Record number :
732851
Title of article :
Culture and characteristics of recombinant protein production of an Escherichia coli strain expressing carboxylesterase B1
Author/Authors :
C.L. Qiao، نويسنده , , B.C. Shen، نويسنده , , J.M. Xing، نويسنده , , J. Huang، نويسنده , , J.L. Zhang، نويسنده , , D.X. Zhao، نويسنده , , B. Yang، نويسنده ,
Issue Information :
روزنامه با شماره پیاپی سال 2006
Pages :
5
From page :
77
To page :
81
Abstract :
High-density culture was achieved through controlling specific growth rate by limiting glucose concentration to <0.2 g L−1. Carboxylesterase B1 capable of hydrolyzing organophosphate esters was purified from Escherichia coli strain BL21 carrying a cloned esterase B1 gene from mosquito. The recombinant strain BL21 carrying pET-ESTB1 was used for the fermentation. Product formation was induced by either a temperature shift from 30 to 42 °C or by feeding a mixture of glucose and lactose. Cell growth and production of detoxifying enzyme were affected by oxygen availability. The maximum biomass of E. coli BL21 (pET-ESTB1) increased from 14.9 to 31.5 g dry cell weight l−1. Using the host strain E. coli BL21 (DE3), detoxifying enzyme was over-expressed at a biomass level of up to 31.5 g dry weight l−1. The enzyme had a molecular mass of 64 kDa, its optimum temperature was approx. 37 °C; at pH 7 the relative activity after 3 h was 85.9% at 28 °C, 64.9% at 34 °C, and 4.5% at 40 °C. The enzyme activity of cells grown at lower temperatures was much higher; at 18 °C it almost twice than at 20 or 22 °C.
Keywords :
recombinant , E. coli , gene expression , Carboxylesterase B1 , High-density culture
Journal title :
International Biodeterioration and Biodegradation
Serial Year :
2006
Link To Document :
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