RESPONSE SHEET

Dear Editor,

Pleased attached herewith the response sheet for all the queries received earlier from the
editorial office for the manuscript entitled Here we are resubmitting our 1rd revised
manuscript entitled with “Isolation, molecular characterization of okara for the
development of synbiotic yogurt”. The response sheet has been prepared after mutual
consensus and incorporating expert opinion of all the research team members. with all the
suggested modifications in yellow highlighted portion up to best of our potential and
knowledge. We have tried our best to remove all the issues highlighted by the
reviewers/editors. Looking forward for your kind considerations!

Reviewer’s Comments Response

Reviewer 1
1) Line 43 Now a day change by Nowadays Thank you for your kind response, The Word Now
a day changed to “Nowadays”. The suggestion has
been incorporated

2) Line 43- 46 This introduction would be if the
dose had been evaluated to have a functional
effect, but it was not the case, delete and leave it as
perspectives to evaluate effect.
Dear reviewer, as per your suggestion leaves the
all COVID-19 perspectives in introduction and
highlights the importance of function foods and
healthy life style. The required suggestion has
been incorporated as per reviewer’s comment.

3) Line 50: animal health ?Or human and animal The suggestion has been incorporatedThank you
health? for your kind response the sentence changed to as
per your suggestion.

4) Line 102: Milk of... complete.
Line 132: You mention the incorporation of 3%,
why was that concentration decided, in a reference
cited by you (Ref 20, mention that 1% is the most
Dear reviewer, The specific sentence has been
rewritten according to your suggestionThe
suggestion has been incorporated at line # 102 as
follows
“Milk of buffalo was purchased from local dairy
farm of Faisalabad.”
Thank you for your kind response, Author
preferred 3 % okara because it contains greater
nutritional, antioxidant profile than 1 % okara
fortification.Our research team observed
significant role of 3% okara incorporation on the
adequate), justify.
Line 138-139: All the physicochemical analyses
were from the same reference...? Verify.
Line 125: In the yogurt preparation section, how
much was the initial inoculum of the yogurt and
probiotic, add it.
Lines 189-190: how many panelists performed the
sensory tests and age ranges.
Lines 203 - 204: remove the values in parentheses
because they are shown in Table 1.
Line 216: put the particle size data, as it how
impacts on the product.
Line 233: put the link C=O correctly.
Line 246: change to Fig. 3. Check in all the
writing: figure, fig or Figure homologate.
nutritional and therapeutic attributes of the end
product while minimally effecting the organoleptic
properties. However at 1% incorporation level
sensory properties were slightly improved by
nutritional and therapeutic features of the end
product were significantly reduced.
Thanks for your valuable suggestion about latest
references. Change the old reference to new
reference and verified.The references have been
updated in the mentioned section of the manuscript
after proper verification.
Thank you for your response, in yogurt preparation
2.5% initial inoculums culture was used in
OSY1and OSY2, on the other hand in control
yogurt only 2 % starter culture are used The
control sample of the yoghurt was inoculated by
2% starter culture of L. Rhamnous while OSY1
and OSY2 were inoculated with 2.5% starter
culture.
Thank you for your kind response, 30 members
included in this panelist and their age varied
between 26 to 45. The panel comprised of 30
members aged 26-45 years
Dear Reviewer, as per user suggestion values are
removed from these lines.The suggestion has been
incorporated
Dear reviewer, we study the morphology of okara
by SEM images, therefore, Okara particles have
wrinkled or aggregative surface and less dispersed.
Our experts on scanning electron microscopy
studied the morphology of okara through SEM
imaging technique and found okara particles to be
wrinkled or having aggregative surface with less
dispersion rate.
Thank you for this correction, C=O indicates
presence of carboxyl group therefore the
correction have been donemade in line # 233.
Thank you for suggestion, All the writing: Figure
are homologateThe changes have been
incorporated.
Lines 254 - 260: what about water holding
capacity. Because it is higher as it is affecting the
added ingredient, in line 260 they mention that the
results match with ref. 26, but they are referring to
okara alone not yogurt. Other yogurts as they are
in composition. It is not valid just to say that the
results coincide, that is validating, it is necessary
to discuss the results better. In Figure 3 modify the
WHC graph, put the days correctly.
Lines: 267-269: reference 27 mentions between
dry and wet okara, but the milk has an impact on
the color, change reference, better to put
application in other yogurts, okara was already
added with respect to those studies as impact on
the color,
Table 1 shows that the Lightness is the same in
