Butyrate Consumption Research Articles

Mixotrophic cultivation of microalgae-bacteria consortia enhances dark fermentation effluent treatment

Dark fermentation (DF) is a waste treatment bioprocess which produces biohydrogen and volatile fatty acids (VFAs) such as acetate or butyrate. DF can be coupled with microalgae cultivation, allowing VFA conversion into valuable biomass. Nevertheless, the process is hindered by slow butyrate consumption. In this study, novel artificial microalgae-bacteria consortia were used as a strategy to accelerate butyrate removal. Three microalgal strains with various trophic metabolisms, Chlorella sorokiniana, Euglena gracilis and Ochromonas danica, were cultivated on DF effluent that was either sterile or contained endogenous bacteria. Bacteria did not impact microalgal biomass production of C. sorokiniana or E. gracilis while accelerating butyrate removal rates 2 to 10-fold. O. danica greatly impacted microbial diversity, probably due to its phagotrophic metabolism. These results show that bacteria in organic rich effluents can greatly aid in substrate removal while allowing microalgal growth, inspiring bioprocesses coupling raw fermentation effluents with microalgae biomass production and valorization.

Transcriptomic evidence for an energetically advantageous relationship between Syntrophomonas wolfei and Methanothrix soehngenii.

Syntrophic interactions are key in anaerobic food chains, facilitating the conversion of complex organic matter into methane. A typical example involves acetogenic bacteria converting fatty acids (e.g., butyrate and propionate), a process thermodynamically reliant on H2 consumption by microorganisms such as methanogens. While most studies focus on H2-interspecies transfer between these groups, knowledge on acetate cross-feeding in anaerobic systems is lacking. This study investigated butyrate oxidation by co-cultures of Syntrophomonas wolfei and Methanospirillum hungatei, both with and without the addition of the acetate scavenger Methanothrix soehngenii. Growth and gene expression patterns of S. wolfei and M. hungatei were followed in the two conditions. Although butyrate consumption rates remained constant, genes in the butyrate degradation pathway of S. wolfei were less expressed in the presence of M. soehngenii, including genes involved in reverse electron transport. Higher expression of a type IV-pili operon in S. wolfei hints to the potential for direct interspecies electron transfer between S. wolfei and M. soehngenii and an energetically advantageous relationship between the two microorganisms. Overall, the presence of the acetate scavenger M. soehngenii positively influenced the energy metabolism of S. wolfei and highlighted the relevance of including acetate scavengers when investigating syntrophic fatty acid degradation.

Metabolic adaptation to consume butyrate under prolonged resource exhaustion.

Bacteria must often survive following the exhaustion of their external growth resources. Fitting with this need, many bacterial species that cannot sporulate, can enter a state known as long term stationary phase (LTSP) in which they can persist for years within spent media. Several recent studies have revealed the dynamics of genetic adaptation of Escherichia coli under LTSP. Yet, the metabolic consequences of such genetic adaptation were not addressed. Here, we characterized the metabolic changes LTSP populations experience, over the first 32 days under LTSP. This allowed us to link genetic adaptations observed in a convergent manner across LTSP populations back to their metabolic adaptive effect. Specifically, we demonstrate that through the acquisition of mutations combinations in specific sets of metabolic genes, E. coli acquires the ability to consume the short chain fatty acid butyrate. Intriguingly, this fatty acid is not initially present within the rich media we used in this study. Instead, it is E. coli itself that produces butyrate during its initial growth within fresh rich media. The mutations that enable butyrate consumption allow E. coli to grow on butyrate. However, the clones carrying these mutations rapidly decrease in frequency, once the butyrate is consumed, likely reflecting an associated cost to fitness. Yet despite this, E. coli populations show a remarkable capability of maintaining these genotypes at low frequency, as standing variation. This in turn allows them to more rapidly re-adapt to consume butyrate, once it again becomes available to them.

