Abstract
Poly gamma glutamic acid (γ-PGA) is an anionic polyamide with numerous applications. Previous studies revealed that L-proline metabolism is implicated in a wide range of cellular processes by increasing intercellular reactive oxygen species (ROS) generation. However, the relationship between L-proline metabolism and γ-PGA synthesis has not yet been analyzed. In this study, our results confirmed that deletion of Δ1-pyrroline-5-carboxylate dehydrogenase gene ycgN in Bacillus licheniformis WX-02 increased γ-PGA yield to 13.91 g L−1, 85.22% higher than that of the wild type (7.51 g L−1). However, deletion of proline dehydrogenase gene ycgM had no effect on γ-PGA synthesis. Furthermore, a 2.92-fold higher P5C content (19.24 μmol gDCW−1) was detected in the ycgN deficient strain WXΔycgN, while the P5C levels of WXΔycgM and the double mutant strain WXΔycgMN showed no difference, compared to WX-02. Moreover, the ROS level of WXΔycgN was increased by 1.18-fold, and addition of n-acetylcysteine (antioxidant) decreased its ROS level, which further reduced γ-PGA synthesis capability of WXΔycgN. Collectively, our results demonstrated that proline catabolism played an important role in maintaining ROS homeostasis, and deletion of ycgN-enhanced P5C accumulation, which induced a transient ROS signal to promote γ-PGA synthesis in B. licheniformis.
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Funding
This work was supported by the National Program on Key Basic Research Project (973 Program, No. 2015CB150505), the Technical Innovation Special Fund of Hubei Province (2018ACA149), and the Science and Technology Program of Wuhan (20160201010086).
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B Li, Z He, and S Chen designed the study. B Li carried out the molecular biology studies and construction of engineering strains. B Li and S Hu carried out the fermentation studies. B Li, D Cai, A Zhu, and S Chen analyzed the data and wrote the manuscript. All authors read and approved the final manuscript.
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Li, B., Cai, D., Hu, S. et al. Enhanced synthesis of poly gamma glutamic acid by increasing the intracellular reactive oxygen species in the Bacillus licheniformis Δ1-pyrroline-5-carboxylate dehydrogenase gene ycgN-deficient strain. Appl Microbiol Biotechnol 102, 10127–10137 (2018). https://doi.org/10.1007/s00253-018-9372-z
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DOI: https://doi.org/10.1007/s00253-018-9372-z