Extracellular heme production method using metabolically engineered microorganism
AssigneeKorea Advanced Institute of Science and Technology
InventorsSang Yup Lee, Xin Rui Zhao, Kyeong Rok Choi
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Abstract
The present invention relates to a microorganism variant having the ability to extracellularly produce heme, and more particularly to a metabolically engineered microorganism variant having the ability to extracellularly produce heme and a method of producing heme using the same. According to the present invention, heme, an organometallic compound which is increasingly used as a health food or food supplement for the treatment of porphyria, can be extracellularly secreted and produced in high yield using the microorganism variant, but not conventional chemical synthesis or enzymatic synthesis.
Claims12
A metabolically engineered variant microorganism that secretes heme extracellularly, wherein the microorganism is modified by the introduction or overexpression of ccmABC genes encoding a heme exporter, wherein the microorganism comprises genes in a C5 biosynthetic pathway that produces 5-aminolevulinate (ALA) from glutamate, and genes in a pathway that synthesizes heme from ALA,
- wherein the genes in the C5 biosynthetic pathway comprise a gltX gene encoding glutamyl-tRNA synthase (GIuRS), a hemA gene encoding glutamyl-tRNA reductase (GluTR), and a hemL gene encoding glutamate-1-semialdehyde 2,1-aminomutase (GSAM),
- wherein the genes in the pathway that synthesizes heme from ALA comprise a hemB gene encoding porphobilinogen synthase (PBGS), a hemC gene encoding porphobilinogen deaminase (PBGD), a hemD gene encoding uroporphyrinogen III synthase (UROS), a hemE gene encoding uroporphyrinogen III decarboxylase (UROD), a hemF gene encoding coproporphyrinogen III oxidase (CPO), a hemG gene encoding protoporphyrinogen oxidase (PPO), and a hemH gene encoding ferrochelatase (FECH), and
- wherein the microorganism is selected from the group consisting of Escherichia coli, Bacillus sp., Corynebacterium sp., Lactobacillus sp., Lactococcus sp., Pseudomonas sp., Anacystis sp., Anabaena sp., Chlorobium sp., Chloroflexus sp., Clostridium sp., Methanobacteria, Propionibacterium sp., Rhodopseudomonas sp., Rhodobacter sp., Rhodovulum sp., Streptococcus sp., Saccharomyces sp., Schizosaccharomyces sp., Yarrowia sp., and Aspergillus sp.
A metabolically engineered variant microorganism that extracellularly secretes heme, wherein the microorganism is modified to overexpress ccmABC genes encoding a heme exporter, wherein the microorganism comprises genes in a C5 biosynthetic pathway that produces 5-aminolevulinate (ALA) from glutamate, and genes in a pathway that synthesizes heme from ALA, wherein at least one of the genes in the C5 biosynthetic pathway that produces 5-aminolevulinate (ALA) from glutamate and at least one of the genes in the pathway that synthesizes heme from ALA, are overexpressed;
- wherein the genes in the C5 biosynthetic pathway comprise at least one gene selected from:
- a gltX gene encoding glutamyl-tRNA synthase (GIuRS), a hemA gene encoding glutamyl-tRNA reductase (GIuTR), and a hemL gene encoding glutamate-1-semialdehyde 2,1-aminomutase (GSAM);
- wherein the genes in the pathway that synthesizes heme from ALA comprise at least one gene selected from:
- a hemB gene encoding porphobilinogen synthase (PBGS), a hemC gene encoding porphobilinogen deaminase (PBGD), a hemD gene encoding uroporphyrinogen III synthase (UROS), a hemE gene encoding uroporphyrinogen III decarboxylase (UROD), a hemF gene encoding coproporphyrinogen III oxidase (CPO), a hemG gene encoding protoporphyrinogen oxidase (PPO), and a hemH gene encoding ferrochelatase (FECH);
- and
- wherein the microorganism is further modified by deletion of a yfeX gene.
Dependencies are inferred from the claim text.