Breakthrough in Recombinant Human Insulin Production Using Novel Promoters in Pseudomonas Fluorescens

Strategic Insights -

Researchers at the Indian Institute of Technology Guwahati have made a significant advancement in the production of recombinant human insulin using novel promoters in Pseudomonas fluorescens. The study aimed to develop an IPTG-free expression system, which is crucial for large-scale production of insulin. The research demonstrated the use of constitutive and native inducible promoters to drive expression in the P. fluorescens host system. The results showed a significant increase in insulin production, with titers reaching up to 54.5 mg/L in a 2 L bioreactor. The study also identified anthranilate and m-toluate as optimal inducers for anthranilate and benzoate promoter-based expression systems, respectively.

Key Takeaways:

  • The study presents a novel IPTG-free expression system for recombinant human insulin production in Pseudomonas fluorescens.
  • The use of constitutive and native inducible promoters increased insulin production titers up to 54.5 mg/L in a 2 L bioreactor.
  • Anthranilate and m-toluate were identified as optimal inducers for anthranilate and benzoate promoter-based expression systems, respectively.
  • The study demonstrates a 29% increase in insulin production compared to shake-flask using a constitutive promoter (P-PsbA)-driven system.
  • The fusion protein and insulin produced through these systems were validated by Western blotting, and insulin integrity was confirmed via MALDI-TOF.
  • The study concludes that the P-Ant- and P-Ben-based expression systems can be used for differential expression of recombinant proteins such as insulin chains and monoclonal antibodies.
  • The research was funded by the Ministry of Education, Government of India, IIT Guwahati, and the Ministry of Education, Government of India.

Statistics:

  • Insulin production titers reached up to 54.5 mg/L in a 2 L bioreactor.
  • The study showed a 29% increase in insulin production compared to shake-flask using a constitutive promoter (P-PsbA)-driven system.
  • Anthranilate (5 mM) and m-toluate (7.5 mM) were identified as optimal inducers for anthranilate (P-Ant) and benzoate (P-Ben) promoter-based expression systems, respectively.

Sources:

  • NewsRx. Findings from Indian Institute of Technology Guwahati Update Knowledge of Pseudomonas (Production of Recombinant Human Insulin Using Constitutive and Non-iptg Inducible Promoters In Pseudomonas Fluorescens and Scale-up Study). Life Science Weekly. November 4, 2025; p 16.
  • Production of Recombinant Human Insulin Using Constitutive and Non-iptg Inducible Promoters In Pseudomonas Fluorescens and Scale-up Study. Process Biochemistry, 2025;158:35-43.