Influence of Growth Conditions on Plasmid DNA Production

• Published in: Journal of microbiology and biotechnology Vol. 19; no. 11; pp. 1408 - 1414
• Main Authors: Silva, Filomena, Universidade da Beira Interior, Avenida Infante D. Henrique, Covilha, Portugal, Passarinha, Luis, Universidade da Beira Interior, Avenida Infante D. Henrique, Covilha, Portugal, Sousa, Fani, Universidade da Beira Interior, Avenida Infante D. Henrique, Covilha, Portugal, Queiroz, Joao A., Universidade da Beira Interior, Avenida Infante D. Henrique, Covilha, Portugal, Domingues, Fernanda C., Universidade da Beira Interior, Avenida Infante D. Henrique, Covilha, Portugal
• Format: Journal Article
• Language: English
• Published: Seoul Korean Society for Applied Microbiology 01-11-2009; 한국미생물·생명공학회
• Subjects: Biological and medical sciences; Biotechnology; Cell Culture Techniques - methods; Culture Media - chemistry; DNA Replication; DNA, Bacterial - biosynthesis; DNA, Bacterial - genetics; Escherichia coli - genetics; Escherichia coli - growth & development; Escherichia coli - metabolism; FERMENTACION; FERMENTATION; filamentous bacteria; Fundamental and applied biological sciences. Psychology; Industrial Microbiology - methods; Methods. Procedures. Technologies; Microbial engineering. Fermentation and microbial culture technology; microbial growth; Nucleic Acid Amplification Techniques - methods; Peptones - chemistry; Plasmid DNA; Plasmids - biosynthesis; Plasmids - genetics; TEMPERATURA; TEMPERATURE; tryptone; Vaccines, DNA - biosynthesis; 생물학; Plasmid; Microorganism growth; Temperature; Growth; DNA; Production; Bacteria; filamentous bacteria; Fermentation; Plasmid DNA; microbial growth; tryptone
• ISSN: 1017-7825; 1738-8872
• DOI: 10.4014/jmb.0805.329

The obtention of high yields of purified plasmid DNA is viewed as an essential issue to be considered towards efficient production of DNA vaccines and therapeutic plasmids. In this work, Escherichia coli DH5α bearing the pVAX1-LacZ plasmid was grown in a developed semi-defined medium at different temperatures and tryptone concentrations. Analysis of pDNA yields and E. coli morphology revealed that at higher temperatures (37 and 40℃), higher specific yields and E. coli filamentation were obtained. However, the best results were achieved when a lower tryptone concentration was used. This approach was shown to be a powerful tool to promote plasmid amplification, keeping the desirable plasmid structure, and favoring the attainment of quality. Our results suggest that by using tryptone alone as an amino acid source, pDNA amplification was improved and a specific yield of 20.43 mg pDNA/g dew was achieved, proving that this strategy can improve pDNA yield even at a small scale.