Strategic metabolic engineering of Escherichia coli for improved ethanol biosynthesis | ||||
| Microbial Biosystems | ||||
| Article 28, Volume 10, Issue 2, June 2025, Page 259-265 PDF (385.93 K) | ||||
| Document Type: Original Article | ||||
| DOI: 10.21608/mb.2025.387913.1328 | ||||
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| Authors | ||||
Kathiresan Subramanian   1; Kagne Suresh2
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| 1Hospira Healthcare Pvt. Ltd, Chennai, Tamil Nadu 600017, India. | ||||
| 2Badrinarayan Barwale Mahavidhyalaya, Jalna 431213, Maharashtra, India. | ||||
| Abstract | ||||
| In this study, E. coli cells were engineered by introducing the pKD46 plasmid using a heat shock method. This allowed for targeted gene editing to knock out the IdhA gene, which plays a role in metabolic pathways. To improve the strain's tolerance to ethanol, the engineered bacteria were then subjected to adaptive laboratory evolution (ALE), gradually exposing them to higher concentrations of ethanol over time. The success of the gene knockouts was confirmed through colony PCR, and BLAST analysis verified that the kanamycin resistance gene had been correctly inserted at both the IdhA and YqhD gene sites. The modified strains not only showed successful genetic changes but also demonstrated significantly enhanced ethanol tolerance. This method proved to be a reliable way to develop E. coli strains capable of efficiently producing ethanol from lignocellulosic sugars. The results highlight the value of genetic engineering and adaptive evolution in creating stable, high-performing microbial platforms for sustainable biofuel production. | ||||
| Keywords | ||||
| Biofuel; carbon flux; fermentation; gene knockout; redox balance; synthetic pathway | ||||
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