OFY0 (control) and OFY2, but they mention that
okara affects the color. You could to change the
parameters a and b by chromaticity and hue value,
would show better if there are differences or not.
Line 141 and Line 270: In the antioxidant analysis
section, why didn't they do it in the okara product,
as part of the characterization or put references on
how much it has as a dry ingredient.
Is the amount of okara in the yogurt product
enough to have a functional effect (antioxidant
potential)? especially if you are mentioning that it
can be an option as a functional food, but in this
section they should discuss or justify better, they
only mention that it is a high activity, but other
studies that have reported functionally to have a
possible benefit.
Thank you for your valuable and critical
comments. We have made the correction
accordingly. The reference was only available for
okara not for yogurt. So the ambiguous statement
has been removed.
The reference was only available for okara,
however the outcomes of the current study
strongly supports improved water holding capacity
of end product after incorporation of okara. The
graph was has been modified accordingly.
Dear reviewer, Aas per your suggestion references
was changed updated with and most relevant
reference. was added, this reference justify the
color variation of yogurt
Thanks for your valuable suggestion about color
parameters. In current study There are no change
in L* and a* color parameters of all yogurts was
observed. On the other hand the color parameter
b* showed significant change of OFY2 as
compared to OFYo and OFY1. Moreover OFYo
and OFY1 showed no change in a* and b*
parameters.
Dear reviewer, Thanks for your valuable
suggestion, we add the more latest data, Dry okara
is a rich source of antioxidants such as TPC
(130mg GAE/100g), FC (2 mg QE/100g), DPPH
(34.7 %) and ABTS (37.84%) radical scavenging
capacity. Therefore, okara could be added in
different food items. It enhances the functional as
well as therapeutic potential of food items and
according to available literature is most suitable to
be used for treatment of different diseases.
Thank you for this comment,
Yes okara hasve functional effect in yogurt due to
its strong antioxidant potential. Please see line#
141 and 270 therefore we added explanations as
“Therefore, okara can be added in different food
items. It enhances the functional as well as
therapeutic potential of food items and used for
treatment of different diseases.”
Line 279: in the microbiological analysis section,
review L. rhamnosus, lactobacillus rhamnosus or
lactobacillus Rhamnosus.
Lines 314-330: these lines are repeated, delete.
The article mentions storage conditions On what
day was the evaluation at the beginning and at the
end? and sensory evaluation of different
parameters, including texture, which is an
important parameter in the finished product,
especially the consistency of the yogurt.
How is the consistency of the control yogurt
The consistency data can be compared with the
sensory texture.
Physicochemically you mention than WHC
increased, this is not seen here, the yogurt 2
presented syneresis?
Physicochemically, how is okara affecting yogurt?
There are studies where the addition of fiber does
not affect the consistency of the yogurt, so what
else could it affect in your developed product? try
to relate all your results.
Thank you for your kind response “in the
microbiological analysis section, L. rhamnosus
homologateThe suggestion has been incorporated
Thank you for your correction and delete this
repeated lines The changes have been made
accordingly.
Thank you dear reviewer for this critical point, we
evaluated the okara fortified yogurt at first day of
storage. All parameters are important in sensory
evaluation of okara fortified yogurt, on the other
hand consistency of okara fortified yogurt was is
rough not smooth as compared to control yogurt.
The consistency of OFY0 and OFY1 are were
smooth.
Thank you for this comment, Control yogurt has
smooth and fine consistency.
Thank you dear reviewer for this critical comment,
In the current study texture was assessed through
sensory evaluation along with we cannot compare
the consistency of yogurt with texture, we only
observed appearance or overall acceptability of
product.
Thank you for your valuable comment, schematic
diagram are made at day 1 of storage, at day 1
there is no syneresis observed and WHC are
increased. The yogurt 2 showed syneresis after
5th day of storage
Thank you dear reviewer for this comment,
oOkara fortification significantly improveds the
fiber contents, nutritional profile as well as
bioactive profile of yogurt. Concisely we can say
that okara acts as prebiotic for probiotics and
improves the gastrointestinal function of human.
Thank you dear reviewer for this comment, okara
is a fine yellow powder. It present in yogurt as a
suspended form therefore yogurt color and
appearance changed The incorporation of Okara
significantly improved the nutritional profile of the
end product, additionally yellow color of okara

