High cell density culture of recombinant E. coli in the miniaturized bubble columns

High cell density culture of recombinant E. coli in the miniaturized bubble columns

Posted on by Laura
Miniaturized bubble columns (MBCs) can present mass switch traits much like stirred tank bioreactors. On this research, a brand new utility was developed for MBCs to analyze the impact of feeding technique and medium sort on the fed-batch tradition of recombinant E. coli. The outcomes confirmed that the exponential feeding technique and outlined M9 medium have been extra appropriate to attain the excessive cell density tradition (HCDC).
The utmost obtained cell focus in exponential feeding technique within the outlined medium with out induction, was at OD600 of 169, whereas glucose focus was maintained beneath 2 g/L. To one of the best of our information, this cell focus can’t be achieved in lab or pilot scale bubble columns. On the finish of the method, adversarial impact of the metabolic burden as a consequence of induction and mass switch limitations decreased the obtained closing cell focus to OD600 of 116.
Lastly, a comparability of the outcomes for fed-batch tradition within the stirred tank bioreactor with these of the MBCs confirmed that their decrease cell concentrations have been as a result of hydrodynamics limitations of MBCs. But, it was discovered that the MBCs are environment friendly instruments in growth of feeding methods and analysis of medium elements for HCDC of recombinant E. coli.

Expression and purification of purposeful recombinant CUL2•RBX1 from E. coli

Cullin-2 (CUL2) based mostly cullin-RING ligases (CRL2s) comprise a household of ubiquitin E3 ligases that exist solely in multi-cellular organisms and are essential for mobile processes akin to embryogenesis and viral pathogenesis. CUL2 is the scaffold protein that binds one of many interchangeable substrate receptor modules, which consists of adaptor proteins and the substrate receptor protein. The VHL protein is a substrate receptor identified to focus on hypoxia-inducible issue α (HIF1α) for ubiquitination and degradation.
Due to its crucial position within the ubiquitination of necessary mobile elements akin to HIF1α, CRL2s have been investigated for his or her organic capabilities and the event of novel therapeutics towards illnesses. Given the significance of CRL2s in organic and biomedical analysis, strategies that effectively produce purposeful CUL2 proteins will vastly facilitate research on the mechanism and regulation of CRL2s.
High cell density culture of recombinant E. coli in the miniaturized bubble columns
Right here, we report two cost-effective programs for the expression and purification of recombinant human CUL2 from E. coli cells. The purified CUL2 proteins have been ~ 95% pure, may bind their substrate receptor modules, and have been enzymatically energetic in transferring ubiquitin or ubiquitin-like protein to the corresponding substrate in in vitro assays. The offered methodological developments will assist advance analysis in CRL2 perform and regulation.

Milligram scale expression, refolding, and purification of Bombyx mori cocoonase utilizing a recombinant E. coli system

Silk is likely one of the most versatile biomaterials with signature properties of excellent mechanical energy and suppleness. A possible avenue for growing extra environmentally pleasant silk manufacturing is to utilize the silk moth (Bombyx mori) cocoonase, this may on the identical time improve the likelihood for utilizing the byproduct, sericin, as a uncooked materials for different purposes. Cocoonase is a serine protease utilized by the silk moth to melt the cocoon to allow its escape after accomplished metamorphosis.
Cocoonase selectively degrades the glue protein of the cocoon, sericin, with out affecting the silk-fiber product of the protein fibroin. Cocoonase will be recombinantly produced in E. coli, nevertheless, it’s solely discovered as insoluble inclusion our bodies. To resolve this drawback and to have the ability to make the most of the advantages related to an E. coli based mostly expression system, we have now developed a protocol that permits the manufacturing of soluble and purposeful protease within the milligram/liter scale. The core of the protocol is refolding of the protein in a buffer with a redox potential that’s optimized for formation of native and intramolecular di-sulfide bridges.
The redox potential was balanced with outlined concentrations of diminished and oxidized glutathione. This E coli based mostly manufacturing protocol will, along with construction dedication, additionally allow modification of cocoonase each by way of catalytic perform and stability. These elements will probably be precious elements within the growth of alternate silk manufacturing methodology.

