title: Enhanced degradation of haloacid by heterologous expression in related Burkholderia species.
abstract: Haloacids are environmental pollutant and can be transformed to non-toxic alkanoic acids by microbial dehalogenase. Bacterium Burkholderia species MBA4 was enriched from soil for its ability to bioremediate haloacids such as mono-chloroacetate (MCA), mono-bromoacetate (MBA), 2-mono-chloropropionate, and 2-mono-bromopropionate. MBA4 produces an inducible dehalogenase Deh4a that catalyzes the dehalogenation process. The growth of MBA4 on haloacid also relies on the presence of a haloacid-uptake system. Similar dehalogenase genes can be found in the genome of many related species. However, wildtype Burkholderia caribensis MWAP64, Burkholderia phymatum STM815, and Burkholderia xenovorans LB400 were not able to grow on MCA. When a plasmid containing the regulatory and structural gene of Deh4a was transformed to these species, they were able to grow on haloacid. The specific enzyme activities in these recombinants ranges from 2- to 30-fold that of MBA4 in similar condition. Reverse transcription-quantitative real-time PCR showed that the relative transcript levels in these recombinant strains ranges from 9 to over 1,600 times that of MBA4 in similar condition. A recombinant has produced nearly five times of dehalogenase that MBA4 could ever achieve. While the expressions of Deh4a were more relaxed in these phylogenetically related species, an MCA-uptake activity was found to be inducible. These metabolically engineered strains are better degraders than the haloacid-enriched MBA4.
authors.affiliation: Molecular Microbiology Laboratory, School of Biological Sciences, The University of Hong Kong, Pokfulam Road, Hong Kong, SAR, China.
authors.forename: Xianbin
authors.initials: X
authors.lastname: Su
authors.forename: Liyu
authors.initials: L
authors.lastname: Deng
authors.forename: Ka Fai
authors.initials: KF
authors.lastname: Kong
authors.forename: Jimmy S H
authors.initials: JS
authors.lastname: Tsang
mesh_terms: D000085:Acetates / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001673:Biodegradation, Environmental; D019117:Burkholderia / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D006867:Hydrolases / Q000235:genetics / Q000378:metabolism*; D060847:Metabolic Engineering / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D000085:Acetates; D001426:Bacterial Proteins; D012333:RNA, Messenger; C006972:chloroacetic acid; D006867:Hydrolases; C034507:2-haloacid dehalogenase
keywords: Burkholderia; biodegradation; dehalogenase; haloacid; heterologous expression