Isolation and characterization of Alicycliphilus denitrificans strain BC, which grows on benzene with chlorate as the electron acceptor.
A bacterium, strain BC, was isolated from a benzene-degrading chlorate-reducing enrichment culture. Strain BC degrades benzene in conjunction with chlorate reduction. Cells of strain BC are short rods that are 0.6 microm wide and 1 to 2 microm long, are motile, and stain gram negative. Strain BC grows on benzene and some other aromatic compounds with oxygen or in the absence of oxygen with chlorate as the electron acceptor. Strain BC is a denitrifying bacterium, but it is not able to grow on benzene with nitrate. The closest cultured relative is Alicycliphilus denitrificans type strain K601, a cyclohexanol-degrading nitrate-reducing betaproteobacterium. Chlorate reductase (0.4 U/mg protein) and chlorite dismutase (5.7 U/mg protein) activities in cell extracts of strain BC were determined. Gene sequences encoding a known chlorite dismutase (cld) were not detected in strain BC by using the PCR primers described in previous studies. As physiological and biochemical data indicated that there was oxygenation of benzene during growth with chlorate, a strategy was developed to detect genes encoding monooxygenase and dioxygenase enzymes potentially involved in benzene degradation in strain BC. Using primer sets designed to amplify members of distinct evolutionary branches in the catabolic families involved in benzene biodegradation, two oxygenase genes putatively encoding the enzymes performing the initial successive monooxygenations (BC-BMOa) and the cleavage of catechol (BC-C23O) were detected. Our findings suggest that oxygen formed by dismutation of chlorite can be used to attack organic molecules by means of oxygenases, as exemplified with benzene. Thus, aerobic pathways can be employed under conditions in which no external oxygen is supplied.
Weelink
Sander A B
SA
Laboratory of Microbiology, Wageningen University, Wageningen, The Netherlands.
Tan
Nico C G
NC
ten Broeke
Harm
H
van den Kieboom
Corné
C
van Doesburg
Wim
W
Langenhoff
Alette A M
AA
Gerritse
Jan
J
Junca
Howard
H
Stams
Alfons J M
AJ
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001554:Benzene / Q000378:metabolism*; D002704:Chlorates / Q000378:metabolism*; D042621:Comamonadaceae / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D049308:Dioxygenases / Q000235:genetics / Q000378:metabolism; D020459:Genes, rRNA; D008124:Locomotion; D006899:Mixed Function Oxygenases / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D009566:Nitrates / Q000378:metabolism; D009573:Nitrites / Q000378:metabolism; D009584:Nitrogen / Q000378:metabolism; D010088:Oxidoreductases / Q000235:genetics / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA; D012689:Sequence Homology, Nucleic Acid
D001426:Bacterial Proteins; D002704:Chlorates; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D009566:Nitrates; D009573:Nitrites; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S; D006899:Mixed Function Oxygenases; D010088:Oxidoreductases; D049308:Dioxygenases; C402794:chlorite dismutase; D001554:Benzene; D009584:Nitrogen; D010100:Oxygen
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