Biotin formation by recombinant strains of Escherichia coli: influence of the host physiology.
Strains of Escherichia coli were transformed with different plasmids bearing the gene clusters bioXWF and bioDAYB isolated from the Gram positive bacterium Bacillus sphaericus. These genes encode for the enzymes involved in the metabolic pathway which synthesizes biotin from the precursor pimelic acid. Transformed E. coli strains were grown in bioreactors to reach a biomass of 18 g l-1 cell dry weight in 1 litre batch culture with substrate feeding and approximately 50 g l-1 in 10 l fed batch culture. Improved yields of total vitamers and biotin formed in these processes were achieved after a comparative analysis of different culture conditions, medium compositions, host strains and expression systems. Production of 27 mg l-1 of biotin and 200 mg l-1 of vitamers was achieved in 1 litre batch culture. Using a 10-1 fed batch process, biotin and vitamer concentrations reached maximum values of 45 mg l-1 and 350 mg l-1, respectively.
Sabatié
J
J
Transgène S.A., Strasbourg, France.
Speck
D
D
Reymund
J
J
Hebert
C
C
Caussin
L
L
Weltin
D
D
Gloeckler
R
R
O'Regan
M
M
Bernard
S
S
Ledoux
C
C
D001407:Bacillus / Q000235:genetics*; D001710:Biotin / Q000096:biosynthesis*; D003470:Culture Media; D004926:Escherichia coli / Q000378:metabolism*; D010957:Plasmids; D014169:Transformation, Bacterial
D003470:Culture Media; D001710:Biotin