Review Reports

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

The study on enzymatic modification of cassava starch using α-amylase is scientifically relevant. However, the manuscript requires improvements in clarity, conciseness and writing style. It contains repetitive content and overly detailed sentences that affect readability and flow. Several comments for improvement are provided below to enhance the overall quality of the manuscript.
Comment 1.
Line 120 & line 372: the abbreviation “NS” is inconsistent in the manuscript. At line 120, it is implied as non-fermented sun dried cassava starch, whereas at line 372 it is used for native starch. This creates confusion for the reader. Please ensure a single, consistent meaning for NS throughout the manuscript.
Comment 2. 
Line 233-235: The reason why 6 U/g α-amylase showed the highest specific volume is not clearly explained. Please provide a proper scientific justification for this trend. 
Comment 3.
Line 273-278: The phrase “this result is consistent with the finding of a work, where was demonstrated…” is repetitive and poorly structured. The wording is reducdant and affects readability. It is recommended to rephrase the sentence to avoid repetition of similar expressions and improve academic flow.
Comment 4.
Lines 434-438: the sentence is too long and complex, which makes it difficult to understand. It is recommended to split it into shorter, clearer sentences to improve readability and ensure better scientific comprehension.
Comment 5.
Lines 476-478: The use of “For example” is unnecessary in a formal discussion section and should be removed or replaced with a more academic transition (e.g., similarly or comparable values were reported by…).
Comment 6.
Line 489: The unit expression “6 U of α-amylase.g” is not written in standard scientific format. It is recommended to revise it for clarity and consistency, e.g., “6 U/g α-amylase”. Please ensure uniform unit formatting throughout the manuscript.
Comment 7.
Line 560: The statement “there is no evidence of similar studies…” is too absolute and should be revised or supported with a literature search.
Comment 8.
Line 593-603: The section on RDS is informative but contains redundancy. It is recommended to merge overlapping interpretations of enzymatic effects on starch digestibility.
Comment 9.
Lines 619-626: The comparison with traditional cassava products is relevant but sentences are long and should simplified for readability.

 

 

 

Comments for author File: Comments.pdf

Comments on the Quality of English Language

The study on enzymatic modification of cassava starch using α-amylase is scientifically relevant and has strong potential for publication. However, the manuscript requires improvements in clarity, conciseness and writing style. It contains repetitive content and overly detailed sentences that affect readability and flow. Several comments for improvement are provided in attachment to enhance the overall quality of the manuscript.

Author Response

We sincerely thank the reviewer for the careful evaluation of our manuscript and for the constructive comments provided. All observations have been carefully addressed, and the manuscript has been revised accordingly. Below, we provide a point-by-point response detailing the modifications made.

Comment 1

Line 120 & line 372: the abbreviation “NS” is inconsistent in the manuscript. At line 120, it is implied as non-fermented sun dried cassava starch, whereas at line 372 it is used for native starch. This creates confusion for the reader. Please ensure a single, consistent meaning for NS throughout the manuscript.

Response: The manuscript has been revised to ensure that “NS” is used consistently and exclusively to denote non-fermented sun-dried cassava starch. All ambiguous instances were corrected to improve clarity and avoid misinterpretation.

Comment 2

Line 233-235: The reason why 6 U/g α-amylase showed the highest specific volume is not clearly explained. Please provide a proper scientific justification for this trend.

Response: The explanation has been strengthened in the revised manuscript to better reflect the underlying mechanism: The highest specific volume was obtained at 6 U/g α-amylase. This behavior is associated with partial hydrolysis of starch chains, which enhances water absorption and promotes granule swelling. Under these conditions, a viscoelastic network forms, which retains gas during baking, thereby contributing to greater product expansion. This interpretation has been clarified and supported in the Discussion section.

Comment 3

Line 273-278: The phrase “this result is consistent with the finding of a work, where was
demonstrated…” is repetitive and poorly structured. The wording is reducdant and affects
readability. It is recommended to rephrase the sentence to avoid repetition of similar expressions and improve academic flow..

Response: The sentence has been rephrased to eliminate redundancy and improve clarity, resulting in a more concise and readable expression.

