PMID- 32876507
OWN - NLM
STAT- MEDLINE
DCOM- 20210719
LR  - 20210719
IS  - 1532-2297 (Electronic)
IS  - 1082-6068 (Linking)
VI  - 51
IP  - 3
DP  - 2021
TI  - Kinetic study and optimization of recombinant human tumor necrosis factor-alpha 
      (rhTNF-alpha) production in Escherichia coli.
PG  - 267-276
LID - 10.1080/10826068.2020.1815056 [doi]
AB  - Tumor necrosis factor-alpha (TNF-alpha) is an inflammatory cytokine that plays a 
      major role in immune regulation, homeostatic function, and cellular organization. 
      The present study was undertaken to overproduce recombinant human TNF-alpha (rhTNF-alpha) 
      in Escherichia coli (E.coli) in high cell density culture. The use of a 
      codon-optimized gene and strong promoter-based (T7) expression system, choice of 
      Terrific Broth (TB) as medium, and subsequent optimization of culture conditions 
      in shake flasks resulted in production of 0.95 g/L insoluble rhTNF-alpha comprising 
      upto 50% of total cellular protein (TCP) The protein yield further increased upto 
      1.26 g/L in 1 L TB medium batch culture in bioreactor with the controlled 
      temperature, pH, and dissolved oxygen. In a series of chemostats operated at 
      dilution rates of 0.2 h(-1), 0.3 h(-1), 0.4 h(-1) and 0.5 h(-1) the specific 
      growth rate (mu) positively correlated with specific yield (Y(p)(/)(x)) and a 
      maximum yield of 164 mg/g DCW was obtained at mu = 0.4 h(-1) within 4 h 
      post-induction. A fed-batch cultivation in TB with an exponential feeding profile 
      (mu =  approximately 0.4 h(-1)) of concentrated feed resulted in an accumulation of 5.5 g/L of 
      rhTNF-alpha within 14 h of cultivation which accounted for  approximately 29% of TCP.
FAU - Singha, Tapan Kumar
AU  - Singha TK
AD  - Department of Microbiology, Maharshi Dayanand University, Rohtak, India.
FAU - Dagar, Vikas Kumar
AU  - Dagar VK
AD  - Department of Microbiology, University of Delhi South Campus, New Delhi, India.
FAU - Gulati, Pooja
AU  - Gulati P
AD  - Department of Microbiology, Maharshi Dayanand University, Rohtak, India.
FAU - Kumar, Sanjay
AU  - Kumar S
AD  - Department of Microbiology, Maharshi Dayanand University, Rohtak, India.
LA  - eng
PT  - Journal Article
DEP - 20200902
PL  - England
TA  - Prep Biochem Biotechnol
JT  - Preparative biochemistry & biotechnology
JID - 9607037
RN  - 0 (Culture Media)
RN  - 0 (Proteins)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Tumor Necrosis Factor-alpha)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - Batch Cell Culture Techniques
MH  - Bioreactors
MH  - Biotechnology/*methods
MH  - Culture Media
MH  - Escherichia coli/*metabolism
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Kinetics
MH  - Oxygen/metabolism
MH  - Promoter Regions, Genetic
MH  - Proteins/chemistry
MH  - Recombinant Proteins/*chemistry
MH  - Solubility
MH  - Temperature
MH  - Tumor Necrosis Factor-alpha/*chemistry
OTO - NOTNLM
OT  - Escherichia coli
OT  - TNF-alpha
OT  - chemostat culture
OT  - expression kinetics
OT  - fed-batch culture
OT  - optimization
OT  - overproduction
EDAT- 2020/09/03 06:00
MHDA- 2021/07/20 06:00
CRDT- 2020/09/03 06:00
PHST- 2020/09/03 06:00 [pubmed]
PHST- 2021/07/20 06:00 [medline]
PHST- 2020/09/03 06:00 [entrez]
AID - 10.1080/10826068.2020.1815056 [doi]
PST - ppublish
SO  - Prep Biochem Biotechnol. 2021;51(3):267-276. doi: 10.1080/10826068.2020.1815056. 
      Epub 2020 Sep 2.