Propionate Production Research Articles

Chia flour combined with a high fat diet increases propionic acid production and improves the microbial richness and diversity in female Wistar rats.

Chia is a functional food because of its positive impact on reducing the risk of metabolic diseases. These benefits are due to its nutritional composition as a source of dietary fiber and bioactive compounds. In our previous study, chia consumption increased the richness of the microbiota and the production of short chain fatty acids (SCFAs) when consumed by male Wistar rats, so, the objective of this study was to assess the effects of the consumption of chia with a high fat diet on gut health in female Wistar rats. 32 adult female Wistar rats were allocated into four groups and received one of the following diets: standard diet (SD), standard diet + chia (SDC), high fat diet (HFD) or high fat diet + chia (HFDC) for 8 weeks. At the end of the study, the intestinal microbiota, SCFA content, cecum content pH, immunoglobulin A (IgA) quantification and brush border membrane functionality were evaluated. There was no difference in the relative abundance of the gut microbiota, but chia consumption increased the microbial richness and diversity, increased the production of acetic and butyric acids in the SDC group and propionic acid in the HFD group, and decreased the pH of cecal content. The HFDC group demonstrated a lower IgA concentration compared to the HFD group. The SDC group increased SI and AP gene expression and decreased SGLT1 and PepT1 compared to the SD group. The consumption of chia can be beneficial for the functionality of the microbiota, improving SCFAs and intestinal pH, and the effects of chia in the microbiota can be more pronounced in HFD.

Chemical composition and prebiotic activity of baru (Dipteryx alata Vog.) pulp on probiotic strains and human colonic microbiota

Little knowledge is available in literature regarding the chemical composition and health-promoting effects of baru (Dipteryx alata Vog.) pulp, a by-product usually discarded by the agro-industry during the processing of baru fruit. This study evaluated the chemical composition of baru pulp and investigated its prebiotic activity on distinct probiotic strains and human colonic microbiota with in vitro assays. Baru pulp had high contents of insoluble dietary fibers and phenolic compounds (mainly hesperidin). Baru pulp stimulated the growth and metabolism of the probiotics Bifidobacterium animalis subsp. lactis BB-12, Lactobacillus acidophilus LA-05, and Lacticaseibacillus casei L-26. In addition, digested baru pulp induced significant benefits on the human colonic microbiota, increasing the relative abundance of Lactobacillus-Enterococcus, Bifidobacterium, and Bacteroides-Prevotella, as well as the production of lactate, acetate, propionate, and butyrate. The results show that baru pulp has potential prebiotic properties to be explored in the formulation of new health-promoting foods.

Prevotella: A Key Player in Ruminal Metabolism.

Ruminants are foregut fermenters that have the remarkable ability of converting plant polymers that are indigestible to humans into assimilable comestibles like meat and milk, which are cornerstones of human nutrition. Ruminants establish a symbiotic relationship with their microbiome, and the latter is the workhorse of carbohydrate fermentation. On the other hand, during carbohydrate fermentation, synthesis of propionate sequesters H, thus reducing its availability for the ultimate production of methane (CH4) by methanogenic archaea. Biochemically, methane is the simplest alkane and represents a downturn in energetic efficiency in ruminants; environmentally, it constitutes a potent greenhouse gas that negatively affects climate change. Prevotella is a very versatile microbe capable of processing a wide range of proteins and polysaccharides, and one of its fermentation products is propionate, a trait that appears conspicuous in P. ruminicola strain 23. Since propionate, but not acetate or butyrate, constitutes an H sink, propionate-producing microbes have the potential to reduce methane production. Accordingly, numerous studies suggest that members of the genus Prevotella have the ability to divert the hydrogen flow in glycolysis away from methanogenesis and in favor of propionic acid production. Intended for a broad audience in microbiology, our review summarizes the biochemistry of carbohydrate fermentation and subsequently discusses the evidence supporting the essential role of Prevotella in lignocellulose processing and its association with reduced methane emissions. We hope this article will serve as an introduction to novice Prevotella researchers and as an update to others more conversant with the topic.

Glucogenic and lipogenic diets affect in vitro ruminal microbiota and metabolites differently.

