TY - JOUR
T1 - RRNPP-Type quorum-sensing systems regulate solvent formation, sporulation and cell motility in Clostridium saccharoperbutylacetonicum
AU - Feng, Jun
AU - Zong, Wenming
AU - Wang, Pixiang
AU - Zhang, Zhong Tian
AU - Gu, Yanyan
AU - Dougherty, Mark
AU - Borovok, Ilya
AU - Wang, Yi
N1 - Publisher Copyright:
© 2020 The Author(s).
PY - 2020/5/8
Y1 - 2020/5/8
N2 - 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.
AB - 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.
KW - Acetone-butanol-ethanol (ABE)
KW - Butanol
KW - CRISPR-Cas9
KW - Cell motility
KW - Clostridium saccharoperbutylacetonicum
KW - RRNPP-Type quorum-sensing systems
KW - Signaling peptide precursor
KW - Sporulation
UR - http://www.scopus.com/inward/record.url?scp=85085253421&partnerID=8YFLogxK
U2 - 10.1186/s13068-020-01723-x
DO - 10.1186/s13068-020-01723-x
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C2 - 32411297
AN - SCOPUS:85085253421
SN - 1754-6834
VL - 13
JO - Biotechnology for Biofuels
JF - Biotechnology for Biofuels
IS - 1
M1 - 84
ER -