scholarly journals RRNPP-type quorum sensing affects solvent formation and sporulation in Clostridium acetobutylicum

Microbiology ◽  
2020 ◽  
Vol 166 (6) ◽  
pp. 579-592 ◽  
Author(s):  
Ann-Kathrin Kotte ◽  
Oliver Severn ◽  
Zak Bean ◽  
Katrin Schwarz ◽  
Nigel P. Minton ◽  
...  

The strictly anaerobic bacterium Clostridium acetobutylicum is well known for its ability to convert sugars into organic acids and solvents, most notably the potential biofuel butanol. However, the regulation of its fermentation metabolism, in particular the shift from acid to solvent production, remains poorly understood. The aim of this study was to investigate whether cell–cell communication plays a role in controlling the timing of this shift or the extent of solvent formation. Analysis of the available C. acetobutylicum genome sequences revealed the presence of eight putative RRNPP-type quorum-sensing systems, here designated qssA to qssH, each consisting of an RRNPP-type regulator gene followed by a small open reading frame encoding a putative signalling peptide precursor. The identified regulator and signal peptide precursor genes were designated qsrA to qsrH and qspA to qspH, respectively. Triplicate regulator mutants were generated in strain ATCC 824 for each of the eight systems and screened for phenotypic changes. The qsrB mutants showed increased solvent formation during early solventogenesis and hence the QssB system was selected for further characterization. Overexpression of qsrB severely reduced solvent and endospore formation and this effect could be overcome by adding short synthetic peptides to the culture medium representing a specific region of the QspB signalling peptide precursor. In addition, overexpression of qspB increased the production of acetone and butanol and the initial (48 h) titre of heat-resistant endospores. Together, these findings establish a role for QssB quorum sensing in the regulation of early solventogenesis and sporulation in C. acetobutylicum .

2017 ◽  
Author(s):  
Ann-Kathrin Kotte ◽  
Oliver Severn ◽  
Zak Bean ◽  
Katrin Schwarz ◽  
Nigel P. Minton ◽  
...  

ABSTRACTThe strictly anaerobic bacteriumClostridium acetobutylicumis well known for its ability to convert sugars into organic acids and solvents, most notably the potential biofuel butanol. However, the regulation of its fermentation metabolism, in particular the shift from acid to solvent production, remains poorly understood. The aim of this study was to investigate whether cell-cell communication plays a role in controlling the timing of this shift or the extent of solvent formation. Analysis of the availableC. acetobutylicumgenome sequences revealed the presence of eight putative RNPP-type quorum sensing systems, here designatedqssAtoqssH, each consisting of RNPP-type regulator gene followed by a small open reading frame encoding a putative signalling peptide precursor. The identified regulator and signal peptide precursor genes were designatedqsrAtoqsrHandqspAtoqspH, respectively. Triplicate regulator mutants were generated in strain ATCC 824 for each of the eight systems and screened for phenotypic changes. TheqsrBmutants showed increased solvent formation during early solventogenesis and hence the QssB system was selected for further characterisation. Overexpression ofqsrBseverely reduced solvent and endospore formation and this effect could be overcome by adding short synthetic peptides to the culture medium representing a specific region of the QspB signalling peptide precursor. In addition, overexpression ofqspBincreased the production of acetone and butanol and the initial (48-hour) titre of heat-resistant endospores. Together, these findings establish a role for QssB quorum sensing in the regulation of early solventogenesis and sporulation inC. acetobutylicum.


2013 ◽  
Vol 63 (Pt_7) ◽  
pp. 2396-2404 ◽  
Author(s):  
Ana Faria Tomás ◽  
Dimitar Karakashev ◽  
Irini Angelidaki

