Lactic Acid Bacteria Isolation from Üçburun Peppers and Comparison of the Different Production Process for Pickled Pepper

Round 1

Reviewer 1 Report

In the work entitled Lactic Acid Bacteria Isolation from Üçburun Peppers and Comparison of the Different Production Process for Pickled Pepper, the isolation of some lactic acid bacteria from the microbiota of Üçburun pepper and the comparison of two methods of obtaining pickled pepper, respectively obtaining these products, were aimed at. by natural fermentation and by vinegar fermentation, were followed.

I recommend the authors carry out additional analyses (e.g. biochemical analysis, sensory analysis) for the two types of products obtained by two types of fermentation, natural and respective with the addition of vinegar, to establish which type of fermentation ensures obtaining products by consumer preferences, especially from the point of view of ensuring safe functional foods.

Author Response

RESPONSE TO REVIEWER 1

Manuscript ID: fermentation-2924219

In the work entitled Lactic Acid Bacteria Isolation from Üçburun Peppers and Comparison of the Different Production Process for Pickled Pepper, the isolation of some lactic acid bacteria from the microbiota of Üçburun pepper and the comparison of two methods of obtaining pickled pepper, respectively obtaining these products, were aimed at. by natural fermentation and by vinegar fermentation, were followed.

I recommend the authors carry out additional analyses (e.g. biochemical analysis, sensory analysis) for the two types of products obtained by two types of fermentation, natural and respective with the addition of vinegar, to establish which type of fermentation ensures obtaining products by consumer preferences, especially from the point of view of ensuring safe functional foods.

Response: Thank you so much for your valuable comments. For this purpose, some chemical analysis results performed within the scope of our study were given as supplementary information (Figure S1, S2, S3, S4 and Table S1) [the salt content, acid content, pH value, flavor, texture, scent and colour of semi-finished products taken on the 10th day and 32nd of fermentation. Microbiological analysis results on the 4th day, 7th month and 14th month of storage]. These graphs were discussed in the material-method and results-discussion section of the manuscript, with reference to them. At the same time, Table 3 has been added to the text. This table contains information about Brine formulations for pickle production.

Annex 1: Supplementary Information

Annex 2: Table 3

Annex 3: Sections added

Annex 1: Supplementary Information

SUPPLEMENTARY INFORMATION

Lactic Acid Bacteria Isolation from Üçburun Peppers and Comparison of the Different Production Process for Pickled Pepper

Ali Nalbant¹, Esra Ersoy Omeroglu¹*

Figure S1. Flow Chart of Assay Production.

Figure S2. Pickled jars of Üçburun peppers pickled.

Figure S3. Results of chemical analysis performed during storage of products produced with semi-finished products taken on the 10th day of fermentation. A: salt content, B: acid content, C: pH value, D: flavor, texture, scent and colour.

Figure S4. Results of chemical analysis performed during storage of products produced with semi-finished products taken on the 32nd day of fermentation. A: salt content, B: acid content, C: pH value, D: flavor, texture, scent and colour.

Table S1. Microbiological analysis results on the 4th day, 7th month and 14th month of storage.

Annex 2: Table 3

Table 3. Brine formulations for pickle production

Annex 3: Sections added

FOR MATERIAL-METHOD:

In 2.4. Preparation of Üçburun pepper pickles section:

To fill the 10th and 32nd day old semi-finished products into jars, 370 cc jars were used. Each jar was filled with semi-finished products and brine formulation specified in Table 3 was added while hot, by heating it to 80 °C. The jars were then sealed and pasteurized at 50-70 °C for 20 min (see supplementary information Figure S1).

In 2.5. Microbiological and Chemical analyses of samples section:

Various chemical (such as salt, acid and pH) and organoleptic (flavour, texture, scent and colour) properties of the products were determined during storage with the semi-finished products taken on the 10th and 32nd days of fermentation (see supplementary information Figure S1).

