Soluble Expression of Recombinant Nerve Growth Factor in Cytoplasm of Escherichia coli

Shamriz, Shabnam; Ofoghi, Hamideh; Amini-Bayat, Zahra

doi:10.15171/ijb.1331

Cited by 4 publications

(3 citation statements)

References 32 publications

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“…Mis-pairing of disulfide bond in cysteines will cause mis-folding during protein synthesis and form the inclusion body. The inclusion body formation in recombinant proteins can result in inactive or non-functional proteins [14,15].…”

Section: Discussionmentioning

confidence: 99%

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Soluble expression and purification of hepatitis B core antigen (HBcAg) subgenotype B3 in Escherichia coli using thioredoxin fusion tag

Waty¹,

Mustopa²,

Suharsono³

et al. 2017

APJTD

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Section: Discussionmentioning

confidence: 99%

“…However, the refolding process may occur randomly which cause low-yield protein [16]. coli [15,17,20]. Fusion partner protein (His) 6tag helped us in the protein purification process.…”

Section: Discussionmentioning

confidence: 99%

Soluble expression and purification of hepatitis B core antigen (HBcAg) subgenotype B3 in Escherichia coli using thioredoxin fusion tag

Waty¹,

Mustopa²,

Suharsono³

et al. 2017

APJTD

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“…Utilizing strong promoters as the routine approach in E. coli expression system is likely to produce great concentrations of desired proteins and formation of insoluble protein aggregates. Since the expression of protein in its soluble format is more desirable considering no need for further refolding steps different approaches have been applied to address this problem ( 26 ). Reducing rate of protein synthesis by decreased inducer concentration, induction at lower temperatures ( 27 ), medium optimization ( 28 ), co-expression of chaperons ( 29 ) and foldases ( 30 ) and the use of fusion tags ( 31 ) have been investigated in several studies.…”

Section: Introductionmentioning

confidence: 99%

Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein

et al. 2018

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BackgroundThe increase of the protein expression via ribosomal manipulation is one of the suggested cellular mechanisms involved in EnBase fed-batch mode of cultivation. However, this system has not been implemented for cytotoxic proteins.ObjectivesHere, the expression pattern of α-Luffin, a ribosome inactivation protein (RIP) with an innate toxicity, was investigated in EnBase system and the effect of low temperature cultivation on the increase of α-Luffin solubility was determined.Materials and MethodsThe encoding cDNA for mature α-Luffin was synthesized and subcloned into pET28a plasmid under the control of T7 promoter. The E. coli expression yield in EnBase® Flo fed-batch system was compared with traditional batch mode at two temperatures: 25 °C and 30 °C. Sampling was performed at several time intervals and solubility of recombinant-protein was checked on SDS-PAGE in pellet and supernatant samples. The purification of recombinant protein was performed by Ni-NTA column.ResultsIn fed-batch cultivation mode, the early incubation time was desirable at 30 °C whereas the maximum amount of soluble α-Luffin was achieved from the extended protein synthesis period (12 and 24h post induction) at 25 °C.ConclusionsOur founding showed that EnBase had a greater efficacy in producing higher soluble protein ratios compared to batch cultivation growth rate, however for cytotoxic proteins, incubation temperature and time need to be optimized. Owing to the advantages of natural toxins from RIP family for producing anticancer immune-conjugates, well optimization of this protein expression is of importance regarding industrial aspects. The optimized condition proposed here is promising in terms of large scale soluble production of α-Luffin without the need for refolding.

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Identification and molecular cloning of Bm86 gene in Boophilus microplus ticks collected from beef cattle in several regions of Indonesia

Anwar,

Mahari,

Lupitasari

et al. 2024

International Conference on Organic and Applied Chemistry (Icoac) 2022

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Soluble Expression of Recombinant Nerve Growth Factor in Cytoplasm of Escherichia coli

Cited by 4 publications

References 32 publications

Soluble expression and purification of hepatitis B core antigen (HBcAg) subgenotype B3 in Escherichia coli using thioredoxin fusion tag

Soluble expression and purification of hepatitis B core antigen (HBcAg) subgenotype B3 in Escherichia coli using thioredoxin fusion tag

Optimization of EnBase Fed-Batch Cultivation to Improve Soluble Fraction Ratio of α-Luffin Ribosome Inactivating Protein

Identification and molecular cloning of Bm86 gene in Boophilus microplus ticks collected from beef cattle in several regions of Indonesia

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