An extremely thermophilic, xylanolytic, spore-forming and strictly anaerobic bacterium, strain DTU01T, was isolated from a continuously stirred tank reactor fed with xylose and household waste. Cells stained Gram-negative and were rod-shaped (0.5–2 µm in length). Spores were terminal with a diameter of approximately 0.5 µm. Optimal growth occurred at 70 °C and pH 7, with a maximum growth rate of 0.1 h−1. DNA G+C content was 34.2 mol%. Strain DTU01T could ferment arabinose, cellobiose, fructose, galactose, glucose, lactose, mannitol, mannose, melibiose, pectin, starch, sucrose, xylan, yeast extract and xylose, but not cellulose, Avicel, inositol, inulin, glycerol, rhamnose, acetate, lactate, ethanol, butanol or peptone. Ethanol was the major fermentation product and a maximum yield of 1.39 mol ethanol per mol xylose was achieved when sulfite was added to the cultivation medium. Thiosulfate, but not sulfate, nitrate or nitrite, could be used as electron acceptor. On the basis of 16S rRNA gene sequence similarity, strain DTU01T was shown to be closely related to Thermoanaerobacter mathranii A3T, Thermoanaerobacter italicus Ab9T and Thermoanaerobacter thermocopriae JT3-3T, with 98–99 % similarity. Despite this, the physiological and phylogenetic differences (DNA G+C content, substrate utilization, electron acceptors, phylogenetic distance and isolation site) allow for the proposal of strain DTU01T as a representative of a novel species within the genus Thermoanaerobacter , for which the name Thermoanaerobacter pentosaceus sp. nov. is proposed, with the type strain DTU01T ( = DSM 25963T = KCTC 4529T = VKM B-2752T = CECT 8142T).


2013 ◽  
Vol 63 (Pt_2) ◽  
pp. 442-448 ◽  
Author(s):  
Samson Viulu ◽  
Kohei Nakamura ◽  
Yurina Okada ◽  
Sakiko Saitou ◽  
Kazuhiro Takamizawa

A novel species of Fe(III)-reducing bacterium, designated strain OSK6T, belonging to the genus Geobacter , was isolated from lotus field mud in Japan. Strain OSK6T was isolated using a solid medium containing acetate, Fe(III)-nitrilotriacetate (NTA) and gellan gum. The isolate is a strictly anaerobic, Gram-negative, motile, straight rod-shaped bacterium, 0.6–1.9 µm long and 0.2–0.4 µm wide. The growth of the isolate occurred at 20–40 °C with optima of 30–37 °C and pH 6.5–7.5 in the presence of up to 0.5 g NaCl l−1. The G+C content of the genomic DNA was determined by HPLC to be 59.7 mol%. The major respiratory quinone was MK-8. The major fatty acids were 16 : 1ω7c and 16 : 0. Strain OSK6T was able to grow with Fe(III)-NTA, ferric citrate, amorphous iron (III) hydroxide and nitrate, but not with fumarate, malate or sulfate as electron acceptors. Among examined substrates grown with Fe(III)-NTA, the isolate grew on acetate, lactate, pyruvate and succinate. Analysis of the near full-length 16S rRNA gene sequence revealed that strain OSK6T is closely related to Geobacter daltonii and Geobacter toluenoxydans with 95.6 % similarity to the type strains of these species. On the basis of phylogenetic analysis and physiological tests, strain OSK6T is described as a representative of a novel species, Geobacter luticola sp. nov.; the type strain is OSK6T ( = DSM 24905T = JCM 17780T).


2020 ◽  
Vol 70 (9) ◽  
pp. 4851-4858 ◽  
Author(s):  
Tiphaine Le Roy ◽  
Patrick Van der Smissen ◽  
Adrien Paquot ◽  
Nathalie Delzenne ◽  
Giulio G. Muccioli ◽  
...  

A strictly anaerobic, Gram-stain-negative, non-spore-forming, non-motile, non-pigmented bacterium, strain J115T, was isolated from human faeces. Cells of strain J115T were straight rods, generally 1.8–3.0 µm, but could be up to 18 µm long. Growth occurred below 2 % (w/v) NaCl and 2 % (v/v) bile. Strain J115T produced acid from myo-inositol but not from d-glucose, d-ribose or d-xylose. Butyric acid was the major end-product from myo-inositol. The genomic DNA G+C content was 58.92 mol%. Phylogenetic analysis based on 16S rRNA gene sequencing indicated that the closest cultivated neighbours of strain J115T were Oscillibacter ruminantium GH1T (95.4 % similarity) and Oscillibacter valericigenes Sjm18-20T (94.1 %). Strain J115T was also related to the not-yet-cultured bacterium Oscillospira guilliermondii (92–93 % similarity). Coherently with the 16S rRNA gene sequence results, the ANI scores don't have units of strain J115T to O. ruminantium GH1T and O. valericigenes Sjm18-20T were 73.37 and 73.24, respectively, while in silico estimations of DNA–DNA hybridization were both 20.4 %, with confidence intervals of 18.2–22.9 % and 18.2–22.8 %, respectively. The major fatty acids were iso-C15 : 0 (24.2 %), C18 : 0 DMA (18.4 %), anteiso-C15 : 0 (15.2 %) and C16 : 0 DMA (7.6 %). No respiratory quinone was detected. Based on phenotypic features and phylogenetic position, it is proposed that this isolate represents a novel species in a new genus, Dysosmobacter welbionis gen. nov., sp. nov. The type strain of Dysosmobacter welbionis is J115T (DSM 106889T=LMG 30601T).