This study was conducted to evaluate the effects of two glucogenic diets (C: ground corn and corn silage; S: steam-flaked corn and corn silage) and a lipogenic diet (L: sugar beet pulp and alfalfa silage) on the ruminal bacterial and archaeal structures, the metabolomic products, and gas production after 48 h in vitro fermentation with rumen fluid of dairy cows. Compared to the C and S diets, the L dietary treatment leaded to a lower dry matter digestibility (DMD), lower propionate production and ammonia-nitrogen concentration. The two glucogenic diets performed worse in controlling methane and lactic acid production compared to the L diet. The S diet produced the greatest cumulative gas volume at any time points during incubation compared to the C and L diet. The metabolomics analysis revealed that the lipid digestion especially the fatty acid metabolism was improved, but the amino acid digestion was weakened in the L treatment than in other treatments. Differences in rumen fermentation characteristics were associated with (or resulting from) changes in the relative abundance of bacterial and archaeal genera. The rumen fluid fermented with L diet had a significantly higher number of cellulolytic bacteria, including the genera of Ruminococcus, Butyrivibrio, Eubacterium, Lachnospira, unclassified Lachnospiraceae, and unclassified Ruminococcaceae. The relative abundances of amylolytic bacteria genera including Selenomonas_1, Ruminobacter, and Succinivibrionaceae_UCG-002 were higher in samples for diets C and S. The results indicated that the two glucogenic diets leaded to a higher relative abundance of bacteria which functions in succinate pathway resulting in a higher propionate production. The steam-flaked corn diet had a higher gas production and lower level of metabolites in fatty acids and amino acids. Most highly abundant bacteria were observed to be not sensitive to dietary alterations of starch and fiber, except for several amylolytic bacteria and cellulolytic bacteria. These finding offered new insights on the digesting preference of ruminal bacteria, which can assist to improve the rumen functioning.

High-Yield Production of Propionate from 1,2-Propanediol by Engineered Pseudomonas putida KT2440, a Robust Strain with Highly Oxidative Capacity.

Bio-based propionate attracts increasing attention owing to its green nature and specific food additive market. To date, the time-consuming and costly fermentation process by strict anaerobes makes propionate production not ideal. In this study, we designed a new route for propionate production, in which 1,2-propanediol was first dehydrated to propionaldehyde and then to propionate by taking advantage of the robust oxidization capacity of the Pseudomonas putida KT2440 strain. The high atom economy (0.97 g/g) in this proposed pathway is more advantageous than the previous l-threonine-derived route (0.62 g/g). The molecular mechanism of the extraordinary oxidation capacity of P. putida KT2440 was first deciphered. The propionate production was realized in P. putida KT2440 by screening suitable glycerol dehydratases and optimizing the expression to eliminate the formation of 1-propanol and the accumulation of the intermediate propionaldehyde. The engineered strain produced propionate with a molar conversion rate of >99% from 1,2-propanediol. A high titer of 46.5 g/L pure propionic acid with a productivity of 1.55 g/L/h and a mass yield of 0.96 g/g was achieved in fed-batch biotransformation. Thus, this study provides another idea for the production of high-purity bio-based propionate from renewable materials with high atom economy.

Effect of brown and green seaweeds on diet digestibility, ruminal fermentation patterns and enteric methane emissions using the rumen simulation technique

Inclusion of the red seaweed Asparagopsis taxiformis as a feed additive, has led to significant reductions in methane (CH4) production from ruminants. However, dietary supplementation with this seaweed is negatively associated with health and environmental concerns mainly due to its bromoform content, a compound with potential carcinogenic properties. Thus, there is renewed focus on ascertaining the anti-methanogenic potential of locally grown brown and green seaweeds, which typically do not contain bromoform. The objective of this study was to investigate the effects of selected brown and green seaweeds on diet digestibility, ruminal fermentation patterns, total gas (TGP) and CH4 production in vitro, using the rumen simulation technique system. In experiment 1, Pelvetia canaliculata (PEC) was examined. In experiment 2, Cystoseira tamariscifolia (CYT), Bifurcaria bifurcata (BIB), Fucus vesiculosus (FUV), Himanthalia elongata (HIM) and Ulva intestinalis (ULI) were analysed. Ascophyllum nodosum (ASC) was included in both experiments. A diet containing A. taxiformis (ASP1; ASP2) and an unsupplemented diet (CON) were included as positive and negative controls, respectively in both experiments. All seaweeds were included at a rate of 10 g/kg dry matter (DM) into a control diet of 50:50 (w:w) forage:concentrate. The seven brown and green seaweeds assessed failed to affect absolute CH4 emissions or alter fermentation patterns. In experiment 1, seaweed treatment had no effect on diet digestibility, CH4%, CH4 mmol/d or CH4 L/d (P>0.1), however ASP1 reduced CH4 mmol/g DOM by 49% (P<0.01) relative to the control. Both ASC and ASP1 tended to increase TGP (P<0.1) relative to the control. In addition to this, the inclusion of seaweed in experiment 1 reduced the production of NH3-N (P<.0001) compared to the control. In experiment 2, seaweed treatment had no effect on diet digestibility or TGP. Both ASP2 and FUV reduced CH4% (P<0.01) but only ASP2 significantly reduced CH4 mmol/d, CH4 L/d and CH4 mmol/g DOM (P<0.05). Daily mMol butyrate was reduced by ASP2 relative to the control and most other seaweeds (P<.0001). In both experiment 1 and 2, seaweed inclusion had no effect on daily total VFA, acetate or propionate production or the acetate:propionate ratio relative to the control. To conclude, including the bromoform-free brown and green seaweeds at 10g/kg DM has no negative effects on diet digestibility or fermentation patterns but also failed to reduce the production of enteric CH4in vitro.