Role of Sodium Butyrate Supplement on Reducing Hepatotoxicity Induced by Lead Acetate in Rats


 
 
 
 Lead has always been a health risk in developing countries. Lead severely affects liver function. Butyrate is effective in treating inflammatory disorders in animals. Thus, this study aimed to determine whether sodium butyrate mitigates lead acetate-induced hepatotoxicity. In this research, 40 adult female albino rats were randomly assigned to one of four treatment groups for a duration of 35 days as follows: group 1 served as a control, group 2 received sodium butyrate (SB) orally at 200 mg/kg daily, group 3 received lead acetate (LA) orally at 50 mg/kg daily, and group 4 received both SB and LA (SB+LA) orally. Blood was collected for complete blood picture (CBC) and some serum biochemical evaluations. Liver samples were collected for histopathological examination. The rats that exposed to lead acetate showed a significant (P<0.05) elevation in globulin, total bilirubin, total serum protein, and total white blood cells with a decrease in total red blood cells, haemoglobin, and packed cell volume, while weight gain shows a significant (P<0.05) decrease in this group. Histologically showed pre-vascular infiltration of the nuclear cell. Body weight of Rat's gavage with sodium butyrate showed a substantial (P<0.05) increase, as well as there, were improvements in red blood cells RBC, haemoglobin, and packed cell volume PCV with the normal histological structure of the liver and no pathological lesion in hepatocyte. The fourth group (SB+LA) showed a significant (P<0.05) decrease in total bilirubin, indirect bilirubin, and total white blood cells, while other tests in this group showed nearly the control group as a result of the effect of SB. In conclusion, sodium butyrate consumption effectively reduces the harmful effects of lead acetate and prevents liver damage.
 
 
 

Intensification of Hydrogen Production by a Co-culture ofSyntrophomonas wolfei and Rhodopseudomonas palustrisEmploying High Concentrations of Butyrate as a Substrate.

The purpose of this study is to present an effective form of developing a sequential dark (DF) and photo (PF) fermentation using volatile fatty acids (VFAs) and nitrogen compounds as bonding components between both metabolic networks of microbial growing in each fermentation. A simultaneous (co-)culture of Syntrophomonas wolfei (with its ability to consume butyrate and produce acetate) and Rhodopseudomonas palustris (that can use the produced acetate as a carbon source) performed a syntrophic metabolism. The former bacteria consumed the acetate/butyrate mixture reducing the butyrate concentration below 2.0g/L, permitting Rhodopseudomonas palustris to produce hydrogen. Considering that the inoculum composition (Syntrophomonas wolfei/Rhodopseudomonas palustris) and the nitrogen source (yeast extract) define the microbial biomass specific productivity and the butyrate consumption, a response surface methodology defined the best inoculum design and yeast extract (YE) yielding to the highest biomass concentration of 1.1g/L after 380.00h. A second culture process (without a nitrogen source) showed the biomass produced in the previous culture process yields to produce a total cumulated hydrogen concentration of 3.4 mmol. This value was not obtained previously with the pure strain Rhodopseudomonas palustris if the culture medium contained butyrate concentration above 2.0g/L, representing a contribution to the sequential fermentation scheme based on DF and PF.

Improvement of Adaptive Saanen Goat Milk Production and Reproduction Fed Diet Supplemented with Indigofera zollingeriana Leaf Meal