Comment 4

Lines 434-438: the sentence is too long and complex, which makes it difficult to understand. It is recommended to split it into shorter, clearer sentences to improve readability and ensure better scientific comprehension.

Response: The sentence has been divided into shorter statements to improve readability and comprehension without altering the scientific content.

Comment 5

Lines 476-478: The use of “For example” is unnecessary in a formal discussion section and should be removed or replaced with a more academic transition (e.g., similarly or comparable values were reported by…).

Response: The expression has been replaced with a more suitable academic transition to maintain formal tone and consistency.

Comment 6

Line 489: The unit expression “6 U of α-amylase.g” is not written in standard scientific format. It is recommended to revise it for clarity and consistency, e.g., “6 U/g α-amylase”. Please ensure uniform unit formatting throughout the manuscript

Response: The unit format has been corrected to “U/g α-amylase” and standardized throughout the manuscript to ensure consistency and adherence to scientific conventions.

Comment 7

Line 560: The statement “there is no evidence of similar studies…” is too absolute and should be revised or supported with a literature search.

Response: The statement has been revised to adopt a more precise and cautious wording, supported by relevant literature.

Comment 8

Line 593-603: The section on RDS is informative but contains redundancy. It is recommended to merge overlapping interpretations of enzymatic effects on starch digestibility

Response: The section has been streamlined, improving conciseness and clarity.

Comment 9

Lines 619-626: The comparison with traditional cassava products is relevant but sentences are long and should simplified for readability.

Response: The text has been simplified and reorganized into shorter sentences to enhance readability.

Reviewer 2 Report

Comments and Suggestions for Authors

The manuscript addresses an interesting and relevant topic regarding the enzymatic modification of cassava starch using α-amylase and its impact on both functional and nutritional properties in breadmaking. The study is well-structured, and the experimental design is generally appropriate. The results provide valuable insights, particularly in identifying the optimal enzyme range (6–9 U/g) to balance technological performance and glycemic response.

However, several aspects should be improved:

Language and clarity:
The manuscript would benefit from careful English editing. There are grammatical errors, inconsistent terminology (e.g., “litterature”), and some sentences that are difficult to follow. Improving clarity and conciseness will significantly enhance readability.

Introduction:
The introduction is comprehensive but could be more concise and better focused. Some sections are repetitive. Additionally, the inclusion of more recent references (last 5 years) would strengthen the scientific context.

Methodology:
Although the methods are generally well described, some details require clarification:

-Justification for the selected enzyme dosages (2–9 U/g) should be better explained.

-Provide more detail on replication and variability (e.g., number of independent experiments vs. technical replicates).

Results and Discussion:
The discussion is thorough but sometimes overly descriptive. The authors are encouraged to:

-Reduce repetition of numerical results already presented in tables.

-Strengthen the interpretation by linking results more explicitly to mechanisms (e.g., structural changes in starch).

-Improve the flow between sections.

Tables and presentation:
Tables are informative but could be simplified for better readability. Consider:

-Reducing redundant information.

-Improving formatting and alignment.

Conclusions:
The conclusions are supported by the results but could be more concise and focused on the main findings and practical implications.

Overall, the manuscript presents valuable findings but requires minor to moderate revisions, particularly in terms of language, clarity, and presentation, before it can be considered for publication.

Comments on the Quality of English Language

The English language in the manuscript is generally understandable; however, it requires improvement to enhance clarity and readability. Several grammatical errors, awkward sentence structures, and inconsistent terminology (e.g., “litterature”) are present throughout the text. In some sections, long and complex sentences make the content difficult to follow.

The authors are encouraged to revise the manuscript carefully, preferably with the assistance of a native English speaker or professional editing service, to improve grammar, consistency, and overall flow of the text.

Author Response

We sincerely thank the reviewer for the careful evaluation of our manuscript and for the constructive comments provided. All observations have been carefully addressed, and the manuscript has been revised accordingly. Below, we provide a point-by-point response detailing the modifications made.

Comment 1

Language and clarity:
The manuscript would benefit from careful English editing. There are grammatical errors, inconsistent terminology (e.g.,“litterature”), and some sentences that are difficult to follow. Improving clarity and conciseness will significantly enhance readability.

