6-Hydroxy--nicotine oxidase (6-HDNO) of , an enzyme inducible by -nicotine, contains FAD covalently bound via an 8α-N(3)His linkage. Expression of the gene encoding 6-HDNO and flavinylation of the protein were studied in . In this heterologous system the following findings were made. 1. An enzymically active covalently flavinylated 6-HDNO of normal size can be expressed in . 2. The natural promoter of the 6-HDNO gene appeared inefficient in . The promoter, when inserted upstream of the promoter, increased 6-HDNO expression >50-fold. 3. Expression of the 6-HDNO gene from plasmids in was, independently of the promoter construct used, stimulated more than fivefold by -nicotine in the growth medium. It is concluded that flavinylation of 6-HDNO is possibly autocatalytic and mediated by factors generally found in bacterial cells.


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