Line 345 - 347: The panelists were asked prior to
the evaluation if they consumed fermented
products?, this is important in the selection of a
population to evaluate the acceptability of a
product. And ref. 20 suggests the addition of 1%
because 3% changes the properties of yogurt.
Lines 352-355. Eliminate lines, only indicate that
the product obtained has good characteristics in
fiber, protein. Put the most representative of what
was generated from Okara, as it influences
nutritionally and functionally.
The amount is sufficient to have a functional
effect. Lines 363-364 why do they mention that,
what results show it? Lines 366-367 do not show
the texture values in the developed product.
370-372 these lines refer to fiber and not to okara.
Reviewer 2
Line 51- The data pulled are too old, please update
the data at least to within nearly five years.
How guaranteed in 2.2.7 to count only the
probiotic (Lactobacillus rhamnosus)? Because S.
thermophilus and L. delbrueckii subsp. bulgaricus
may also grow in MRS medium.
also effected the color of the end product.
Thank you dear reviewer for this comment,
panelist did not know the okara and its nutritional
profile, they just suggest or evaluate the yogurt on
appearance, color or aroma. Therefore panelist
more prefers the simple yogurt because they know
this product. The panelist only graded the product
on the basis of appearance, color, aroma etc.
However they had no idea about the nutritional
profile of okara, therefore simple yoghurt was
preferred by the panelists.
In ref. 20 1 % addition more preferable because
this apparently produced less changed in yogurt
In view of ref. 20 1% incorporation of okara in
yoghurt improved the nutritional status of yoghurt
while minimally effecting its sensorial properties.
Thank you dear reviewer for this comment, The
mentioned sentence has now been removed, The
specific line has been rewritten according to your
suggestion. The changes have been made as per
reviewer’s comment.
Thank you dear reviewer for this comment, The
specific line has been rewritten according to your
suggestion. The mentioned line has been improved
as per provided guideline.
The mentioned sentence has now been
improved/rephrased and completed The needful
has been done in the specific section of the
manuscript.
Thank you for this correction, In line 51 placed the
latest data with latest reference The data has been
updated along with latest reference available on
specific study.
Dear reviewer, thank you critical comment, 2.7
showed only probiotics because we use selective
media for L. Rhamnosus count. This media only
give the specific colonies growth. Before the
initiation of the microbiological studies, on the
recommendations of our expert on microbiologh
sslective medium was used for the growth of L.
Rhamnous. So that to prevent the growth of
undesired microorganisms and to further

Line 129- “and” Italics were used
How soluble is okara in milk? dissolution?
Suspension? Precipitate? Please specify.
Line 34- Please state what properties of yogurt are
significantly affected. Because the properties do
not permit a determination of how much.
Line 37- Why would the number of probiotics be
higher than the total viable count ? “probiotic
count” should be in plural form.
The conclusion paragraph is too lengthy, please
summarize the full text. And speculative content
should not appear in the conclusion
harmonize the outcomes of this study.
In line 129 “and” changed to “and”Suggestion has
been incorporated
Dear reviewer okara is not soluble or dissolve or
not precipitate but it suspended in the yogurt. Due
to presence of high fiber content Okara is
suspendable in aqueous medium, and its
distribution was further made uniform due to
natural stabilizing properties of yoghurt
Thank you for your suggestion nutritional and
antioxidant properties of yogurt significantly
effects. During the storage period the research
team observed a significant improvement in
nutritional as well as storage stability of yoghurt
after incorporation of Okara. Please see results of
talble ----- clearly elucidating improvement in
nutritional properties of the end product.
Dear reviewer, thank you for your critical
comment, Our research team has observed during
the current study that the Okara dietary fiber
possesses promising prebiotic potential Okara has
dietary fibers, this dietary fiber acts as a prebiotic
source for probiotic L. Rhamnosus. Therefore
probiotic growth was increased during storage.
Moreover probiotic is a singular word.
Dear reviewer, as per your suggestion cConclusion
has been revised as per provided guidelines
incorporating the key outcomes of the current
study summarized