BIOTIN BIOSYNTHESIS IN HIGHER-PLANT CELLS - IDENTIFICATION OF INTERMEDIATES

Biotin biosynthesis was investigated in lavender cell cultures (Lavandula vera L.). Two different biological assays and two different HPLC procedures were used to identify all the intermediates involved in biotin biosynthesis. The pathway for biotin biosynthesis could be analyzed starting with [H-3]pimelic acid as precursor, leading to labelled biotin and even to labelled biotinylated enzymes. Intermediates known from the bacterial pathway (7-oxo-8-amino-pelargonic acid, 7,8-diamino-pelargonic acid, dethiobiotin) were present in detectable amounts. Pimelic acid activation to pimeloyl-CoA could be observed. In contrast to bacterial cells, an unknown stable labelled intermediate, named compound A, accumulated. This compound coeluted with an authentic sample of 9-mercaptodethiobiotin from HPLC with an anion-exchange column and was as effective as biotin in supporting the growth of the strain bioB105 of Escherichia coli. When H-3-labelled compound A was added to the growth medium of the lavender cells it was incorporated in an acidomycin-sensitive manner into biotin. [H-3]Dethiobiotin was incorporated into both compound A and biotin. These results strongly suggest that, in higher plant cells, the reaction catalysed by biotin synthase may proceed in two distinct steps involving mercaptodethiobiotin (9-mercaptodethiobiotin ?) as an intermediate.