Lactic Acid Bacteria Co-Encapsulated with Lactobionic Acid: Probiotic Viability during In Vitro Digestion
, Francesca Passannanti 2,3,*,†, Marianna Gallo 2,3,4,*, Rosa Colucci Cante 2, Federica Nigro 3, Andrea Luigi Budelli 2,5, Manuel Rendueles 1, Roberto Nigro 2 and Mario DÃaz 1Round 1
Reviewer 1 Report
The manuscript is well written and clearly presented. The co-encapsulation is well established method, so that originality of work is not great, but the benefit in expanding current knowledge is evident.
Minor spell check is required, but manuscript can be accepted in this form.
Author Response
We thank the referee for all the comments
Reviewer 2 Report
This MS is relevant for LAB co-encapsulated with lactobionic acid: Probiotic viability during in-vitro digestion. In this MS, use different conditions to evaluate resistant capsules in the article.
Microencapsulation technology in the most cross-linked capsules, coated with alginate and chitosan, had a useful microbial concentration to be considered probiotics and preservation.
Although some related literatures have been discussed. This MS showed the efficacy of double-coating microencapsulation for preserving LAB properties and more resistant freeze- and thermal-dried, survival rate, and demonstrates its potential for probiotic application.
Therefore, I do recommend this manuscript for publication in Applied Sciences.
Author Response
We thank the referee for all the comments
Reviewer 3 Report
After reviewing the manuscript “Lactic Acid Bacteria co-encapsulated with Lactobionic Acid: Probiotic Viability during in-vitro Digestion” submitted to the journal of Applied Sciences, I consider that the manuscript is well presented and the structure is of great merit. However, I have some observations and comments that need to be attained.
1.-page 3, L139: Please specify the type of thermal-dryer employed and conditions used such as vacuum pressure, air flow, how were the capsules distributed in the dryer.
2. page 6, L249: Can you please give a discussion about why is that LBA as a carbon source gave better cell viability and lactic acid production after 48 h when compared to the glucose as carbon source experiments.
3. page 7, L284: Missing a point after reference [37]
- page 7, L302: Can you explain what is the highlight, for a process industries , the observation of no difference on viability over the capsule drying method used.
Author Response
After reviewing the manuscript “Lactic Acid Bacteria co-encapsulated with Lactobionic Acid: Probiotic Viability during in-vitro Digestion” submitted to the journal of Applied Sciences, I consider that the manuscript is well presented and the structure is of great merit.
We thank the referee for the comment
However, I have some observations and comments that need to be attained.
1.-page 3, L139: Please specify the type of thermal-dryer employed and conditions used such as vacuum pressure, air flow, how were the capsules distributed in the dryer.
As written in the manuscript the thermal dryer was a CL252, Trevi. We did not apply a vacuum pressure, the air flow was 0.1-0.3 m/s. The capsules were evenly distributed on the surface to ensure homogeneous drying. As suggested by the referee we have added more informations in the manuscript.
- page 6, L249: Can you please give a discussion about why is that LBA as a carbon source gave better cell viability and lactic acid production after 48 h when compared to the glucose as carbon source experiments.
Although growth appears better at 48 hours for the LBA samples, the differences are not statistically significant. On the other hand, statistically significant differences are noted for the production of lactic acid. This may probably be due to substrate inhibition (considering glucose as the "direct substrate" to obtain lactic acid). This was not observed when lactobionic acid (requiring a more complex metabolic pathway) was used as a substrate; the same for the LBA-glucose mixture, for which the glucose concentration was halved. Furthermore this can be also due for the prebiotic effect of the LBA.
- page 7, L284: Missing a point after reference [37]
We thank the referee for the attention paid
- page 7, L302: Can you explain what is the highlight, for a process industries , the observation of no difference on viability over the capsule drying method used.
Between the two drying technologies there are several differences, not only from a technological point of view, but also in terms of time/costs. The heat drying, altough is not the golden method for probiotic drying (as the freezedrying is considered) is less expensive and faster, and this is a key factor to consider for an industrial process.
Author Response File: 
Author Response.docx
Reviewer 4 Report
Here are some minor changes/suggestions that might be needed.
Page 3 line 109: Change “equipped with a column C18” to “equipped with a C18 column”
Page 3 line 112: Was the lactic acid quantitively determined by HPLC? What internal standard/external standard was used?
Page 4 line 175 “LBA is a fiber resistant to human digestive enzymes”. LBA is resistant to human digestive enzyme, but it is a sugar acid, not fiber.
Typo:
Page 7 line 297: Change “Cacl2” to “CaCl2”
Page 10 line 407: should it be “(B-3 and B-5)” instead of (B-3 and C-5)?
Author Response
Here are some minor changes/suggestions that might be needed.
Page 3 line 109: Change “equipped with a column C18” to “equipped with a C18 column”
We thank the referee for the suggestion, and we have adjusted the sentence
Page 3 line 112: Was the lactic acid quantitively determined by HPLC? What internal standard/external standard was used?
Yes, lactic acid was quantitatively determined by HPLC, using pure lactic acid as an external standard. In particular we have built a calibration curve with 5 different concentrations of lactic acid. Before each analysis session we first analyzed the lactic acid standard, at the concentration closest to what we expected in our samples, to verify the method
Page 4 line 175 “LBA is a fiber resistant to human digestive enzymes”. LBA is resistant to human digestive enzyme, but it is a sugar acid, not fiber.
We thank the referee for the attention paid and we have modified the sentence
Typo:
Page 7 line 297: Change “Cacl2” to “CaCl2”
Page 10 line 407: should it be “(B-3 and B-5)” instead of (B-3 and C-5)?
We thank the referee for the great attention paid
Author Response File: Author Response.docx
