Enzymatic Carbocycle Formation in Microbial Secondary Metabolism. The Mechanism of the 2-Deoxy-scyllo-inosose Synthase Reaction as a Crucial Step in the 2-Deoxystreptamine Biosynthesis in Streptomyces fradiae

Noriaki Yamauchi, Katsumi Kakinuma

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53 Citations (Scopus)

Abstract

The mechanism of 2-deoxy-scyllo-inosose synthase reaction, a carbocycle formation step from D-glucose-6-phosphate in the biosynthesis of a major 2-deoxystreptamine aglycon of clinically important aminoglycoside antibiotics, was investigated with a partially purified enzyme fraction from neomycin-producing Streptomyces fradiae IFO 13147. Singly and doubly labeled d-[4-2H]- and D-[4-2H,3-18O]glucose-6-phosphate were used for chase experiments, and the 2-deoxy-scyllo-inosose product was analyzed by 2H-NMR and GC-MS. The deuterium label at C-4 of the substrate appeared to be retained at C-6 of the product without scrambling the doubly-labeled isotopes. Since the oxidative process with the aid of NAD+ is essential, which was reported previously, the hydride abstraction and returning appear to take place within the same glucose molecule. These results strongly suggest that this carbocycle formation is catalyzed by a single 2-deoxy-seyllo-inosose synthase enzyme with a catalytic requirement of NAD cofactor, the mechanism of which is closely resembled to the dehydroquinate synthase in the shikimate pathway.

Original languageEnglish
Pages (from-to)5614-5619
Number of pages6
JournalJournal of Organic Chemistry
Volume60
Issue number17
DOIs
Publication statusPublished - Sep 1 1995
Externally publishedYes

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Glucose-6-Phosphate
Biosynthesis
NAD
Glucose
Neomycin
Deuterium
Aminoglycosides
Enzymes
Hydrides
Isotopes
Labels
Nuclear magnetic resonance
Anti-Bacterial Agents
Molecules
Substrates
Experiments
2-deoxystreptamine
2-deoxy-scyllo-inosose synthase
Secondary Metabolism
inosose

All Science Journal Classification (ASJC) codes

  • Organic Chemistry

Cite this

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title = "Enzymatic Carbocycle Formation in Microbial Secondary Metabolism. The Mechanism of the 2-Deoxy-scyllo-inosose Synthase Reaction as a Crucial Step in the 2-Deoxystreptamine Biosynthesis in Streptomyces fradiae",
abstract = "The mechanism of 2-deoxy-scyllo-inosose synthase reaction, a carbocycle formation step from D-glucose-6-phosphate in the biosynthesis of a major 2-deoxystreptamine aglycon of clinically important aminoglycoside antibiotics, was investigated with a partially purified enzyme fraction from neomycin-producing Streptomyces fradiae IFO 13147. Singly and doubly labeled d-[4-2H]- and D-[4-2H,3-18O]glucose-6-phosphate were used for chase experiments, and the 2-deoxy-scyllo-inosose product was analyzed by 2H-NMR and GC-MS. The deuterium label at C-4 of the substrate appeared to be retained at C-6 of the product without scrambling the doubly-labeled isotopes. Since the oxidative process with the aid of NAD+ is essential, which was reported previously, the hydride abstraction and returning appear to take place within the same glucose molecule. These results strongly suggest that this carbocycle formation is catalyzed by a single 2-deoxy-seyllo-inosose synthase enzyme with a catalytic requirement of NAD cofactor, the mechanism of which is closely resembled to the dehydroquinate synthase in the shikimate pathway.",
author = "Noriaki Yamauchi and Katsumi Kakinuma",
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AB - The mechanism of 2-deoxy-scyllo-inosose synthase reaction, a carbocycle formation step from D-glucose-6-phosphate in the biosynthesis of a major 2-deoxystreptamine aglycon of clinically important aminoglycoside antibiotics, was investigated with a partially purified enzyme fraction from neomycin-producing Streptomyces fradiae IFO 13147. Singly and doubly labeled d-[4-2H]- and D-[4-2H,3-18O]glucose-6-phosphate were used for chase experiments, and the 2-deoxy-scyllo-inosose product was analyzed by 2H-NMR and GC-MS. The deuterium label at C-4 of the substrate appeared to be retained at C-6 of the product without scrambling the doubly-labeled isotopes. Since the oxidative process with the aid of NAD+ is essential, which was reported previously, the hydride abstraction and returning appear to take place within the same glucose molecule. These results strongly suggest that this carbocycle formation is catalyzed by a single 2-deoxy-seyllo-inosose synthase enzyme with a catalytic requirement of NAD cofactor, the mechanism of which is closely resembled to the dehydroquinate synthase in the shikimate pathway.

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