Secretion of bacterial xenobiotic-degrading enzymes from transgenic plants by an apoplastic expressional system: an applicability for phytoremediation.
In search of an effective method for phytoremediation of wastewater contaminated with organic compounds, we investigated the application of an apoplastic expressional system that secretes useful bacterial enzymes from transgenic plants into hydroponic media through the addition of a targeting signal. We constructed transgenic Arabidopsis expressing the aromatic-cleaving extradiol dioxygenase (DbfB), which degrades 2,3-dihydroxybiphenyl (2,3-DHB), and transgenic tobacco expressing haloalkane dehalogenase (DhaA), which catalyzes hydrolytic dechlorination of 1-chlorobutane (1-CB). Although crude leaf extracts of transgenic plants expressing cytoplasm-targeted degradative enzymes showed higher activity than did those from transgenic plants expressing apoplast-targeted enzymes, the hydroponic media of the latter showed 23.2 times (DbfB) and 76.4 times (DhaA) higher activity than plants containing the cytoplasm-targeted enzymes. Addition of crystalline 2,3-DHB to 100 mL of the hydroponic medium of transgenic or wild-type seedlings revealed that only medium from the transgenic Arabidopsis expressing apoplast-targeted DbfB showed rapid ring cleavage of 2,3-DHB. Transgenic tobacco expressing apoplast-targeted DhaA also resulted in the accumulation of the dehalogenation product 1-butanol in the hydroponic medium and showed a higher tolerance to 1-CB than wild-type or transgenic plants expressing cytoplasm-targeted DhaA. These results demonstrate the usefulness of the apoplastic expression of bacterial recombinant proteins in phytoremediation.
Uchida
E
E
Biotechnology Research Center and Department of Applied Biological Chemistry, University of Tokyo, 1-1-I Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
Ouchi
T
T
Suzuki
Y
Y
Yoshida
T
T
Habe
H
H
Yamaguchi
I
I
Omori
T
T
Nojiri
H
H
D000192:Actinomycetales / Q000201:enzymology*; D017360:Arabidopsis / Q000201:enzymology / Q000235:genetics* / Q000382:microbiology; D001673:Biodegradation, Environmental; D001713:Biphenyl Compounds / Q000378:metabolism; D015153:Blotting, Western; D002073:Butanes / Q000378:metabolism; D002396:Catechols / Q000378:metabolism; D017931:DNA Primers; D005822:Genetic Vectors; D006867:Hydrolases / Q000378:metabolism; D010105:Oxygenases / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000201:enzymology* / Q000382:microbiology; D011994:Recombinant Proteins / Q000378:metabolism*; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics* / Q000382:microbiology; D018675:Toxicity Tests; D014874:Water Pollutants, Chemical / Q000378:metabolism*; D018508:Water Purification / Q000379:methods*; D015262:Xenobiotics / Q000378:metabolism*
D001713:Biphenyl Compounds; D002073:Butanes; D002396:Catechols; D017931:DNA Primers; D011994:Recombinant Proteins; D014874:Water Pollutants, Chemical; D015262:Xenobiotics; C454813:2,3-dihydroxybiphenyl; D010105:Oxygenases; C439587:extradiol dioxygenase; D006867:Hydrolases; C046137:haloalkane dehalogenase; C006854:butyl chloride