The objective of this research was to examine the effect of the addition of Indigofera zollingeriana leaf meal (IZLM) in diets on milk production and fermentation products of adaptive Saanen goat. The research was conducted at Balai Besar Pembibitan Ternak Unggul dan Hijauan Pakan Ternak (BBPTUHPT) or or the Center for Superior Animal Breeding and Forage Animal Feed, Baturraden, Central Java. A total of 18 first lactating adaptive Saanen goats with a body weight of 34.83±7.13 and aged 20-24 months were used in this study. The goats were kept in individual pens and grouped into 6 groups based on body weight and each group were randomized to receive three kinds of concentrate substitution with IZLM of 0%, 10% and 20 % of dry matter (DM) concentrates, for A, B and C treatments, respectively. Therefore, this research was designed according to randomized block design. Dry matter intake (DMI) of each goat was 4.5% of live weight with dry matter (DM) ratio of forage and concentrate were 60:40. The variables measured were DMI, partial volatile fatty acid (VFA), energy conversion efficiency of glucose into VFA (ECEVFA), methane gas, total protozoa, milk production and the first estrus after kidding. Analysis of variance showed that the treatment had a significant effect (P <0.05) on propionate, milk production and fat, but no significant effect (P> 0.05) on consumption of DM, acetate, butyrate, EKVFA, methane gas, lactose and protein milk. Increasing the IZLM substitution level enhanced linearly (P<0.05) on production of milk and milk fat, while the propionate concentrate responded to quadratic (P<0.05). Milk production increased at IZLM level of 20%, while goat in this group resulted only 16.70% estrous after birth compared to goat group received 10% IZLM level resulting 50% of estrus goat. The results of this study concluded that the recommended level of using IZLM as a concentrate substitute was only 10%.

Stimulating Effect of Trichococcus flocculiformis on a Coculture of Syntrophomonas wolfei and Methanospirillum hungatei.

ABSTRACTSyntrophic anaerobic consortia comprised of fatty acid-degrading bacteria and hydrogen/formate-scavenging methanogenic archaea are of central importance for balanced and resilient natural and manufactured ecosystems: anoxic sediments, soils, and wastewater treatment bioreactors. Previously published studies investigated interaction between the syntrophic bi-cultures, but little information is available on the influence of fermentative bacteria on syntrophic fatty acid oxidation, even though fermentative organisms are always present together with syntrophic partners in the above-mentioned ecosystems. Here, we present experimental observations of stimulated butyrate oxidation and methane generation by a coculture of Syntrophomonas wolfei with any of the following methanogens: Methanospirillum hungatei, Methanobrevibacter arboriphilus, or Methanobacterium formicicum due to the addition of a fermentative Trichococcus flocculiformis strain ES5. The addition of T. flocculiformis ES5 to the syntrophic cocultures led to an increase in the rates of butyrate consumption (120%) and volumetric methane production (150%). Scanning electron microscopy of the most positively affected coculture (S. wolfei, M. hungatei, and T. flocculiformis ES5) revealed a tendency of T. flocculiformis ES5 to aggregate with the syntrophic partners. Analysis of coculture’s proteome with or without addition of the fermentative bacterium points to a potential link with signal transducing systems of M. hungatei, as well as activation of additional butyryl coenzyme A dehydrogenase and an electron transfer flavoprotein in S. wolfei.IMPORTANCE Results from the present study open doors to fascinating research on complex microbial cultures in anaerobic environments (of biotechnological and ecological relevance). Such studies of defined mixed populations are critical to understanding the highly intertwined natural and engineered microbial systems and to developing more reliable and trustable metabolic models. By investigating the existing cultured microbial consortia, like the ones described here, we can acquire knowledge on microbial interactions that go beyond “who feeds whom” relations but yet benefit the parties involved. Transfer of signaling compounds and stimulation of gene expression are examples of indirect influence that members of mixed communities can exert on each other. Understanding such microbial relationships will enable development of new sustainable biotechnologies with mixed microbial cocultures and contribute to the general understanding of the complex natural microbial interactions.

Nivolumab/ipilimumab with or without CBM588 in metastatic renal cell carcinoma: A randomized phase Ib study and the evolution of the functionality of microbial communities with treatment.

