HETEROLOGOUS EXPRESSION OF THERMOSTABLE ESTERASE GENE FROM GEOBACILLUS THERMOLEOVORANS YN UNDER DIFFERENT EXPRESSION PROMOTERS

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ABDEL MOHSEN SOLIMAN NADIA; REFAT ABDE FATTAH YASSER; MOSTAFA HESHAM E.; GABALLA AHMED A.

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Journal Paper

Paper Information

Journal: INTERNATIONAL JOURNAL OF ENVIRONMENTAL SCIENCE AND TECHNOLOGY (IJEST) Year:2014 | Volume:11 | Issue:1 Page(s): 119-126

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Title

HETEROLOGOUS EXPRESSION OF THERMOSTABLE ESTERASE GENE FROM GEOBACILLUS THERMOLEOVORANS YN UNDER DIFFERENT EXPRESSION PROMOTERS

Author(s)

ABDEL MOHSEN SOLIMAN NADIA | REFAT ABDE FATTAH YASSER | MOSTAFA HESHAM E. | GABALLA AHMED A.

Keywords

BIOFLO III BIOREACTOR

ENCODING GENE

ESCHERICHIA COLI

INDUCTION

SIX-HISTIDINE

Abstract

The gene encoding extracellular thermostable carboxylesterase (estA) from Geobacillus thermoleovorans YN was fused to SIX-HISTIDINE and expressed under different promoters. Three different clones carrying this gene were constructed in host cell ESCHERICHIA COLI BL21 (DE2) using three expression systems pET-17b, pBluescript ® II KS (+) and pCYTEX-P1 (pET-estA T7- promoter; pBlue-estA T7-promoter and pBlue-estA T7 promoter & Lambda pL promoter). The efficiency of expression of the three constructs was evaluated, where the highest esterase expression (589 u/ ml) using p-Nitrophenyl laurate as substrate (pNP-laurate: C18H27NO4) was measured under the control of T7-promoter in expression vector pET-17b after 4 h of the INDUCTION. A 1.5 fold increase in enzyme activity was measured with specific activity 1043 u/ mg protein on growing the clone expressed the target gene under T7-promoter (pET-estAT7) in 2-L working volume BIOFLO III BIOREACTOR under optimal conditions. Recombinant expression of the tested thermostable esterase was monitored by sodium dodecyl sulphate polyacrylamid gel electrophoresis, zymogram and activity measurements with a--Naphthyl acetate (C12H10O2) and Fast Red. The SDS-PAGE analysis of E. coli BL21 (DE3)/pET-estA lysates indicated the presence of a protein band (29 kDa) related to the targeted expressed gene.

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APA: Copy

ABDEL MOHSEN SOLIMAN, NADIA, REFAT ABDE FATTAH, YASSER, MOSTAFA, HESHAM E., & GABALLA, AHMED A.. (2014). HETEROLOGOUS EXPRESSION OF THERMOSTABLE ESTERASE GENE FROM GEOBACILLUS THERMOLEOVORANS YN UNDER DIFFERENT EXPRESSION PROMOTERS. INTERNATIONAL JOURNAL OF ENVIRONMENTAL SCIENCE AND TECHNOLOGY (IJEST), 11(1), 119-126. SID. https://sid.ir/paper/285625/en

Vancouver: Copy

ABDEL MOHSEN SOLIMAN NADIA, REFAT ABDE FATTAH YASSER, MOSTAFA HESHAM E., GABALLA AHMED A.. HETEROLOGOUS EXPRESSION OF THERMOSTABLE ESTERASE GENE FROM GEOBACILLUS THERMOLEOVORANS YN UNDER DIFFERENT EXPRESSION PROMOTERS. INTERNATIONAL JOURNAL OF ENVIRONMENTAL SCIENCE AND TECHNOLOGY (IJEST)[Internet]. 2014;11(1):119-126. Available from: https://sid.ir/paper/285625/en

IEEE: Copy

NADIA ABDEL MOHSEN SOLIMAN, YASSER REFAT ABDE FATTAH, HESHAM E. MOSTAFA, and AHMED A. GABALLA, “HETEROLOGOUS EXPRESSION OF THERMOSTABLE ESTERASE GENE FROM GEOBACILLUS THERMOLEOVORANS YN UNDER DIFFERENT EXPRESSION PROMOTERS,” INTERNATIONAL JOURNAL OF ENVIRONMENTAL SCIENCE AND TECHNOLOGY (IJEST), vol. 11, no. 1, pp. 119–126, 2014, [Online]. Available: https://sid.ir/paper/285625/en

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