Expression and Purification of Recombinant Human Granulocyte Colony-Stimulating Factor in Fed-Batch Culture of Escherichia coli

2014 ◽  
Vol 172 (5) ◽  
pp. 2425-2435 ◽  
Author(s):  
Chang-Kyu Kim ◽  
Jun-Ha Choi ◽  
Seung-Bae Lee ◽  
Sang-Mahn Lee ◽  
Jae-Wook Oh
Keyword(s):  
Escherichia Coli ◽  
Batch Culture ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Fed Batch ◽  
Expression And Purification ◽  
Fed Batch Culture ◽  
Stimulating Factor

Suitable Signal Peptides for Secretory Production of Recombinant Granulocyte Colony Stimulating Factor in Escherichia coli

2020 ◽  
Vol 14 (4) ◽  
pp. 269-282
Author(s):  
Sadra S. Tehrani ◽  
Golnaz Goodarzi ◽  
Mohsen Naghizadeh ◽  
Seyyed H. Khatami ◽  
Ahmad Movahedpour ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Signal Peptide ◽  
Fusion Proteins ◽  
Inclusion Bodies ◽  
Clinical Aspects ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
E Coli ◽  
Secretory Production ◽  
Stimulating Factor

Background: Granulocyte colony-stimulating factor (G-CSF) expressed in engineered Escherichia coli (E. coli) as a recombinant protein is utilized as an adjunct to chemotherapy for improving neutropenia. Recombinant proteins overexpression may lead to the creation of inclusion bodies whose recovery is a tedious and costly process. To overcome the problem of inclusion bodies, secretory production might be used. To achieve a mature secretory protein product, suitable signal peptide (SP) selection is a vital step. Objective: In the present study, we aimed at in silico evaluation of proper SPs for secretory production of recombinant G-CSF in E. coli. Methods: Signal peptide website and UniProt were used to collect the SPs and G-CSF sequences. Then, SignalP were utilized in order to predict the SPs and location of their cleavage site. Physicochemical features and solubility were investigated by ProtParam and Protein-sol tools. Fusion proteins sub-cellular localization was predicted by ProtCompB. Results: LPP, ELBP, TSH, HST3, ELBH, AIDA and PET were excluded according to SignalP. The highest aliphatic index belonged to OMPC, TORT and THIB and PPA. Also, the highest GRAVY belonged to OMPC, ELAP, TORT, BLAT, THIB, and PSPE. Furthermore, G-CSF fused with all SPs were predicted as soluble fusion proteins except three SPs. Finally, we found OMPT, OMPF, PHOE, LAMB, SAT, and OMPP can translocate G-CSF into extracellular space. Conclusion: Six SPs were suitable for translocating G-CSF into the extracellular media. Although growing data indicate that the bioinformatics approaches can improve the precision and accuracy of studies, further experimental investigations and recent patents explaining several inventions associated to the clinical aspects of SPs for secretory production of recombinant GCSF in E. coli are required for final validation.

The effect of rare codons following the ATG start codon on expression of human granulocyte-colony stimulating factor in Escherichia coli

2015 ◽  
Vol 114 ◽  
pp. 108-114 ◽  
Author(s):  
Zeinab Karimi ◽  
Navid Nezafat ◽  
Manica Negahdaripour ◽  
Aydin Berenjian ◽  
Shiva Hemmati ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Start Codon ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Rare Codons ◽  
Stimulating Factor

scholarly journals Kinetics of High Cell Density Fed-batch Culture of Recombinant Escherichia coli Producing Human-like Collagen

2006 ◽  
Vol 14 (2) ◽  
pp. 242-247 ◽  
Author(s):  
Xiufu HUA ◽  
Daidi FAN ◽  
Yan'e LUO ◽  
Xi ZHANG ◽  
Huijuan SHI ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Density ◽  
Batch Culture ◽  
High Cell Density ◽  
Recombinant Escherichia Coli ◽  
High Cell ◽  
Fed Batch ◽  
Fed Batch Culture ◽  
Kinetics Of

Analysis of metabolic flux in Escherichia coli expressing human-like collagen in fed-batch culture

2007 ◽  
Vol 30 (4) ◽  
pp. 637-643 ◽  
Author(s):  
Yan E. Luo ◽  
Dai D. Fan ◽  
Long A. Shang ◽  
Hui J. Shi ◽  
Xiao X. Ma ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Batch Culture ◽  
Metabolic Flux ◽  
Fed Batch ◽  
Fed Batch Culture
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