RRNPP-Type quorum-sensing systems regulate solvent formation, sporulation and cell motility in Clostridium saccharoperbutylacetonicum

Jun Feng, Wenming Zong, Pixiang Wang, Zhong Tian Zhang, Yanyan Gu, Mark Dougherty, Ilya Borovok*, Yi Wang

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Background: Clostridium saccharoperbutylacetonicum N1-4 (HMT) is a strictly anaerobic, spore-forming Gram-positive bacterium capable of hyper-butanol production through the well-known acetone-butanol-ethanol fermentation process. Recently, five putative RRNPP-Type QSSs (here designated as QSS1 to QSS5) were predicted in this bacterial strain, each of which comprises a putative RRNPP-Type regulator (QssR1 to QssR5) and a cognate signaling peptide precursor (QssP1 to QssP5). In addition, both proteins are encoded by the same operon. The functions of these multiple RRNPP-Type QSSs are unknown. Results: To elucidate the function of multiple RRNPP-Type QSSs as related to cell metabolism and solvent production in N1-4 (HMT), we constructed qssR-deficient mutants ΔR1, ΔR2, ΔR3 and ΔR5 through gene deletion using CRISPR-Cas9 and N1-4-dcas9-R4 (with the QssR4 expression suppressed using CRISPR-dCas9). We also constructed complementation strains by overexpressing the corresponding regulator gene. Based on systematic characterization, results indicate that QSS1, QSS2, QSS3, and QSS5 positively regulate the sol operon expression and thus solvent production, but they likely negatively regulate cell motility. Consequently, QSS4 might not directly regulate solvent production, but positively affect cell migration. In addition, QSS3 and QSS5 appear to positively regulate sporulation efficiency. Conclusions: Our study provides the first insights into the roles of multiple RRNPP-Type QSSs of C. saccharoperbutylacetonicum for the regulation of solvent production, cell motility, and sporulation. Results of this study expand our knowledge of how multiple paralogous QSSs are involved in the regulation of essential bacterial metabolism pathways.

Original languageEnglish
Article number84
JournalBiotechnology for Biofuels
Volume13
Issue number1
DOIs
StatePublished - 8 May 2020

Funding

FundersFunder number
USDA‑NIFA
U.S. Department of Energy
Office of Energy Efficiency and Renewable EnergyDE‑EE0008483
Office of Energy Efficiency and Renewable Energy
Auburn UniversityALA014‑1017025
Auburn University
Alabama Agricultural Experiment Station

    Keywords

    • Acetone-butanol-ethanol (ABE)
    • Butanol
    • CRISPR-Cas9
    • Cell motility
    • Clostridium saccharoperbutylacetonicum
    • RRNPP-Type quorum-sensing systems
    • Signaling peptide precursor
    • Sporulation

    Fingerprint

    Dive into the research topics of 'RRNPP-Type quorum-sensing systems regulate solvent formation, sporulation and cell motility in Clostridium saccharoperbutylacetonicum'. Together they form a unique fingerprint.

    Cite this