Journal article
Identification, Characterization, and Application of a Highly Sensitive Lactam Biosensor from Pseudomonas putida
Caprolactam is an important polymer precursor to nylon traditionally derived from petroleum and produced on a scale of 5 million tons per year. Current biological pathways for the production of caprolactam are inefficient with titers not exceeding 2 mg/L, necessitating novel pathways for its production.
As development of novel metabolic routes often require thousands of designs and result in low product titers, a highly sensitive biosensor for the final product has the potential to rapidly speed up development times. Here we report a highly sensitive biosensor for valerolactam and caprolactam from Pseudomonas putida KT2440 which is >1000× more sensitive to an exogenous ligand than previously reported sensors.
Manipulating the expression of the sensor oplR (PP_3516) substantially altered the sensing parameters, with various vectors showing Kd values ranging from 700 nM (79.1 μg/L) to 1.2 mM (135.6 mg/L). Our most sensitive construct was able to detect in vivo production of caprolactam above background at ~6 μg/L.
The high sensitivity and range of OplR is a powerful tool toward the development of novel routes to the biological synthesis of caprolactam.
Language: | English |
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Publisher: | American Chemical Society |
Year: | 2020 |
Pages: | 53-62 |
ISSN: | 21615063 |
Types: | Journal article |
DOI: | 10.1021/acssynbio.9b00292 |
ORCIDs: | 0000-0002-4556-9685 , 0000-0002-6513-7425 and 0000-0003-4170-6088 |