Molecular characteristics and transcription of the gene encoding a multifunctional alcohol dehydrogenase in relation to the deactivation of pyruvate formate-lyase in the ruminal bacterium Streptococcus bovis.
To clarify the deactivation mechanism of pyruvate formate-lyase (PFL) and its role in the regulation of fermentation in Streptococcus bovis, the molecular properties and genetic expression of multifunctional alcohol dehydrogenase (ADHE) were investigated. S. bovis was found to have ADHE, which was deduced to consist of 872 amino acids with a molecular mass of 97.4 kDa. The ADHE was shown to harbor three enzyme activities: (1) alcohol dehydrogenase, (2) coenzyme-A-linked acetaldehyde dehydrogenase that catalyzes the conversion of acetyl-CoA to ethanol, and (3) PFL deactivase. Similar to Escherichia coli ADHE, S. bovis ADHE required Fe2+ for its activity. The gene encoding ADHE ( adhE) was shown to be monocistronic. The level of adhE mRNA changed in parallel with the mRNA levels of the genes encoding PFL (pfl) and PFL-activating enzyme (act) as the growth conditions changed, although these genes are independently transcribed. Synthesis of ADHE, PFL-activating enzyme, and PFL appears to be regulated concomitantly. Overexpression of ADHE did not cause a change in the formate-to-lactate ratio. It is conceivable that ADHE is not significantly involved in the reversible inactivation of active PFL under anoxic conditions. Partition of the flow from pyruvate appears to be mainly regulated by the activities of lactate dehydrogenase and PFL.
Asanuma
Narito
N
Department of Life Science, College of Agriculture, Meiji University, Higashimita, Tamaku, 214-8571 Kawasaki, Japan. asanuma@isc.meiji.ac.jp
Yoshii
Takahiro
T
Hino
Tsuneo
T
D000123:Acetyltransferases / Q000235:genetics / Q000378:metabolism*; D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D000445:Aldehyde Oxidoreductases / Q000378:metabolism; D000693:Anaerobiosis; D000818:Animals; D002264:Carboxylic Acids / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000302:isolation & purification; D020536:Enzyme Activators / Q000032:analysis; D029968:Escherichia coli Proteins; D005285:Fermentation; D015964:Gene Expression Regulation, Bacterial; D005947:Glucose / Q000378:metabolism; D007501:Iron; D007785:Lactose / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism; D009876:Operon; D010100:Oxygen / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D012417:Rumen / Q000382:microbiology; D017422:Sequence Analysis, DNA; D016985:Streptococcus bovis / Q000201:enzymology* / Q000235:genetics*; D014158:Transcription, Genetic
D002264:Carboxylic Acids; D004269:DNA, Bacterial; D020536:Enzyme Activators; D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; D012333:RNA, Messenger; D007501:Iron; D000426:Alcohol Dehydrogenase; C068383:adhE protein, E coli; D000445:Aldehyde Oxidoreductases; C018699:aldehyde dehydrogenase (NAD(P)+); D000123:Acetyltransferases; C022560:formate C-acetyltransferase; D005947:Glucose; D007785:Lactose; D010100:Oxygen