Open AccessExpression and Flavinylation of Arthrobacter oxydans 6-Hydroxy-D-nicotine Oxidase in Bacillus subtilis
Author(s) -
Roderich Brandsch,
Lars Hederstedt
Publication year - 1989
Publication title -
microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.352
H-Index - 35
eISSN - 1465-2080
pISSN - 1350-0872
DOI - 10.1099/00221287-135-5-1093
Subject(s) - bacillus subtilis , gene , heterologous expression , biochemistry , biology , oxidase test , plasmid , promoter , heterologous , enzyme , arthrobacter , chemistry , microbiology and biotechnology , levansucrase , lantibiotics , gene expression , recombinant dna , bacteria , genetics , nisin
6-Hydroxy-d-nicotine oxidase (6-HDNO) of Arthrobacter oxydans, an enzyme inducible by dl-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 Bacillus subtilis. In this heterologous system the following findings were made. 1. An enzymically active covalently flavinylated 6-HDNO of normal size can be expressed in B. subtilis. 2. The natural promoter of the 6-HDNO gene appeared inefficient in B. subtilis. The B. subtilis sdh promoter, when inserted upstream of the A. oxydans promoter, increased 6-HDNO expression >50-fold. 3. Expression of the 6-HDNO gene from plasmids in B. subtilis was, independently of the promoter construct used, stimulated more than fivefold by dl-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