Combined Inclusion of Former Foodstuff and Distiller Grains in Dairy Cows Ration: Effect on Milk Production, Rumen Environment, and Fiber Digestibility.

The aim of the present study was to investigate the effect of the substitution, in dairy cow rations, of traditional protein and starch sources with more sustainable "circular" feeds to increase the sustainability of dairy production. For this purpose, eight multiparous mid-lactating cows were blocked and assigned to one of four treatments and were used in a replicated 4 × 4 Latin squares design with 21-days periods (14 days of adaptation and 7 of data collection). Two different circular feedstuffs were tested: a bakery's former foodstuff (FF) and a wheat distiller's grain with solubles (WDGS). These ingredients were used, alone and in combination, in three experimental diets (FF, WDGS; FF + WDGS) and compared to a standard ration (CTR). Dry matter intake and rumination time were not influenced by these diets. Conversely, dietary treatments partially influenced the milk yield, rumen pH, Volatile Fatty Acids (VFA) production, and fibre digestibility. In particular, the combined inclusion of FF and WDGS increased milk production (37.39 vs. 36.92, 35.48, 35.71 kg/day, for FF, WDGS and CTR diets, respectively) and reduced milk urea content (13.14 vs. 16.19, 15.58, 16.95 mg/dL for FF, WDGS, and CTR diets, respectively). No effects of this association were found in the milk composition, acetic and propionic production, and fibre digestibility. These results suggest that the association of former foodstuff and wheat distillers' grains could be safely included in dairy cow rations to increase the sustainability of cow nutrition and improve milk production without impairing animal health, dry matter intake, and fibre digestibility.

Kombuchas from Green and Black Tea Modulate the Gut Microbiota and Improve the Intestinal Health of Wistar Rats Fed a High-Fat High-Fructose Diet.

The Western diet can negatively affect the gut microbiota and is associated with metabolic disorders. Kombucha, a tea fermented by a symbiotic culture of bacteria and yeast (SCOBY), is known for its bioactive properties and has become popular in the last years. In this study, we evaluated the effects of regular kombucha consumption on the gut microbiota and on outcomes related to the intestinal health of Wistar rats fed a high-fat high-fructose diet. After eight weeks receiving a standard diet (AIN-93M) (n = 10) or a high-fat and high-fructose diet (HFHF) (n = 30) to induce metabolic disorders, the animals were subdivided into four groups: AIN-93M (n = 10); HFHF (n = 10); GTK (HFHF + green tea kombucha (n = 10); and BTK (HFHF + black tea kombucha; n = 10) for 10 weeks. Although body composition did not differ among the groups, the HFHF diet was associated with metabolic alterations, and stimulated the growth of gram-negative bacteria such as Proteobacteria and Bacteroides. Kombucha ingestion could somewhat modulate the gut microbiota, attenuating the effects of a Western diet by increasing propionate production and favoring the growth of beneficial bacteria, such as Adlercreutzia in the GTK group. Our results suggest that regular kombucha consumption may be beneficial to intestinal health, which can be mostly attributed to its high content and diversity of phenolic compounds.

A shift towards succinate-producing Prevotella in the ruminal microbiome challenged with monensin.

