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Because of the high temperature, traditional butyl acetate synthesis by Fischer esterification of butanol and acetic acid using catalytic strong acids is not environmentally friendly. Therefore, a butyl acetate manufacturing approach through fermentation of renewable resources would lead to a more eco friendly approach to butyl acetate production. To synthesize butanol, a modified Clostridium CoA-dependent butanol production pathway was used, synthesize butanol, which was then condensed with acetyl-CoA by an alcohol acetyltransferase. This research found a strain of E. coli capable of extracting butyl acetate from renewable sources at ambient temperatures.
Source link: https://doi.org/10.1186/s12934-022-01755-y
Background: CRISPR-Cas conversion of bacterial genomes has evolved to a common laboratory procedure to recombine bacterial genomes. In one round of editing, we first developed a CRISPR-Cas genome editing kit in the Escherichia coli BL21 strain and successfully deleted 10 kb of DNA from the genome in this study. Our technique greatly reduced the time taken and reduced the cost of homology arm preparations relative to conventional homology arms that are based on overlapping PCR, cloning into a plasmid, or annealing synthetic DNA fragments. In this report, a noteworthy, gRNA assembly design is crucial for the CRISPR-Cas system, and a general heuristic gRNA layout has been suggested. We targeted 16 genes and iteratively deleted 7 genes from the BL21 genome to implement our established protocol. Any E. coli strain in which genome engineering may be able to raise metabolite production could be beneficial to further improve metabolite production.
Source link: https://doi.org/10.1186/s12934-022-01746-z
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