期刊
CHEMOSPHERE
卷 186, 期 -, 页码 510-518出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.chemosphere.2017.08.027
关键词
Naphthalene; Whole-cell bioreporter; Acinetobacter baylyi; Gibson cloning; Polycyclic aromatic hydrocarbons (PAHs)
资金
- National Natural Science Foundation of China [51178048, 51378064, 41301331]
- EPSRC [EP/M002403/1] Funding Source: UKRI
- Engineering and Physical Sciences Research Council [EP/M002403/1] Funding Source: researchfish
A new naphthalene bioreporter was designed and constructed in this work. A new vector, pVVH1274_Nah, was constructed by the Gibson isothermal assembly fused with a 9 kb naphthalene-degrading gene nahAD (nahAa nahAb nahAc nahAd nahB nahF nahC nahQ nahE nahD) and cloned into Acinetobacter ADPWH_lux as the host, capable of responding to salicylate (the central metabolite of naphthalene). The ADPWH_Nah bioreporter could effectively metabolize naphthalene and evaluate the naphthalene in natural water and soil samples. This whole-cell bioreporter did not respond to other polycyclic aromatic hydrocarbons (PAHs; pyrene, anthracene, and phenanthrene) and demonstrated a positive response in the presence of 0.01 mu M naphthalene, showing high specificity and sensitivity. The bioluminescent response was quantitatively measured after a 4 h exposure to naphthalene, and the model simulation further proved the naphthalene metabolism dynamics and the salicylate-activation mechanisms. The ADPWH_Nah bioreporter also achieved a rapid evaluation of the naphthalene in the PAH-contaminated site after chemical spill accidents, showing high consistency with chemical analysis. The engineered Acinetobacter variant had significant advantages in rapid naphthalene detection in the laboratory and potential in situ detection. The state-of-the-art concept of cloning PAHs-degrading pathway in salicylate bioreporter hosts led to the construction and assembly of high-throughput PAH bioreporter array, capable of crude oil contamination assessment and risk management. (C) 2017 Elsevier Ltd. All rights reserved.
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