• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.722-725
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• 1999

Filamentation-suppressed recombinant Escherichia coli strain harboring the Alcaligenes latus polyhydroxyalkanoate (PHA) biosynthesis genes and the E. coli ftsZ gene was constructed and cultivated for the production of poly(3-hydroxybutyrate) [P(3HB)] with high concentration and high content. By the pH-stat fed-batch culture of this recombinant E. coli strain XL1-Blue(pJC5), the final cell concentration and P(3HB) concentration obtained in 44.25h were 172.2g cell dry weight/l and 141.9g P(3HB)/l, respectively, resulting in productivity of 3.21g P(3HB)/l-h. More importantly, the P(3HB) content obtained was 82.4 wt %, which was significantly higher than that obtained with the recombinant E. coli harboring only the PHA biosynthesis genes.

• Journal of Microbiology and Biotechnology
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• v.6 no.2
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• pp.147-149
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• 1996

Poly(3-hydroxybutyrate) (PHB) was synthesized and accumulated intracellularly to a high concentration (7 g/l) by cultivating recombinant Escherichia coli XL1-Blue (pSYLl05) in a complex medium containing 20 g/l glucose. The morphology of PHB granules was examined by transmission electron microscopy. The PHB granules synthesized in recombinant E. coli were much larger than reported values for wild type microorganisms, and were often irregularly shaped. Some cells were apparently extruding PHB into the medium, which suggests that PHB granules maintain some fluidity and cells become fragile due to PHB accumulation.

• Journal of Microbiology and Biotechnology
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• v.14 no.1
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• pp.1-14
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• 2004

More than twenty years have passed since the approval of the first recombinant DNA product for therapeutic use (recombinant human insulin, 1982). However, the biotechnology industry is still facing a shortage of manufacturing capacity due to the increasing demand of therapeutic proteins. This demand has prompted the search for a growing number of biological production systems but, nevertheless, the Gram-negative bacterium Escherichia coli remains one of the most attractive production hosts. This review highlights the most important features and developments of plasmid vector design, emphasizing the different reported strategies for improving the expression and secretion of heterologous proteins using the cellular machinery of E. coli.

• KSBB Journal
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• v.10 no.3
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• pp.249-256
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• 1995

${\gamma}$-Glutamylcysteine production by the immobilized microbial system of recombinant Escherichia coli and yeast was investigated. ${\gamma}$-Glutamylcysteine was synthesized from L-glutamic acid and L-cysteine in the presence of ATP by the reaction catalyzed by ${\gamma}$-glutamylcysteine synthetase. An immobilized microbial cell system was developed for the efficient ${\gamma}$-glutamylcysteine production. Recombinant Escherichia coli and yeast were immobilized by alginate. Production of ${\gamma}$-glutamylcysteine was better with the recombinant Escherichia coli for both the synthesis of ${\gamma}$-glutamylcysteine and the ATP regeneration than the mixed system of recombinant Escherichia coli and yeast. The proper radio of recombinant Escherichia coli to yeast was experimentary observed to be 1:4 in the mixed system. Although the immobi1ized system had the slower reaction rate, its reaction stability was increased by 10%.

• Journal of Microbiology and Biotechnology
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• v.4 no.1
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• pp.1-6
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• 1994

Continuous culture was carried out with a recombinant Escherichia coli W3110/pCR185, which encodes trp-operon enzymes when the temperature is shifted from $37^{circ}C\;t;42^{\circ}C$. Under derepressed condition of $42^{\circ}C$. plasmlid stability and gene expression were analysed as function of the dilution rate. The stability of plasmid increased with the dilution rate, but maximal levels of gene expression (tryptophan concentration) and plasmid DNA content were obtained at the lowest dilution rate, $0.075\;hr^{-1}$. The plasmid instability, observed at low dilution rates, could be explained by the unbalanced biosynthetic state of the recombinant cell harboring a high copy number of plasmid.

