Detoxification of the plant toxin fluoroacetate by a genetically modified rumen bacterium.
We isolated the fluoroacetate dehalogenase gene (H1), from Moraxella species strain B, and placed it under the transcriptional control of a 154 bp fragment of the erm gene promoter. The promoter/gene construct was attached to the Butyrivibrio fibrisolvens shuttle vector pBHerm, and the resulting dehalogenase expression plasmid (pBHf) was transferred to B. fibrisolvens OB156 by electroporation. The erm gene promoter directed expression of dehalogenase activity in both E. coli and B. fibrisolvens OB156. Cell-free lysates of the genetically modified OB156 defluorinated 10.6 nmol fluoroacetate/min/mg protein. Growing cultures of OB156 were able to detoxify fluoroacetate in the culture medium, at the rate of 9.9 nmol/min/mg. Plasmid pBHf was retained by 100% of OB156 cells after 500 generations of non-selective culture. The restriction pattern of pBHf remained unchanged after extensive non-selective growth and host bacteria continued to produce active dehalogenase. The construction of rumen bacteria that are able to detoxify an important natural poison supports the feasibility of using genetically modified rumen bacteria to aid animal production.
Gregg
K
K
Institute of Biotechnology, University of New England, Armidale, Australia.
Cooper
C L
CL
Schafer
D J
DJ
Sharpe
H
H
Beard
C E
CE
Allen
G
G
Xu
J
J
D000818:Animals; D001421:Bacteria, Anaerobic / Q000201:enzymology / Q000235:genetics; D001483:Base Sequence; D018274:Electroporation; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005463:Fluoroacetates / Q000378:metabolism*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D006090:Gram-Negative Bacteria / Q000201:enzymology / Q000235:genetics; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism; D008658:Inactivation, Metabolic; D008969:Molecular Sequence Data; D009016:Moraxella / Q000201:enzymology / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D012417:Rumen / Q000382:microbiology*
D005463:Fluoroacetates; C014815:fluoroacetic acid; D006867:Hydrolases; C023098:haloacetate dehalogenase