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Invention Publication
CN105331569A 一株重组大肠杆菌及其在发酵生产[2Fe2S]铁氧还蛋白中的应用 失效 - 权利终止

一株重组大肠杆菌及其在发酵生产[2Fe2S]铁氧还蛋白中的应用
Abstract:
本发明公开了一株重组大肠杆菌，所述菌命名为大肠杆菌(Escherichia coli)pCodonPlus/pRKISC/pETDuet-S33[2Fe2S]Fd，该菌株含有来源于根癌农杆菌(Agrobacterium tumefaciens)S33的[2Fe2S]铁氧还蛋白基因，其核苷酸序列如SEQ ID NO.1所示；所述重组大肠杆菌是将[2Fe2S]铁氧还蛋白基因连接入pETDuet-1载体，制得重组质粒命名为pETDuet-[2Fe2S]，并将制得的重组质粒转入含有pCodonPlus和pRKISC质粒的E.coli C41(DE3)中获得。本发明还公开了所述重组大肠杆菌在发酵生产[2Fe2S]铁氧还蛋白中的应用，实验证实：每升发酵培养物最终可以获得23mg纯化后的[2Fe2S]铁氧还蛋白，远大于之前报道的从野生菌中纯化出来的0.03mg的产量，预示本发明提供的重组大肠杆菌在工业化生产中具有很好的应用前景。
Abstract(English):

The invention discloses recombinant Escherichia coli, which is named as Escherichia coli pCodonPlus/pRKISC/pETDuet-S33[2Fe2S]Fd, the strain contains [2Fe2S] ferredoxin genes sourced from agrobacterium tumefaciens S33, and a nucleotide sequence thereof is shown as SEQ ID NO.1; the recombinant Escherichia coli is obtained by inoculating the [2Fe2S] ferredoxin into a pETDuet-1 carrier to prepare recombinant plasmids named as pETDuet-[2Fe2S] and transferring the prepared recombinant plasmids into Escherichia coli C41(DE3) containing pCodonPlus and pRKISC plasmids. The invention also discloses an application of the recombinant Escherichia coli in fermenting and producing [2Fe2S] ferredoxin. The experiment proves that 23mg of pure [2Fe2S] ferredoxin can be obtained from every liter of fermented culture, which is far higher than the reported yield 0.03mg of [2Fe2S] purified from wild mushrooms, so that the recombinant Escherichia coli has good application prospect in the industrialized production.

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