Advancements in Metabolic Engineering: Enhancing Biofuel Production Through Escherichia coli and Saccharomyces cerevisiae Models

Round 1

Reviewer 1 Report

Comments and Suggestions for Authors

According to the manuscript, “A Review on Role of Metabolic Engineering in Enhancing Biofuel Production”. The article is a review on the role of metabolic engineering in enhancing biofuel production, focusing on microbial pathways and innovative strategies to optimize biofuel yields from lignocellulosic biomass and advanced biofuels. My suggestions would be as follows:

• Keywords: it could be reduced with searchable word.
• The manuscript includes no assessment of the environmental consequences of the proposed biofuel production methods, such as waste production or resource usage.
• While the article describes biofuel manufacturing technologies, it does not go into great detail about their economic viability or cost-effectiveness for industrial-scale applications.
• The article does not address the political or regulations that might limit biofuel commercialization.
• The paper does not include examples of effective practical applications or case studies for the metabolic engineering methodologies addressed.
• Although the manuscript emphasizes model organisms such as E. coli and S. cerevisiae, it does not investigate the possibilities of other developing microbial hosts or systems.
• Line 679, “Dal’Molin et al.” please recheck the citation style.

Author Response

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Author Response File: Author Response.pdf

Reviewer 2 Report

Comments and Suggestions for Authors

This review manuscript addresses the biochemical engineering processes involved in biofuel production utilizing microorganisms, specifically the model organisms Escherichia coli and Saccharomyces cerevisiae. The overall presentation of the manuscript is sound and provides a solid foundation for understanding the subject matter. However, minor revisions are necessary to enhance clarity and depth in certain areas, as outlined below.

• The title suggested be change to “Advancements in Metabolic Engineering: Enhancing Biofuel Production through Escherichia coli and Saccharomyces cerevisiae Models”
• Line 128: furfual change to “furfural”.
• Add the full name of “PP pathway” in Line 147.
• A careful proofreading of the manuscript is essential to resolve any language, phrasing, or stylistic issues that may undermine its quality.

Author Response

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Author Response File: Author Response.pdf

Reviewer 3 Report

Comments and Suggestions for Authors

The review explores the pivotal role of metabolic engineering in advancing biofuel production as a sustainable alternative to fossil fuels. It highlights how genetic and synthetic biology tools are used to optimize microbial pathways for the efficient conversion of lignocellulosic biomass and other substrates into advanced biofuels. The manuscript is well organized with appropriate visualization to help audience better follow the flow. This manuscript is recommended for publication.

Author Response

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Author Response File: Author Response.pdf

Reviewer 4 Report

Comments and Suggestions for Authors

This review article explores the significant advances in metabolic engineering strategies for improving biofuel production. The authors detail how engineered microbes, particularly Escherichia coli and Saccharomyces cerevisiae, are leveraged to increase yields of biofuels such as n-butanol, iso-butanol, isoprenoids, and fatty-acid-derived fuels. The review emphasizes synthetic biology tools (e.g., CRISPR/Cas9, MAGE), metabolic flux analysis, and microbial tolerance mechanisms. The manuscript’s main strength lies in its breadth of coverage and inclusion of recent advances in microbial engineering and synthetic biology.
1. Section 5.2 and 5.3 are dense and highly technical. Consider breaking down into subsections with summary boxes or comparative tables for clarity.
2. How do the authors envision the translation of these metabolic engineering strategies to industrial-scale biofuel production?
3. Why were E. coli and S. cerevisiae given primary focus over others like Corynebacterium glutamicum or Bacillus subtilis?
4. What are the main limitations of CRISPR/Cas9 and MAGE in microbial systems that the review should address?
5. Could the authors include a comparative summary table of biofuel yields (g/L) across the discussed host systems and pathways?
6. Can the authors discuss challenges related to byproduct formation or metabolic burden during biofuel production in engineered microbes?

Author Response

Please see the attachment.

Author Response File: Author Response.pdf