Response: The manuscript has been thoroughly edited to improve grammar, clarity, and consistency. Sentences were simplified, redundant expressions removed, and terminology standardized.

Comment 2

Introduction:
The introduction is comprehensive but could be more concise and better focused. Some sections are repetitive. Additionally, including more recent references (the last 5 years) would strengthen the scientific context.

Response: The Introduction has been streamlined and updated with recent literature (the last 5 years), strengthening the scientific context of enzymatic starch modification.

Comment 3

Methodology:
Although the methods are generally well described, some details require clarification:
-Justification for the selected enzyme dosages (2–9 U/g) should be better explained.

Response: The selection of α-amylase dosages was based on preliminary experimental trials guided by previous findings by Maldonado-Alvarado et al. (2023) [1]. Initial assays focused on intermediate enzyme levels (10, 20, and 30 U/g α-amylase), which were directly evaluated in breadmaking through specific volume measurements. These trials revealed a consistent decrease in bread expansion as enzyme concentration increased, confirming the system's sensitivity to enzymatic dosage under processing conditions.

Based on this response, a lower and narrower range of α-amylase concentrations (2, 4, 6, 8, and 9 U/g α-amylase) was selected to more accurately characterize the dose–response behavior under controlled hydrolysis conditions. This approach enabled a more precise evaluation of the relationship among enzymatic modification, starch functionality, and breadmaking performance.

Comment 4

Provide more detail on replication and variability (e.g., number of independent experiments vs. technical replicates).

Response: It has been clarified that all experiments were conducted in triplicate using independent replicates, and results are expressed as mean ± standard deviation.

Comment 5

Results and Discussion:
The discussion is thorough but sometimes overly descriptive. The authors are encouraged to:
-Reduce repetition of numerical results already presented in tables.

Response: The discussion has been revised to reduce repetition and improve interpretation, emphasizing mechanisms rather than numerical repetition.

Comment 6

Strengthen the interpretation by linking results more explicitly to mechanisms (e.g., structural changes in starch).

Response: The discussion has been revised to more clearly connect the observed results with the structural changes induced by α-amylase. The interpretation now emphasizes that intermediate enzyme levels promote controlled depolymerization of amylose and amylopectin, enhancing water–starch interactions, granule swelling, and formation of a cohesive viscoelastic matrix.

At higher dosages, the discussion explains the effects in terms of excessive hydrolysis, leading to low-molecular-weight dextrins, reduced structural integrity, and lower viscosity and stability.

These additions provide a more mechanistic and integrated interpretation of the functional, rheological, and nutritional results, supported by recent literature.

Comment 7

Improve the flow between sections

Response: The manuscript has been reorganized to improve coherence and logical progression between sections.

Comment 8

Tables and presentation:
Tables are informative but could be simplified for better readability. Consider: Reducing redundant information.

Response: Tables have been revised to reduce redundancy and improve clarity and formatting.

Comment 9

Improving formatting and alignment.

Response: The format was standardized, and the presentation of the tables was improved

 

Comment 10

Conclusions:
The conclusions are supported by the results but could be more concise and focused on the main findings and practical implications.

Response: The Conclusions section has been critically revised to improve precision, focus, and scientific impact. Redundant and descriptive statements were removed, and the section was restructured to clearly emphasize the main findings, particularly the identification of an optimal α-amylase range (6–9 U/g) that balances functional performance and starch digestibility.

In addition, the revised conclusions more explicitly highlight the practical relevance of the results, positioning controlled enzymatic modification as a viable and reproducible strategy for tailoring cassava starch functionality for gluten-free breadmaking and modulating glycemic response.

Reviewer 3 Report

Comments and Suggestions for Authors

The research undertaken and the results described in the publication are undoubtedly interesting. Despite numerous attempts to find new carbohydrate sources, especially for use in gluten-free products, the topic remains relevant. However, the work submitted for review contains many shortcomings and requires numerous corrections.

The introduction is rather chaotically written and requires streamlining and possibly shortening.