371 Background: The role of gut microbial composition as a determinant of clinical outcomes has been well established in several cancers, including metastatic renal cell carcinoma (mRCC) (Routy et al Science 2018). A growing body of evidence suggests that examining the metabolic function of microbial communities may provide a more insightful understanding of these associations (Helmink et al Nature Medicine 2019). Herein, we aimed to examine the effect of nivolumab/ipilimumab with or without CBM588 on clinical outcomes and gut microbiome functionality. Methods: Treatment naïve mRCC pts with clear cell and/or sarcomatoid histology and IMDC intermediate/high risk disease were enrolled and randomized into receiving nivolumab/ipilimumab or nivolumab/ipilimumab with CBM588 in 1:2 fashion. Whole metagenome sequencing was performed on stool samples collected at baseline and week 12. Generated MetaPhlan 3.0 data was run through HUMAnN 3.0 to identify differentially expressed metabolic pathways between two timepoints in each arm and with respect to treatment response. Results: A total of 30 pts were enrolled and randomized, and 29 pts were eligible for analysis as one patient was excluded as tumor tissue next-generation sequencing revealed genomic alterations pathognomonic for sarcoma after initiation of protocol-based therapy. Median age of the participants was 66 years, 21 pts (72%) were male, 10 pts (34%) had sarcomatoid features, and 29 pts (45%) had prior nephrectomy. Objective response was achieved in 58% and 20% of the pts in nivolumab/ipilimumab/CBM-588 and nivolumab/ipilimumab arm, respectively. Significant changes in 40 metabolic pathways (37 with upregulation and 3 with downregulation) in nivolumab/ipilimumab arm and 52 metabolic pathways (49 with downregulation and 3 with upregulation) in nivolumab/ipilimumab with CBM588 arm were identified. In detail, dTDP-β-L-rhamnose biosynthesis, L-lysine biosynthesis II and superpathway of pyrimidine ribonucleosides degradation pathways were found upregulated while O-antigen building blocks biosynthesis (E. coli) pathway was found downregulated after treatment with nivolumab/ipilimumab and CBM588 (p = 0.001, p = 0.007, p = 0.037, p = 0.005 respectively). Heatmaps detailing the dynamics of metabolic pathway expressions in regard to response in each arm will be presented. Conclusions: We observed an increase in the activity of the pathways associated with butyrate consumption and a resultant decrease in glycolytic dependence. Further, suppression of the pathogenic E. coli function was observed, suggesting a role for CBM588 in protection from pathogenic species. Our findings provide mechanistic evidence for the effect of the addition of CBM588 to nivolumab/ipilimumab on gut microbiome function and resultant improvement in clinical outcomes in mRCC. Clinical trial information: NCT03829111.

Regulating light, oxygen and volatile fatty acids to boost the productivity of purple bacteria biomass, protein and co-enzyme Q10

Purple non‑sulfur bacteria (PNSB) possess significant potential for bioresource recovery from wastewater. Effective operational tools are needed to boost productivity and direct the PNSB biomass towards abundant value-added substances (e.g., protein and co-enzyme Q10, CoQ10). This study aimed to investigate the impact of light, oxygen and volatile fatty acids (VFAs) on PNSB growth (i.e., Rhodobacter sphaeroides) and productivity of protein and CoQ10. Overall, the biomass yields and specific growth rates of PNSB were in the ranges of 0.57–1.08 g biomass g−1 CODremoved and 0.48–0.71 d−1, respectively. VFAs did not influence the biomass yield, yet acetate and VFA mixtures enhanced the specific growth rate with a factor of 1.2–1.5 compared to propionate and butyrate. The most PNSB biomass (1.08 g biomass g−1 CODremoved and 0.71 d−1) and the highest biomass quality (protein content of 609 mg g−1 dry cell weight (DCW) and CoQ10 content of 13.21 mg g−1 DCW) were obtained in the presence of VFA mixtures under natural light and microaerobic (low light alternated with darkness; dissolved oxygen (DO) between 0.5 and 1 mg L−1) conditions (vs. light anaerobic and dark aerobic cultivations). Further investigation on VFAs dynamics revealed that acetate was most rapidly consumed by PNSB in the individual VFA feeding (specific uptake rate of 0.76 g COD g−1 DCW d−1), while acetate as a co-substrate in the mixed VFAs feeding might accelerate the consumption of propionate and butyrate through providing additional cell metabolism precursor. Enzymes activities of succinate dehydrogenase and fructose-1,6-bisphosphatase as well as the concentration of photo pigments confirmed that light, oxygen and VFAs regulated the key enzymes in the energy metabolism and biomass synthesis to boost PNSB growth. These results provide a promising prospect for utilization of fermented waste stream for the harvest of PNSB biomass, protein and CoQ10.

