Software for Matching Standard Activity Enzyme Biosensors for Soil Pollution Analysis

Round 1

Reviewer 1 Report

Software for enzyme biosensors for soil pollution analysis: The experimental work appears to have been carried out well. However, a few points deserve attention for further publication. I suggest that it is accepted for publication after the following revisions:

- ABSTRACT: This work is dedicated to developing enzyme biosensor software to solve problems regarding soil pollution analysis: What parameters were optimized? Authors must include numbers with the results found. Stability? How much protein (mg) was utilized to enzymes process? Furthermore, what are the conditions of enzymatic reactions? Temperature, pH, ionic strength, for example. This information should be included in the abstract.

- INTRODUCTION:

- Soil pollution, this information needs to be explained in the introduction of the manuscript.

- A paragraph describing the properties, application mechanism of actuation of enzyme to biosensors must be included in the manuscript.

- Another point that must be considered, in this study the authors is the inhibitory activity and specific affinity to enzymes onto biosensors. This information must be clear in the introduction.

- In this study, we describe the development of a specialized enzymatic biosensor software: The contribution and importance of these studies in the work performed must be explained in the introduction of the manuscript.

- RESULTS AND DISCUSSION:

- The stability of enzyme biosensors for soil pollution in organic solvents, metal ions, or detergent enables its wide application in synthesis processes which nowadays are in great demand from the point of view of industrial enzymes. This information should be included.

- Enzyme biosensors for soil pollution: With respect to maintenance of the enzymatic activity was slow/ fast at all studied pH values? A significant decrease in enzyme activity? This information must be clear in the manuscript.

- Was determined the full loading of Enzyme biosensors for soil pollution prepared under the optimized immobilization conditions? This information must be clear in the manuscript.

- Enzyme biosensors for soil pollution: The optimization of enzyme preparation process, the preparations shown having diffusion limitations? Considering the strategy presented in this manuscript. Please, this should be explained in the manuscript. What were the optimum conditions?

- Enzyme biosensors for soil pollution: The loading capacity of lipase immobilized prepared and the amount of attached Protein: What were the optimum conditions? Please use numbers, no qualitative statements (high, good, higher, etc).

- The reusability of enzyme biosensors for soil pollution: After cycles, please, an explanation of these results. What other factors can influence the results achieved? In addition, the results should be compared with other works of the literature in the same line of application.

- The thermal stability of enzyme biosensors for soil pollution prepared is one of the most important application criteria for different applications. This stability depends on the enzyme preparation strategy. It also depends on the stabilization of the enzyme. This discussion could be improved. Please include it in the manuscript.

- Enzyme biosensors for soil pollution: The storage stability of the immobilized enzyme, other factors that cause the loss of durability and stability of the biocatalysts should be explained in the manuscript.

- CONCLUSIONS: The main contributions to the accomplishment of this work must be included in the conclusion. Please, authors must use numbers.

- Please, check all references according to the author's instructions.

- Include more details in the figures (error bars) and table captions.

- The manuscript must be formatted according to the journal's standards.

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Reviewer 2 Report

The article by Kratasyuk et al. is devoted to an important area of soil ecotoxicity assessment and Risk management. Despite the promising topic, this article should be improved significantly before publication.

At first, the title, abstract, and introduction do not match with the presented data. It looks that the authors give a technical overview of a new JavaScript-based program. Simultaneously, the methodology part and the result covered data, which includes only standard soil testing results without any pollution. So it's not clear does this test is suitable for soil pollution assessment or not. Moreover, there is no data and links where this program could be downloaded and tested (arXiv link or etc).

Second, the list of parameters used in this test is not supported (Table 1). It's not clear why authors measured the parameters like "residual activity of BChE Residual luminescence value of Red + Luc system"," Residual luminescence value of LDH + Red + Luc system", and "D250". Probably they proposed and approved these parameters in previous publications, but relevant references are also not listed.
It's not supported by references to how the "soil pH" and "Clay content" should be measured. For example, should I calculate the soil pH after KCL extraction or in H2O?
Notably, "soil humus" is highly controversial in literature terms, so I recommend using it in high-quality journals. More than five years later, Lehmann and Kleber postulated in Nature, 2015 that there is less evidence that humic substances and 'humus" are existing. It is probably better to use the terms "soil organic matter", but the authors should clarify the methodology. The presented COST's in references (17-19) in the Russian language, please provide links to international standards.

The authors have not presented the chemical properties of tested soils, which is essential for soil studies.

Presented figures definitely should be improved from the side of the color scheme and also in Titles size.
Figure 4 includes a strange legend. One box "reference" is crossed out. Is it an existing PrintScreen in the program, or is it a photoshop?

Author Response

Please see the attachment

Author Response File: Author Response.pdf

Round 2

Reviewer 2 Report

The manuscript was enhanced significantly and the authors modified most of the requested changes and they answers most of the comments.

I think the paper can be accepted for publishing in Sensors journal in the present form.