Heat-Induced Production of Human Growth Hormone by High Cell Density Cultivation of Recombinant Escherichia Coli

Tabandeh, Fatemeh; Shojaosadati, Seyed Abbas; Zomorodipour, Alireza; Khodabandeh, Mahvash; Sanati, Mohammad Hossein; Yakhchali, Bagher

doi:10.1023/b:bile.0000013714.88796.5f

Cited by 34 publications

(22 citation statements)

References 13 publications

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“…Nevertheless, the activity in aggregates inversely correlates to the temperature [16]. In thermoinducible systems, the IB formation has also been attributed to the increase in recombinant protein synthesis rate and mRNA overexpression [43], recombinant protein amount [42][43][44][45][46], and activation of some heat shock proteins that could favor the disorder in folding reactions [3,42,43,47]. Furthermore, IB formation is favored by shake flask conditions using chemical [13] or thermo-inducible [48] recombinant strains.…”

Section: Introductionmentioning

confidence: 99%

Influence of pH control in the formation of inclusion bodies during production of recombinant sphingomyelinase-D in Escherichia coli

et al. 2014

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Background: Inclusion bodies (IBs) are aggregated proteins that form clusters when protein is overexpressed in heterologous expression systems. IBs have been considered as non-usable proteins, but recently they are being used as functional materials, catalytic particles, drug delivery agents, immunogenic structures, and as a raw material in recombinant therapeutic protein purification. However, few studies have been made to understand how culture conditions affect the protein aggregation and the physicochemical characteristics that lead them to cluster. The objective of our research was to understand how pH affects the physicochemical properties of IBs formed by the recombinant sphingomyelinase-D of tick expressed in E. coli BL21-Gold (DE3) by evaluating two pH culture strategies.

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Section: Introductionmentioning

confidence: 99%

Influence of pH control in the formation of inclusion bodies during production of recombinant sphingomyelinase-D in Escherichia coli

et al. 2014

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“…Bacterial Strains and Plasmids E. coli JM109 (endA1, gyrA96, thi, hsdR17, supE44, relA1, Δ(lac-proAB), recA1, F′[traD36, proAB + , lacl q , lacZΔM15]) was used as the host strain for pMKAK1-2 [2]. The recombinant plasmid pMKAK1-2 contains a gene encoding the cDNA of porcine muscle ADK under the control of the E. coli tac promoter [2] (Fig.…”

Section: Methodsmentioning

confidence: 99%

“…The recombinant plasmid pMKAK1-2 contains a gene encoding the cDNA of porcine muscle ADK under the control of the E. coli tac promoter [2] (Fig. 1).…”

Section: Methodsmentioning

confidence: 99%

“…This enzyme has been regarded as the potential enzyme for ATP regeneration in cell-free protein synthesis [1]. In a previous study, we achieved high expression levels of porcine ADK in recombinant Escherichia coli harboring pMKAK1-2 with a pUC18-based replicative origin [2]. However, an efficient production system in a bioreactor was necessary, while there have been no such report for the porcine ADK production.…”

Section: Introductionmentioning

confidence: 99%

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Optimized Culture Conditions for the Efficient Production of Porcine Adenylate Kinase in Recombinant Escherichia coli

Matsui

Togari

Misawa

et al. 2010

Appl Biochem Biotechnol

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Temperature shift cultivations with amino acid supplementation were optimized to produce porcine adenylate kinase (ADK) in recombinant Escherichia coli harboring a pUC-based recombinant plasmid under the control of the trp promoter. With regard to temperature control, the culture condition was initially maintained at 35 degrees C for cellular growth, but ADK expression was suppressed until the late logarithmic growth phase; subsequently, a temperature shift was applied (from 35 degrees C to 42 degrees C), which resulted in maximal ADK production. In addition, supplementation of amino acids, especially valine and leucine, during the temperature shift stimulated ADK expression from 3.5% to 9.2% and 8.6% of the total protein, respectively. After optimization, 1 g ADK per liter was produced within 16 h of cultivation with a dry cell weight of 21.8 g/l. In this system, there was no loss of the recombinant plasmid during cultivation without selective pressure.

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“…It appears that the overly high protein expression rate is another reason for the formation of inclusion bodies. It has been confirmed that prokaryotic expression systems are preferred in the production of non-glycosylated r-hGH with biological activity [27,28], and that the r-hGH in inclusion bodies can be easily purified from the culture broth and undergo renaturation to bioactive forms [29]. Therefore, all fed-batch cultures were conducted at 37°C for enhancing the cell growth and the expression of r-hGH.…”

Section: Expression Of R-hgh At Different Temperaturesmentioning

confidence: 98%

Effects of Oxygen Supply Modes on the Production of Human Growth Hormone in Different Scale Bioreactors

Shang¹,

Tian²,

Kim³

et al. 2009

Chem Eng & Technol

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Recombinant Escherichia coli has been studied as a main host for recombinant protein productions, but it is still difficult to cultivate E. coli in a large industrialscale process due to the oxygen supply limitation. In this study, E. coli BL(21) harboring a new constructed plasmid (pEHUb-hGH) was used for producing recombinant human growth hormone (r-hGH) in 5-L and 30-L scale fermentors by supplying air and high purity oxygen, respectively, where the high purity oxygen was produced from a vacuum pressure swing adsorption (PSA). The impact of oxygen supply modes, i.e., air and high purity oxygen, on cell growth and r-hGH production was investigated in different scale fermentors. In the case of high purity oxygen supply, the final cell density and r-hGH concentrations were 63.0 and 4.8 g/L in the 5-L fermentor, 51.6 and 4.0 g/L in the 30-L fermentor, respectively. In addition, the productivity of r-hGH was doubled in the 5-L fermentor, and increased 4-fold in the 30-L fermentor, compared to the results obtained in the case of the air supply. The supply of high purity oxygen eliminated the oxygen limitation and acetate formation effectively, and apparently, did not affect the degradation of r-hGH. This shows that the recombinant E. coli cultivation with high purity oxygen produced from PSA may provide an effective method for large-scale industrial production of recombinant proteins.

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Heat-Induced Production of Human Growth Hormone by High Cell Density Cultivation of Recombinant Escherichia Coli

Cited by 34 publications

References 13 publications

Influence of pH control in the formation of inclusion bodies during production of recombinant sphingomyelinase-D in Escherichia coli

Influence of pH control in the formation of inclusion bodies during production of recombinant sphingomyelinase-D in Escherichia coli

Optimized Culture Conditions for the Efficient Production of Porcine Adenylate Kinase in Recombinant Escherichia coli

Effects of Oxygen Supply Modes on the Production of Human Growth Hormone in Different Scale Bioreactors

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