2014 ◽  
Vol 64 (Pt_9) ◽  
pp. 2986-2991 ◽  
Author(s):  
Xiao-Li Su ◽  
Qi Tian ◽  
Jie Zhang ◽  
Xian-Zheng Yuan ◽  
Xiao-Shuang Shi ◽  
...  

A strictly anaerobic, mesophilic, carbohydrate-fermenting, hydrogen-producing bacterium, designated strain RL-CT, was isolated from a reed swamp in China. Cells were Gram-stain-negative, catalase-negative, non-spore-forming, non-motile rods measuring 0.7–1.0 µm in width and 3.0–8.0 µm in length. The optimum temperature for growth of strain RL-CT was 37 °C (range 25–40 °C) and pH 7.0–7.5 (range pH 5.7–8.0). The strain could grow fermentatively on yeast extract, tryptone, arabinose, glucose, galactose, mannose, maltose, lactose, glycogen, pectin and starch. The main end products of glucose fermentation were acetate, H2 and CO2. Organic acids, alcohols and amino acids were not utilized for growth. Yeast extract was not required for growth; however, it stimulated growth slightly. Nitrate, sulfate, sulfite, thiosulfate, elemental sulfur and Fe(III) nitrilotriacetate were not reduced as terminal electron acceptors. Aesculin was hydrolysed but not gelatin. Indole and H2S were produced from yeast extract. The G+C content of the genomic DNA was 51.2 mol%. The major cellular fatty acids were iso-C15 : 0, anteiso-C15 : 0 and C16 : 0. The most abundant polar lipid of strain RL-CT was phosphatidylethanolamine. 16S rRNA gene sequence analysis revealed that the isolate belongs to the uncultured Blvii28 wastewater-sludge group (http://www.arb-silva.de/) in the family Rikenellaceae of the phylum Bacteroidetes, and shared low sequence similarities with the related species Alistipes shahii WAL 8301T (81.8 %), Rikenella microfusus ATCC 29728T (81.7 %) and Anaerocella delicata WN081T (80.9 %). On the basis of these data, a novel species in a new genus of the family Rikenellaceae is proposed, Acetobacteroides hydrogenigenes gen. nov., sp. nov. The type strain of the type species is RL-CT ( = JCM 17603T = DSM 24657T = CGMCC 1.5173T).


2014 ◽  
Vol 64 (Pt_9) ◽  
pp. 3081-3086 ◽  
Author(s):  
Daisuke Suzuki ◽  
Zhiling Li ◽  
Xinxin Cui ◽  
Chunfung Zhang ◽  
Arata Katayama

