Anaerobic growth of Escherichia coli on glycerol by importing genes of the dha regulon from Klebsiella pneumoniae.
The dha regulon of Klebsiella pneumoniae specifying fermentative dissimilation of glycerol was mobilized by the broad-host-range plasmid RP4:mini Mu and introduced conjugatively into Escherichia coli. The recipient E. coli was enabled to grow anaerobically on glycerol without added hydrogen acceptors, although its cell yield was less than that of K. pneumoniae. The reduced cell yield was probably due to the lack of the coenzyme-B12-dependent glycerol dehydratase of the dha system. This enzyme initiates the first step in an auxiliary pathway for disposal of the extra reducing equivalents from glycerol. The lack of this enzyme would also account for the absence of 1,3-propanediol (a hallmark fermentation product of glycerol) in the spent culture medium. In a control experiment, a large quantity of this compound was detected in a similar culture medium following the growth of K. pneumoniae. The other three known enzymes of the dha system, glycerol dehydrogenase, dihydroxyacetone kinase and 1,3-propanediol oxidoreductase, however, were synthesized at levels comparable to those found in K. pneumoniae. Regulation of the dha system in E. coli appeared to follow the same pattern as in K. pneumoniae: the three acquired enzymes were induced by glycerol, catabolite repressed by glucose, and glycerol dehydrogenase was post-translationally inactivated during the shift from anaerobic to aerobic growth. The means by which the E. coli recipient can achieve redox balance without formation of 1,3-propanediol during anaerobic growth on glycerol remains to be discovered.
Sprenger
G A
GA
Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, MA 02115.
Hammer
B A
BA
Johnson
E A
EA
Lin
E C
EC
D000429:Alcohol Oxidoreductases / Q000235:genetics; D000693:Anaerobiosis; D001426:Bacterial Proteins / Q000235:genetics; D003227:Conjugation, Genetic; D004734:Energy Metabolism; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D005285:Fermentation; D005798:Genes, Bacterial; D005990:Glycerol / Q000378:metabolism; D006836:Hydro-Lyases / Q000235:genetics / Q000502:physiology; D007711:Klebsiella pneumoniae / Q000235:genetics*; D010084:Oxidation-Reduction; D010770:Phosphotransferases / Q000235:genetics; D017853:Phosphotransferases (Alcohol Group Acceptor); D010957:Plasmids; D013401:Sugar Alcohol Dehydrogenases / Q000235:genetics
D001426:Bacterial Proteins; D000429:Alcohol Oxidoreductases; D013401:Sugar Alcohol Dehydrogenases; C020672:glycerol dehydrogenase; C021494:lactaldehyde reductase; D010770:Phosphotransferases; D017853:Phosphotransferases (Alcohol Group Acceptor); C023132:glycerone kinase; D006836:Hydro-Lyases; C020671:glycerol dehydratase; D005990:Glycerol