FOR RESULTS AND DISCUSSION:

In 3.4. Processing of Üçburun peppers pickles section:

Pepper pickles taken from the P0F and P0A storage barrels on days 10 and 32 were pickled, sorted, washed and packed in brines suitable for consumption and final product jars.  When the acid analysis results of the P10A coded pickles were analyzed, it was found that the acid values were in the range of 0.42% - 0.90%, while the acid values of the P10F coded pickles produced by fermentation were in the range of 0.54% - 0.90%.  The results of the 32-day semi-finished pickles and the total acid analyses of the final products obtained after composition equivalence were in the range of 0.72% to 0.99%. When the results of the salt analyses were analyzed, it was found that the products coded P10A were saltier than the product coded P10F because more salt was added in the brine. The salt values of the products coded P32A, P32F1 and P32F2 during the shelf life were between 2.93% and 4.1% (see supplementary information Figure S3 and S4).

In a study conducted by Çetinyokuş (1991), the brine titration acidity of commercial pickles collected from different provinces was investigated and found to be 0.50-3.70% in pepper. The salt content values of pickled peppers were determined as 1.73-10.24%. Akbaş (2006) determined the titration acidity values of pepper pickles to be 0.68-1.44% at storage periods of 0, 2 and 4 months at 4 ºC and 20±2 ºC. Salt content changes of pickles were determined as 5.01-6.19 %.

Sensory analysis was carried out throughout the shelf life of the products, looking at changes in taste, odour, texture and colour. Tasting tests were carried out with at least 5 panellists at intervals throughout the shelf life of all samples obtained. It was found that the fastest changing difference of all parameters during shelf life was the colour criterion. The final jars of P10A and P10F products started to differ in terms of the taste criterion at month 11. In terms of taste, odour and colour parameters, the P10A application maintained the highest score until the 9th month and as a result of this evaluation it was considered appropriate to end the shelf life at around the 9th month. When the products were evaluated in terms of structural parameters, it was found that the jars with P10A application were stable up to 14 months. It was observed that the fastest changing difference in terms of all parameters during the shelf life of the final product jars of P32A, P32F1 and P32F2, obtained with the semi-finished products taken on the 32nd day of storage tanks, was the taste and colour criterion. Compared to the other treatments in terms of taste, odour and colour parameters, the P32A application maintained the highest score until the 11th day of storage (see supplementary information Figure S3 and S4).

In additions microbiological analyses were conducted on samples taken from pasteurized pickle jars coded P10A, P10F, P32A, P32F1, and P32F2 using various methods. Microorganisms were detected in the range of 3.0x10¹ to 8.5x10¹ CFU/g in the TVC analyses of P10A, P10F, P32F1 and P32F2 products on the 4th day. No yeast or lactic acid bacteria were detected in the yeast analyses of the P10A, P10F, and P32A samples. However, yeast in the range of 2.5x10⁴ - 2.9x10⁴ CFU/g was detected in the 1st and 10th month yeast analyses of P32F1 and P32F2 samples. Additionally, no mold was detected in the shelf life analyses of the samples. Furthermore, the 10th month lactic acid bacteria count analyses of P32F1 and P32F2 samples detected lactic acid bacteria within the range of 5.0x10³-1.3x10⁴ CFU/g (see supplementary information Table S1). It was found that this increase in microorganisms in the jars was due to the increase in biota during the fermentation process of the products and that the pasteurization recipe used was not sufficient to eliminate the microorganisms in the product. On the other hand, it was found that the initial acid content of the P32A coded jars obtained by the acidification method was higher than that of the jars obtained by the fermentation method. In this context, it can be said that the pasteurization prescription applied is sufficient for acidified.