A strictly anaerobic, mesophilic, sulfate-reducing bacterial strain (DST), isolated from river sediment contaminated with volatile organic compounds, was characterized phenotypically and phylogenetically. Cells were Gram-reaction-negative, non-motile short rods. For growth, optimum NaCl concentration was 0.9 g l−1, optimum temperature was 30 °C and optimum pH was 7.2. Strain DST utilized phenol, benzoate, 4-hydroxybenzoate, 4-methylphenol, 4-chlorophenol, acetate, butyrate and pyruvate as electron donors for sulfate reduction. Electron donors were completely oxidized. Strain DST did not utilize sulfite, thiosulfate or nitrate as electron acceptors. The genomic DNA G+C content of strain DST was 58.9 mol%. Major cellular fatty acids were iso-C14 : 0, anteiso-C15 : 0 and C18 : 1ω7c. Phylogenetic analyses based on the 16S rRNA gene indicated its closest relatives were strains of Desulfobacterium anilini (about 98–99 % sequence similarity) but the DNA–DNA hybridization value with Desulfobacterium anilini Ani1T was around 40 %. Although strain DST and its relatives shared most phenotypic and chemotaxonomic characteristics, the utilization of 4-chlorophenol, the range of electron acceptors and the optimum growth conditions differed. Strain DST is closely related to strains of Desulfobacterium anilini , but constitutes a different species within the genus. Based on phylogeny, phenotypic characteristics and chemotaxonomic characteristics, strain DST and Desulfobacterium anilini were clearly different from strains of other species of the genus Desulfobacterium . We thus propose the reclassification of Desulfobacterium anilini within a new genus, Desulfatiglans gen. nov., as Desulfatiglans anilini comb. nov. We also propose Desulfatiglans parachlorophenolica sp. nov. to accommodate strain DST. The type strain is DST ( = JCM 19179T = DSM 27197T).


2013 ◽  
Vol 63 (Pt_1) ◽  
pp. 109-113 ◽  
Author(s):  
Zhao-Ming Gao ◽  
Xin Liu ◽  
Xi-Ying Zhang ◽  
Ling-Wei Ruan

A moderately thermophilic and strictly anaerobic bacterium, designated HS1T, was isolated from offshore hot spring sediment in Xiamen, China. Cells were Gram-negative, catalase-positive, oxidase-negative, slender and flexible rods without flagella. The strain could grow at 35–55 °C (optimum at 50 °C) and in 1–8 % NaCl (w/v; optimum 2–4 %). Phylogenetic analysis based on 16S rRNA gene sequences showed that strain HS1T was affiliated with the family Marinilabiliaceae and shared a distant relationship with the previously described genera. The isolate was most closely related to Anaerophaga thermohalophila Fru22T with 16S rRNA gene sequence similarity of 92.4 %, followed by the other members of the family Marinilabiliaceae with 88.7–91.1 % similarity. The dominant cellular fatty acids were iso-C15 : 0 and anteiso-C15 : 0. The predominant quinone was MK-7. The major polar lipids were phosphatidylethanolamine (PE) and an unknown polar lipid. The genomic DNA G+C content was 38.7 mol%. Besides the phylogenetically distant relationship, strain HS1T was obviously distinguished from the most closely related genera in several phenotypic properties including colony colour and pigment production, optimal temperature, optimal NaCl, relation to O2, bicarbonate/carbonate requirement, catalase activity, nitrate reduction, fermentation products and cellular fatty acid profile. Based on the phenotypic and phylogenetic data, strain HS1T represents a novel species of a new genus, for which the name Thermophagus xiamenensis gen. nov., sp. nov. is proposed. The type strain of the type species is HS1T ( = DSM 19012T = CGMCCC 1.5071T).


2013 ◽  
Vol 63 (Pt_6) ◽  
pp. 1947-1950 ◽  
Author(s):  
Belkis Fraj ◽  
Wajdi Ben Hania ◽  
Anne Postec ◽  
Moktar Hamdi ◽  
Bernard Ollivier ◽  
...  

A strictly anaerobic, moderately thermophilic, halotolerant rod, designated BELH25T, was isolated from a water sample of a Tunisian hot spring. Cells were non-motile, 2–6 µm long and 0.4–0.6 µm wide, appearing singly or in pairs. The isolate grew at 45–70 °C (optimum 55 °C), at pH 6.2–8.0 (optimum pH 7.0) and with 0–4 % NaCl (optimum 0–2.0 %). Sulfate, thiosulfate, elemental sulfur, sulfite, nitrate and nitrite were not used as terminal electron acceptors. Strain BELH25T used cellobiose, fructose, galactose, glucose, maltose, mannose, sucrose, starch and yeast extract as electron donors. The main fermentation products from glucose metabolism were formate, acetate, ethanol and CO2. The predominant cellular fatty acids were iso-C15 : 0, iso-C17 : 0 and anteiso-C15 : 0. The DNA G+C content was 37.2 mol%. Phylogenetic analysis of the 16S rRNA gene sequence indicated that strain BELH25T was most closely related to Caloramator viterbiensis JW/MS-VS5T and Fervidicella metallireducens AeBT (92.2 and 92.1 % sequence similarity, respectively), and the isolate was positioned approximately equidistantly between these genera. Based on phenotypic, phylogenetic and chemotaxonomic characteristics, strain BELH25T is proposed to be a member of a novel species of a novel genus within the order Clostridiales , family Clostridiaceae , for which the name Fonticella tunisiensis gen. nov., sp. nov. is proposed. The type strain of the type species is BELH25T ( = DSM 24455T = JCM 17559T).