The time-resolved impact of monensin on the active rumen microbiome was studied in a rumen-simulating technique (Rusitec) with metaproteomic and metabolomic approaches. Monensin treatment caused a decreased fibredegradation potential that was observed by the reduced abundance of proteins assigned to fibrolytic bacteria and glycoside hydrolases, sugar transporters and carbohydrate metabolism. Decreased proteolytic activities resulted in reduced amounts of ammonium as well as branched-chain fatty acids. The family Prevotellaceae exhibited increased resilience in the presence of monensin, with a switch of the metabolism from acetate to succinate production. Prevotella species harbour a membrane-bound electron transfer complex, which drives the reduction of fumarate to succinate, which is the substrate for propionate production in the rumen habitat. Besides the increased succinate production, a concomitant depletion of methane concentration was observed upon monensin exposure. Our study demonstrates that Prevotella sp. shifts its metabolism successfully in response to monensin exposure and Prevotellaceae represents the key bacterial family stabilizing the rumen microbiota during exposure to monensin.

Enhancing propionic acid production in the acidogenic fermentation of food waste facilitated by a fungal mash under neutral pH

Fungal mash derived from Aspergillus spp. is a green enzymatic additive for food waste (FW) valorization. In this study, the production of volatile fatty acids (VFAs) and the proportion of propionic acid (PA) in VFAs were increased by utilizing a complex enzyme (CE) obtained from Aspergillus oryzae. Results showed that CE addition significantly promoted SCOD concentration in the hydrolysis at a wide pH range from 4 to 9. In contrast, the production of VFAs was influenced by pH, and the highest yields of VFAs and PA were found at pH 7. At the CE dosage of 0.2 g/g VSS, the concentration of VFAs in the FW fermentation liquid reached 38.1 g COD/L with the PA proportion up to 42.7%, which increased by 107.9% and 63.7%, respectively, relative to that in the zero-dosage group. With CE continuing to be added, the C/N ratio declined, and the primary metabolic pathway was converted from acetic acid-type to PA-type. Further investigation of the dominant microbial communities and their metabolic capacities showed that the acrylate-mediated pathway was the potential metabolic reaction in PA-type fermentation. These results indicated that CE pretreatment was a feasible strategy to enhance the PA-rich fermentation of FW under neutral pH conditions.

Clostridium butyricum alleviates weaned stress of piglets by improving intestinal immune function and gut microbiota

Clostridium butyricum (C. butyricum) has been reported to improve the intestinal health, whereas the mechanism is still uncertain. In this study, weaned piglets were treated with C. butyricum SLZX19-05 to test its effects and mechanisms on growth and intestinal functions. The major findings were that this probiotic reduced diarrhea rates and serum cortisol contents and improved growth performance of weaned piglets. In small intestine, its supplementation inhibited the inflammation and improved the morphology of piglets. Meanwhile, the ileal microbiota was remodeled and propionate production increased. In colon, its supplementation enhanced the expressions of barrier proteins and changed the expression model of host defense peptides, whereas minor changes occurred in microbiota and butyrate production increased. These evidences hint C. butyricum SLZX19-05 can alleviate weaned stress of piglets by improving the morphology, immune defense capacity and barrier function of gut via regulating microbiota and short chain fatty acids.

Red seaweed extracts reduce methane production by altering rumen fermentation and microbial composition in vitro.

A series of in vitro batch culture incubations were carried out to investigate changes in rumen fermentation characteristics, methane (CH4) production, and microbial composition in response to supplementation with five different red seaweed species (Amphiroa anceps, AANC; Asparagopsis taxiformis, ATAX; Chondracanthus tenellus, CTEN; Grateloupia elliptica, GELL; and Gracilaria parvispora, GPAR). Prior to the incubations, the total flavonoid and polyphenol content of the red seaweed extracts was quantified. The incubated substrate consisted of timothy hay and corn grain [60:40 dry matter (DM) basis]. Treatments were substrate mixtures without seaweed extract (CON) or substrate mixtures supplemented with 0.25 mg/mL of red seaweed extract. Samples were incubated for 6, 12, 24, 36, and 48 h. Each sample was incubated in triplicates in three separate runs. In vitro DM degradability, fermentation parameters (i.e., pH, volatile fatty acids, and ammonia nitrogen), total gas production, and CH4 production were analyzed for all time points. Microbial composition was analyzed using 16S rRNA amplicon sequencing after 24 h of incubation. The highest CH4 reduction (mL/g DM, mL/g digested DM, and % of total gas production) was observed in ATAX (51.3, 50.1, and 51.5%, respectively, compared to CON; P < 0.001) after 12 h of incubation. The other red seaweed extracts reduced the CH4 production (mL/g DM; P < 0.001) in the range of 4.6-35.0% compared to CON after 24 h of incubation. After 24 h of incubation, supplementation with red seaweed extracts tended to increase the molar proportion of propionate (P = 0.057) and decreased the acetate to propionate ratio (P = 0.033) compared to the CON. Abundances of the genus Methanobrevibacter and total methanogens were reduced (P = 0.050 and P = 0.016) by red seaweed extract supplementation. The linear discriminant analysis effect size (P < 0.05, LDA ≥ 2.0) showed that UG Succinivibrionaceae, Anaeroplasma, and UG Ruminococcaceae, which are associated with higher propionate production, starch degradation, and amylase activity were relatively more abundant in red seaweed extracts than in the CON. Our results suggest that supplementation with red seaweed extracts altered the microbiota, leading to the acceleration of propionate production and reduction in CH4 production.

