Gene combination for expressing and producing terrequinone A in Escherichia coli and use thereof

A preparation method of a recombinant Escherichia coli strain capable of producing terrequinone A, comprising the following steps:

• step S1:
• optimizing a tdiA gene, a tdiB gene, a tdiC gene, a tdiD gene, a tidE gene, an sfp gene, an ScCK gene, and an AtIPK gene according to an Escherichia coli expression pattern to obtain eight genes, namely, a tdiAS gene, a tdiBS gene, a tdiCS gene, a tdiDS gene, a tdiES gene, an sfpS gene, an ScCKS gene, and an AtIPKS gene with nucleotide sequences set forth in SEQ ID NOS:
• 1 to 8, ligating each of the eight genes to a T7 promoter and a terminator to construct gene expression cassettes T7 tdiAS, T7 tdiBS, T7 tdiCS, T7 tdiDS, T7 tdiES, T7 sfpS, T7 ScCKS, and T7 AtIPKS;
• step S2:
• sequentially linking and ligating six gene expression cassettes obtained in step S1, namely, the T7 tdiAS, the T7 tdiBS, the T7 tdiCS, the T7 tdiDS, the T7 tdiES, and the T7 sfpS, to an Escherichia coli expression vector to obtain a recombinant plasmid pC02 comprising six gene expression cassettes tdiAS, tdiBS, tdiCS, tdiDS, tdiES, and sfpS;
• step S3:
• sequentially linking and ligating two gene expression cassettes obtained in step S1, namely, the T7 ScCKS and the T7 AtIPKS, to an Escherichia coli expression vector to obtain a recombinant plasmid pU03 comprising two gene expression cassettes ScCKS and AtIPKS;
• and
• step S4:
• simultaneously transforming the recombinant plasmid pC02 obtained in step S2 and the recombinant plasmid pU03 obtained in step S3 into Escherichia coli to obtain the recombinant Escherichia coli strain capable of producing the terrequinone A.

A recombinant Escherichia coli strain capable of producing terrequinone A obtained by the following steps:

• step S1:
• optimizing a tdiA gene, a tdiB gene, a tdiC gene, a tdiD gene, a tidE gene, an sfp gene, an ScCK gene, and an AtIPK gene according to an Escherichia coli expression pattern to obtain eight genes, namely, a tdiAS gene, a tdiBS gene, a tdiCS gene, a tdiDS gene, a tdiES gene, an sfpS gene, an ScCKS gene, and an AtIPKS gene with nucleotide sequences set forth in SEQ ID NOS:
• 1 to 8, ligating each of the eight genes to a T7 promoter and a terminator to construct gene expression cassettes T7 tdiAS, T7 tdiBS, T7 tdiCS, T7 tdiDS, T7 tdiES, T7 sfpS, T7 ScCKS, and T7 AtIPKS;
• step S2:
• sequentially linking and ligating six gene expression cassettes obtained in step S1, namely, the T7 tdiAS, the T7 tdiBS, the T7 tdiCS, the T7 tdiDS, the T7 tdiES, and the T7 sfpS, to an Escherichia coli expression vector to obtain a recombinant plasmid pC02 comprising six gene expression cassettes tdiAS, tdiBS, tdiCS, tdiDS, tdiES, and sfpS;
• step S3:
• sequentially linking and ligating two gene expression cassettes obtained in step S1, namely, the T7 ScCKS and the T7 AtIPKS, to an Escherichia coli expression vector to obtain a recombinant plasmid pU03 comprising two gene expression cassettes ScCKS and AtIPKS;
• and
• step S4:
• simultaneously transforming the recombinant plasmid pC02 obtained in step S2 and the recombinant plasmid pU03 obtained in step S3 into Escherichia coli to obtain the recombinant Escherichia coli strain capable of producing the terrequinone A.