Labrys portucalensis: a newly described PFAS degrading strain

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+ What is the orthogonal replication?

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Experiments associated with labrys

1- PFOS degradation

1- PFOS degradation

Labrys portucalensis combined with an improved dehalogenase

Directed evolution for improving DeHa2

Orthogonal replication

Design of the OrthoRep system

1- Creation of the replication operon

Figure 1: Structure of a typical PFAS molecule. Pass the mouse or click on the different components to see their characteristics.

TP dnap ssb dsb

Terminal protein gene: expresses proteins binding to ITR sequences therefore functioning as origins for replication for PRD1 DNAP. PRD1 DNA polymerase gene : encodes a DNA polymerase derived from phage PRD1, for which terminal protein (TP) bound to inverted terminal repeats (ITR) naturally serve as replication origins in the phage genome.This DNAP was mutated by the authors to make it error-prone. Single-stranded DNA-binding protein gene: early-expressed gene in phage PRD1, hypothesized to be necessary for PRD1 replication. Double-stranded DNA-binding protein gene: early-expressed gene in phage PRD1, hypothesized to be necessary for replication for PRD1 replication.

2- Implementation of the replication operon in E. coli DH10B

3- Creation of the linear replicon

4- Maintaining the linear replicon

Cell sorting by fluorescence: FluorMango aptamer and microfluidics

Proof of concept for the screening method

1- Production of FluorMango

2- Assessing FluorMango’s sensitivity

3- Assessing FluorMango’s specificity

4- Test with a monofluorinated compound

5- Microfluidics trials

Improving P. putida’s response to fluoride stress

Overexpressing FluC in P. putida KT2440

1- Creation of pSEVA438-fluC

2- Testing of P. putida WT against P.putida pSEVA438-fluC

Design of the industrial process

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