Pseudomonas putida KT2440
A certified safety-strain soil bacterium with exceptional solvent/stress tolerance and diverse metabolism. The premier chassis for environmental biosensing of pollutants and heavy metals in soil and water.
Identity
- Species
- Pseudomonas putida
- Strain
- KT2440
- NCBI taxid
- 160488
- Genome
- NC_002947.4
Safety
- Biosafety level
- BSL-1
- Pathogenic
- No
- GRAS
- No
- Biocontainment
- Well-developed containment circuits; the first Gram-negative soil bacterium certified as a safe cloning host (HV1).
- BSL-1 basis
- ABSA Risk Group 1; KT2440 is a certified host-vector biosafety (HV1) strain (US Federal Register, 1982).
Rifampicin-resistant plasmid-cured derivative of P. putida mt-2; metabolically versatile, robust outdoors.
Traits
- Gram
- negative
- Oxygen
- aerobe
- Optimal temp
- 30 °C
- Doubling time
- ~40 min
- Spore-forming
- No
- Transformable
- Yes
Engineering
- CRISPR tooling
- dCas9/CRISPRi and CRISPR base-editing established in P. putida.
- Common promoters
- Pm/XylS, Pu, lac-derived, Anderson series
- Selection markers
- KanR, GmR, TcR
- Toolkits
- SEVA (Standard European Vector Architecture), Marburg Collection-style parts
Biosensors using this chassis
Alkane/oil sensor (AlkS + CRISPRa)
Detects medium-chain alkanes (diesel / crude-oil contamination). The AlkS regulator gates a CRISPRa circuit driving a fluorescent reporter, in the solvent-tolerant chassis P. putida KT2440.
Benzoate sensor (BenM + CRISPRi)
Detects benzoate, a common aromatic pollutant and degradation intermediate. The BenM LysR-family regulator gates a CRISPRi circuit driving a visible pigment, in P. putida KT2440.
Copper soil sensor (CueR + CRISPRi)
Detects bioavailable copper Cu(I)/Cu(II) in soil and water. The CueR copper-responsive regulator gates a CRISPRi circuit driving a visible chromoprotein, in the robust environmental chassis P. putida KT2440.
Crown-gall sensor (TraR/3OC8-HSL + CRISPRi)
Detects the plant pathogen Agrobacterium tumefaciens via its 3-oxo-C8-HSL quorum signal. The TraR receptor gates a CRISPRi circuit driving a visible pigment, in the soil chassis P. putida KT2440 — for agricultural crown-gall surveillance.
Cuminate sensor (CymR + CRISPRa)
Detects cuminate (p-isopropylbenzoate), a terpenoid-degradation marker. The native P. putida CymR sensor gates a CRISPRa circuit driving a fluorescent reporter, in P. putida KT2440.
DAPG biocontrol sensor (PhlF + CRISPRi)
Detects 2,4-diacetylphloroglucinol (DAPG), an antibiotic made by biocontrol pseudomonads in soil. The Marionette PhlF sensor gates a CRISPRi circuit driving a visible pigment, in the soil chassis P. putida KT2440 — useful for monitoring soil biocontrol activity.
Iron sensor (Fur + CRISPRi)
Reports bioavailable iron via the Fur ferric-uptake regulator, gating a CRISPRi circuit driving a fluorescent reporter, in the robust environmental chassis P. putida KT2440. Useful for soil/water iron and process monitoring.
Mercury soil/water sensor (MerR + CRISPRi)
Detects ionic mercury Hg(II) in soil and water. The MerR/PmerT sensing module gates a CRISPRi circuit driving a visible chromoprotein, in the rugged environmental chassis P. putida KT2440.
P. aeruginosa sensor (PQS/PqsR + CRISPRi)
Detects Pseudomonas aeruginosa via its quinolone signal (PQS). The PqsR (MvfR) receptor gates a CRISPRi circuit driving a visible pigment, in the safe Pseudomonas relative P. putida KT2440.
Phenol sensor (DmpR + CRISPRa)
Detects phenol and methylphenols in water/soil. The DmpR aromatic sensor gates a CRISPRa circuit driving a fluorescent reporter, in the phenol-tolerant chassis P. putida KT2440.
Toluene/BTEX sensor (XylR + CRISPRi)
Detects toluene and related BTEX aromatic pollutants. The native XylR/Pu regulator gates a CRISPRi circuit driving a visible pigment, in P. putida KT2440 — a natural toluene-degrader and rugged environmental chassis.