Cysteamine Supplementation In Vitro Remarkably Promoted Rumen Fermentation Efficiency towards Propionate Production via Prevotella Enrichment and Enhancing Antioxidant Capacity.

Cysteamine (CS) is a vital antioxidant product and nutritional regulator that improves the productive performance of animals. A 2 × 4 factorial in vitro experiment was performed to determine the effect of the CS supplementation levels of 0, 20, 40, and 60 mg/g, based on substrate weight, on the ruminal fermentation, antioxidant capacity, and microorganisms of a high-forage substrate (HF, forage:corn meal = 7:3) in the Statistical Analysis System Institute. After 48 h of incubation, the in vitro dry matter disappearance and gas production in the LF group were higher when compared with a low-forage substrate (LF, forge hay:corn meal = 3:7), which was analyzed via the use of the MIXED procedure of the HF group, and these increased linearly with the increasing CS supplementation (p &lt; 0.01). With regard to rumen fermentation, the pH and acetate were lower in the LF group compared to the HF group (p &lt; 0.01). However, the ammonia N, microbial crude protein, total volatile fatty acids (VFA), and propionate in the LF group were greater than those in the HF group (p &lt; 0.05). With the CS supplementation increasing, the pH, ammonia N, acetate, and A:P decreased linearly, while the microbial crude protein, total VFA, and propionate increased linearly (p &lt; 0.01). Greater antioxidant capacity was observed in the LF group, and the increasing CS supplementation linearly increased the superoxide dismutase, catalase, glutathione peroxidase, total antioxidant capacity, glutathione, and glutathione reductase, while it decreased the malondialdehyde (p &lt; 0.05). No difference occurred in the ruminal bacteria alpha diversity with the increasing CS supplementation, but it was higher in the LF group than in the HF group (p &lt; 0.01). Based on the rumen bacterial community, a higher proportion of Bacteroidota, instead of Firmicutes, was in the LF group than in the HF group. Furthermore, increasing the CS supplementation linearly increased the relative abundance of Prevotella, norank_f_F082, and Prevotellaceae_UCG-001 under the two substrates (p &lt; 0.05). Prevotella, norank_f_F082, and Prevotellaceae_UCG-001 were positively correlated with gas production, rumen fermentation, and antioxidant capacity in a Spearman correlation analysis (r &gt; 0.31, p &lt; 0.05). Overall, a CS supplementation of not less than 20 mg/g based on substrate weight enhanced the rumen fermentation and rumen antioxidant capacity of the fermentation system, and it guided the rumen fermentation towards glucogenic propionate by enriching the Prevotella in Bacteroidetes.

Optimising bioreactor processes with in-situ product removal using mathematical programming: A case study for propionate production

Designing and operating bioreactors with in-situ product removal (ISPR) can be challenging, particularly in discontinuous systems, where the ISPR and substrate feeding need to be effectively scheduled. Mathematical models can help assess different scheduling regimes in the fermentation medium and provide a means to optimise the process. Focusing on a propionate production case study, a model of a co-culture batch fermentation with electrodialysis (the ISPR system), was developed. Using this model, the product yield and/or the productivity were maximised by 1) single objective optimisation maximising the product yield (0.49 gpropionate /gglucose) or productivity (0.75 gpropionate/L/h), 2) multi objective optimisation to pursue trade-off solutions between the yield and productivity and 3) a stochastic optimisation maximising the productivity robustly (0.64 gpropionate/L/h) to account for uncertainties associated to the model parameters. With this contribution it is demonstrated that, through mathematical models, ISPR can be implemented and adapted to the user's objectives.

