Expression and translocation of glucose isomerase as a fusion protein in E. coli

Sarıyar, Berna; Özkan, Pınar; Kırdar, Betül; Hortaçsu, Amable

doi:10.1016/j.enzmictec.2003.10.021

Cited by 5 publications

(3 citation statements)

References 33 publications

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“…Therefore, by considering technical innovations in the area of molecular biotechnology, for efficient production of thermostable GI by mesophiles, it will be more feasible to develop recombinant strains encoding the xylA gene of thermophiles. Production of recombinant GI has been achieved in several mesophilic host organisms including Bacillus subtilis , Bacillus brevis , Streptomyces lividans , Schizosaccharomyces pombe , Saccharomyces cerevisiae and different E. coli strains . In fact, application of bacterial expression systems for over‐production of heterologous non‐glycosylated proteins has several benefits such as attaining high cell density and expression level of proteins via cultivation of recombinant cells on inexpensive media, availability of well‐characterized genetic information, and accessibility to a variety of cloning vectors and genetically modified host strains.…”

Section: Introductionmentioning

confidence: 99%

Beet molasses based exponential feeding strategy for thermostable glucose isomerase production by recombinant Escherichia coli BL21 (DE3)

Angardi

Çalı́k

2012

J of Chemical Tech & Biotech

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BACKGROUND: The effects of pretreated beet molasses based feeding strategies on thermostable glucose isomerase (GI) production by recombinant Escherichia coli BL21 (DE3) pLysS were investigated. RESULTS: The thermostable GI encoding gene of Thermus thermophilus (xylA) was recombined with pRSETA vector, and the pRSETA::xylA obtained was transferred into E.coli BL21 (DE3) pLysS and used for GI production. The highest soluble GI activity was obtained at t = 30 h, as A = 16 400 U L−1 (20.6 U mg−1 protein) under molasses based fed‐batch operation, with a specific growth rate µ = 0.1 h−1 (M‐0.1); on the other hand, the highest cell concentration was obtained at µ = 0.15 h−1 operation as 9.6 g L−1 at t = 32 h. The highest oxygen uptake was 4.57 mol m−3 s−1 at M‐0.1 operation. CONCLUSIONS: Molasses based fed‐batch operations were more successful in terms of cell concentration and thermostable enzyme production due to the existence of a natural sugar inducer, galactose, in the molasses composition. This study demonstrates the significance of proper feeding strategy development for over‐production of enzymes by recombinant E. coli strains. © 2012 Society of Chemical Industry

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Section: Introductionmentioning

confidence: 99%

Beet molasses based exponential feeding strategy for thermostable glucose isomerase production by recombinant Escherichia coli BL21 (DE3)

Angardi

Çalı́k

2012

J of Chemical Tech & Biotech

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“…Several studies have also reported that low efficiency in active recombinant proteins recovery occurred when culture temperature was unchanged after induction due to formation of inclusion bodies, the latter is related to the high growth rate during the expression (Strandberg and Enfors, 1991;Garcí a-Junceda et al, 1995;Georgiou and Valax, 1996). Thus, lowering the culture temperature during expression of the protein may increase cellular responses to induction (Sariyar et al, 2004). In this study, cultures were induced with optimal concentration of 1.0 mM IPTG at OD600 1.5.…”

Section: Selection Of Post-induction Temperaturementioning

confidence: 99%

“…Comparison of the two temperatures using one-way ANOVA showed significant difference in the results (p ≤ 0.05). Post-induction temperature reduction has led to higher volumetric recombinant protein production by redirecting the cell's metabolic system to expressing the foreign gene, rather than performing the natural cellular responses of multiplication growth, while also suppressing the expression of other cellular proteins, and represses protease activity (Sariyar et al, 2004).…”

Section: Selection Of Post-induction Temperaturementioning

confidence: 99%

Production of Brugia malayi BmSXP recombinant protein expressed in Escherichia coli

Khoo¹,

Agilan²,

Noordin³

et al. 2010

MJM

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A rapid antibody detection test is very useful for detection of lymphatic filariasis, especially for certification and surveillance of post-mass drug administration. One such kit, panLF Rapid TM (commercialized by Malaysian BioDiagnostic Research Sdn. Bhd.) had been developed in our laboratory for the detection of all species of filarial infections. It is based on the detection of anti-filarial IgG4 antibodies that react with recombinant Brugia malayi antigens, BmR1 and BmSXP. In this study, the growth of recombinant bacteria that produce BmSXP was optimized under shake flask fermentation for high yield of the recombinant antigen. The optimizations involved selection of suitable growth medium, IPTG concentration and induction time. The medium that yielded the highest biomass as well as total protein was Terrific Broth (TB) medium, which is an undefined medium. Initiation of induction of protein expression was found to be best at mid-log phase (OD600 = 1.5), with IPTG concentration of 1.0 mM, and harvest time at 9 h post-induction. This study showed that under the optimized conditions, the shake flask culture produced 4 g/L biomass (dry cell weight) of recombinant Escherichia coli BmSXP/pPROEXHTa/TOP10F', which yielded 2.42 mg/L of purified BmSXP recombinant antigen. The purified antigen was analyzed by SDS-PAGE and the antigenicity of protein was confirmed by Western blot.

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Glycine and Triton X-100 enhanced secretion of recombinant α-CGTase mediated by OmpA signal peptide in Escherichia coli

Wang

et al. 2012

Biotechnol Bioproc E

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Expression and translocation of glucose isomerase as a fusion protein in E. coli

Cited by 5 publications

References 33 publications

Beet molasses based exponential feeding strategy for thermostable glucose isomerase production by recombinant Escherichia coli BL21 (DE3)

Beet molasses based exponential feeding strategy for thermostable glucose isomerase production by recombinant Escherichia coli BL21 (DE3)

Production of Brugia malayi BmSXP recombinant protein expressed in Escherichia coli

Glycine and Triton X-100 enhanced secretion of recombinant α-CGTase mediated by OmpA signal peptide in Escherichia coli

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