Genetically engineered bacterium using glucose as substrate for de novo synthesis of vanillin and application thereof

A genetically engineered bacterium that uses glucose as a substrate for de novo synthesis of vanillin, wherein the genetically engineered bacterium is a recombinant Corynebacterium glutamicum comprising a vanillin synthesis module and a methyl cyclic regeneration module;

• wherein the recombinant Corynebacterium glutamicum is a variant of a Corynebacterium glutamicum ATCC13032 cell obtained by knocking out the pcaHG, van, vdh and fud genes in said Corynebacterium glutamicum ATCC13032 cell, wherein the fud gene is an alcohol dehydrogenase (NADP+) gene, the alcohol dehydrogenase (NADP+) gene being a Zn-dependent alcohol dehydrogenase gene;
• wherein the vanillin synthesis module comprises a transketolase gene, a 3-deoxy-7-phosphoheptulonate synthase gene, an O-methyltransferase gene, a carboxylic acid reductase gene and a 4′-phosphopantetheinyl transferase gene;
• wherein the methyl cyclic regeneration module comprises a 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase gene, an S-adenosylmethionine synthase gene, a homoserine O-acetyltransferase gene and an adenosylhomocysteinase gene;
• wherein the recombinant Corynebacterium glutamicum expresses any one of the combinations of genes selected from:
• (a) an Escherichia coli tktA gene, an Escherichia coli K12 3-deoxy-7-phosphoheptulonate synthase gene having the sequence of SEQ ID NO:
• 45, a Rattus norvegicus O-methyltransferase gene having the sequence of SEQ ID NO:
• 46, a Nocardia iowensis carboxylic acid reductase gene having the sequence of SEQ ID NO:
• 47, a Mycobacterium marinum 4′-phosphopantetheinyl transferase gene having the sequence of SEQ ID NO:
• 48, an endogenous Corynebacterium glutamicum 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase gene having the sequence of SEQ ID NO:
• 49, an endogenous Corynebacterium glutamicum S-adenosylmethionine synthase gene having the sequence of SEQ ID NO:
• 50, an endogenous Corynebacterium glutamicum homoserine O-acetyltransferase gene having sequence of SEQ ID NO:
• 51, and an endogenous Corynebacterium glutamicum adenosylhomocysteinase gene having the sequence of SEQ ID NO:
• 52;
• (b) an endogenous Corynebacterium glutamicum transketolase gene having the sequence of SEQ ID NO:
• 83, an endogenous Corynebacterium glutamicum 3-deoxy-7-phosphoheptulonate synthase gene having the sequence of SEQ ID NO:
• 84, an Arabidopsis thaliana O-methyltransferase gene having the sequence of SEQ ID NO:
• 85, a Mycobacterium marinum carboxylic acid reductase gene having the sequence of SEQ ID NO:
• 86, a Nocardia iowensis 4′-phosphopantetheinyl transferase gene having the sequence of SEQ ID NO:
• 87, an Escherichia coli K12 metE gene, an Escherichia coli K12 metK gene, a Saccharomyces cerevisiae S288C met2 gene, and an endogenous Corynebacterium glutamicum adenosylhomocysteinase gene having the sequence of SEQ ID NO:
• 52;
• (c) a Saccharomyces cerevisiae S288C tkl1 gene, a Saccharomyces cerevisiae S288C aro4 gene, s Homo sapiens O-methyltransferase gene having the sequence of SEQ ID NO:
• 105, a Nocardia iowensis carboxylic acid reductase gene having the sequence of SEQ ID NO:
• 47, a Bacillus subtilis 4′-phosphopantetheinyl transferase gene having the sequence of SEQ ID NO:
• 106, a Saccharomyces cerevisiae S288C met6 gene, a Saccharomyces cerevisiae S288C SAM2 gene, a Saccharomyces cerevisiae S288C met2 gene, and an endogenous Corynebacterium glutamicum adenosylhomocysteinase gene having the sequence of SEQ ID NO:
• 52;
• and
• (d) an Escherichia coli tktA gene, an Escherichia coli K12 3-deoxy-7-phosphoheptulonate synthase gene having the sequence of SEQ ID NO:
• 45, a Mus musculus O-methyltransferase gene having the sequence of SEQ ID NO:
• 137, a Nocardia iowensis carboxylic acid reductase gene having the sequence of SEQ ID NO:
• 47, a Mycobacterium marinum 4′-phosphopantetheinyl transferase gene having the sequence of SEQ ID NO:
• 48, an endogenous Corynebacterium glutamicum 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase gene having the sequence of SEQ ID NO:
• 49, an endogenous Corynebacterium glutamicum S-adenosylmethionine synthase gene having the sequence of SEQ ID NO:
• 50, an endogenous Corynebacterium glutamicum homoserine O-acetyltransferase gene having sequence of SEQ ID NO:
• 51, and an endogenous Corynebacterium glutamicum adenosylhomocysteinase gene having the sequence of SEQ ID NO:
• 52.