Expression of Bacillus amyloliquefaciens transglutaminase in recombinant E. coli beneath the management of a bicistronic plasmid system in DO-stat fed-batch bioreactor cultivations

We studied the expression of Bacillus amyloliquefaciens transglutaminase cloned in Escherichia coli BL21(DE3)pLysS harboring the plasmid pBAD/3C/bTGase, a bicistronic expression system, in bioreactor cultivation. Batch and fed-batch managed as DO-stat methods have been employed for the manufacturing of the recombinant enzyme. In 30 h-batch cultivations utilizing Terrific broth (TB), 6 g/L of biomass and three.12 U/mgprotein of transglutaminase exercise have been obtained. DO-stat fed-batch cultivations beneath the management of oxygen focus (DO-stat) utilizing TB as medium however fed with glucose allowed the increment in biomass formation (17.5 g/L) and enzyme exercise (6.43 U/mgprotein).

Recombinant Human Trefoil Factor-1
MBS143342-1mg 1mg
EUR 2880

Recombinant Human Trefoil Factor-1
MBS143342-5x1mg 5x1mg
EUR 12630

Recombinant Human Trefoil Factor 1
AP60455 100ug
EUR 896

Recombinant Human Trefoil factor 1
MBS9420733-002mg 0.02mg
EUR 285

Recombinant Human Trefoil factor 1
MBS9420733-01mg 0.1mg
EUR 470

Recombinant Human Trefoil factor 1
MBS9420733-1mg 1mg
EUR 1785

Recombinant Human Trefoil factor 1
MBS9420733-5x1mg 5x1mg
EUR 7885

Recombinant Human Trefoil factor 1
MBS9422520-002mg 0.02mg
EUR 320

Recombinant Human Trefoil factor 1
MBS9422520-01mg 0.1mg
EUR 540

Recombinant Human Trefoil factor 1
MBS9422520-05mg 0.5mg
EUR 1360

Recombinant Human Trefoil factor 1
MBS9422520-5x05mg 5x0.5mg
EUR 5965

Recombinant Human Trefoil Factor 1
P1615-.005 5ug
EUR 108.8
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 1
P1615-.1 100ug
EUR 900
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 1
P1615-.5 500ug
EUR 2021.6
Description: Peptides & Proteins|Recombinant Proteins

Trefoil Factor-1 Human Recombinant
rAP-0742 Inquiry Ask for price

Recombinant Human Trefoil factor 1 (TFF1)
CSB-YP023431HU 6033 mg Ask for price

Recombinant Human Trefoil factor 1(TFF1)
AP70837 1mg
EUR 1978

Recombinant Human Trefoil factor 1(TFF1)
AP72810 each Ask for price

Recombinant Human Trefoil factor 1 (TFF1)
CSB-EP023431HU 2315 mg Ask for price

Recombinant Human Trefoil Factor 1(TFF1)
HEOPP-2009 5ug Ask for price

Recombinant Human Trefoil factor 1 (TFF1)
RPC22102-100ug 100ug
EUR 460.9

Recombinant Human Trefoil factor 1 (TFF1)
RPC22102-1mg 1mg
EUR 2237.8

Recombinant Human Trefoil factor 1 (TFF1)
RPC22102-20ug 20ug
EUR 258.7

Recombinant Human Trefoil factor 1 (TFF1)
MBS950844-002mgEColi 0.02mg(E-Coli)
EUR 255

Recombinant Human Trefoil factor 1 (TFF1)
MBS950844-002mgYeast 0.02mg(Yeast)
EUR 270

Recombinant Human Trefoil factor 1 (TFF1)
MBS950844-01mgEColi 0.1mg(E-Coli)
EUR 405

Recombinant Human Trefoil factor 1 (TFF1)
MBS950844-01mgYeast 0.1mg(Yeast)
EUR 450

Recombinant Human Trefoil factor 1 (TFF1)
MBS950844-05mgYeast 0.5mg(Yeast)
EUR 1065