Tolerance of photo-fermentative biohydrogen production system amended with biochar and nanoscale zero-valent iron to acidic environment

Anaerobic fermentation system is easy to become acidic due to the generation of small molecular acids, which will affect the metabolism of bacteria. Therefore, it is necessary to improve the acid resistance of system. In this work, the tolerance of photo-fermentative biohydrogen production system amended with biochar, nanoscale zero-valent iron (nZVI) and biochar + nZVI to acidic environment was studied. Results showed that additives improved the stability and performance of the photo fermentation. The best increment of biohydrogen from 0 to 286.83 ± 2.77 mL was obtained by adding biochar and nZVI together at the original pH of 4.5. The additive reduced the oxidation–reduction potential and promoted the consumption of acetate and butyrate. At initial pH of 5, 6 and 7, the highest biohydrogen yield of 361.02 ± 10.11, 419.36 ± 10.70 and 382.67 ± 25.08 mL was obtained by adding nZVI, respectively, representing 42%–44.45% increase compared with the control group under the same conditions.

An efficient system for intestinal on-site butyrate production using novel microbiome-derived esterases

Short-chain fatty acids, especially butyrate, play beneficial roles in sustaining gastrointestinal health. However, due to limitations associated with direct consumption of butyrate, there has been interest in using prodrugs of butyrate. Tributyrin (TB), a triglyceride composed of three butyrate molecules and a glycerol, is a well-studied precursor of butyrate. We screened a metagenome library consisting of 5760 bacterial artificial chromosome clones, with DNA inserts originating from mouse microbiomes, and identified two clones that efficiently hydrolyse TB into butyrate. Nucleotide sequence analysis indicated that inserts in these two clones are derived from unknown microbes. BLASTp analysis, however, revealed that each insert contains a gene homologous to acetylesterase or esterase genes, from Clostridium spp. and Bacteroides spp., respectively. Predicted structures of these two proteins both contain serine-histidine-aspartate catalytic triad, highly conserved in the family of esterases. Escherichia coli host expressing each of the two candidate genes invariably produced greater amounts of butyrate in the presence of TB. Importantly, administration of TB together with cloned E. coli cells alleviated inflammatory symptoms in a mouse model of acute colitis. Based on these results, we established an efficient on-site and real-time butyrate production system that releases butyrate in a controlled manner inside the intestine.

Production of acetoin from renewable resources under heterotrophic and mixotrophic conditions

This study aimed to reveal whether Cupriavidus necator H16 is suited for the production of acetoin based on the carboxylic acids acetate, butyrate and propionate under heterotrophic and mixotrophic conditions. The chosen production strain, lacking the polyhydroxybutyrate synthases phaC1 and phaC2, was revealed to be beneficiary for autotrophic acetoin production. Proteomic analysis of the strain determined that the deletions do indeed have a significant impact on pyruvate formation and its subsequent direction towards the introduced acetoin-synthesis pathway. Moreover, the strain was tested for its ability to use typical dark fermentation products under hetero- and mixotrophic conditions. Growth with butyrate and acetate led to low efficiencies, while 46.54% ±0.78 of the added propionate was converted into acetoin. Interestingly, mixotrophic conditions led to simultaneous consumption of acetate and butyrate with the gaseous substrates and lowered efficiency. In contrast, mixotrophic propionate consumption led to diauxic behavior and high carbon efficiency of 71.2% ±0.64.