The most significant concerns concern the methodology. This chapter is described very vaguely. From the description, one might assume that starch determination primarily involves extraction, centrifugation, and drying. Despite the references to the literature, it would be worthwhile to provide more details regarding the nature of the determination. I also have serious doubts about the methodology for determining the glycemic index, and the obtained GI results are rather controversial. Can the authors clarify these issues?

The authors did not address the toxicity of modified cassava at all. What is the content of cyanogenic glycosides in the final product?

The discussion also requires revision, and above all, organization.

Author Response

We sincerely thank the reviewer for the careful evaluation of our manuscript and for the constructive comments provided. All observations have been carefully addressed, and the manuscript has been revised accordingly. Below, we provide a point-by-point response detailing the modifications made.

Comment 1

The introduction is rather chaotically written and requires streamlining and possibly shortening.

Response: The Introduction has been revised to improve structure, focus, and readability. Repetitive statements were removed, and the text was reorganized to present the scientific rationale more clearly: first, the technological limitations of native cassava starch; second, the relevance of sour cassava starch in gluten-free breadmaking; and finally, the potential of controlled α-amylase treatment as a reproducible modification strategy.

Recent literature was also incorporated to strengthen the scientific context and better position the study's novelty

Comment 2.

Doubts about glycemic index methodology and controversial GI results

Response: The methodology for the estimated glycemic index has been clarified in the revised manuscript. We now explicitly state that the reported values correspond to an estimated glycemic index (eGI) derived from in vitro starch hydrolysis kinetics, not to a direct in vivo glycemic index.

The revised Methods section explains that hydrolysis curves were constructed using pancreatic α-amylase and amyloglucosidase, followed by calculation of the hydrolysis index and conversion to eGI using an empirical equation. This approach is widely used to compare relative starch digestibility among treatments under controlled conditions. Recent studies continue to apply in vitro hydrolysis methods to estimate glycemic response, while recognizing that these values should be interpreted as approximations rather than direct physiological measurements.

To avoid overinterpretation, the Results and Discussion section was also revised. The term “glycemic index” was replaced or clarified as “estimated glycemic index” where appropriate.

Comment 3

What is the content of cyanogenic glycosides in the final product?

Response: Cyanogenic glycoside content was not determined in the present study. The raw material corresponded to the INIAP 651 variety, for which an initial hydrocyanic acid (HCN) content of 0.230 mg·g⁻¹ in fresh roots has been reported by Castro-Moreira et al. (2021) [2]. This value decreases to 0.040 mg·g⁻¹ after processing steps—including peeling, grating, washing, sedimentation, and drying—representing an overall reduction of approximately 85–88%.

Given that the study focused on the functional and nutritional properties of already processed starch, the determination of cyanogenic glycosides was not included in the experimental design.

1. Maldonado-Alvarado, P.; Abad-Quevedo, V.; Maldonado-Alvarado, P.; Abad-Quevedo, V. Evaluation of a Low-Glucose Gluten-Free Bread Made from Hydrolyzed Cassava Starch and Lupine Flour. Biol. Life Sci. Forum 2023, Vol. 25, 2023, 25, 1, doi:10.3390/BLSF2023025001.
2. Castro, Y.; Cristellot, F.; Murgueitio, N.; Gómez, Y.; Rosero, E. Efecto del procesamiento tradicional de la yuca (Manihot esculenta) y derivados sobre el contenido de glucósidos cianogénicos | Revista Científica INGENIAR: Ingeniería, Tecnología e Investigación. ISSN: 2697-3693. Ing. Tecnol. e Investig. 2021, 157–170, doi:https://doi.org/10.46296/ig.v4i8.0033.

Round 2

Reviewer 3 Report

Comments and Suggestions for Authors

I would like to thank the authors for their quick and specific responses to my questions and concerns. I believe the introduction has been significantly improved and provides a much better explanation of the research undertaken. The authors have clarified my doubts regarding the glycemic index; the term "estimated glycemic index" used in this version of the publication is much more precise. I also accept the explanation for omitting the cyanogenic glycoside content label; the explanation is reasonable and convincing. The authors have corrected the shortcomings I reported and convincingly explained the ambiguities.