Recombinant Human Trefoil Factor-1 His Tag
7-01585 5µg Ask for price

Recombinant Human Trefoil Factor-1 His Tag
7-01586 20µg Ask for price

Recombinant Human Trefoil Factor-1 His Tag
7-01587 1mg Ask for price

Recombinant Human Trefoil Factor-1 His Tag
cyt-610-1mg 1mg
EUR 3000

Recombinant Human Trefoil Factor-1 His Tag
cyt-610-20g 20µg
EUR 145

Recombinant Human Trefoil Factor-1 His Tag
cyt-610-5g 5µg
EUR 60

Recombinant Human Trefoil Factor-1 His Tag
MBS143343-0005mg 0.005mg
EUR 240

Recombinant Human Trefoil Factor-1 His Tag
MBS143343-002mg 0.02mg
EUR 310

Recombinant Human Trefoil Factor-1 His Tag
MBS143343-1mg 1mg
EUR 4175

Recombinant Human Trefoil Factor-1 His Tag
MBS143343-5x1mg 5x1mg
EUR 18455

Recombinant Human Trefoil Factor-2
7-01588 5µg Ask for price

Recombinant Human Trefoil Factor-2
7-01589 20µg Ask for price

Recombinant Human Trefoil Factor-2
7-01590 1mg Ask for price

Recombinant Human Trefoil Factor-3
7-01594 5µg Ask for price

Recombinant Human Trefoil Factor-3
7-01595 20µg Ask for price

Recombinant Human Trefoil Factor-3
7-01596 1mg Ask for price

Recombinant Human Trefoil Factor-2
cyt-612-1mg 1mg
EUR 2700

Recombinant Human Trefoil Factor-2
cyt-612-20g 20µg
EUR 145

Recombinant Human Trefoil Factor-2
cyt-612-5g 5µg
EUR 60

Recombinant Human Trefoil Factor-3
cyt-005-1mg 1mg
EUR 2700

Recombinant Human Trefoil Factor-3
cyt-005-20g 20µg
EUR 145

Recombinant Human Trefoil Factor-3
cyt-005-5g 5µg
EUR 60

Recombinant Human Trefoil Factor-2
MBS143344-0005mg 0.005mg
EUR 240

Recombinant Human Trefoil Factor-2
MBS143344-002mg 0.02mg
EUR 310

Recombinant Human Trefoil Factor-2
MBS143344-1mg 1mg
EUR 2880

Recombinant Human Trefoil Factor-2
MBS143344-5x1mg 5x1mg
EUR 12630

Recombinant Human Trefoil Factor-3
MBS144182-0005mg 0.005mg
EUR 240

Recombinant Human Trefoil Factor-3
MBS144182-002mg 0.02mg
EUR 310

Recombinant Human Trefoil Factor-3
MBS144182-1mg 1mg
EUR 2880

Recombinant Human Trefoil Factor-3
MBS144182-5x1mg 5x1mg
EUR 12630

Recombinant Human Trefoil Factor 2
AP60456 100ug
EUR 896

Recombinant Human Trefoil Factor 3
AP60458 100ug
EUR 896

Recombinant Human Trefoil factor 3
MBS9018625-02mg 0.2mg
EUR 505

Recombinant Human Trefoil factor 3
MBS9018625-05mg 0.5mg
EUR 810

Recombinant Human Trefoil factor 3
MBS9018625-1mg 1mg
EUR 1260

Recombinant Human Trefoil factor 3
MBS9018625-5x1mg 5x1mg
EUR 5620

Recombinant Human Trefoil factor 3
MBS969622-02mg 0.2mg
EUR 505

Recombinant Human Trefoil factor 3
MBS969622-05mg 0.5mg
EUR 810

Recombinant Human Trefoil factor 3
MBS969622-1mg 1mg
EUR 1260

Recombinant Human Trefoil factor 3
MBS969622-5x1mg 5x1mg
EUR 5620

Recombinant Human Trefoil Factor 2
P1616-.005 5ug
EUR 108.8
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 2
P1616-.1 100ug
EUR 900
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 2
P1616-.5 500ug
EUR 2021.6
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 3
P1618-.005 5ug
EUR 108.8
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 3
P1618-.1 100ug
EUR 900
Description: Peptides & Proteins|Recombinant Proteins

Recombinant Human Trefoil Factor 3
P1618-.5 500ug
EUR 2021.6
Description: Peptides & Proteins|Recombinant Proteins

Trefoil Factor-3 Human Recombinant
rAP-0741 Inquiry Ask for price

Trefoil Factor-2 Human Recombinant
rAP-0745 Inquiry Ask for price

Trefoil Factor-1 Human Recombinant, His Tag
rAP-0743 Inquiry Ask for price

Recombinant Human Trefoil factor 1 protein (TFF1)
MBS9423196-0005mg 0.005mg
EUR 210

Recombinant Human Trefoil factor 1 protein (TFF1)
MBS9423196-002mg 0.02mg
EUR 365

Recombinant Human Trefoil factor 1 protein (TFF1)
MBS9423196-01mg 0.1mg
EUR 1135

Recombinant Human Trefoil factor 1 protein (TFF1)
MBS9423196-025mg 0.25mg
EUR 1990

Recombinant Human Trefoil factor 1 protein (TFF1)
MBS9423196-05mg 0.5mg
EUR 2570

TFF1 Trefoil Factor-1 Human Recombinant Protein
PROTP04155 Regular: 20ug
EUR 380.4
Description: TFF-1 Human Recombinant produced in E.Coli is a homodimer, non-glycosylated, polypeptide chain containing 2 x 60 amino acids which includes a 40 amino acid trefoil motif containing 3 conserved intramolecular disulfide bonds and having a total molecular mass of 13.2 kDa. ;TFF-1 Human Recombinant is purified by proprietary chromatographic techniques.

