Co-Immobilization and Co-Localization of Oxidases and Catalases: Catalase from Bordetella Pertussis Fused with the Zbasic Domain
Round 1
Reviewer 1 Report
The paper is interesting although not entirely new, the same group has already proposed similar approached with various methodologies.
The paper is well written and interesting.
I have few general comments:
1) the product of the reaction is also a reactive aldehyde, how is this managed within the system? Is the aldehyde interacting with the lysines of the Zbasic domain and interfering with the rigidity of the cross-linking?
2) Did the authors test the reusability of the system?
3) the application to degradation of amines in wine is surely interesting, but are the by products (aldehyde or if oxidised carboxylic acids) compatible with the organolectic properties of wine after treatment?
As a minor comment the Bordetella pertussis is indicated in the title and at line 444 in a font which is not italics, as commonly used. I don't know if this is to be edited or if ti is a jpurnal format, but I would recomment the use od italics for such scientific names.
Author Response
The paper is interesting although not entirely new, the same group has already proposed similar approached with various methodologies.
The paper is well written and interesting.
I have few general comments:
1) the product of the reaction is also a reactive aldehyde, how is this managed within the system? Is the aldehyde interacting with the lysines of the Zbasic domain and interfering with the rigidity of the cross-linking?
the reaction between amino groups of Zbasic and glyoxyl groups of the supports yields unstable Schiff´s bases. However, the final borohydride reduction of the immobilized derivative transforms Schiff´s bases into very stable secondary amino bonds which are not affected by the presence of aldehydes in the reaction media
2) Did the authors test the reusability of the system?
In a previous paper, we had tested the good reusability of a similar bi-enzymatic biocatalyst with the same oxidase and other catalase. The new immobilized catalase is very stable and the biocatalyst stability (depending on oxidase stability) is supposed to be similar.
3) the application to degradation of amines in wine is surely interesting, but are the by products (aldehyde or if oxidised carboxylic acids) compatible with the organoleptic properties of wine after treatment?
It has been reported that the reaction products are not toxic and they are compatible with the organoleptic properties of fermented drinks
As a minor comment the Bordetella pertussis is indicated in the title and at line 444 in a font which is not italics, as commonly used. I don't know if this is to be edited or if ti is a jpurnal format, but I would recomment the use od italics for such scientific names.
Bordetella pertussis is now written in italics
Reviewer 2 Report
Please see attached a separate PDF file.
Comments for author File: 
Comments.pdf
Author Response
Comments on the Manuscript of “Co-immobilization and co-localization of oxidases and catalases: Catalase from Bordetella pertussis fused with the Zbasic domain” (Manuscript ID: catalysts-856659)
The present manuscript reported the preparation of a catalase from Bordetella pertussis fused with a small domain (Zbasic) and the subsequent co-immobilization and co-localization of this enzyme as an auxiliary enzyme with an amine oxidase from Pisum sativum to form a bi-enzymatic biocatalyst on the glyoxyl-agarose supports. The authors first characterized and optimized the conditions for the capability, stability, immobilization, and localization of the catalase, and then characterized and optimized the conditions for the co-immobilization and co-localization of both the catalase and oxidase. The authors also measured the catalytic activities of the prepared bi-enzymatic biocatalyst and showed some advantages of using such co-immobilized and co-localized bi-enzymes in some industrial processes (e.g., in wine).
The experiments in this manuscript were well designed and performed. The prepared enzymes and bienzymatic biocatalyst in this manuscript were well characterized as well. The bi-enzymatic biocatalyst prepared also showed good catalytic activities compared to free enzymes. This manuscript is recommended for publication in Catalysts after a minor revision, as commented below.
- Introduction section could be more
Introduction has been shortened
- In Figure 2: the manuscript mentioned that “The soluble protein extract that was adsorbed to SPagarose appears in lane 2.”, but Figure 2 doesn’t show anything in lane
- Formatting of the manuscript main text: on one hand, it seems that extra space sometimes exists between words, e.g., in lines 117, 124, 127, 159, 188, and 214. On the other hand, there is no space at all between some words, e.g., in lines 148 and
All these mistakes have been revised
- Line 444: “Bordetella pertussis” should be OK
- Conclusion section could be improved: the current format of this section looks like a lab
Conclusion has been summarized.
