An Escherichia coli Strain for Plasmid DNA Production with a Low Endotoxin Level

IF 1 4区生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Applied Biochemistry and Microbiology Pub Date : 2024-12-18 DOI:10.1134/S0003683824604517

D. D. Kharlampieva, P. A. Bobrovsky, E. N. Grafskaia, V. N. Lazarev, V. A. Manuvera

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引用次数: 0

Abstract

Plasmid DNA (pDNA) is one of the possible types of vectors used in gene therapy. Escherichia coli is the most common bacterium for the production of pDNA. Lipopolysaccharides are the main component of the outer membrane of E. coli. Lipopolysaccharides are strong endotoxins, and their presence in pharmacological substances is highly undesirable. One of the approaches to eliminating endotoxin contamination of target substances is knocking out the genes of the lipopolysaccharide biosynthesis pathway. Using the CRISPR/Cas9 genome editing technology, we obtained a mutant form of the E. coli strain SCS110. We knocked out the genes kdsD, gutQ, lpxM, lpxL, lpxP, pagP, and eptA. As a result, lipopolysaccharide contamination of pDNA samples isolated by different methods from the mutant E. coli SCS 110 was significantly lower (19 to 295 times) than in pDNA samples from the original strain.

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来源期刊

Applied Biochemistry and Microbiology 生物-生物工程与应用微生物

CiteScore

1.70

自引率

12.50%

发文量

审稿时长

6-12 weeks

期刊介绍： Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.