Recombinant Human Trefoil Factor 1/TFF1 (C-6His)
C659-10ug 10ug
EUR 169.2
Description: Lyophilized from a 0.2 μm filtered solution of PBS, pH7.4.

Recombinant Human Trefoil Factor 1/TFF1 (C-6His)
C659-1mg 1mg
EUR 2739.6
Description: Lyophilized from a 0.2 μm filtered solution of PBS, pH7.4.

Recombinant Human Trefoil Factor 1/TFF1 (C-6His)
C659-500ug 500ug
EUR 1765.2
Description: Lyophilized from a 0.2 μm filtered solution of PBS, pH7.4.

Recombinant Human Trefoil Factor 1/TFF1 (C-6His)
C659-50ug 50ug
EUR 333.6
Description: Lyophilized from a 0.2 μm filtered solution of PBS, pH7.4.

Recombinant Human Trefoil Factor 1 protein (C-6His)
CD00769-10ug 10ug
EUR 212
Description: Recombinant Human Trefoil Factor 1 is produced by our Mammalian expression system and the target gene encoding Glu25-Phe84 is expressed with a 6His tag at the C-terminus.

Recombinant Human Trefoil Factor 1 protein (C-6His)
CD00769-50ug 50ug
EUR 529
Description: Recombinant Human Trefoil Factor 1 is produced by our Mammalian expression system and the target gene encoding Glu25-Phe84 is expressed with a 6His tag at the C-terminus.

Human Trefoil Factor 1 Recombinant Protein Lyophilized
IHUTFF1RLY5UG each
EUR 341
Description: Human Trefoil Factor 1 Recombinant Protein Lyophilized

Human Trefoil Factor 1 Recombinant Protein Lyophilized
MBS8433531-INQUIRE INQUIRE Ask for price

Recombinant Human Trefoil Factor-2 His Tag
7-01591 5µg Ask for price

Recombinant Human Trefoil Factor-2 His Tag
7-01592 20µg Ask for price

Recombinant Human Trefoil Factor-2 His Tag
7-01593 1mg Ask for price

Recombinant Human Trefoil Factor-3 His Tag
7-01597 5µg Ask for price

Recombinant Human Trefoil Factor-3 His Tag
7-01598 20µg Ask for price

Recombinant Human Trefoil Factor-3 His Tag
7-01599 1mg Ask for price

Recombinant Human Trefoil Factor-2 His Tag
cyt-611-1mg 1mg
EUR 2900

Recombinant Human Trefoil Factor-2 His Tag
cyt-611-20g 20µg
EUR 145

Recombinant Human Trefoil Factor-2 His Tag
cyt-611-5g 5µg
EUR 60

Recombinant Human Trefoil Factor-3 His Tag
cyt-616-1mg 1mg
EUR 2900

Recombinant Human Trefoil Factor-3 His Tag
cyt-616-20g 20µg
EUR 145

Recombinant Human Trefoil Factor-3 His Tag
cyt-616-5g 5µg
EUR 60

Recombinant Human Trefoil Factor-2 His Tag
MBS143345-0005mg 0.005mg
EUR 240

Recombinant Human Trefoil Factor-2 His Tag
MBS143345-002mg 0.02mg
EUR 310

Recombinant Human Trefoil Factor-2 His Tag
MBS143345-1mg 1mg
EUR 4175

Recombinant Human Trefoil Factor-2 His Tag
MBS143345-5x1mg 5x1mg
EUR 18455

Recombinant Human Trefoil Factor-3 His Tag
MBS143346-0005mg 0.005mg
EUR 240

Recombinant Human Trefoil Factor-3 His Tag
MBS143346-002mg 0.02mg
EUR 310

Recombinant Human Trefoil Factor-3 His Tag
MBS143346-1mg 1mg
EUR 4175

Recombinant Human Trefoil Factor-3 His Tag
MBS143346-5x1mg 5x1mg
EUR 18455

Recombinant Human Trefoil factor 1 protein(TFF1) (Active)
AP73611 1mg
EUR 3939

Recombinant Human Trefoil factor 1 protein (TFF1) (Active)
CSB-AP000341HU 8514 mg Ask for price

