High cell density fermentation of recombinant Escherichia coli expressing human interferon alpha 1

Riesenberg, D.; Menzel, Klaus; Schulz, Volkmar; Schümann, Klaus; G, Veith; Zuber, Georg; Knorre, W. A.

doi:10.1007/bf00170927

Cited by 119 publications

(56 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At the lower fed-batch growth rate, the specific CAT activity remained elevated for a longer time period. Both of these expression characteristics are similar to what has been observed for E. coli MG1655 under similar conditions (Hoffman et al, 2002;Koo and Park, 1999;Levisauskas et al, 2003;Riesenberg et al, 1990).…”

Section: Fed-batch Fermentationsupporting

confidence: 85%

“…Thus, it is possible that the lower observed specific CAT activity in the high fed-batch growth rate fermentation cultures could have been due to redirection of the carbon resources from the recombinant protein to acetate, due to overflow metabolism, as seen with E. coli MG1655. The effect of growth rate on recombinant protein expression has been addressed in literature for strains related to E. coli MG1655, and the E. coli MDS40 strain behavior was consistent with respect to productivity and growth rates (Hoffman et al, 2002;Koo and Park, 1999;Levisauskas et al, 2003;Riesenberg et al, 1990). …”

Section: Fed-batch Fermentationmentioning

confidence: 93%

See 1 more Smart Citation

Recombinant protein production in an Escherichia coli reduced genome strain

Sharma

Blattner

Harcum

2007

Metabolic Engineering

View full text Add to dashboard Cite

Recently, efforts have been made to improve the properties of E. coli as a recombinant host by 'genomic surgery' -deleting large segments of the E. coli K12 MG1655 genome without scars.These excised segments included K-islands, which contain a high proportion of transposons, insertion sequences, cryptic phage, damaged, and unknown-function genes. The resulting multiple-deletion strain, designated E. coli MDS40, has a 14% (about 700 genes) smaller genome than the parent strain, E. coli MG1655. The multiple-deletion and parent E. coli strains were cultured in fed-batch fermenters to high cell densities on minimal medium to simulate industrial conditions for evaluating growth and recombinant protein production characteristics. Recombinant protein production and by-product levels were quantified at different controlled growth rates. These results indicate that the multiple-deletion strain's growth behavior and recombinant protein productivity closely matched the parent stain. Thus, the multiple-deletion strain E. coli MDS40 provides a suitable foundation for further genomic reduction.

show abstract

Section: Fed-batch Fermentationsupporting

confidence: 85%

Section: Fed-batch Fermentationmentioning

confidence: 93%

Recombinant protein production in an Escherichia coli reduced genome strain

Sharma

Blattner

Harcum

2007

Metabolic Engineering

View full text Add to dashboard Cite

show abstract

“…Nevertheless, the specific growth rate, not the feeding method, has been regarded to be the intrinsic variable affecting recombinant protein synthesis [5]. The literature has well documented that the productivity of recombinant proteins is dependent on specific growth rate [6][7][8][9]; however, this is not universal [10]. In this work, we investigated the influence of specific growth rate on the nuclease production by the recombinant strain, E. coli DH5␣ (pSI026); the information obtained was then employed to design the feeding strategy in fed-batch cultivation.…”

Section: Introductionmentioning

confidence: 99%

Effect of specific growth rate on the production of a recombinant nuclease by Escherichia coli

Cheng

Hor

et al. 2003

Biochemical Engineering Journal

View full text Add to dashboard Cite

“…The M9 minimal medium and R medium (11) were chosen for enrichment experiments because of their popular use in metabolic engineering (1, 2, 7) and in high-cell-density fermentation (8,10,11). After 11 serial transfers of the transformants in the M9 medium, and 27 transfers in the R medium, cultured cells were diluted and plated onto LB agar for single-colony isolation.…”

mentioning

confidence: 99%

Restoration of Growth Phenotypes of Escherichia coli DH5α in Minimal Media through Reversal of a Point Mutation in purB

Jung

Smith

Lee

et al. 2010

Appl Environ Microbiol

View full text Add to dashboard Cite

A point mutation (E115K) resulting in slower growth of Escherichia coli DH5␣ and XL1-Blue in minimal media was identified in the purB gene, coding for adenylosuccinate lyase (ASL), through complementation with an E. coli K-12 genomic library and serial subcultures. Chromosomal modification reversing the mutation to the wild type restored growth phenotypes in minimal media.The Escherichia coli DH5␣ strain possesses many beneficial genotypes (recA, deoR, gyrA, and endA1) and has been widely used for many purposes, such as gene cloning and protein production (5). However, E. coli DH5␣ also exhibits inferior growth phenotypes, especially in minimal media, compared to other E. coli strains. As such, the utilization of this bacterium has been limited to the laboratory despite its numerous advantages. We can assume that these inferior growth phenotypes have resulted from unknown accumulated mutations during the strain development process (5). Some of those mutations, which might impact growth in minimal media, have been characterized, including the phenotypes for thiamine requirement and relaxed amino acid synthesis (5). Still, there may be other uncharacterized mutations whose interactions hamper the growth of E. coli DH5␣ in minimal media.Based on successful identifications (6, 7) of gene targets for metabolic engineering (3), we performed serial subcultures of E. coli DH5␣ transformants with an E. coli K-12 genomic library based on a multicopy plasmid (9) to isolate genes that improve growth phenotypes in minimal media. The M9 minimal medium and R medium (11) were chosen for enrichment experiments because of their popular use in metabolic engineering (1, 2, 7) and in high-cell-density fermentation (8, 10, 11). After 11 serial transfers of the transformants in the M9 medium, and 27 transfers in the R medium, cultured cells were diluted and plated onto LB agar for single-colony isolation. Although more than 10 colonies were picked, only three distinctive plasmids, containing different inserts, were isolated from the transformants enriched in M9 medium. In the case of R medium enrichment, all isolated plasmids were identical. Sequencing of the isolated plasmids revealed the exact genome coordinates of each insert. A diagram of the inserts in the context of the E. coli genome sequence is shown in Fig. 1. Interestingly, all of the isolated plasmids contained similar regions of genomic DNA. mnmA (tRNA 5-methylaminomethyl-2-thiouridylate-methyltransferase), purB (adenylosuccinate lyase), and hflD (lysogenization regulator) were the annotated genes in the overlapping region among distinctive isolated fragments. However, since the N-terminal portions of mnmA and hflD were truncated in some of the inserts, we selected only the M3 and R1 plasmids for further experimentation. These two plasmids were retransformed into E. coli DH5␣ for confirmation of their beneficial effects on growth of E. coli in minimal media. The newly transformed strains showed growth phenotypes almost identical to those of the previously isolated transfor...

show abstract

High cell density fermentation of recombinant Escherichia coli expressing human interferon alpha 1

Cited by 119 publications

References 17 publications

Recombinant protein production in an Escherichia coli reduced genome strain

Recombinant protein production in an Escherichia coli reduced genome strain

Effect of specific growth rate on the production of a recombinant nuclease by Escherichia coli

Restoration of Growth Phenotypes of Escherichia coli DH5α in Minimal Media through Reversal of a Point Mutation in purB

Contact Info

Product

Resources

About