4.7 Article

Indigoidine biosynthesis triggered by the heavy metal-responsive transcription regulator: a visual whole-cell biosensor

Journal

APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
Volume 105, Issue 14-15, Pages 6087-6102

Publisher

SPRINGER
DOI: 10.1007/s00253-021-11441-5

Keywords

Whole-cell biosensor; Pigment biosynthesis; Mercury; Lead; Visual detection

Funding

  1. National Natural Science Foundation of China [82073517]
  2. Natural Science Foundation of Guangdong Province [2019A1515011989, 2021A1515012472]
  3. S c i e n c e a n d T e c h n o l o g y P r o g r am o f S h e n z h e n [JCYJ20180306170237563, JCYJ20190808175205480]
  4. Shenzhen Fund for Guangdong Provincial High-level Clinical Key Specialties [SZGSP015]
  5. Shenzhen Key Medical Discipline Construction Fund [SZXK068]

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Whole-cell biosensors using natural pigments show great potential in indicating the presence of bioavailable heavy metals, with good selectivity and high sensitivity. The color change and absorbance measurement at 600 nm can quantify the target metal ions, showing dose-response relationships between exposure concentrations and production of indigoidine.
During the last few decades, whole-cell biosensors have attracted increasing attention for their enormous potential in monitoring bioavailable heavy metal contaminations in the ecosystem. Visual and measurable output signals by employing natural pigments have been demonstrated to offer another potential choice to indicate the existence of bioavailable heavy metals in recent years. The biosynthesis of the blue pigment indigoidine has been achieved in E. coli following heterologous expression of both BpsA (a single-module non-ribosomal peptide synthetase) and PcpS (a PPTase to activate apo-BpsA). Moreover, we demonstrated herein the development of the indigoidine-based whole-cell biosensors to detect bioavailable Hg(II) and Pb(II) in water samples by employing metal-responsive transcriptional regulator MerR and PbrR as the sensory elements, and the indigoidine biosynthesis gene cluster as a reporter element. The resulting indigoidine-based biosensors presented a good selectivity and high sensitivity to target metal ions. High concentration of target metal exposure could be clearly recognized by the naked eye due to the color change by the secretion of indigoidine, and quantified by measuring the absorbance of the culture supernatants at 600 nm. Dose-response relationships existed between the exposure concentrations of target heavy metals and the production of indigoidine. Although fairly good linear relationships were obtained in a relatively limited concentration range of the concentrations of heavy metal ions, these findings suggest that genetically controlled indigoidine biosynthesis triggered by the MerR family transcriptional regulator can enable a sensitive, visual, and qualitative whole-cell biosensor for bioindicating the presence of bioaccessible heavy metal in environmental water samples.

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