Recombinant Human Trefoil factor 1 protein (TFF1) (Active)
MBS969152-0005mg 0.005mg
EUR 210

Recombinant Human Trefoil factor 1 protein (TFF1) (Active)
MBS969152-01mg 0.1mg
EUR 970

Recombinant Human Trefoil factor 1 protein (TFF1) (Active)
MBS969152-05mg 0.5mg
EUR 2045

Recombinant Human Trefoil factor 1 protein (TFF1) (Active)
MBS969152-5x05mg 5x0.5mg
EUR 8935

Trefoil Factor-2 Human Recombinant, His Tag
rAP-0744 Inquiry Ask for price

Trefoil Factor-3 Human Recombinant, His Tag
rAP-0746 Inquiry Ask for price

Recombinant Human Trefoil Factor 1/TFF1 Protein (His Tag)
PKSH033494-10ug 10ug
EUR 111
Description: Human

Recombinant Human Trefoil Factor 1/TFF1 Protein (His Tag)
PKSH033494-50ug 50ug
EUR 268
Description: Human

Recombinant Human Trefoil factor 3 (TFF3), partial
CSB-YP023433HU 6034 mg Ask for price

Recombinant Human Trefoil factor 3 (TFF3), partial
CSB-EP023433HU 2316 mg Ask for price

Recombinant Human Trefoil factor 3 (TFF3) ,partial
RPC25717-100ug 100ug
EUR 569.8

Recombinant Human Trefoil factor 3 (TFF3) ,partial
RPC25717-20ug 20ug
EUR 353.4

Recombinant Human Trefoil factor 3 (TFF3) ,partial
RPC28215-100ug 100ug
EUR 528

Recombinant Human Trefoil factor 3 (TFF3) ,partial
RPC28215-1mg 1mg
EUR 2412.5

Recombinant Human Trefoil factor 3 (TFF3) ,partial
RPC28215-20ug 20ug
EUR 295.7

Recombinant Human Trefoil Factor 3 (TFF3), Partial
MBS7115263-002mgEColi 0.02mg(E-Coli)
EUR 275

Recombinant Human Trefoil Factor 3 (TFF3), Partial
MBS7115263-002mgYeast 0.02mg(Yeast)
EUR 320

Recombinant Human Trefoil Factor 3 (TFF3), Partial
MBS7115263-01mgEColi 0.1mg(E-Coli)
EUR 460

Recombinant Human Trefoil Factor 3 (TFF3), Partial
MBS7115263-01mgYeast 0.1mg(Yeast)
EUR 515

Recombinant Human Trefoil Factor 3 (TFF3), Partial
MBS7115263-1mgEColi 1mg(E-Coli)
EUR 1615

Recombinant Human Trefoil factor 2 protein (TFF2)
MBS9423197-0005mg 0.005mg
EUR 210

Recombinant Human Trefoil factor 2 protein (TFF2)
MBS9423197-002mg 0.02mg
EUR 365

Recombinant Human Trefoil factor 2 protein (TFF2)
MBS9423197-01mg 0.1mg
EUR 1135

Recombinant Human Trefoil factor 2 protein (TFF2)
MBS9423197-025mg 0.25mg
EUR 1990

Recombinant Human Trefoil factor 2 protein (TFF2)
MBS9423197-05mg 0.5mg
EUR 2570

Recombinant Human Trefoil factor 3 protein (TFF3)
MBS9423198-0005mg 0.005mg
EUR 210

Recombinant Human Trefoil factor 3 protein (TFF3)
MBS9423198-002mg 0.02mg
EUR 365
DO-stat fed-batch utilizing mineral medium (M9) and fed with glucose beneath the identical circumstances produced even greater enzymatic exercise (9.14 U/mgprotein). The pH impact was investigated, and one of the best enzymatic exercise may very well be noticed at pH 8. In all cultivations, the bicistronic system remained secure, with 100% of plasmid-bearing cells. These outcomes present that E. coli bearing bicistronic plasmid constructs to precise recombinant TGase may very well be cultivated in bioreactors beneath DO-stat fed-batch utilizing mineral medium and it’s a promising technique in future optimizations to provide this necessary enzyme.

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