• Journal of Microbiology and Biotechnology
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• v.25 no.10
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• pp.1709-1713
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• 2015

Sepiapterin is a precursor for the synthesis of tetrahydrobiopterin (BH4), which is a wellknown cofactor for aromatic amino acid hydroxylation and nitric oxide synthesis in higher mammals. In this study, a recombinant Escherichia coli BL21(DE3) strain harboring cyanobacterial guanosine 5’-triphosphate cyclohydrolase 1 (GCH1) and human 6-pyruvoyltetrahydropterin synthase (PTPS) genes was constructed to produce sepiapterin. The optimum conditions for T₇ promoter–driven expression of GCH1 and PTPS were 30℃ and 0.1 mM isopropyl-β-D-thioglucopyranoside (IPTG). The maximum sepiapterin concentration of 88.1 ± 2.4 mg/l was obtained in a batch cultivation of the recombinant E. coli, corresponding to an 18-fold increase in sepiapterin production compared with the control condition (37℃ and 1 mM IPTG).

• Journal of Microbiology and Biotechnology
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• v.24 no.12
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• pp.1685-1689
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• 2014

Baeyer-Villiger (BV) oxidation of cyclohexanone to ${\varepsilon}$-caprolactone in a microbial system expressing cyclohexanone monooxygenase (CHMO) can be influenced by not only the efficient regeneration of NADPH but also a sufficient supply of oxygen. In this study, the bacterial hemoglobin gene from Vitreoscilla stercoraria (vhb) was introduced into the recombinant Escherichia coli expressing CHMO to investigate the effects of an oxygen-carrying protein on microbial BV oxidation of cyclohexanone. Coexpression of Vhb allowed the recombinant E. coli strain to produce a maximum ${\varepsilon}$-caprolactone concentration of 15.7 g/l in a fed-batch BV oxidation of cyclohexanone, which corresponded to a 43% improvement compared with the control strain expressing CHMO only under the same conditions.

• Korean Journal of Agricultural Science
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• v.41 no.3
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• pp.245-249
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• 2014

Among dozens of DNA polymerases cloned from thermophilic bacteria, Taq DNA polymerase from Thermus aquaticus has been most frequently used in polymerase chain reaction (PCR) that is being applied to gene cloning, DNA sequencing, gene expression analysis, and detection of infectious and genetic diseases. Since native Taq DNA polymerase is expressed at low level in T. aquaticus, recombinant Escherichia coli system was used to produce Taq DNA polymerase in a large amount. Taq DNA polymerase was expressed as a soluble form under the control of tac promoter in E. coli, and purified by heat treatment and ion exchange chromatographies. The purified Taq DNA polymerase was nearly homogeneous and exhibited a similar DNA amplification activity with a commercial Taq DNA polymerase.

• Microbiology and Biotechnology Letters
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• v.32 no.3
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• pp.212-217
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• 2004

The gene coding for mannanase from Bacillus subtilis WL-7, a number of glycosyl hydrolase family 26, was hyperexpressed in Escherichia coli. Two recombinant plasmids, pE7MAN and pENS7, were constructed by introducing the complete mannanase gene and the mature mannanase gene lacking N-terminal signal peptide region into a expression vector pET24a(＋), respectively. The level of mannanase produced by E. coli BL21 (DE3) carrying pENS7, which included the mature mannanase gene, was considerably higher than that by E. coli BL21 (DE3)/pE7MAN. Almost mannanase produced by the recombinant E. coli carrying pENS7 at growth temperature of $37^{\circ}C$ existed as inactive enzyme of insoluble form. Growth at temperature below $31^{\circ}C$ increased the soluble fraction of mannanase having catalytic activity in the recombinant E. coli cells. The highest productivity of active mannanase was observed in cell-free extract of the recombinant E. coli grown at growth temperature ranging from $25^{\circ}C$ to $28^{\circ}C$, while mannanase activity per soluble protein of the cell-free extract was highest in the cells grown at $^31{\circ}C$.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.4
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• pp.353-356
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• 2005

The effect of silkworm hemolymph on the expression of recombinant protein in Escherichia coli was investigated. The addition of silkworm hemolymph to the culture medium in­creased the production of recombinant $\beta$-galactosidase in E. coli. The production was dependent on the concentration of the added silkworm hemolymph, which increased 2-, 5-, and 8-fold in media supplemented with 1, 3, and $5\%$ silkworm hemolymph, respectively. To identify the effective component, the silkworm hemolymph was fractionated by gel filtration column chromatography. A fraction, with a molecular weight of about 30 K was identified as the effective component.