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KMID : 0545120130230050668
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Journal of Microbiology and Biotechnology
2013 Volume.23 No. 5 p.668 ~ p.673
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Effect of Gene Amplifications in Porphyrin Pathway on Heme Biosynthesis in a Recombinant Escherichia coli
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Lee Min-Ju
Kim Hye-Jung
Lee Ju-Young
Kwon An-Sung
Jun Soo-Youn
Kang Sang-Hyeon
Kim Pil
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Abstract
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A recombinant E. coli co-expressing ALA synthase (hemA), NADP-dependent malic enzyme (maeB), and dicarboxylic acid transporter (dctA) was reported to synthesize porphyrin derivatives including iron-containing heme. To enhance the synthesis of bacterial heme, five genes of the porphyrin biosynthetic pathway [pantothenate kinase (coaA), ALA dehydratase (hemB), 1-hydroxymethylbilane synthase (hemC), uroporphyrinogen III synthase (hemD), and uroporphyrinogen III decarboxylase (hemE)] were amplified in the recombinant E. coli co-expressing hemA-maeB-dctA. Pantothenate kinase expression enabled the recombinant E. coli to accumulate intracellular CoA. Intracellular ALA was the most enhanced by uroporphyrinogen III synthase expression, porphobilinogen by ALA dehydratase expression, and uroporphyrin and coproporphyrin by 1- hydroxymethylbilane synthase expression. The strain coexpressing coaA, hemA, maeB, and dctA produced heme of 0.49 μmol/g-DCW, which was twice as much from the strain without coaA expression. Further strain improvement for the porphyrin derivatives is discussed based on the results.
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KEYWORD
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porphyrin pathway, bacterial heme, pantothenate kinase, uroporphyrinogen III synthase, uroporphyrinogen III decarboxylase
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