Health-Promoting Role of Fermented Pigeon Pea (Cajanus cajan L (Mill)) Milk Enriched with γ-aminobutyric Acid (GABA) Using Probiotic Lactiplantibacillus plantarum Dad-13
and Dietmar Haltrich 6Round 1
Reviewer 1 Report
Although this work is presented in a systematic way with explained obtained results that have been compared with the literature data, some things have to be revised so the manuscript would gain clarity. In the attached file there are several things listed that have to be revised.
Comments for author File: 
Comments.pdf
Author Response
Lines 5 and 6: remove free space between author’s last name and superscript of the affiliations
Comment: Done
Line 23: replace one gadB genes with one gadB gene
Comment: Done, the change is mark in red, thank you
Line 43: replace 46. 7 with 46.7
Comment: Done
Line 45: replace in with In
Comment: Done
Line 46: replace 26. 4 with 26.4
Comment: Done
Line 51: replace coma with dot at the end of the sentence
Comment: Done
Line 53: It should be either “or their GRAS status” or “due to GRAS status of the LAB”. Rephrase it.
Comment: Thank you, we have rephrased it.
Line 54: For what Y stands for?
Comment: we’re sorry, it is just typo and we have deleted it.
Line 57: This sentence is unclear. Should it be that biosynthesized GABA are mainly secreted to the…?
Comment: Thank you for your revision, yes GABA are secreted to the extracellular environment and we have corrected the sentences to make it clear.
Line 61: replace L pentosus with L. pentosus; L.plantarum replace with L. plantarum and there is at least one free space to much between lactis and [13,21].
Comment: Done.
Line 63: This sentence should be revised. If the authors are counting the effect they should be written as nouns e.g. resistant to high bile salts should be replaced with resistance to high bile salts and so on.
Comment: Thank you, we have corrected the sentences.
Line 78: remove it
Comment: Done
Line 89: Which isolates?
Comment: It was L. plantarum Dad-13, we have corrected the sentences
Line 97: Please add the country of origin for Philips.
Comment: Done, we have added the country
Line 105: Please add the country of origin for Oxoid.
Comment: Done.
Line 106: Please add the producer’s name and country for the pH meter. Also, there is lack of reference
for the titratable method.
Comment:
Lines 111-113: This is really unclear written. Please rephrase it.
Comment: Done, we have added the missing information in this section.
Line 115: Remove dot at the end of the title
Comment: Done.
Lines 116-117: This sentence in unclear. Please rephrase it.
Comment: Done
Line 123: It should be 350°C.
Comment: Done.
Line 130: Please add the country of origin for Thermo Scientific.
Comment: Done
Line 152: There should at least a few sentences before the actual results so that a reader has a slight introduction.
Comment: Thank you, we have added a few sentences
Line 174-178: These sentences are unclear. Please rephrase them. What d isolate soy bean?
Comment: Done, we have corrected the sentences
Line 178: Remove ) at the end of the sentence.
Comment: Done
Line 199: replace et al with et al.
Comment: Done
Line 201: remove free space between ] and .
Comment: Done
Line 211: replace in with In
Comment: Done
Line 221: This sentence is not finished.
Comment: thank you, we have added some sentences.
Line 225: replace et al with et al.
Comment: Done
Line 240: replace strain with strains
Comment: Done
Line 242: replace “which is may activate” with “which may activate” and “located in” replace with “is located in”
Comment: Thank you, we have changed the sentences.
Line 244: remove in (gene is located between)
Comment: Done, thank you
Line 247: Put a dot at the end of the sentence
Comment: Done
Line 249: remove one to
Comment: Done
Line 255: replace in with In; replace et al with et al.
Comment: Done
Line 256: replace one genes with one gene
Comment: Done
Line 257: replace {42} with [42]; put a dot at the end of the sentence
Comment: Done
Line 263: finish the legend
Comment: Done
Line 291: replace {48] with [48]
Comment: Done
Line 311: remove a dot at the end of the title
Comment: Done
Line 340: remove a dot at the end of the title
Comment: Done
Line 425: remove free space between ( and M
Comment: Done
Reviewer 2 Report
The topic that the authors deal with - the addition of GABA-producing strains to food - is very popular and this research will be of interest to readers. The research is well designed. These are suggestions by which the authors can improve the manuscript:
L60 Several LAB and probiotic strains are able to produce GABA namely, L. pentosus, L. brevis, L. plantarum and L. lactis.
There are some other GABA-producing strains such as L. reuteri. The authors should consider changing that sentence: Several LAB and probiotic strains are able to produce GABA including L. pentosus, L. brevis, L. plantarum and L. lactis
L100 The effects of nutrients such as MSG (0-1%), whey isolate (0-4%) and sucrose (0-3%) were determined.
Did you add those nutrients during sample preparation? Written like this, it sounds as if they were already in the germinated beans, not that they were added. Explain in more detail.
L152 GABA production in CCM can be enhanced by the addition of nutrients (Fig 1)
The authors should consider adding which nutrients, i.e. (GABA production in CCM can be enhanced by nutrient supplementation, MSG, whey, sucrose (Figure 1-3)
L240 L. plantarum WCFS1 and L. brevis ATCC 367
Please include these strains in the Materials section (where you obtained them, are they part of a microbiology collection)
L249 next to to gadC
L339 Figure 8. Instead of TNF alpha, write TNF-α, the fermentation time should be in bold on each graph, mark the graphs with a), b), c)
Author Response
L60 Several LAB and probiotic strains are able to produce GABA namely, L. pentosus, L. brevis, L. plantarum and L. lactis.
There are some other GABA-producing strains such as L. reuteri. The authors should consider changing that sentence: Several LAB and probiotic strains are able to produce GABA including L. pentosus, L. brevis, L. plantarum and L. lactis
Comments: Thank you, the sentences were already written in line 60.
L100 The effects of nutrients such as MSG (0-1%), whey isolate (0-4%) and sucrose (0-3%) were determined.
Did you add those nutrients during sample preparation? Written like this, it sounds as if they were already in the germinated beans, not that they were added. Explain in more detail.
Comments: yes we add the nutrients during preparation of fermented CCM. the nutrients such as MSG (0-1%), whey isolate (0-4%) and sucrose (0-3%) were added into CCM drink. The mixtures were then sterilized at 121oC for 5 min, prior to fermentation. Freshly prepared CCM milk was inoculated with L. plantarum Dad-13 (5% v/v) with approximately final cell density of 9 log CFU/mL. Fermentation was carried out under a close container at 30oC for 48 h without shaking. The GABA production in fermented CCM was further determined
L152 GABA production in CCM can be enhanced by the addition of nutrients (Fig 1)
The authors should consider adding which nutrients, i.e. (GABA production in CCM can be enhanced by nutrient supplementation, MSG, whey, sucrose (Figure 1-3)
Comment: thank you, we have added a few sentences.
L240 L. plantarum WCFS1 and L. brevis ATCC 367
Please include these strains in the Materials section (where you obtained them, are they part of a microbiology collection)
Comment: Actually, we do not use these strains in our experiments, these strains we merely used as a comparison for the whole genome sequence (WGS) of our strain. The WGS of these strains were automatically generated and annotated using RAST system (https://rast.nmpdr.org/).
L249 next to to gadC
Comment: done, thank you.
L339 Figure 8. Instead of TNF alpha, write TNF-α, the fermentation time should be in bold on each graph, mark the graphs with a), b), c)
Comment: Thank you, we have corrected the graph.
