High-cell density fermentation studies of recombinant Escherichia coli strains expressing human interleukin-1β

Jung, Gérard; Denèfle, Patrice; Becquart, Jérôme; Mayaux, Jean‐François

doi:10.1016/0769-2609(88)90100-7

Cited by 71 publications

(31 citation statements)

References 20 publications

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“…These results indicated that the recombinant S. marcescens strains requires a proteinaceous substrates such as casein for a high level expression of SMP. The requirement of organic nitrogen sources for the production of recombinant proteins in E. coli was also observed by Tsai et aL(1987), Jung et al(1988), and Shimizu et al(1987. Although the role of proteinaceous substrate in SMP production from recombinant S. marcescens strains remains to be discovered, we presumed that the proteinaceous substrate might protect SMP from proteolytic degradation of host cells or induce some specific proteins that play a crucial role in the production of SMP.…”

Section: Methodssupporting

confidence: 58%

Overproduction of Serratia marcescens metalloprotease (SMP) from the recombinant Serratia marcescens strains

et al. 1995

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To overproduce Serratia marcescens metalloprotease(SMP), various recombinant plasmids encoding SMP gene were constructed and the SMP productivities from the recombinant S. marcescens strains were examined. The recombinant S. marcescens strains indispensably required proteinaceous substrates such as casein for the extracellutar production of SMP. We obtained maximum 9,100U/ml of SMP from the culture supernatant of S. marcescens ATCC27117 containing a regulatory plasmid pTSP2 encoding SMP gene fused with a strong trc99a promoter and its repressor gene lacI q, which is about 23 times higher than that of wild t3q3e strain. SMP produced from the recombinant S. marcescens(pTSP2) was 88.3% of total extracellular proteins.

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Section: Methodssupporting

confidence: 58%

Overproduction of Serratia marcescens metalloprotease (SMP) from the recombinant Serratia marcescens strains

et al. 1995

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“…In fed-batch cultures jz was determined by the feeding rate of the limiting carbon source (Gleiser and Bauer 1981 ;Jung et al 1988). Therefore, a specific fedbatch strategy was developed on the basis of the designed mass culture medium.…”

Section: Introductionmentioning

confidence: 99%

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

Riesenberg¹,

Menzel²,

Schulz³

et al. 1990

Appl Microbiol Biotechnol

119

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A defined medium was developed which, by means of a specific fed-batch mode, allows growth of the recombinant Escherichia coli strain TG1 (pBB210) up to a cell density of 60 g dry weight/l. Apart from glucose and aqueous ammonia fed as carbon and nitrogen sources, it was necessary to supply other nutrients or O2-enriched air. Aqueous ammonia also served for pH control. The pO2 level was kept at 20% saturation via closed-loop controls operating the two output variables of stirrer speed and glucose feeding rate. This fed-batch method prevented significant accumulation of acetate and other metabolic by-products. The recombinant E. coli expressed interferon alpha 1 more efficiently at a lower specific growth rate (muPr approximately 0.15 h-1) than at the maximum specific growth rate (mu max = 0.45 h-1). Therefore, fermentation in the batch phase at mu max was only allowed to continue up to a medium cell density. In the succeeding fed-batch phase, the specific growth rate was reduced to muPr by increasing the stirrer speed according to an empirically developed time scale.

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“…maintain E. coli and B. lactofermentum plasmids, respectively. Conditions for the expression of heterologous expression in E. coli have been described previously (7,11). B. lactofermentum RP2 is a spontaneous rifampin-resistant isolate of B. lactofermentum ATCC 21086, the endogenous plasmids of which were cured after extensive protoplastisation and regeneration of the cell wall (27a).…”

mentioning

confidence: 99%

Purification, cloning, and primary structure of a new enantiomer-selective amidase from a Rhodococcus strain: structural evidence for a conserved genetic coupling with nitrile hydratase

et al. 1991

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A new enantiomer-selective amidase active on several 2-aryl propionamides was identified and purified from a newly isolated Rhodococcus strain. The characterized amidase is an apparent homodimer, each molecule of which has an Mr of 48,554; it has a specific activity of 16.5 g,mol of S(+)-2-phenylpropionic acid formed per min per mg of enzyme from the racemic amide under our conditions. An oligonucleotide probe was deduced from limited peptide information and was used to clone the corresponding gene, named amdA. As expected, significant homologies were found between the amino acid sequences of the enantiomer-selective amidase of Rhodococcus sp., the corresponding enzyme from Brevibacterium sp. strain R312, and several known amidases, thus confirming the existence of a structural class of amidase enzymes. Genes probably coding for the two subunits of a nitrite hydratase, albeit in an inverse order, were found 39 bp downstream of amdA, suggesting that such a genetic organization might be conserved in different microorganisms. Although we failed to express an active Rhodococcus amidase in Escherichia coli, even in conditions allowing the expression of an active R312 enzyme, the high-level expression of the active recombinant enzyme could be demonstrated in Brevibacterium lactofermentum by using a pSRl-derived shuttle vector.After a systematic search for soil microorganisms able to carry out the enantiomer-selective hydrolysis of racemic nitriles or amides, especially 2-aryl propionamides, we recently reported the purification and cloning of such an amidolytic activity from Brevibacterium sp. strain R312 (15). We found that the gene coding for the enantiomer-selective amidase, amdA, was closely linked to the genes coding for the two subunits, a and ,B, of nitrile hydratase, the enzyme responsible for the hydration of various nitriles to the corresponding amides (10). The structure of this region led to the hypothesis that the two genes could be translated from a polycistronic mRNA, thus supporting the view that there is a nitrile utilization operon, at least in this bacterium. These results have been confirmed by another group (9). Moreover, it was found that the identified amidase displays a striking homology to other known amidases, such as the acetamidase from Aspergillus nidulans and the indoleacetamide hydrolases from Pseudomonas syringae (savastanoi) and Agrobacterium tumefaciens. Interestingly, the homology was not scattered through the whole amino acid sequence but was mainly focused in a portion of the sequence around amino acids 150 to 220 of the R312 sequence (47 to 65% strict identity), strongly suggesting that this conserved sequence could be part of the active site of this class of enzymes.We now report the identification of a new Rhodococcus strain, obtained during a microbiological screening of soil samples, which is also characterized by efficient stereospecific amidase activity on 2-aryl propionamides. The enzyme responsible for this activity was purified, cloned, and sequenced. It is significantly ...

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High-cell density fermentation studies of recombinant Escherichia coli strains expressing human interleukin-1β

Cited by 71 publications

References 20 publications

Overproduction of Serratia marcescens metalloprotease (SMP) from the recombinant Serratia marcescens strains

Overproduction of Serratia marcescens metalloprotease (SMP) from the recombinant Serratia marcescens strains

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

Purification, cloning, and primary structure of a new enantiomer-selective amidase from a Rhodococcus strain: structural evidence for a conserved genetic coupling with nitrile hydratase

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