EC Number | Application | Comment | Organism |
---|---|---|---|
1.14.99.50 | synthesis | the ergothioneine biosynthetic pathway (EgtA-EgtE catalysis) provides an opportunity for ergothioneine production through metabolic engineering | Mycolicibacterium smegmatis |
1.14.99.52 | synthesis | the ergothioneine biosynthetic pathway (EgtA-EgtE catalysis) provides an opportunity for ergothioneine production through metabolic engineering, but the kinetic properties and the presence of side-reaction render OvoA not suitable for use in ergothioneine production through metabolic engineering | Erwinia tasmaniensis |
1.21.3.10 | synthesis | the ergothioneine biosynthetic pathway (EgtA-EgtE catalysis) provides an opportunity for ergothioneine production through metabolic engineering | Neurospora crassa |
4.4.1.36 | synthesis | the ergothioneine biosynthetic pathway (EgtA-EgtE catalysis) provides an opportunity for ergothioneine production through metabolic engineering | Neurospora crassa |
EC Number | Cloned (Comment) | Organism |
---|---|---|
1.14.99.50 | contruction of an EgtB homologue sequence similarity network using a BLAST e value cutoff | Mycolicibacterium smegmatis |
1.21.3.10 | gene egt1, Egt1 recombinant overexpression from vector pASK-IBA3+ in Escherichia coli | Neurospora crassa |
4.4.1.36 | expression in Escherichia coli | Neurospora crassa |
4.4.1.36 | gene NCU11365, genome mining, cloning from a sequence determined by screening of an EgtB homologue sequence similarity network from Mycobacterum smegmatis EgtB, recombinant expression of enzyme EgtE in Escherichia coli strain BL21(DE3) | Neurospora crassa |
EC Number | KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.21.3.10 | 0.39 | - |
hercynine | pH and temperature not specified in the publication, with gamma-L-glutamyl-L-cysteine | Neurospora crassa | |
1.21.3.10 | 0.436 | - |
hercynine | pH and temperature not specified in the publication, with L-Cys | Neurospora crassa | |
1.21.3.10 | 0.603 | - |
L-cysteine | pH and temperature not specified in the publication | Neurospora crassa | |
1.21.3.10 | 7.68 | - |
gamma-L-glutamyl-L-cysteine | pH and temperature not specified in the publication | Neurospora crassa | |
4.4.1.36 | 0.1947 | - |
S-(hercyn-2-yl)-L-cysteine S-oxide | pH and temperature not specified in the publication | Neurospora crassa | |
4.4.1.36 | 0.195 | - |
S-(hercyn-2-yl)-L-cysteine S-oxide | pH not specified in the publication, temperature not specified in the publication | Neurospora crassa |
EC Number | Metals/Ions | Comment | Organism | Structure |
---|---|---|---|---|
1.14.99.50 | Fe2+ | non-heme iron, required for catalysis | Mycolicibacterium smegmatis | |
1.14.99.52 | Fe2+ | non-heme iron, required for catalysis | Erwinia tasmaniensis | |
1.21.3.10 | Fe2+ | required for catalysis, the enzyme contains about 0.90 Fe per Egt1 monomer | Neurospora crassa |
EC Number | Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.99.50 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | Mycolicibacterium smegmatis | - |
gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.50 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | Mycolicibacterium smegmatis ATCC 700084 | - |
gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.52 | hercynine + L-cysteine + O2 | Erwinia tasmaniensis | reaction of EC 1.14.99.51 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.52 | L-histidine + L-cysteine + O2 | Erwinia tasmaniensis | - |
S-(L-histidin-5-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | Neurospora crassa | reaction of EC 1.14.99.50, low activity | gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | Neurospora crassa ATCC 24698 | reaction of EC 1.14.99.50, low activity | gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + L-cysteine + O2 | Neurospora crassa | - |