Author Response File: Author Response.pdf

Reviewer 2 Report

The authors conducted research aimed at isolating LAB from Üçburun peppers and comparing two different methods of producing pickles using this pepper in terms of suitability for industrial production. The authors collected samples from two different locations and isolated LAB. In turn, the obtained pickles were subjected to microbiological analysis, salt content, acidity and pH control. The data was presented in an accessible way (appropriate tables, figures and photos), described and briefly discussed. They were concluded with a summary indicating that pickles obtained by acidification were more suitable for production in the pickle industry. This includes indications for the practical use of the proposed pepper processing. My comment concerns only the "Conclusions" section. The first part of this section should rather be moved to the "Introduction" section and placed before the purpose of the work.

Author Response

RESPONSE TO REVIEWER 2

Manuscript ID: fermentation-2924219

The authors conducted research aimed at isolating LAB from Üçburun peppers and comparing two different methods of producing pickles using this pepper in terms of suitability for industrial production. The authors collected samples from two different locations and isolated LAB. In turn, the obtained pickles were subjected to microbiological analysis, salt content, acidity and pH control. The data was presented in an accessible way (appropriate tables, figures and photos), described and briefly discussed. They were concluded with a summary indicating that pickles obtained by acidification were more suitable for production in the pickle industry. This includes indications for the practical use of the proposed pepper processing. My comment concerns only the "Conclusions" section. The first part of this section should rather be moved to the "Introduction" section and placed before the purpose of the work.

Response: Thank you so much for your valuable comments. The first part of the conclusion section was removed and placed before the sentence in the introduction section stating the purpose of the study. At the same time, supplementary information regarding some chemical parameters determined within the scope of the study has been added to the publication and stated in the text. Additionally, Table 3 has been added to the publication and information about “Brine formulations for pickle production” was given.

Annex 1: Supplementary Information

Annex 2: Table 3

Annex 3: Sections added

Annex 1: Supplementary Information

SUPPLEMENTARY INFORMATION

Lactic Acid Bacteria Isolation from Üçburun Peppers and Comparison of the Different Production Process for Pickled Pepper

Ali Nalbant¹, Esra Ersoy Omeroglu¹*

Figure S1. Flow Chart of Assay Production.

Figure S2. Pickled jars of Üçburun peppers pickled.

Figure S3. Results of chemical analysis performed during storage of products produced with semi-finished products taken on the 10th day of fermentation. A: salt content, B: acid content, C: pH value, D: flavor, texture, scent and colour.

Figure S4. Results of chemical analysis performed during storage of products produced with semi-finished products taken on the 32nd day of fermentation. A: salt content, B: acid content, C: pH value, D: flavor, texture, scent and colour.

Table S1. Microbiological analysis results on the 4th day, 7th month and 14th month of storage.

Annex 2: Table 3

Table 3. Brine formulations for pickle production

Annex 3: Sections added

FOR MATERIAL-METHOD:

In 2.4. Preparation of Üçburun pepper pickles section:

To fill the 10th and 32nd day old semi-finished products into jars, 370 cc jars were used. Each jar was filled with semi-finished products and brine formulation specified in Table 3 was added while hot, by heating it to 80°C. The jars were then sealed and pasteurized at 50-70°C for 20 min (see supplementary information Figure S1).

In 2.5. Microbiological and Chemical analyses of samples section:

Various chemical (such as salt, acid and pH) and organoleptic (flavour, texture, scent and colour) properties of the products were determined during storage with the semi-finished products taken on the 10th and 32nd days of fermentation (see supplementary information Figure S1).

FOR RESULTS AND DISCUSSION:

In 3.4. Processing of Üçburun peppers pickles section:

Pepper pickles taken from the P0F and P0A storage barrels on days 10 and 32 were pickled, sorted, washed and packed in brines suitable for consumption and final product jars.  When the acid analysis results of the P10A coded pickles were analyzed, it was found that the acid values were in the range of 0.42% - 0.90%, while the acid values of the P10F coded pickles produced by fermentation were in the range of 0.54% - 0.90%.  The results of the 32-day semi-finished pickles and the total acid analyses of the final products obtained after composition equivalence were in the range of 0.72% to 0.99%. When the results of the salt analyses were analyzed, it was found that the products coded P10A were saltier than the product coded P10F because more salt was added in the brine. The salt values of the products coded P32A, P32F1 and P32F2 during the shelf life were between 2.93% and 4.1% (see supplementary information Figure S3 and S4).

