Isolation of a new butanol-producing Clostridium strain: high level of hemicellulosic activity and structure of solventogenesis genes of a new Clostridium saccharobutylicum isolate.
New isolates of solventogenic bacteria exhibited high hemicellulolytic activity. They produced butanol and acetone with high selectivity for butanol (about 80% of butanol from the total solvent yield). Their 16S rDNA sequence was 99% identical to that of Clostridium saccharobutylicum. The genes responsible for the last steps of solventogenesis and encoding crotonase, butyryl-CoA dehydrogenase, electron-transport protein subunits A and B, 3-hydroxybutyryl-CoA dehydrogenase, alcohol dehydrogenase, CoA-transferase (subunits A and B), acetoacetate decarboxylase, and aldehyde dehydrogenase were identified in the new C. saccharobutylicum strain Ox29 and cloned into Escherichia coli. The genes for crotonase, butyryl-CoA dehydrogenase, electron-transport protein subunits A and B, and 3-hydroxybutyryl-CoA dehydrogenase composed the bcs-operon. A monocistronic operon containing the alcohol dehydrogenase gene was located downstream of the bcs-operon. Genes for aldehyde dehydrogenase, CoA-transferase (subunits A and B), and acetoacetate decarboxylase composed the sol-operon. The gene sequences and the gene order within the sol- and bcs-operons of C. saccharobutylicum Ox29 were most similar to those of Clostridium beijerinckii. The activity of some of the bcs-operon genes, expressed in heterologous E. coli, was determined.
Berezina
Oksana V
OV
State Research Institute of Genetics and Selection of Industrial Microorganisms, 1st Dorojniy pr. 1, 117545 Moscow, Russian Federation. mashchenko@yandex.ru
Brandt
Agnieszka
A
Yarotsky
Sergey
S
Schwarz
Wolfgang H
WH
Zverlov
Vladimir V
VV
D000096:Acetone / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics; D000440:Butanols / Q000378:metabolism*; D003001:Cloning, Molecular; D003013:Clostridium / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D016000:Cluster Analysis; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D023061:Gene Order; D005796:Genes; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009876:Operon; D010802:Phylogeny; D011134:Polysaccharides / Q000378:metabolism*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D017385:Sequence Homology
D001426:Bacterial Proteins; D000440:Butanols; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D011134:Polysaccharides; D012336:RNA, Ribosomal, 16S; D000096:Acetone; C007916:hemicellulose