Cloning and expression of a Ralstonia eutropha HF39 gene mediating indigo formation in Escherichia coli.
On complex medium Escherichia coli strains carrying hybrid plasmid pBEC/EE:11.0, pSKBEC/BE:9.0, pSKBEC/PP:3.3, or pSKBEC/PP:2.4 harboring genomic DNA of Ralstonia eutropha HF39 produced a blue pigment characterized as indigo by several chemical and spectroscopic methods. A 1,251-bp open reading frame (bec) was cloned and sequenced. The deduced amino acid sequence of bec showed only weak similarities to short-chain acyl-coenzyme A dehydrogenases, and the gene product catalyzed formation of indoxyl, a reactive preliminary stage for production of indigo.
Drewlo
S
S
Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, Corrensstrasse 3, D-48149 Münster, Germany.
Brämer
C O
CO
Madkour
M
M
Mayer
F
F
Steinbüchel
A
A
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D020118:Cupriavidus necator / Q000235:genetics* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics*; D007203:Indigo Carmine; D007211:Indoles / Q000302:isolation & purification / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013050:Spectrometry, Fluorescence; D014368:Tryptophanase / Q000378:metabolism
D001426:Bacterial Proteins; D007211:Indoles; D011994:Recombinant Proteins; C030374:indole; D007203:Indigo Carmine; D006899:Mixed Function Oxygenases; C432944:Bec protein, Ralstonia eutropha; D014368:Tryptophanase