S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + L-cysteine + O2 | Neurospora crassa ATCC 24698 | - |
S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
4.4.1.36 | 2-(hydroxysulfanyl)hercynine + reduced acceptor | Neurospora crassa | - |
ergothioneine + acceptor + H2O | - |
? | |
4.4.1.36 | 2-(hydroxysulfanyl)hercynine + reduced acceptor | Neurospora crassa DSM 1257 | - |
ergothioneine + acceptor + H2O | - |
? | |
4.4.1.36 | 2-(hydroxysulfanyl)hercynine + reduced acceptor | Neurospora crassa ATCC 24698 | - |
ergothioneine + acceptor + H2O | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | Neurospora crassa | - |
2-(hydroxysulfanyl)hercynine + pyruvate + ammonia | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | Neurospora crassa DSM 1257 | - |
2-(hydroxysulfanyl)hercynine + pyruvate + ammonia | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | Neurospora crassa ATCC 24698 | - |
2-(hydroxysulfanyl)hercynine + pyruvate + ammonia | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | Neurospora crassa | - |
ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | Neurospora crassa | overall reaction | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | Neurospora crassa DSM 1257 | - |
ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | Neurospora crassa DSM 1257 | overall reaction | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | Neurospora crassa ATCC 24698 | - |
ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | Neurospora crassa ATCC 24698 | overall reaction | ergothioneine + pyruvate + ammonia + acceptor | - |
? |
EC Number | Organism | UniProt | Comment | Textmining |
---|---|---|---|---|
1.14.99.50 | Mycolicibacterium smegmatis | A0R5N0 | - |
- |
1.14.99.50 | Mycolicibacterium smegmatis ATCC 700084 | A0R5N0 | - |
- |
1.14.99.52 | Erwinia tasmaniensis | - |
- |
- |
1.21.3.10 | Neurospora crassa | Q7RX33 | - |
- |
1.21.3.10 | Neurospora crassa ATCC 24698 | Q7RX33 | - |
- |
4.4.1.36 | Neurospora crassa | A7UX13 | - |
- |
4.4.1.36 | Neurospora crassa ATCC 24698 | A7UX13 | - |
- |
4.4.1.36 | Neurospora crassa DSM 1257 | A7UX13 | - |
- |
EC Number | Purification (Comment) | Organism |
---|---|---|
4.4.1.36 | recombinant enzyme from Escherichia coli strain BL21(DE3) | Neurospora crassa |
EC Number | Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|---|
1.14.99.50 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | - |
Mycolicibacterium smegmatis | gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.50 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | - |
Mycolicibacterium smegmatis ATCC 700084 | gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.50 | additional information | enzyme EgtB is extremely specific in terms of substrate specificity. Enzyme activity determination using : a 1H NMR assay of chemical shift of the imidazole hydrogen atoms | Mycolicibacterium smegmatis | ? | - |
? | |
1.14.99.50 | additional information | enzyme EgtB is extremely specific in terms of substrate specificity. Enzyme activity determination using : a 1H NMR assay of chemical shift of the imidazole hydrogen atoms | Mycolicibacterium smegmatis ATCC 700084 | ? | - |
? | |
1.14.99.52 | hercynine + L-cysteine + O2 | reaction of EC 1.14.99.51 | Erwinia tasmaniensis | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.52 | L-histidine + L-cysteine + O2 | - |
Erwinia tasmaniensis | S-(L-histidin-5-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.14.99.52 | additional information | when OvoA's native substrate histidine is replaced by hercynine, OvoA can catalyze a one-step transformation reaction of hercynylcysteine S-oxide synthase, EC 1.14.99.51, resulting in kobs changes by 2fold for hercynine, while Km for cysteine increases from 0.3 mM to 3.19 mM. OvoA preferentially uses histidine and cysteine. OvoA also catalyzes the oxidation of cysteine to cysteine sulfinic acid, EC 1.13.11.20. Enzyme activity determination using : a 1H NMR assay of chemical shift of the imidazole hydrogen atoms | Erwinia tasmaniensis | ? | - |