Effects of a proinflammatory response on metabolic function of cultured, primary ruminal epithelial cells

Inflammation of ruminal epithelium may occur during ruminal acidosis as a result of translocation and interaction of ruminal epithelial cells (REC) with molecules such as lipopolysaccharide (LPS). Such inflammation has been reported to alter cellular processes such as nutrient absorption, metabolic regulation, and energy substrate utilization in other cell types but has not been investigated for REC. The objectives of this study were to investigate the effects of LPS on metabolism of short-chain fatty acids by primary REC, as well as investigating the effects of media containing short-chain fatty acids on the proinflammatory response. Ruminal papillae from 9 yearling Speckle Park beef heifers were used to isolate and culture primary REC. Cells were grown in minimum essential medium (MEM) for 12 d before use and then reseeded in 24-well culture plates. The study was conducted as a 2 × 2 factorial, where cells were grown in unaltered MEM (REG) or medium containing 2 mM butyrate and 5 mM propionate (SCFA) with (50,000 EU/mL; +LPS) or without LPS (-LPS) for 24 h. Supernatant samples were collected for analysis of glucose and SCFA consumption. Cells were collected to determine the expression of mRNA for genes associated with inflammation (TNF, IL1B, CXCL2, CXCL8, PTGS2, and TLR4), purinergic signaling (P2RX7, ADORAB2, and CD73), nutrient transport [SLC16A1 (MCT1), SLC16A3 (MCT4), SLC5A8, and MCU], and cell metabolism [ACAT1, SLC2A1 (GLUT1), IGFBP3, and IGFBP5]. Protein expression of TLR4 and ketogenic enzymes (BDH1 and HMGCS1) were also analyzed using flow cytometry. Statistical analysis was conducted with the MIXED model of SAS version 9.4 (SAS Institute Inc., Cary, NC) with medium, LPS exposure, and medium × LPS interaction as fixed effects and animal within plate as a random effect. Cells tended to consume more glucose when exposed to LPS as opposed to no LPS exposure (31.8 vs. 28.7 ± 2.7), but consumption of propionate and butyrate was not influenced by LPS. Expression of TNF and IL1B was upregulated when exposed to LPS, and expression of CXCL2 and CXCL8 increased following LPS exposure with SCFA (medium × LPS). For cells exposed to LPS, we found a downregulation of ACAT1 and IGFBP5 and an upregulation of SLC2A1, SLC16A3, MCU, and IGFBP3. Medium with SCFA led to greater expression of MCU. SLC16A1 was upregulated in cells incubated with SCFA and without LPS compared with the other groups. Protein expression of ketogenic enzymes was not affected; however, BDH1 mean fluorescence intensity (MFI) expression tended to be less in cells exposed to LPS. These data are interpreted to indicate that when REC are exposed to LPS, they may increase glucose metabolism. Moreover, transport of solutes was affected by SCFA in the medium and by exposure to LPS. Overall, the results suggest that metabolic function of REC in vitro is altered by a proinflammatory response, which may lead to a greater glucose requirement.

Effects of oral butyrate and inulin supplementation on inflammation-induced pyroptosis pathway in type 2 diabetes: A randomized, double-blind, placebo-controlled trial

PurposePyroptosis, a form of inflammatory programmed cell death, is activated in diabetic patients. This study was conducted to investigate the effects of daily consumption of sodium butyrate (NaBut) and high-performance (HP) inulin supplementation, individually or in combination, on the expression of pyroptosis-related genes, microRNA (miR) 146a-5p, miR-9-5p and biomarkers of oxidative stress in patients with type 2 diabetes (T2DM). MethodsIn this study, we conducted a randomized, double-blinded, placebo-controlled clinical involving sixty patients with type 2 diabetes. Participants received 600 mg/d of NaBut (group A), 10 g/d of HP inulin (group B), 600 mg/d of NaBut + 10 g/d of HP inulin (group C) or placebo (group D) for 45 consecutive days. We assessed the pyroptosis-related genes mRNA expression in peripheral blood mononuclear cells (PBMCs), as well as the plasmatic levels of miR-146a and miR-9 before and after the intervention. Moreover, blood samples of the patients at baseline and following the intervention were tested for total antioxidant capacity (TAC), superoxide dismutase (SOD) and catalase levels using enzyme-linked immunosorbent assay (ELISA). This study was registered on the Iranian Registry of Clinical Trials website (identifier: IRCT201605262017N29; https://www.irct.ir/). ResultsFollowing butyrate supplementation, the relative expression levels of TLR2/4, NF-κB1, Caspase-1, NLRP3, IL-1β & IL-18 were significantly downregulated (p < 0.05). Furthermore, butyrate and concomitant use of butyrate and inulin caused a significant increase in the fold change of miR-146a and miR-9 compared with the placebo group (p < 0.05). Interestingly, the changes in total antioxidant capacity (p = 0.047) and superoxide dismutase (p = 0.006) were significantly increased after butyrate and concomitant use of butyrate and inulin supplement, respectively. ConclusionIn summary, the change in expression level of miR-146a-5p and miR-9-5p due to butyrate supplementation may have a pivotal role in alleviating of diabetes via inhibiting pyroptosis by targeting TLR2 and NF-κB1. These microRNAs might be considered as potential therapeutic targets in the treatment of type 2 diabetes but further researches is required to prove the link.

