2021
DOI: 10.1007/s00253-021-11441-5
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Indigoidine biosynthesis triggered by the heavy metal-responsive transcription regulator: a visual whole-cell biosensor

Abstract: 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 sin… Show more

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Cited by 27 publications
(20 citation statements)
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“…The whole dose-response of TOP10/pPb-CACD toward Pb(II) was shown in Figure 4C, which was similar to that of the previously constructed PbrR-based bacterial biosensor with the blue pigment indigoidine as the output signal (Hui et al, 2021a). The CACD-derived signal was significantly increased from 0 to 12.5 μM and gradually stabilized from 25 to 200 μM (Figure 4C).…”
Section: Response Properties Of Metabolic Engineered Bacterial Biosen...supporting
confidence: 78%
“…The whole dose-response of TOP10/pPb-CACD toward Pb(II) was shown in Figure 4C, which was similar to that of the previously constructed PbrR-based bacterial biosensor with the blue pigment indigoidine as the output signal (Hui et al, 2021a). The CACD-derived signal was significantly increased from 0 to 12.5 μM and gradually stabilized from 25 to 200 μM (Figure 4C).…”
Section: Response Properties Of Metabolic Engineered Bacterial Biosen...supporting
confidence: 78%
“…The detection limit could be defined as the lowest metal concentration that evokes a significant increase in biosensing signal (background + 3 × SD) 7 , 11 . The detection limit using bacterial biosensor in the present investigation was about 0.049 μM (Fig.…”
Section: Resultsmentioning
confidence: 99%
“…To validate the capability of the biosensor to detect bioavailable Cd(II) in environmental water samples, engineered TOP10/pPcad-vio was cultured in LB medium prepared with purified water, tap water, and two kinds of surface water as described previously 11 . The early exponential-phase culture were spiked with 0, 0.39, 0.78, 1.56, 3.125, 6.25, and 12.5 μM Cd(II) using a double dilution method.…”
Section: Methodsmentioning
confidence: 99%
“…In-gel fluorescent protein detection can be of value if a non-precise molecular mass determination, yet confirmation of difference in size from control is needed. The ability to use fluorescence for plate-level rapid colony screening and target recombinant protein visualization in gel allowed determination of the expression of two new transgene targets after integration into the algal nuclear genome: the nonribosomal peptide synthase (NRPS) A from Streptomyces lavendulae (BpsA, NCBI: AB240063.1, (Hui et al, 2021)) and the 4′phosphopantetheinyl transferase (4'-PPTase) from Pseudomonas aeruginosa (PaPcpS, Protein Data Bank ID: 7BCZ_A, (Carivenc et al, 2021)). BpsA expression was chosen for several reasons: it is the first example of heterologous NRPS expression in a eukaryotic algal host, it is a validation of C-terminal fusion to mVenus using the current iteration of intron spreading, and the fusion protein is ~170 kDa making it one of the larger heterologous proteins to be expressed from the nuclear genome of this alga.…”
Section: Selective Filter Sets Allow In Vitro Protein Visualization I...mentioning
confidence: 99%