Improving the Functionality of Lentil–Casein Protein Complexes through Structural Interactions and Water Kefir-Assisted Fermentation
, Thuan-Chew Tan 1,3,*, Azhar Mat Easa 1, Muhammad H. Alu’datt 4, Carole C. Tranchant 5, Ali Madi Almajwal 6, Sana Gammoh 4, Sofyan Maghaydah 4,7, Mohammed Ali Dheyab 8, Mahmood S. Jameel 8 and Ali Al-Qaisi 9Round 1
Reviewer 1 Report
The research presented in the manuscript „fermentation-2216176” aims to enhance the solubility and protein quality of lentil proteins through the application of combined techniques, protein complexation of lentil proteins (LP) and casein proteins (CP) with water kefir-assisted fermentation. The authors extracted the lentil proteins and then obtained several protein complexes (9:1, 2:1, and 1:1) using lentil and casein proteins. Water solubility and composition of protein complexes were performed. Various methods include Fourier-Transform Infrared Spectroscopy (FTIR), spectrofluorometry, surface hydrophobicity, UV-VIS spectroscopy, a Zeta potential analyser, and scanning electron microscopy (SEM). The selected lentil-casein proteins complex (LCPC) was then subjected to fermentation using fermented water kefir. The following properties: pH, total soluble solids, protein digestibility and nutrients: phenolic compounds, total phenolic content, total saponin content, sugars, and yeasts, lactic acid bacteria, and acetic acid bacteria in the unfermented and fermented protein complex were determined. The results are presented in tables and graphics, statistically analyzed, discussed and compared to the literature.
The manuscript is organised according to the journal’s requirements. It is also well written, and the results support the conclusions.
However, a few shortcomings were detected and presented below.
Shortcomings of the manuscript
There is a difference between the LPCP complex presented in figure 1 as composed of 90% LP and 10% CP, and the selected LPCP complex at a ratio of 1.0:01 to study the effect of water kefir-assisted fermentation on its pH, composition, protein quality and microbiological quality.
References
The references are not numbered in order of appearance in the text. Also, they are not listed in this order at the end of the manuscript.
Some errors occur, even citing the name of the first author et al. or the name of two authors and the year. For instance, some citations contain the name of the first three authors:
L40-41 (Alrosan, Tan, Easa, et al., 2021a) and L54 (Alrosan et al., 2021a)
L36 (Alrosan, Tan, Easa, et al., 2021b)
L37 (Alrosan, Tan, Mat Easa, et al., 2021c)
Other citations contain the initials of the first author:
L41, 332, 351 (R. Wang et al., 2019)
L41, 45, 282, 291, 333, 366 (T. Wang et al., 2019)
L45, 477 (Y. H. Wang et al., 2020)
The differentiation should be done using „a”, „b”, etc., after the year when two or more references have the same first author and year.
Author Response
Manuscript Fermentation – 2216176– Submission of R1
Title: “Improving the functionality of lentil-casein protein complexes through structural interaction and water kefir-assisted fermentation”
Author's Response to Reviewers’ Comments – Feb 2023
Response to Reviewer 1
Dear Reviewer,
Thank you for your thorough assessment of our work. Your constructive comments are appreciated. They have all been considered to improve the manuscript, as indicated below. All amendments are highlighted (in yellow) in the revised manuscript.
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Reviewer’s Comment/Authors’ response |
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Comment |
The research presented in the manuscript „fermentation-2216176” aims to enhance the solubility and protein quality of lentil proteins through the application of combined techniques, protein complexation of lentil proteins (LP) and casein proteins (CP) with water kefir-assisted fermentation. The authors extracted the lentil proteins and then obtained several protein complexes (9:1, 2:1, and 1:1) using lentil and casein proteins. Water solubility and composition of protein complexes were performed. Various methods include Fourier-Transform Infrared Spectroscopy (FTIR), spectrofluorometry, surface hydrophobicity, UV-VIS spectroscopy, a Zeta potential analyser, and scanning electron microscopy (SEM). The selected lentil-casein proteins complex (LCPC) was then subjected to fermentation using fermented water kefir. The following properties: pH, total soluble solids, protein digestibility and nutrients: phenolic compounds, total phenolic content, total saponin content, sugars, and yeasts, lactic acid bacteria, and acetic acid bacteria in the unfermented and fermented protein complex were determined. The results are presented in tables and graphics, statistically analyzed, discussed and compared to the literature. The manuscript is organised according to the journal’s requirements. It is also well written, and the results support the conclusions. However, a few shortcomings were detected and presented below. Shortcomings of the manuscript |
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Response |
Thank you for your comments. We have further improved the manuscript based on your comments and suggestions. |
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Comment |
There is a difference between the LPCP complex presented in figure 1 as composed of 90% LP and 10% CP, and the selected LPCP complex at a ratio of 1.0:01 to study the effect of water kefir-assisted fermentation on its pH, composition, protein quality and microbiological quality. |
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Response |
Thank you for your comment. We have changed Fig. 1 to make it more understandable Line (104). |
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Comment |
References
The references are not numbered in order of appearance in the text. Also, they are not listed in this order at the end of the manuscript.
Some errors occur, even citing the name of the first author et al. or the name of two authors and the year. For instance, some citations contain the name of the first three authors:
L40-41 (Alrosan, Tan, Easa, et al., 2021a) and L54 (Alrosan et al., 2021a)
L36 (Alrosan, Tan, Easa, et al., 2021b)
L37 (Alrosan, Tan, Mat Easa, et al., 2021c)
Other citations contain the initials of the first author:
L41, 332, 351 (R. Wang et al., 2019)
L41, 45, 282, 291, 333, 366 (T. Wang et al., 2019)
L45, 477 (Y. H. Wang et al., 2020) |
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Response |
Thank you for your comment. We have revised the references following the journal's reference format. |
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Author Response File: 
Author Response.pdf
Reviewer 2 Report
Mohammad Alrosan and co-authors describes a new and successful method for overcoming low solubility of lentil proteins through structural interaction and water kefir-assisted fermentation. The work is interesting and scientifically sound, but there are still some minor points need focused .