2021 ◽  
Vol 3 (3) ◽  
Author(s):  
Aya Ahmad Elnegery ◽  
Wafaa Kamel Mowafy ◽  
Tarek Ahmed Zahra ◽  
Noha Tharwat Abou El-Khier

Background. Pseudomonas aeruginosa (P. aeruginosa) is an opportunistic pathogen responsible for burn-wound infection. High incidence, infection severity and increasing resistance characterize P. aeruginosa -induced burn infection. Purpose. To estimate quorum-sensing (QS)-dependent virulence factors of P. aeruginosa isolates from burn wounds and correlate it to the presence of QS genes. Methods. A cross-sectional descriptive study included 50 P . aeruginosa isolates from burn patients in Mansoura University Plastic and Burn Hospital, Egypt. Antibiotic sensitivity tests were done. All isolates were tested for their ability to produce biofilm using a micro-titration assay method. Protease, pyocyanin and rhamnolipid virulence factors were determined using skimmed milk agar, King’s A medium and CTAB agar test, respectively. The identity of QS lasR and rhlR genes was confirmed using PCR. Results. In total, 86 % of isolates had proteolytic activity. Production of pyocyanin pigment was manifested in 66 % of isolates. Altogether, 76 % of isolates were rhamnolipid producers. Biofilm formation was detected in 96 % of isolates. QS lasR and rhlR genes were harboured by nearly all isolates except three isolates were negative for both lasR and rhlR genes and two isolates were positive for lasR gene and negative for rhlR gene. Forty-nine isolates were considered as extremely QS-proficient strains as they produced QS-dependent virulence factors. In contrast, one isolate was a QS deficient strain. Conclusions. QS affects P. aeruginosa virulence-factor production and biofilm in burn wounds. Isolates containing lasR and rhlR seem to be a crucial regulator of virulence factors and biofilm formation in P. aeruginosa whereas the lasR gene positively regulates biofilm formation, proteolytic activity, pyocyanin production and rhamnolipid biosurfactant synthesis. The QS regulatory RhlR gene affects protease and rhamnolipid production positively.


2020 ◽  
Vol 70 (8) ◽  
pp. 4767-4773 ◽  
Author(s):  
Jong-Shian Liou ◽  
Chien-Hsun Huang ◽  
Nao Ikeyama ◽  
Ai-Yun Lee ◽  
I-Ching Chen ◽  
...  

A strictly anaerobic predominant bacterium, designated as strain gm001T, was isolated from a freshly voided faecal sample collected from a healthy Taiwanese adult. Cells were Gram-stain-negative rods, non-motile and non-spore-forming. Strain gm001T was identified as a member of the genus Prevotella , and a comparison of 16S rRNA and hsp60 gene sequences revealed sequence similarities of 98.5 and 93.3 %, respectively, demonstrating that it was most closely related to the type strain of Prevotella copri . Phylogenomic tree analysis indicated that the gm001T cluster is an independent lineage of P. copri DSM 18205T. The average nucleotide identity, digital DNA‒DNA hybridization and average amino acid identity values between strain gm001T and P. copri DSM 18205T were 80.9, 28.6 and 83.8 %, respectively, which were clearly lower than the species delineation thresholds. The species-specific genes of this novel species were also identified on the basis of pan-genomic analysis. The predominant menaquinones were MK-11 and MK-12, and the predominant fatty acids were anteiso-C15 : 0, C15 : 0 and iso-C15 : 0. Acetate and succinate were produced from glucose as metabolic end products. Taken together, the results indicate that strain gm001T represents a novel species of the genus Prevotella , for which the name Prevotella hominis sp. nov. is proposed. The type strain is gm001T (=BCRC 81118T=JCM 33280T).


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