Tags

Type your tag names separated by a space and hit enter

Enhanced plasmid DNA production by enzyme-controlled glucose release and an engineered Escherichia coli.
Biotechnol Lett. 2016 Apr; 38(4):651-7.BL

Abstract

OBJECTIVES

To evaluate the combination of a culture medium employing glucoamylase-mediated glucose reléase from a gluco-polysaccharide and an E. coli strain engineered in its glucose transport system for improving plasmid DNA (pDNA) production.

RESULTS

The production of pDNA was tested using E. coli DH5α grown in shake-flasks and the recently developed VH33 Δ(recA deoR)-engineered strain, which utilizes glucose more efficiently than wild type strains. Three glucoamylase concentrations for releasing glucose from the polysaccharide carbon source were used: 1, 2 and 3 U l(-1). Both strains reached similar cell densities ranging from 5 to 8.8 g l(-1) under the different conditions. The highest pDNA yields on biomass (YpDNA/X) for both strains were obtained when 3 U enzyme l(-1)were used. Under these conditions, 35 ± 3 mgof pDNA l(-1) were produced by DH5α after 24 h of culture. Under the same conditions, the engineered strain produced 66 ± 1 mgpDNAl(-1) after 20 h. pDNA supercoiled fractionswere close to 80 % for both strains.

CONCLUSIONS

The pDNA concentration achieved by the engineered E. coli was 89 % higher than that of DH5α. The combination of the engineered strain and enzyme-controlled glucose release is an attractive alternative for pDNA production in shake-flasks.

Authors+Show Affiliations

Plan de estudios en Ingeniería Biológica, Universidad Autónoma Metropolitana-Cuajimalpa, México, DF, Mexico.Plan de estudios en Ingeniería Biológica, Universidad Autónoma Metropolitana-Cuajimalpa, México, DF, Mexico.Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Av. Vasco de Quiroga 4871, Col. Santa Fe, 05348, México, DF, Mexico. alara@correo.cua.uam.mx.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26696535

Citation

Ramírez, Elisa A., et al. "Enhanced Plasmid DNA Production By Enzyme-controlled Glucose Release and an Engineered Escherichia Coli." Biotechnology Letters, vol. 38, no. 4, 2016, pp. 651-7.
Ramírez EA, Velázquez D, Lara AR. Enhanced plasmid DNA production by enzyme-controlled glucose release and an engineered Escherichia coli. Biotechnol Lett. 2016;38(4):651-7.
Ramírez, E. A., Velázquez, D., & Lara, A. R. (2016). Enhanced plasmid DNA production by enzyme-controlled glucose release and an engineered Escherichia coli. Biotechnology Letters, 38(4), 651-7. https://doi.org/10.1007/s10529-015-2017-8
Ramírez EA, Velázquez D, Lara AR. Enhanced Plasmid DNA Production By Enzyme-controlled Glucose Release and an Engineered Escherichia Coli. Biotechnol Lett. 2016;38(4):651-7. PubMed PMID: 26696535.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Enhanced plasmid DNA production by enzyme-controlled glucose release and an engineered Escherichia coli. AU - Ramírez,Elisa A, AU - Velázquez,Daniela, AU - Lara,Alvaro R, Y1 - 2015/12/22/ PY - 2015/08/17/received PY - 2015/12/07/accepted PY - 2015/12/24/entrez PY - 2015/12/24/pubmed PY - 2016/12/15/medline KW - Galactose permease KW - Glucoamylase (for glucose release) KW - Glucose release KW - Metabolic engineering KW - pDNA vaccines SP - 651 EP - 7 JF - Biotechnology letters JO - Biotechnol. Lett. VL - 38 IS - 4 N2 - OBJECTIVES: To evaluate the combination of a culture medium employing glucoamylase-mediated glucose reléase from a gluco-polysaccharide and an E. coli strain engineered in its glucose transport system for improving plasmid DNA (pDNA) production. RESULTS: The production of pDNA was tested using E. coli DH5α grown in shake-flasks and the recently developed VH33 Δ(recA deoR)-engineered strain, which utilizes glucose more efficiently than wild type strains. Three glucoamylase concentrations for releasing glucose from the polysaccharide carbon source were used: 1, 2 and 3 U l(-1). Both strains reached similar cell densities ranging from 5 to 8.8 g l(-1) under the different conditions. The highest pDNA yields on biomass (YpDNA/X) for both strains were obtained when 3 U enzyme l(-1)were used. Under these conditions, 35 ± 3 mgof pDNA l(-1) were produced by DH5α after 24 h of culture. Under the same conditions, the engineered strain produced 66 ± 1 mgpDNAl(-1) after 20 h. pDNA supercoiled fractionswere close to 80 % for both strains. CONCLUSIONS: The pDNA concentration achieved by the engineered E. coli was 89 % higher than that of DH5α. The combination of the engineered strain and enzyme-controlled glucose release is an attractive alternative for pDNA production in shake-flasks. SN - 1573-6776 UR - https://www.unboundmedicine.com/medline/citation/26696535/Enhanced_plasmid_DNA_production_by_enzyme_controlled_glucose_release_and_an_engineered_Escherichia_coli_ L2 - https://doi.org/10.1007/s10529-015-2017-8 DB - PRIME DP - Unbound Medicine ER -