In a study conducted by Çetinyokuş (1991), the brine titration acidity of commercial pickles collected from different provinces was investigated and found to be 0.50-3.70% in pepper. The salt content values of pickled peppers were determined as 1.73-10.24%. Akbaş (2006) determined the titration acidity values of pepper pickles to be 0.68-1.44% at storage periods of 0, 2 and 4 months at 4 ºC and 20±2 ºC. Salt content changes of pickles were determined as 5.01-6.19 %.

Sensory analysis was carried out throughout the shelf life of the products, looking at changes in taste, odour, texture and colour. Tasting tests were carried out with at least 5 panellists at intervals throughout the shelf life of all samples obtained. It was found that the fastest changing difference of all parameters during shelf life was the colour criterion. The final jars of P10A and P10F products started to differ in terms of the taste criterion at month 11. In terms of taste, odour and colour parameters, the P10A application maintained the highest score until the 9th month and as a result of this evaluation it was considered appropriate to end the shelf life at around the 9th month. When the products were evaluated in terms of structural parameters, it was found that the jars with P10A application were stable up to 14 months. It was observed that the fastest changing difference in terms of all parameters during the shelf life of the final product jars of P32A, P32F1 and P32F2, obtained with the semi-finished products taken on the 32nd day of storage tanks, was the taste and colour criterion. Compared to the other treatments in terms of taste, odour and colour parameters, the P32A application maintained the highest score until the 11th day of storage (see supplementary information Figure S3 and S4).

In additions microbiological analyses were conducted on samples taken from pasteurized pickle jars coded P10A, P10F, P32A, P32F1, and P32F2 using various methods. Microorganisms were detected in the range of 3.0x10¹ to 8.5x10¹ CFU/g in the TVC analyses of P10A, P10F, P32F1 and P32F2 products on the 4th day. No yeast or lactic acid bacteria were detected in the yeast analyses of the P10A, P10F, and P32A samples. However, yeast in the range of 2.5x10⁴ - 2.9x10⁴ CFU/g was detected in the 1st and 10th month yeast analyses of P32F1 and P32F2 samples. Additionally, no mold was detected in the shelf life analyses of the samples. Furthermore, the 10th month lactic acid bacteria count analyses of P32F1 and P32F2 samples detected lactic acid bacteria within the range of 5.0x10³-1.3x10⁴ CFU/g (see supplementary information Table S1). It was found that this increase in microorganisms in the jars was due to the increase in biota during the fermentation process of the products and that the pasteurization recipe used was not sufficient to eliminate the microorganisms in the product. On the other hand, it was found that the initial acid content of the P32A coded jars obtained by the acidification method was higher than that of the jars obtained by the fermentation method. In this context, it can be said that the pasteurization prescription applied is sufficient for acidified.

Author Response File: Author Response.pdf

Round 2

Reviewer 1 Report

In the work entitled Lactic Acid Bacteria Isolation from Üçburun Peppers and Comparison of the Different Production Process for Pickled Pepper, the isolation of some lactic acid bacteria from the microbiota of Üçburun pepper and the comparison of two methods of obtaining pickled pepper, respectively obtaining these products, were aimed at. by natural fermentation and by vinegar fermentation, were followed.

The novelty, the significance of the content, the quality of the presentation, the scientific validity, the interest of the readers, and the general merit can be greatly improved by carrying out new experiments (for example, the natural fermentation of peppers with lactic acid bacteria isolated from their microbiota) with the statistical analysis of the obtained data.