Butyrate consumption of differentiated colonocytes in the upper crypt promotes homeostatic proliferation of stem and progenitor cells near the crypt base.

The mammalian intestinal tract epithelium is renewed every ~4–6 days, which is arguably the most rapidly turned over tissue in the body. Cellular turnover is facilitated by the intestinal epithelium being organized as crypts with highly proliferative stem cells and progenitor/transit-amplifying cells located near the crypt base and quiescent, terminally-differentiated cell types such as enterocytes, goblet cells, and enteroendocrine cells located above that undergo apoptosis and exfoliate into the lumen ( Figure 1A ). In recent years, tremendous progress has been made identifying Lgr5+ stem cells and crucial factors within the stem-cell niche such as Wnts, R-spondins, BMPs, Noggin, and the Notch pathway (1). This knowledge has been exploited to create 3D organoid culture systems that recapitulate the crypt with respect to stem cell renewal and their ability to give rise to descendants that represent each differentiated cell type.

Peat: home to novel syntrophic species that feed acetate- and hydrogen-scavenging methanogens.

Syntrophic bacteria drive the anaerobic degradation of certain fermentation products (e.g., butyrate, ethanol, propionate) to intermediary substrates (e.g., H2, formate, acetate) that yield methane at the ecosystem level. However, little is known about the in situ activities and identities of these syntrophs in peatlands, ecosystems that produce significant quantities of methane. The consumption of butyrate, ethanol or propionate by anoxic peat slurries at 5 and 15 °C yielded methane and CO2 as the sole accumulating products, indicating that the intermediates H2, formate and acetate were scavenged effectively by syntrophic methanogenic consortia. 16S rRNA stable isotope probing identified novel species/strains of Pelobacter and Syntrophomonas that syntrophically oxidized ethanol and butyrate, respectively. Propionate was syntrophically oxidized by novel species of Syntrophobacter and Smithella, genera that use different propionate-oxidizing pathways. Taxa not known for a syntrophic metabolism may have been involved in the oxidation of butyrate (Telmatospirillum-related) and propionate (unclassified Bacteroidetes and unclassified Fibrobacteres). Gibbs free energies (ΔGs) for syntrophic oxidations of ethanol and butyrate were more favorable than ΔGs for syntrophic oxidation of propionate. As a result of the thermodynamic constraints, acetate transiently accumulated in ethanol and butyrate treatments but not in propionate treatments. Aceticlastic methanogens (Methanosarcina, Methanosaeta) appeared to outnumber hydrogenotrophic methanogens (Methanocella, Methanoregula), reinforcing the likely importance of aceticlastic methanogenesis to the overall production of methane. ΔGs for acetogenesis from H2 to CO2 approximated to -20 kJ mol(-1) when acetate concentrations were low, indicating that acetogens may have contributed to the flow of carbon and reductant towards methane.

