Development of an E-coli expression cassette for the efficient production of a harmful protein

作者：Kim OS; Kwak HJ; Lee JH; Ha JM; Ha BJ; Lee SH^*

来源：Biotechnology and Bioprocess Engineering, 2004, 9(5): 389-392.

DOI：10.1007/BF02933063

• colicin promoter
• expression cassette
• E. coli W3110 cells
• carboxypeptidase Taq
• mitomycin C

In order to produce a harmful protein more efficiently, this expression cassette, dubbed pCol-MICT, is directed by the colicin promoter, and was constructed by the insertion of a rrnBT(1)T(2) fragment of pEXP7, and a MxeInteinCBD fragment of pTXB3, into pSH375. To test whether harmful proteins, including proteolytic enzymes, could be effectively produced by this cassette, the carboxypeptidase (CPase) Taq gene was inserted into the pCol-MICT cassette to yield pCol-CPaseTaq-MICT. E. coli W3110 cells harboring pCol-CPaseTaq-MICT produced a large quantity of this enzyme, as much as 47.2 mg of purified from per liter of culture, when cultured in the presence of mitomycin C (0.4 mug/mL). This indicates that the colicin promoter-controlled E. coli expression cassette was able to produce almost 8 times of protein than the conventional tac promoter-based system, and that this cassette may be useful in the synthesis of other harmful proteins.

• 出版日期2004-10