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Significant expression of a Chinese scorpion peptide, BmK1, in Escherichia coli through promoter engineering and gene dosage strategy.

Wang J, Xiong Z, Yang Y, Zhao N, Wang Y - Biotechnol. Appl. Biochem. (2013)

Bottom Line: Heterologous expression is an efficient alternative to conventional extraction to produce a specific Buthus martensii Karsch (BmK) peptide.The yield of the purified BmK1 achieved 196.74 mg L(-1) in E. coli BL21(DE3) pJF431, which was improved 2.09-fold compared with the control.This was the highest reported production of scorpion peptides in E. coli.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, People's Republic of China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People's Republic of China.

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Codon optimization and expression cassette construction. (A) DNA sequence alignment between original and optimized BmK1 gene. (B) The entire open reading frame of the recombinant BmK1 gene.
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fig01: Codon optimization and expression cassette construction. (A) DNA sequence alignment between original and optimized BmK1 gene. (B) The entire open reading frame of the recombinant BmK1 gene.

Mentions: A DNA sequence of BmK1 deposited in EMBL database (accession number: AAD39510) was optimized for E. coli expression by a Web-based program optimizer 18. Rare codons of the BmK1 gene were optimized, including nine codons for glycine and six codons for arginine (Fig.1A). Additionally, to simplify the purification process, the optimized BmK1 gene was fused upstream with a 6× His tag. To easily remove the His tag after purification, an enterokinase recognition site DDDDK was inserted between a 6× His tag and BmK1 peptide. The entire open reading frame of the recombinant BmK1 gene is shown in Fig.1B.


Significant expression of a Chinese scorpion peptide, BmK1, in Escherichia coli through promoter engineering and gene dosage strategy.

Wang J, Xiong Z, Yang Y, Zhao N, Wang Y - Biotechnol. Appl. Biochem. (2013)

Codon optimization and expression cassette construction. (A) DNA sequence alignment between original and optimized BmK1 gene. (B) The entire open reading frame of the recombinant BmK1 gene.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4269186&req=5

fig01: Codon optimization and expression cassette construction. (A) DNA sequence alignment between original and optimized BmK1 gene. (B) The entire open reading frame of the recombinant BmK1 gene.
Mentions: A DNA sequence of BmK1 deposited in EMBL database (accession number: AAD39510) was optimized for E. coli expression by a Web-based program optimizer 18. Rare codons of the BmK1 gene were optimized, including nine codons for glycine and six codons for arginine (Fig.1A). Additionally, to simplify the purification process, the optimized BmK1 gene was fused upstream with a 6× His tag. To easily remove the His tag after purification, an enterokinase recognition site DDDDK was inserted between a 6× His tag and BmK1 peptide. The entire open reading frame of the recombinant BmK1 gene is shown in Fig.1B.

Bottom Line: Heterologous expression is an efficient alternative to conventional extraction to produce a specific Buthus martensii Karsch (BmK) peptide.The yield of the purified BmK1 achieved 196.74 mg L(-1) in E. coli BL21(DE3) pJF431, which was improved 2.09-fold compared with the control.This was the highest reported production of scorpion peptides in E. coli.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Synthetic Biology, Institute of Plant Physiology and Ecology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, People's Republic of China; State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, People's Republic of China.

No MeSH data available.


Related in: MedlinePlus