Photoheterotrophic microalgal hydrogen production using acetate- and butyrate-rich wastewater effluent

Microalgal (Micractinium reisseri YSW05) photo-heterotrophic hydrogen production was investigated with using volatile fatty acids (acetate and butyrate) from dark-fermentation effluent. Dilution ratio (effluent: deionized water) of 1:0 at pH 8.0 supported the maximum hydrogen and biomass production, and the highest acetate and butyrate consumption. Microalgal hydrogen production was higher with continuous light condition compared to alternating dark/light cycle. Hydrogen production correlated with both, biomass production and hydrogenase activity under all experimental conditions. Hydrogenase activity and apparent hydrogen production decreased significantly with increasing headspace oxygen. Interestingly substantial hydrogenase activity was observed at almost atmospheric partial pressure of oxygen (19%), indicating that the enzyme could be oxygen tolerant. The present study demonstrated the application of conventional microbial fermentation effluent for microalgae cultivation and hydrogen production, and showed the presence of oxygen tolerant hydrogenase in M. reisseri which might help to establish a novel strategy for biohydrogen technology in atmospheric condition.

Use of fermentative metabolites for heterotrophic microalgae growth: Yields and kinetics

The growth of two lipid-producing Chlorella species on fermentative end-products acetate, butyrate and lactate, was investigated using a kinetic modeling approach. Chlorella sorokiniana and Auxenochlorella protothecoides were grown on synthetic media with various (acetate:butyrate:lactate) ratios. Both species assimilated efficiently acetate and butyrate with yields between 0.4 and 0.5g carbon of biomass/g carbon of substrate, but did not use lactate. The highest growth rate on acetate, 2.23d−1, was observed for C. sorokiniana, and on butyrate, 0.22d−1, for A. protothecoides. Butyrate removal started after complete acetate exhaustion (diauxic effect). However, butyrate consumption may be favored by the increase of biomass concentration induced by the initial use of acetate. A model combining Monod and Haldane functions was then built and fitted the experimental data well for both species. Butyrate concentration and (acetate:butyrate) ratios were identified as key parameters for heterotrophic growth of microalgae on fermentative metabolites.

Enhanced cutinase production of Thermobifida fusca by a two-stage batch and fed-batch cultivation strategy

In this study, cutinase production by Thermobifida fusca WSH03-11 was investigated with mixed short-chain organic acids as co-carbon sources to demonstrate the possibility of producing high value-added products from organic wastes. T. fusca WSH03-11 was cultured with different combinations of butyrate, acetate, and lactate with a purpose of increasing cutinase activity. The optimum proportion of butyrate, acetate, and lactate was 4:1:3. In batch cultivation, acetate and lactate were consumed quickly, while the consumption of butyrate was depressed in the presence of acetate with a concentration higher than 0.5 g/L. Based on these results, a two-stage batch and fed-batch cultivation strategy was proposed: a batch culture with acetate and lactate as the co-carbon sources in the first 10 h, and then a fed-batch culture with a constant butyrate feeding rate of 12 mL/h during 11∼20 h. By this two-stage cultivation strategy, cutinase activity, dry cell weight, and consumption rate of butyrate were increased by 70%, 103.4%, and 4.3-fold, respectively, compared to those of the batch cultivation. These results provided a novel and efficient way to produce high value-added products from organic wastes.

Photofermentative hydrogen production from volatile fatty acids present in dark fermentation effluents

Abstract In the present study, the growth and hydrogen production of Rhodobacter sphaeroides O.U. 001, was investigated in media containing five different volatile fatty acids (VFA) individually (malate, acetate, propionate, butyrate and lactate) and in media containing mixtures of these acids that reflect the composition of dark fermentation effluents. The highest hydrogen production rate was obtained in malate (24 ml hydrogen /l reactor h) and the highest biomass concentration was obtained in acetate containing media (1.65 g/l). The substrate conversion efficiencies for different volatile fatty acids were found to vary between 14 and 50%. The malate and butyrate consumption rates were first order with consumption rate constants of 0.026 h −1 and 0.015 h −1 , respectively. In the case of substrate mixtures, it was observed that the bacteria consumed acetate first, followed by propionate and then butyrate. It was also found that the consumption rate of the main substrate significantly increased when the minor substrates were depleted.