High‐level soluble expression of <i><scp>S</scp>erratia marcescens</i><scp>H</scp>30 lipase in <i><scp>E</scp>scherichia coli</i>

Su, Erzheng; Xu, Jingjing; Wu, Xiaoyan

doi:10.1002/bab.1248

Cited by 11 publications

(1 citation statement)

References 28 publications

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“…Many Bacillus lipases had been cloned, expressed, purified and characterised to an extent until date [20][21][22]. Various studies has been carried out to optimise the production of recombinant proteins in E. coli by changing media composition [23][24][25]. Some studies have also been carried out to optimise the expression system and host cells for the production of recombinant proteins in heterologous system [11,26,27].…”

Section: Introductionmentioning

confidence: 99%

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Kaur¹

2017

IJMBOA

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Recombinant protein production is a vast field growing larger in each passing year due to continuous emergence of new host, vectors and techniques for cloning and expression. Earlier, a Bacillus lipase gene was cloned in pGEMT/ E. coli DH5α with 12 units/ml lipase production. In the present investigation, attempt has been made to optimize the production of recombinant lipase by E. coli by varying different parameters. Amongst the different strains of E. coli, M46 cells were observed to be the best host with 19 units/ml lipase production. Addition of surfactant and changing media to nutrient broth lead to 2.8 fold and 1.3 fold respective increase in lipase production. Amongst different vectors, pET 28a vector showed the highest total lipase production but it proved toxic for the cells, while pQE30-UA vector gives 5 fold enhanced total lipase production with more stable expression. With the combination of optimized host, vector, surfactant and media, 18 fold increases in lipase production (214 units/ml) was achieved.

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Section: Introductionmentioning

confidence: 99%

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Kaur¹

2017

IJMBOA

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Production of a bioactive recombinant chicken matrix metalloproteinase‐11 peptide in Escherichia coli

Paul

Kumar

Kataria

2017

Biotech and App Biochem

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Matrix metalloproteinase-11 (MMP-11) is known to be highly expressed in metastatic and most invasive forms of tumors. Being selectively expressed in tumor tissues, MMP-11 is a promising target for immunotherapy against tumors. Here, we report the production of a thioredoxin-tagged bioactive recombinant chicken MMP-11 (cMMP-11) peptide excluding the secretory signal and propeptide in Escherichia coli T7 Express lysY using pET32b(+) vector. High-level expression and purification of the bioactive peptide were achieved by induction with 1.0 mM isopropyl-β-d-thiogalactopyranoside for 4 H at 37 °C followed by affinity chromatography under denaturing condition and slow dialysis. The recombinant peptide exhibited both caseinolytic and gelatinase activities without requiring activation by 4-aminophenylmercuric acetate. The antisera raised against the peptide in rabbits showed a strong reaction with the whole recombinant peptide as well as 37 kDa cMMP-11 mature peptide and cross-reactivity with a 43 kDa protein in murine breast tumor of 4T1 origin in Western blot analysis. The 43 kDa protein in the tumor homogenate showed immunoreactivity with a monoclonal antibody against human MMP-11, suggesting it to be murine MMP-11 having cross-reactivity with the antisera raised against cMMP-11 peptide. Altogether, the study characterized the production of a bioactive and immunogenic recombinant cMMP-11 peptide in E. coli.

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Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration

Lopes

Santos

Dupont

et al. 2019

Biotech and App Biochem

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Green fluorescent protein (GFP) is a globular protein used as biosensor and biomarker in medical and industrial fields. However, due to the expensive production costs of expressing proteins using high-cost inducers like isopropyl-β-D-1thiogalactopyranoside (IPTG), the number of GFP applications are still scarce. This work studied the production of enhanced GFP (EGFP) using Escherichia coli BL21 (DE3) [pLysS; pET28(a)], aiming to increase its yield and reduce costs. First, the influence of agitation rate, induction time, and concentration of IPTG in the production of EGFP was evaluated, but only the first two parameters were significant. Subsequently, aiming to reduce costs related to the use of inducer, the IPTG concentration (0.005, 0.010, and 0.025 mM) was decreased and, interestingly, the production levels were maintained or increased. These results show that a proper choice of production conditions, particularly through the decrease of inducer concentration, is effective to reduce the upstream production costs and guarantee high EGFP expression.

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High‐level soluble expression of Serratia marcescensH30 lipase in Escherichia coli

Cited by 11 publications

References 28 publications

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Studies on Recombinant Lipase Production by E. Coli: Effect of Media And Bacterial Expression System Optimization

Production of a bioactive recombinant chicken matrix metalloproteinase‐11 peptide in Escherichia coli

Improving the cost effectiveness of enhanced green fluorescent protein production using recombinant Escherichia coli BL21 (DE3): Decreasing the expression inducer concentration

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