Expression and production optimization of sucrose isomerase from Pantoea dispersa in Escherichia coli / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1070-1080, 2016.
Article
em Zh
| WPRIM
| ID: wpr-242273
Biblioteca responsável:
WPRO
ABSTRACT
To improve the yield of sucrose isomerase from Pantoea dispersa UQ68J, we studied the effect of different signal peptides and fermentation conditions on sucrose isomerase expression in Escherichia coli. The gene of sucrose isomerase was optimized and expressed in E. coli BL21 (DE3) with native signal peptide which was named as ORI strain. The total and extracellular enzyme activity was 85 and 65 U/mL in the flask, respectively. The mature protein, which started from the 22th amino acid, was connected with the PelB and OmpA signal peptide to construct P22 and O22 strain, respectively. The total activity of P22 reached 138 U/mL, which was 1.6 times of ORI strain. The total activity of O22 strain was similar to that of ORI strain. Induced by 3.0 g/L lactose, the total activity of P22 strain increased to 168 U/mL. In 3 L fermentor, the effects of glycine concentration and induction time were studied. Induction when the DCW reached 18 g/L (OD₆₀₀=30), with 0.5% glycine, the extracellular enzyme activity reached 1 981 U/mL, and the total enzyme activity reached 2 640 U/mL, which is the highest activity of sucrose isomerase that was expressed in recombinant E. coli.
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Texto completo:
1
Índice:
WPRIM
Assunto principal:
Proteínas de Bactérias
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Proteínas Recombinantes
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Sinais Direcionadores de Proteínas
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Expressão Gênica
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Reatores Biológicos
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Pantoea
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Escherichia coli
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Fermentação
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Glucosiltransferases
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Lactose
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2016
Tipo de documento:
Article