The missing link in petrobactin biosynthesis: asbF encodes a (-)-3-dehydroshikimate dehydratase

David T. Fox; Kinya Hotta; Chu Young Kim; Andrew T. Koppisch

doi:10.1021/bi801876q

The missing link in petrobactin biosynthesis: asbF encodes a (-)-3-dehydroshikimate dehydratase

David T. Fox, Kinya Hotta, Chu Young Kim, Andrew T. Koppisch

Chemistry and Biochemistry

Research output: Contribution to journal › Article › peer-review

44 Scopus citations

Abstract

The siderophore petrobactin harbors unique 3,4-dihydroxybenzoyl iron-liganding groups. These moieties are known to be synthesized from shikimate pathway precursors, but no reports of the biosynthetic enzymes responsible for this conversion have been published. The gene encoding AsbF from Bacillus thuringiensis 97-27 was overexpressed in an Escherichia coli host. AsbF rapidly and efficiently transforms (-)-3-dehydroshikimate (DHS) into 3,4-dihydroxybenzoate (k_cat^DHS = 217 ± 10 min ^-1; K_m^DHS = 125 ± 14 μM) at 37°C and has an absolute requirement for divalent metal. Finally, the pH versus k_cat^DHS profile revealed two ionizable groups (pK _a1 = 7.9 ± 0.1, and pK_a2 = 9.3 ± 0.1).

Original language	English (US)
Pages (from-to)	12251-12253
Number of pages	3
Journal	Biochemistry
Volume	47
Issue number	47
DOIs	https://doi.org/10.1021/bi801876q
State	Published - Nov 25 2008

ASJC Scopus subject areas

Biochemistry

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title = "The missing link in petrobactin biosynthesis: asbF encodes a (-)-3-dehydroshikimate dehydratase",

abstract = "The siderophore petrobactin harbors unique 3,4-dihydroxybenzoyl iron-liganding groups. These moieties are known to be synthesized from shikimate pathway precursors, but no reports of the biosynthetic enzymes responsible for this conversion have been published. The gene encoding AsbF from Bacillus thuringiensis 97-27 was overexpressed in an Escherichia coli host. AsbF rapidly and efficiently transforms (-)-3-dehydroshikimate (DHS) into 3,4-dihydroxybenzoate (kcatDHS = 217 ± 10 min -1; KmDHS = 125 ± 14 μM) at 37°C and has an absolute requirement for divalent metal. Finally, the pH versus kcatDHS profile revealed two ionizable groups (pK a1 = 7.9 ± 0.1, and pKa2 = 9.3 ± 0.1).",

author = "Fox, {David T.} and Kinya Hotta and Kim, {Chu Young} and Koppisch, {Andrew T.}",

year = "2008",

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T1 - The missing link in petrobactin biosynthesis

T2 - asbF encodes a (-)-3-dehydroshikimate dehydratase

AU - Fox, David T.

AU - Hotta, Kinya

AU - Kim, Chu Young

AU - Koppisch, Andrew T.

PY - 2008/11/25

Y1 - 2008/11/25

N2 - The siderophore petrobactin harbors unique 3,4-dihydroxybenzoyl iron-liganding groups. These moieties are known to be synthesized from shikimate pathway precursors, but no reports of the biosynthetic enzymes responsible for this conversion have been published. The gene encoding AsbF from Bacillus thuringiensis 97-27 was overexpressed in an Escherichia coli host. AsbF rapidly and efficiently transforms (-)-3-dehydroshikimate (DHS) into 3,4-dihydroxybenzoate (kcatDHS = 217 ± 10 min -1; KmDHS = 125 ± 14 μM) at 37°C and has an absolute requirement for divalent metal. Finally, the pH versus kcatDHS profile revealed two ionizable groups (pK a1 = 7.9 ± 0.1, and pKa2 = 9.3 ± 0.1).

AB - The siderophore petrobactin harbors unique 3,4-dihydroxybenzoyl iron-liganding groups. These moieties are known to be synthesized from shikimate pathway precursors, but no reports of the biosynthetic enzymes responsible for this conversion have been published. The gene encoding AsbF from Bacillus thuringiensis 97-27 was overexpressed in an Escherichia coli host. AsbF rapidly and efficiently transforms (-)-3-dehydroshikimate (DHS) into 3,4-dihydroxybenzoate (kcatDHS = 217 ± 10 min -1; KmDHS = 125 ± 14 μM) at 37°C and has an absolute requirement for divalent metal. Finally, the pH versus kcatDHS profile revealed two ionizable groups (pK a1 = 7.9 ± 0.1, and pKa2 = 9.3 ± 0.1).

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The missing link in petrobactin biosynthesis: asbF encodes a (-)-3-dehydroshikimate dehydratase

Abstract

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