Production of Xylooligosaccharide from Cassava Pulp’s Waste by Endo-β-1,4-D-Xylanase and Characterization of Its Prebiotic Effect by Fermentation of Lactobacillus acidophilus

No

Review

Revisions & Rebuttals

1

Do  the hydrolysis condition the author used optimally？ It is encouraged to provide more details related to the production of xylooligosaccharide or add the reference the author referred to.

Additional reference from the same author about the production of xylooligosaccharide has been added [15]

2

Do the authors have an idea of the xylooligosaccharides yields?

from 1 g  cassava pulp constitute about 2.370 mg/ml of XOS (line 193)

3

What is the difference between samples 5 and 6 in Fig. 1?

Sample s3 & 4 as well as samples 5 & 6, were actually the same, they are only replicate of one another, to ensure Their feasibility.

4

The results of the growth of L. acidophilus obtained from Fig. 2 should be concise. The description on lines 206 and 214 are repeated.

Revision has been (lines 225-243)

5

 Line 217: The sentence is not clear. What is “all other media with the 0% mean?

The correction has been made (lines 233-243)

6

The caption of Fig. 5 is confused with some information missing. The author should check it carefully.

Fig 5 has been replaced with a better one

7

The author must check the punctuation (superfluous or absent) throughout the manuscript. For example: line 109, 145, 192, 204, 209, 258.

Revision has been  (Thank you)

8

The English of the manuscript must be improved. Most sentences contain grammatical or spelling mistakes or are not complete sentences.

Yes, Thank you for your suggestion, I have sent my revised manuscript to professional proof read service, I hope it is now much better.

No

Review

Revisions & Rebuttals

1

The topic is interesting, the manuscript of general interest treat a subject quite well documented by the authors. The subject of current investigation and method carried out are within the scope of the Fermentation Journal.

However, a major revision is required before the paper can be considered for publication as follows:

Thank you very much, I will try my best to comply with the necessity.

2

Generally, the English grammar is poor, several words have incorrect spelling, so the application of English is not acceptable and the writing style needs to be improved. A scan by a native English speaker or a Professional English Language Editing Service is advisable in order to avoid error as "and the its effectiveness for...", "was analyzed by gas chromatography for the SCFA profile by gas chromatography" etc.

Yes, Thank you for your suggestion, I have sent my revised manuscript to a professional proofreading service, and I hope it is now much better.

3

- Keywords: adding xylooligosaccharide is necessary.

Revision has been made

4

 Introduction: authors are asked to underline the originality of this paper.

Please see, lines 108-109 to fulfill the suggestion  

5

Materials and methods: precise the type of recipe for the fermentation of L. acidophillus and the media volume; necessary to precise clearly the objectives of TLC and HPLC analysis (qualitative/quantitative analysis of xylan after hydrolysis etc.)

Please see line 163 for revision on L. acidophile fermentation condition.

Line 146 explained the necessity of TLC while line 152 described analysis of HPLC

6

- Authors are asked to explains why choosing Bacillus strain that was isolated from abdomen of soil invertebrates as termite and not other host. Are they anaerobic or microaerophilic bacteria?

 added some information about this in lines 110-111

Bacillus strain from abdominal termite is microaerophilic bacteria

7

 Figures: the quality of figure 1, 2 and 5 should be improve. The symbols' presented in the title of figure 5 are not readable.

Fig 1, 2 and 5 have beed replace with a better one

8

Results and discussion: the information's  presented in the first paragraph of chapter 3.1 and chapter 4 are most appropriate in Introduction chapter.

Revision has been made

No

Review

Revisions & Rebuttals

1

Terminology needs a thorough revision, especially microbiology. There are problems with the basics of microbiology.

Revision has been made

2

XOS production and fermentation processes need more detail. 

Please see line 136

3

Analytical methods must be completed with validation aspects (standard buffers reliable, reference materials, accuracy to the second decimal of pH, etc., and other parameters). There is missing information about Lactobacillus acidophilus used for the study.

All data are displayed and explained as obtained from each analysis, by using SD for replicates.

4

Information about data processing is missing in Materials and methods.

Revision has been made

5

Results and discussion parts have been revised and supplemented.

