Heterologous expression, purification, immobilization and characterization of recombinant α-amylase AmyLa from Laceyella sp. DS3

Heterologous expression, purification, immobilization and characterization of recombinant α-amylase AmyLa from Laceyella sp. DS3

AmyLa α-amylase gene from Laceyella sp. DS3 was heterologously expressed in E. coli BL21. E. coli BL21 maximally expressed AmyLa after 4 h of adding 0.02 mM IPTG at 37 °C. The recombinant AmyLa α-amylase was purified 2.19-fold through gel filtration and ion exchange chromatography. We immobilized th...

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Veröffentlicht in:International journal of biological macromolecules 2019-07, Vol.132, p.1274-1281
Hauptverfasser: El-Sayed, Ahmed K.A., Abou-Dobara, Mohamed I., El-Fallal, Amira A., Omar, Noha F.
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Heterologous expression
Immobilization
α-Amylase
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container_title International journal of biological macromolecules
container_volume 132
creator El-Sayed, Ahmed K.A.
Abou-Dobara, Mohamed I.
El-Fallal, Amira A.
Omar, Noha F.
description AmyLa α-amylase gene from Laceyella sp. DS3 was heterologously expressed in E. coli BL21. E. coli BL21 maximally expressed AmyLa after 4 h of adding 0.02 mM IPTG at 37 °C. The recombinant AmyLa α-amylase was purified 2.19-fold through gel filtration and ion exchange chromatography. We immobilized the purified recombinant AmyLa α-amylase on four carriers; chitosan had the best efficiency. The recombinant free and the immobilized AmyLa α-amylase showed optimum activity in the pH ranges of 6.0–7.0 and 4.0–7.0, respectively and possessed an optimum temperature of 55 °C. The free enzyme had activation energy, Km, and Vmax of 291.5 kJ, 1.5 mg/ml, and 6.06 mg/min, respectively. The immobilized enzyme had activation energy, Km, and Vmax of 309.74 kJ, 6.67 mg/ml, and 50 mg/min, respectively. The immobilized enzyme was calcium-independent and insensitive (relative to the free enzyme) to metals. It could also be reused for seven cycles.
doi_str_mv 10.1016/j.ijbiomac.2019.04.010
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The immobilized enzyme was calcium-independent and insensitive (relative to the free enzyme) to metals. 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The immobilized enzyme had activation energy, Km, and Vmax of 309.74 kJ, 6.67 mg/ml, and 50 mg/min, respectively. The immobilized enzyme was calcium-independent and insensitive (relative to the free enzyme) to metals. It could also be reused for seven cycles.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>30953727</pmid><doi>10.1016/j.ijbiomac.2019.04.010</doi><tpages>8</tpages></addata></record>
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subjects Heterologous expression
Immobilization
α-Amylase
title Heterologous expression, purification, immobilization and characterization of recombinant α-amylase AmyLa from Laceyella sp. DS3
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