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A Tryptophan 6‐Halogenase and an Amidotransferase Are Involved in Thienodolin Biosynthesis
Author(s) -
Milbredt Daniela,
Patallo Eugenio P.,
van Pée KarlHeinz
Publication year - 2014
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201400016
Subject(s) - biosynthesis , glutamine amidotransferase , orfs , gene , gene cluster , biology , heterologous expression , biochemistry , streptomyces , enzyme , chlamydomonas reinhardtii , genetics , mutant , amino acid , open reading frame , peptide sequence , bacteria , recombinant dna , glutamine
The biosynthetic gene cluster for the plant growth‐regulating compound thienodolin was identified in and cloned from the producer organism Streptomyces albogriseolus MJ286‐76F7. Sequence analysis of a 27 kb DNA region revealed the presence of 21 ORFs, 14 of which are involved in thienodolin biosynthesis. Three insertional inactivation mutants were generated in the sequenced region to analyze their involvement in thienodolin biosynthesis and to functionally characterize specific genes. The gene inactivation experiments together with enzyme assays with enzymes obtained by heterologous expression and feeding studies showed that the first step in thienodolin biosynthesis is catalyzed by a tryptophan 6‐halogenase and that the last step is the formation of a carboxylic amide group catalyzed by an amidotransferase. The results led to a hypothetical model for thienodolin biosynthesis.