Details
| Original language | English |
|---|---|
| Pages (from-to) | 1203-1213 |
| Number of pages | 11 |
| Journal | Applied Microbiology and Biotechnology |
| Volume | 91 |
| Issue number | 4 |
| Early online date | 23 Jun 2011 |
| Publication status | Published - Aug 2011 |
Abstract
Protein production under the control of lac operon regulatory elements using autoinduction is based on diauxic growth of Escherichia coli on lactose after consumption of more preferred carbon substrates. A novel simple and cost-effective defined autoinduction medium using a mixture of glucose, glycerol, and lactose as carbon substrate and NH 4 + as sole nitrogen source without any supplementation of amino acids and vitamins was developed for T7-based E. coli expression systems. This medium was successfully employed in 96-well microtiter plates, test tubes, shake flasks, and 15-L bioreactor cultivations for production of different types of proteins achieving an average yield of 500 mg L-1 product. Cell-specific protein concentrations and solubility were similar as during conventional isopropyl β-d-1-thiogalactopyranoside induction using Luria-Bertani broth. However, the final yield of target proteins was about four times higher, as a higher final biomass was achieved using this novel defined autoinduction broth.
Keywords
- Autoinduction, Defined medium, Escherichia coli, Recombinant protein production
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Applied Microbiology and Biotechnology, Vol. 91, No. 4, 08.2011, p. 1203-1213.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Simple defined autoinduction medium for high-level recombinant protein production using T7-based Escherichia coli expression systems
AU - Li, Zhaopeng
AU - Kessler, Wolfgang
AU - Van Den Heuvel, Joop
AU - Rinas, Ursula
PY - 2011/8
Y1 - 2011/8
N2 - Protein production under the control of lac operon regulatory elements using autoinduction is based on diauxic growth of Escherichia coli on lactose after consumption of more preferred carbon substrates. A novel simple and cost-effective defined autoinduction medium using a mixture of glucose, glycerol, and lactose as carbon substrate and NH 4 + as sole nitrogen source without any supplementation of amino acids and vitamins was developed for T7-based E. coli expression systems. This medium was successfully employed in 96-well microtiter plates, test tubes, shake flasks, and 15-L bioreactor cultivations for production of different types of proteins achieving an average yield of 500 mg L-1 product. Cell-specific protein concentrations and solubility were similar as during conventional isopropyl β-d-1-thiogalactopyranoside induction using Luria-Bertani broth. However, the final yield of target proteins was about four times higher, as a higher final biomass was achieved using this novel defined autoinduction broth.
AB - Protein production under the control of lac operon regulatory elements using autoinduction is based on diauxic growth of Escherichia coli on lactose after consumption of more preferred carbon substrates. A novel simple and cost-effective defined autoinduction medium using a mixture of glucose, glycerol, and lactose as carbon substrate and NH 4 + as sole nitrogen source without any supplementation of amino acids and vitamins was developed for T7-based E. coli expression systems. This medium was successfully employed in 96-well microtiter plates, test tubes, shake flasks, and 15-L bioreactor cultivations for production of different types of proteins achieving an average yield of 500 mg L-1 product. Cell-specific protein concentrations and solubility were similar as during conventional isopropyl β-d-1-thiogalactopyranoside induction using Luria-Bertani broth. However, the final yield of target proteins was about four times higher, as a higher final biomass was achieved using this novel defined autoinduction broth.
KW - Autoinduction
KW - Defined medium
KW - Escherichia coli
KW - Recombinant protein production
UR - http://www.scopus.com/inward/record.url?scp=80052628175&partnerID=8YFLogxK
U2 - 10.1007/s00253-011-3407-z
DO - 10.1007/s00253-011-3407-z
M3 - Article
C2 - 21698378
AN - SCOPUS:80052628175
VL - 91
SP - 1203
EP - 1213
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
SN - 0175-7598
IS - 4
ER -