Expression, Characterisation and Homology Modelling of a Novel Hormone-Sensitive Lipase (HSL)-Like Esterase from Glaciozyma antarctica
, Adam Thean Chor Leow 1,4 and Mohd Shukuri Mohamad Ali 1,2,*Round 1
Reviewer 1 Report
Biggest loophole is not highlighting the novelty of the work. How this work is advancing science compared to other reports which are already in public domain. Rigorous revision is required to improve the writing part. Rest details are very generic and routine cloning and expression experiments with no discussion on novelty, technological advancements, filling of knowledge-gap existing in the field of research. Manuscript required serious reorganization.
Abstract
Line 17-19: which structure or characterization?
Name those computer-aided software
No information is given on the isolation and characterization of the yeast
How it is related to Antartica or cold env. as discussed in abstract, not explained in Result or Discussion
Author Response
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Author Response File: 
Author Response.pdf
Reviewer 2 Report
The manuscript of M. Ali and co-authors describes the isolation of a new hormone-sensitive esterase. It was expressed in E. coli and purified followed by isolation with affinity chromatography. The authors conducted full biochemical characterisation of the enzyme, which showed higher activity and even stability at higher temperatures as compared with other enzymes from this family. The authors also carried out homology modeling using Robetta. Overall, the work delivers a discovery of a new enzyme and its full characterisation. I don't fill confident to assess the literature references because of lack of experience in this type of enzymes, but the works seems to be done at high level and I can recommend it for publishing.
Author Response
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Reviewer 3 Report
The authors isolated novel hormone-sensitive lipase (GlaEst12) from Glaciozyma antarctica. Enzyme was fully biochemically characterized (optimal temperature, optimal pH, substrate specificity, addition of ions and solvents). This publication seems to be within the scope of journal. However it needs several corrections to be more acceptable for publication.
Publication would gain on value if introduction has contained a information on the potential applications of an isolated enzyme. Manuscript contains numerous editorial errors, for example: line 58 and 66: name of strain should be in italic; line 62: „Glaciozyma antarctic” instead of „ antarctica”; line 84: please remove (De Simone et al., 2004); in line 393 please remove (Laemmli,1970) and add appropriate reference number; line 104: „Glaciozyma antarctica” instead of „G. antarctica”. In all manuscript compound names should be only written in lowercase. References should be formatted according to the instructions of the journal. Figure 1 should be of better quality because the amino acid symbols are illegible. What was the composition of the reaction mixture during studying the effect of temperature, pH, metal ions, etc. If 10 mM p-nitrophenyl decanoate was used as the substrate, in section 3.8 should be placed sentence that the measurement was made analogically to the point 3.3. In section: Effect of Substrate Specificity information on substrate concentrations is missing. Has it always been 10 mM? Please verify and add.Author Response
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Author Response File: Author Response.pdf
Round 2
Reviewer 1 Report
Previously raised concerns are rectified
