of ammonium. Thus, it is undoubted that hydrogen was produced from lipid fermentation effluent or organic acids presented in the effluent by Rhodobacter. Although authors studied hydrogen production from the sole organic acids and combination of organic acids as well as fermentation effluent under different concentrations and pHs, the absence of analysis of combination
effect of organic acids on hydrogen production really decreasing its interest. More
analysis and discussions on this should be given.

1. Hydrogen yield and hydrogen production rate should be given.

2. P5, L57 and P8 L29, Hydrogen production by PNS bacteria can be inhibited by
ammonium (under non-nitrogen-fixing conditions). Authors should explain why the
ammonium was added to media.

3. What gas atmosphere was used for entire experiments?

4. Why did the authors not use a fermentative hydrogen producing bacteria instead of
Tichosponron to enhance hydrogen yield.

5. Basically medium pH increases when PNS grew. Authors should discuss why pH
was stable (around 7) in Fig1..

6. The error bars should be shown based on testing in triple in the figures.

7. Introduction: The authors made a wrong description about categories of
biohydrogen production (P3, 7-21). Wording modification is needed. Photosynthetichydrogen producing bacteria also include algae and Cyanobacteria.

8. Some recent related literatures on hydrogen production by Rhodobacter are
missed.

Reviewer #2: There are a number of grammatical errors and instances of badly
constructed sentences. Much more discussion on the experimental data is necessary.
Specific comments:
1. P3L5-8: "digestion" is not the only method of biohydrogen production!
2. P3L8-16: Clostridium butyricum is not the only microbe responsible for dark
fermentation!
3. P3L16-22: Rhodobacter sphaeroides is not the only microbe responsible for photo
fermentation!
4. P4L29-38: meaning?
5. P4L40-49: This is an experimental method!
6. P4L52-58: meaning?
7. P7L21-24: meaning?
8. P7L54-57: It is nitrogen concentration, not nitrogen!
9. P9L32-43: You do not need to describe the experimental methods again!
10. P9L43-44: meaning?
11. P10L7-11: How did you know the cell growth?
12. P11L21-27: meaning?
13. P11L27, 38, 49: It is "concentrations", not concentration!
14. P12L16-19: meaning!
15. P12L30, P13L49: "lipid fermentation"?
16. P12L52: "R sphaeroides ZX-5 fermentation"?
17. P13L46-50: meaning?
18. P13L57: Did you measure "residual sugars"?
19. P14L10-14: meaning?
20. P14L35: "organic acid was low"?
21. Tables 1 and 2: Were the OD values before or after fermentation? For "Time", you used different units in both tables. It was not "Total H2"!
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mandolin

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Originally posted by jonan at 2009-8-9 23:06:
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Reviewers' comments:
Reviewer # ...

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5Â¥2009-08-09 23:59:16
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