? | |
1.21.3.10 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | reaction of EC 1.14.99.50, low activity | Neurospora crassa | gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + gamma-L-glutamyl-L-cysteine + O2 | reaction of EC 1.14.99.50, low activity | Neurospora crassa ATCC 24698 | gamma-L-glutamyl-S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + L-cysteine + O2 | - |
Neurospora crassa | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | hercynine + L-cysteine + O2 | - |
Neurospora crassa ATCC 24698 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
? | |
1.21.3.10 | additional information | three different assay methods for enzyme activity determination: a 1H NMR assay of chemical shift of the imidazole hydrogen atoms, an oxygen consumption assay, and a 13C NMR assay to monitor the presence of other cysteine related oxygen consumption. Egt1 from Neurospora crassa prefers cysteine as substrate compared to gamma-L-glutamyl-L-cysteine, Egt1 has a 62fold greater specificity (kobs/Km) for L-cysteine relative to gamma-Glu-Cys | Neurospora crassa | ? | - |
? | |
1.21.3.10 | additional information | three different assay methods for enzyme activity determination: a 1H NMR assay of chemical shift of the imidazole hydrogen atoms, an oxygen consumption assay, and a 13C NMR assay to monitor the presence of other cysteine related oxygen consumption. Egt1 from Neurospora crassa prefers cysteine as substrate compared to gamma-L-glutamyl-L-cysteine, Egt1 has a 62fold greater specificity (kobs/Km) for L-cysteine relative to gamma-Glu-Cys | Neurospora crassa ATCC 24698 | ? | - |
? | |
4.4.1.36 | 2-(hydroxysulfanyl)hercynine + reduced acceptor | - |
Neurospora crassa | ergothioneine + acceptor + H2O | - |
? | |
4.4.1.36 | 2-(hydroxysulfanyl)hercynine + reduced acceptor | - |
Neurospora crassa DSM 1257 | ergothioneine + acceptor + H2O | - |
? | |
4.4.1.36 | 2-(hydroxysulfanyl)hercynine + reduced acceptor | - |
Neurospora crassa ATCC 24698 | ergothioneine + acceptor + H2O | - |
? | |
4.4.1.36 | additional information | enzyme activity determination by a 1H NMR assay of chemical shift of the imidazole hydrogen atoms and a colorimetric assay by coupling the C-S lyase reaction with lactate dehydrogenas, product determination by high-resolution mass spectrometry and NMR | Neurospora crassa | ? | - |
? | |
4.4.1.36 | additional information | enzyme activity determination by a 1H NMR assay of chemical shift of the imidazole hydrogen atoms and a colorimetric assay by coupling the C-S lyase reaction with lactate dehydrogenas, product determination by high-resolution mass spectrometry and NMR | Neurospora crassa DSM 1257 | ? | - |
? | |
4.4.1.36 | additional information | enzyme activity determination by a 1H NMR assay of chemical shift of the imidazole hydrogen atoms and a colorimetric assay by coupling the C-S lyase reaction with lactate dehydrogenas, product determination by high-resolution mass spectrometry and NMR | Neurospora crassa ATCC 24698 | ? | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
Neurospora crassa | 2-(hydroxysulfanyl)hercynine + pyruvate + ammonia | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
Neurospora crassa DSM 1257 | 2-(hydroxysulfanyl)hercynine + pyruvate + ammonia | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + H2O | - |
Neurospora crassa ATCC 24698 | 2-(hydroxysulfanyl)hercynine + pyruvate + ammonia | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | - |
Neurospora crassa | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | overall reaction | Neurospora crassa | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | - |
Neurospora crassa DSM 1257 | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | overall reaction | Neurospora crassa DSM 1257 | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | - |
Neurospora crassa ATCC 24698 | ergothioneine + pyruvate + ammonia + acceptor | - |
? | |
4.4.1.36 | S-(hercyn-2-yl)-L-cysteine S-oxide + reduced acceptor | overall reaction | Neurospora crassa ATCC 24698 | ergothioneine + pyruvate + ammonia + acceptor | - |
? |
EC Number | Synonyms | Comment | Organism |
---|---|---|---|
1.14.99.50 | EgtB | - |
Mycolicibacterium smegmatis |
1.14.99.50 | hercynine oxygenase | UniProt | Mycolicibacterium smegmatis |
1.14.99.52 | OvoA | - |
Erwinia tasmaniensis |
1.21.3.10 | Egt1 | - |
Neurospora crassa |
4.4.1.36 | Egt2 | - |
Neurospora crassa |
4.4.1.36 | egtE | - |
Neurospora crassa |
4.4.1.36 | hercynylcysteine sulfoxide lyase | - |
Neurospora crassa |
4.4.1.36 | hercynylcysteine sulfoxide lyase | UniProt | Neurospora crassa |
4.4.1.36 | NCU01256 protein | - |
Neurospora crassa |
4.4.1.36 | NCU11365 | - |
Neurospora crassa |
4.4.1.36 | PLP-dependent C-S lyase | - |
Neurospora crassa |
EC Number | Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|---|
1.21.3.10 | 0.461 | - |
gamma-L-glutamyl-L-cysteine | pH and temperature not specified in the publication | Neurospora crassa | |
1.21.3.10 | 0.461 | - |
hercynine | pH and temperature not specified in the publication, with gamma-L-glutamyl-L-cysteine | Neurospora crassa | |
1.21.3.10 | 2.267 | - |
L-cysteine | pH and temperature not specified in the publication | Neurospora crassa | |
1.21.3.10 | 2.267 | - |
hercynine | pH and temperature not specified in the publication, with L-Cys | Neurospora crassa |
EC Number | Cofactor | Comment | Organism | Structure |
---|---|---|---|---|
4.4.1.36 | pyridoxal 5'-phosphate | - |
Neurospora crassa |
EC Number | General Information | Comment | Organism |
---|---|---|---|
1.14.99.50 | evolution | OvoA, EC 1.14.99.52, and EgtB are related in sequence, while they are biochemically distinct | Mycolicibacterium smegmatis |
1.14.99.50 | metabolism | the mononuclear non-heme iron enzyme EgtB catalyzes an oxidative C-S bond formation in the ergothioneine biosynthesis. One of the EgtB substrates is gamma-Glu-Cys, which is part of the glutathione biosynthesis, resulting in competition between ergothioneine and glutathione biosyntheses | Mycolicibacterium smegmatis |
1.14.99.52 | evolution | OvoA and EgtB, EC 1.14.99.50, are related in sequence, while they are biochemically distinct | Erwinia tasmaniensis |
1.14.99.52 | metabolism | enzyme OvoA is responsible for the oxidative C-S bond formation in ovothiol biosynthesis. OvoA in ovothiol biosynthesis has a relaxed substrate specificity | Erwinia tasmaniensis |
1.21.3.10 | malfunction | Egt1 knockout in Neurospora crassa makes the mutant much more sensitive to oxidative stress compared to wild-type | Neurospora crassa |
1.21.3.10 | metabolism | the fungus Neurospora crassa has a concise ergothioneine biosynthetic pathway, its nonheme iron enzyme Egt1 makes use of cysteine instead of gamma-Glu-Cys (like EgtB, EC 1.14.99.50) as the substrate. Such a change of substrate preference eliminates the competition between ergothioneine and glutathione biosyntheses. In Neuropsora crassa ergothioneine biosynthesis, there are reactions distinct from the pathway via EgtB: the oxidative C-S bond formation and the subsequent C-S bond cleavage reaction, which result in a net transfer of the sulfur atom form cysteine to histidine imidazole side-chain | Neurospora crassa |
4.4.1.36 | metabolism | the enzyme catalyzes a pyridoxal 5'-phosphate-mediated C-S lyase reaction in the ergothioneine biosynthesis | Neurospora crassa |