Molecular characterization of the 1,3-propanediol (1,3-PD) operon of Clostridium butyricum.
The genes encoding the 1,3-propanediol (1,3-PD) operon of Clostridium butyricum VPI1718 were characterized from a molecular and a biochemical point of view. This operon is composed of three genes, dhaB1, dhaB2, and dhaT. When grown in a vitamin B12-free mineral medium with glycerol as carbon source, Escherichia coli expressing dhaB1, dhaB2, and dhaT produces 1,3-PD and high glycerol dehydratase and 1,3-PD dehydrogenase activities. dhaB1 and dhaB2 encode, respectively, a new type of glycerol dehydratase and its activator protein. The deduced proteins DhaB1 and DhaB2, with calculated molecular masses of 88,074 and 34,149 Da, respectively, showed no homology with the known glycerol dehydratases that are all B12 dependent but significant similarity with the pyruvate formate lyases and pyruvate formate lyases activating enzymes and their homologues. The 1,158-bp dhaT gene codes for a 1,3-PD dehydrogenase with a calculated molecular mass of 41,558 Da, revealing a high level of identity with other DhaT proteins from natural 1,3-PD producers. The expression of the 1,3-PD operon in C. butyricum is regulated at the transcriptional level, and this regulation seems to involve a two-component signal transduction system DhaASDhaA, which may have a similar function to DhaR, a transcriptional regulator found in other natural 1,3-PD producers. The discovery of a glycerol dehydratase, coenzyme B12 independent, should significantly influence the development of an economical vitamin B12-free biological process for the production of 1,3-PD from renewable resources.
Raynaud
Céline
C
Laboratoire de Biotechnologies-Bioprocédés, UMR-CNRS 5504, Unité de Recherche Institut National de la Recherche Agronomique 792, Centre de Bioingénierie Gilbert Durand, Institut National des Sciences Appliquées, Toulouse, France.
Sarçabal
Patricia
P
Meynial-Salles
Isabelle
I
Croux
Christian
C
Soucaille
Philippe
P
D000426:Alcohol Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D003013:Clostridium / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D005798:Genes, Bacterial; D006836:Hydro-Lyases / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009876:Operon; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic
D000429:Alcohol Oxidoreductases; D000426:Alcohol Dehydrogenase; C052370:1,3-propanediol dehydrogenase; D006836:Hydro-Lyases; C020671:glycerol dehydratase