Quorum Sensing Systems Engineering for Enhanced iso-Butylamine Production in Escherichia coli

Abstract

Quorum sensing (QS), characterized by pathway-independence and autonomous control, has been applied in bio-manufacturing, while the lack of versatile and functional regulatory components limits its broader applications. To address this issue, a series of efficient QS systems with diverse properties were established in Escherichia coli. Firstly, combinatorial optimization, including element selection and promoter replacement, led to an improvement of 8.82- and 3.03-fold in output range and response threshold, respectively. Then, a library of LuxR mutants was constructed for screening novel variants with decreased sensitivity to acyl-homoserine lactone through the high-throughput screening technique. Notably, the optimal variant V36E/H89L/P97L exhibited a decrease of 266-fold in the sensitivity. As a proof-of-concept, iso-butylamine biosynthesis was tested by re-directing pyruvate catabolism using QS circuits, and in particular, a total of 15.4 g/L iso-butylamine was generated in strain IB21 during the fed-batch culture, marking a 2.96-fold increase over the static control. Finally, the generated bioproduct reached 44.23 g/L in a bioreactor, representing the highest reported titer so far. In summary, this study not only enriches the genetic toolbox of QS systems, but also facilitates industrial applications in value-added chemical production.

Keywords

Quorum sensing / Regulatory elements / Signal sensitivity / Synthetic biology / iso-Butylamine / Escherichia coli

Supplementary Materials

The following supporting information can be found at: https://www.sciepublish.com/article/pii/535, Table S1. The plasmids used in this study; Table S2. The strains used in this study; Table S3. The primers used in this study; Table S4. Amino acid sequences of heterologous genes; Figure S1. Dynamic range of strain QMI2 at different temperatures. Figure S2. The characterization of LuxR mutants failing to self-induce; Figure S3. Growth profiles of strain IB01 with exogenous pyruvate addition; Figure S4. The characterization of selected QS systems for regulation of iso-butylamine synthesis; Figure S5. Effects of increasing pyruvate supply by downregulating TCA cycle or reconstructing non-PTS glucose transportation system on the titer.

Author Contributions

Conceptualization, Y.L., M.L. and J.W.; Methodology, Y.L. and M.L.; Validation, M.L. and Y.L.; Formal Analysis,M.L. and Y.L.; Investigation, M.L. and Y.L.; Resources, Y.L., M.L., P.Y. and J.W.; Data Curation, M.L.; Writing—Original Draft Preparation, M.L.; Writing—Review & Editing, Y.L., M.L., H.F. and J.W.; Visualization, M.L.; Supervision, J.W.; Project Administration, J.W.; Funding Acquisition, J.W.

Ethics Statement

Not applicable.

Informed Consent Statement

Not applicable.

Data Availability Statement

Data will be made available on request.

Funding

This research was supported by the National Natural Science Foundation of China (22178133) and Guangdong Provincial Key Research and Development Program (2024B1111160006).

Declaration of Competing Interest

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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