1. References should be updated. Some references are missing volume or page numbers.
2. Fig 3 is not clear enough; the ordinate of Fig 6 is irregular.
Author Response
Manuscript Fermentation – 2216176– Submission of R1
Title: “Improving the functionality of lentil-casein protein complexes through structural interaction and water kefir-assisted fermentation”
Author's Response to Reviewers’ Comments – Feb 2023
Response to Reviewer 2
Dear Reviewer,
Thank you for your thorough assessment of our work. Your constructive comments are appreciated. They have all been considered to improve the manuscript, as indicated below. All amendments are highlighted (in yellow) in the revised manuscript.
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Reviewer’s Comment/Authors’ response |
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Comment |
Mohammad Alrosan and co-authors describes a new and successful method for overcoming low solubility of lentil proteins through structural interaction and water kefir-assisted fermentation. The work is interesting and scientifically sound, but there are still some minor points need focused.
1. References should be updated. Some references are missing volume or page numbers. |
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Response |
Thank you for your comment. We have revised the references following the journal's reference format. |
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Comment |
2. Fig 3 is not clear enough; the ordinate of Fig 6 is irregular
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Response |
Thank you for your comment. We have revised Fig. 3 to be clear for readers to view. |
Author Response File: Author Response.pdf
Reviewer 3 Report
The application of protein-protein interaction has been used in various industries, such as food, nutraceutical, and pharmaceutical. The design of new structures with improved functional properties, e.g solubility, emulsion, gelation, has been a priority for many research groups.
Water kefir is a sparkling fermented beverage prepared with water kefir grains. The slightly acidic drink is produced by fermenting a solution of sucrose, to which dried fruits have been added, with water kefir grains. Beside acid production, LAB species produce the homoexopolysaccharide matrix from which the kefir grain is formed, while yeast, through their proteolytic activity, assists the bacteria by providing nitrogen in a form that can be assimilated.
Fermentation is a viable processing technique to transform whole grains in edible foods since it is an affordable, less complicated technique, which not only transforms whole grains but also increases nutrient bioavailability and positively alters the levels of health-promoting components in derived whole grain products.
This study describes a new and successful method improving the nutritional-functional properties of lentil protein. By combining protein complexation with water kefir -assisted fermentation, the authors have studied the mechanism of the formation of novel compounds.
The authors of the study have investigated the occurrence of lentil-casein protein complexes as well as structure-function relationships for the novel compounds.
The study has involved many experimental steps, which have been suggestively presented in a diagram. The research procedure was developed in order to extract lentil proteins, perform protein complexation, select a protein complex and monitor fermentation. Protein structure and properties were evaluated using various techniques, such as determining water solubility, FTIR, spectrofluorimetry, non-covalent forces, surface hydrophobicity, UV-spectroscopy, zeta potential, SEM.
For the fermented lentil-casein protein complex, the authors have determined the pH, composition, protein quality and microbiological qualities. The style of presentation and the clarity of the results and discussion section are remarkable.
Accept in present form
Author Response
Manuscript Fermentation – 2216176– Submission of R1
Title: “Improving the functionality of lentil-casein protein complexes through structural interaction and water kefir-assisted fermentation”
Author's Response to Reviewers’ Comments – Feb 2023
Response to Reviewer 3
Dear Reviewer,
Thank you for your thorough assessment of our work. Your constructive comments are appreciated. They have all been considered to improve the manuscript, as indicated below. All amendments are highlighted (in yellow) in the revised manuscript.
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Reviewer’s Comment/Authors’ response |
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Comment |
The application of protein-protein interaction has been used in various industries, such as food, nutraceutical, and pharmaceutical. The design of new structures with improved functional properties, e.g solubility, emulsion, gelation, has been a priority for many research groups.
Water kefir is a sparkling fermented beverage prepared with water kefir grains. The slightly acidic drink is produced by fermenting a solution of sucrose, to which dried fruits have been added, with water kefir grains. Beside acid production, LAB species produce the homoexopolysaccharide matrix from which the kefir grain is formed, while yeast, through their proteolytic activity, assists the bacteria by providing nitrogen in a form that can be assimilated.
Fermentation is a viable processing technique to transform whole grains in edible foods since it is an affordable, less complicated technique, which not only transforms whole grains but also increases nutrient bioavailability and positively alters the levels of health-promoting components in derived whole grain products.
This study describes a new and successful method improving the nutritional-functional properties of lentil protein. By combining protein complexation with water kefir -assisted fermentation, the authors have studied the mechanism of the formation of novel compounds.
The authors of the study have investigated the occurrence of lentil-casein protein complexes as well as structure-function relationships for the novel compounds.
The study has involved many experimental steps, which have been suggestively presented in a diagram. The research procedure was developed in order to extract lentil proteins, perform protein complexation, select a protein complex and monitor fermentation. Protein structure and properties were evaluated using various techniques, such as determining water solubility, FTIR, spectrofluorimetry, non-covalent forces, surface hydrophobicity, UV-spectroscopy, zeta potential, SEM.
For the fermented lentil-casein protein complex, the authors have determined the pH, composition, protein quality and microbiological qualities. The style of presentation and the clarity of the results and discussion section are remarkable. |
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Response |
Thank you for your kind comments. |
Author Response File: Author Response.pdf
