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Transcriptomics-guided rational engineering in Bacillus licheniformis for enhancing poly-γ-glutamic acid biosynthesis

Rui Han¹, Qian Zhong¹, Yifan Yan¹

¹College of Food Science and Light Industry, Nanjing Tech University, Nanjing 211816, China; State Key Laboratory of Materials-oriented Chemical Engineering, Nanjing Tech University, Nanjing 211816, China.

International Journal of Biological Macromolecules

|November 12, 2024

Summary

This study engineered Bacillus licheniformis to efficiently produce poly-γ-glutamic acid (γ-PGA) from untreated molasses. Metabolic modifications significantly boosted γ-PGA yield, demonstrating cost-effective bioconversion for industrial applications.

Keywords:

Poly-γ-glutamic acid Transcriptomic analysis Untreated molasses

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Area of Science:

Biotechnology
Microbial Synthesis
Metabolic Engineering

Background:

Poly-γ-glutamic acid (γ-PGA) is a biodegradable polymer with diverse applications.
Utilizing non-food raw materials like molasses for γ-PGA production offers a sustainable alternative.
Molasses's complex composition can hinder microbial conversion efficiency.

Purpose of the Study:

To enhance Bacillus licheniformis's ability to convert untreated molasses into γ-PGA.
To guide metabolic engineering strategies using transcriptomic analysis.
To achieve cost-effective and high-yield γ-PGA biosynthesis.

Main Methods:

Transcriptomic analysis of Bacillus licheniformis grown on molasses.
Metabolic engineering strategies including gene knockouts (AlsS, CcpA) and gene overexpression.
Fermentation optimization using untreated molasses as the sole carbon source.

Main Results:

Transcriptomic data revealed altered gene expression in substrate utilization, by-product synthesis, and precursor synthesis modules.
Knockout of AlsS and CcpA reduced by-product formation and alleviated carbon catabolite repression.
Tandem overexpression of precursor genes, combined with knockouts, resulted in a γ-PGA titer of 48.26 g/L.
Achieved a 3.12-fold increase in γ-PGA production compared to the wild-type strain.

Conclusions:

Metabolic engineering guided by transcriptomics can significantly improve γ-PGA production from untreated molasses.
The developed strain and process offer a cost-effective route for industrial γ-PGA synthesis.
This study provides valuable insights for optimizing microbial bioconversion of complex, non-food feedstocks.