Thank you

6

Lactobacillus acidophilus grows very well in MRS media supplemented with prebiotics or without. It seems to the reviewer that studied XOS do not significantly change the growth pattern of L.acidophilus in the selected substrate. I suggest to make an additional experiment comparing your results with commercial XOS.

The difference in Lactobacillus growth at 0% XOS was different from 3 & 5% XOS (Fig.3) after 24 hours and this data was confirmed by data on total reducing sugars (Fig.4)

We have revised explain of the growth of bacteria and total reducing sugars and discussion (lines 217-264 and lines 305-345)

7

Conclusion part needs to revise.

Revision has been made

8

Some comments are given in the manuscript (attached).

Thank you. Revision has been made

No

Review

Revisions & Rebuttals

1

The title must be concise and meaningful 

We consider the title to represent our research

We consider the title to represent our research. If it has to be changed, we ask for improvements

2

no italicized some of the bacterial names. Spacing is major error often 

Revision has been made

3

The introduction did not include information related to the challenges of synthesizing XOS from plant biomass, technological barriers, and the market potential of the existing XOS market and their major sources for commercial production.  

Please see line 77-79

4

Line nos. 113-117: . Protein content in crude enzyme was determined by the Bradford method [32] using BSA (bovine serum albumin) as a standard while the activity was measured by the determination of reducing sugars by DNS (di-nitrosalicylic acid) by the Miller method The probiotic and media used was L. acidophilus MRSB (merck) and MRSm (MRS modified) respectively. Sentence framing is terrible, no proper punctuation.  Poor presentation where is protein part where is carbohydrate...all mixed. 

Revision has been made

5

Line No 126-127: Specific activity of endo-β-1,4-D-xylanase is 31.30 U/mg with total protein content is 0.454 mg 125 uL cassava pulp xylan substrate 1.1% in buffer (b/v dry base) and 125 uL 126 endo-β-1,4-D-xylanase were mixed. No Idea what to read until when? punctuation is a major issue here.

Revision has been made

6

pH measurement was done by pH meter.  Way of scientific writing must be improved. 

Revision has been made

7

Image quality is extremely poor.

All figures have been replace with a better one

8

The reducing sugar profile of XOS concentrations of 1% declined, Must be simple as XOS was consumed in line with the microbial growth. 

Revision has been made in discussion (lines 305-334)

9

LN 274: is massively digested, not a good term

Revision has been made (line 329)

10

LN 295: "trophic" effect on the epithelium? what is trophic defined here please elaborate ?

I deleted the above statement

11

LN 307-308: Bacterial  community maintains its pH for their growth and survival through managing the SCFA metabolism??? I dont think bacteria has control over pH regulation. Its a response to the consumption and release of nutrients and bioproducts

Revision has been made (lines 283-288)

12

Poor and no significance of discussion 

Revision has been made

13

LN 323: SCFA can  protect digestive system again diseases such as colectal cancer, chron's disease,: several typo mistakes in a small sentence

Revision has been made

14

Conclusion: growing rates? improper use of technical terms. MRSB must be consistent throughout the MS.  

Revision has been made

15

All the graphs and Table data must be statistically analyzed and must show the significant difference (p<0.05) between values with English letters. 

All data are displayed and explained as obtained from each analysis, by using SD for replicates.

Yes, Thank you for your suggestion, I have sent my revised manuscript to a professional proofreading service, and I hope it is now much better.

No

Review

Revisions & Rebuttals

1

Still, the write-up is poor. The author is unable to understand the exact technical term.  for example,  in Line number 333

The correction has been made

2

SCFA can protect the digestive system again diseases such as colectal cancer, chron's disease, and ulcerative colitis [22]. Here 'again' must be 'against' and 'colectal cancer' must be colorectal cancer.

Revision has been

3

The author is advised to take help from a technical writer however scientific content is still to be checked by an experienced researcher. 

Yes, Thank you for your suggestion, I have sent my revised manuscript to a professional proofreading service, and I hope it is now much better.

4

Line number 127: L.acidophilus spacing issue

Revision has been

5

Line number 266: The line 5 and 6 of TLC must be lane 5 and 6 of TLC

Revision has been

6

The graph 5 is not statistically analyzed to show the significant difference (p<0.05) between values.

All data are displayed and explained as obtained from each analysis, by using SD for replicates.

SD data is so small that it is not visible.

I send the data below