Shatianyu (Citrus grandis L. Osbeck) Flavonoids and Dietary Fiber in Combination Are More Effective Than Individually in Alleviating High-Fat-Diet-Induced Hyperlipidemia in Mice by Altering Gut Microbiota

This study was aimed at exploring the separate and combined anti-hyperlipidemic effect of Shatianyu (Citrus grandis L. Osbeck) flavonoids (SPFEs) and DF (SPDF) on HFD-fed mice after 14-week administration in diet, together with the possible microbiota-mediated mechanisms. SPFEs and SPDF were more effective together than separately in improving serum lipid profiles, decreasing hepatic lipid accumulation, and upregulating the expression of hepatic CPT1a, CYP7A1, ABCG5, and ABCG8. Butyrate has been previously proved to have an anti-hyperlipidemic effect. The fecal butyrate contents were negatively correlative with serum/liver lipid but positively correlated with fecal total bile acids levels, and SPDF + SPFEs had the most fecal butyrate in this study. SPDF or SPFEs enriched microbiota related to acetic and propionic acids production, while SPDF + SPFEs also bloomed norank_f_Muribaculaceae, Dubosiella, Lachnoclostridium, and norank_f_Eubacterium_coprostanoligenes_group, which were positively correlated to fecal butyrate contents. Thus, SPFEs and SPDF might alleviate hyperlipidemia synergistically by regulating microbiota to produce butyrate, thereby regulating lipid metabolism.

Effects of acetate, propionate, and pH on volatile fatty acid thermodynamics in continuous cultures of ruminal contents.

To investigate the effects of acetate, propionate, and pH on thermodynamics of volatile fatty acids (VFA) in the rumen, a dual-flow continuous culture study was conducted to quantify production of major VFA, interconversions among the VFA, and H2 and CH4 emissions in a 4 × 4 Latin square design. The 4 treatments were (1) control: pH buffered to an average of 6.75; (2) control plus 20 mmol/d of infused acetate (InfAc); (3) control plus 7 mmol/d of infused propionate (InfPr); and (4) a 0.5-unit decline in pH elicited by adjustment of the buffer (LowpH). All fermentors were fed 40 g of a pelleted diet containing whole alfalfa pellets and concentrate mix pellets (50:50) once daily. After 7 d of treatment, sequential, continuous infusions of [2-13C] sodium acetate (3.5 mmol/d), [U-13C] sodium propionate (2.9 mmol/d), and [1-13C] sodium butyrate (0.22 mmol/d) were carried out from 12 h before feeding for 36 h. Filtered liquid effluent (4 mL) was sampled at 0, 2, 4, 6, 8, 12, 16, and 22 h after feeding, and assessed for VFA concentrations, with another filtered sample (20 mL) used to quantify aqueous concentrations of CH4 and H2. Headspace CH4 and H2 gases were monitored continuously. Ruminal microbes were isolated from the mixed effluent samples, and the microbial community structure was analyzed using the 16S rRNA amplicon sequencing technique. The digestibility of neutral detergent fiber, acid detergent fiber, and starch and microbial C sequestrated from VFA were not affected by treatments. The LowpH treatment increased net propionate production and decreased H2 and CH4 headspace emissions, primarily due to shifts in metabolic pathways of VFA formation, likely due to the observed changes in bacterial community structure. Significant interconversions occurred between acetate and butyrate, whereas interconversions of other VFA with propionate were relatively small. The InfAc and InfPr treatments increased net acetate and propionate production, respectively; however, interconversions among VFA were not affected by pH, acetate, or propionate treatments, suggesting that thermodynamics might not be a primary influencer of metabolic pathways used for VFA formation.

Sustainable Process Design of Propionic Acid Production from Glycerol: A Comparative Study of Bio-Based and Petroleum-Based Technologies

Sustainable Process Design of Propionic Acid Production from Glycerol: A Comparative Study of Bio-Based and Petroleum-Based Technologies

Altered gut microbiome diversity and function in patients with propionic acidemia.

Propionic acidemia (PA) is an inherited metabolic disorder of propionate metabolism, where the gut microbiota may play a role in pathophysiology and therefore, represent a relevant therapeutic target. Little is known about the gut microbiota composition and activity in patients with PA. Although clinical practice varies between metabolic treatment centers, management of PA requires combined dietary and pharmaceutical treatments, both known to affect the gut microbiota. This study aimed to characterize the gut microbiota and its metabolites in fecal samples of patients with PA compared with healthy controls from the same household. Eight patients (aged 3-14y) and 8 controls (4-31y) were recruited from Center 1 (UK) and 7 patients (11-33y) and 6 controls (15-54y) from Center 2 (Austria). Stool samples were collected 4 times over 3 months, alongside data on dietary intakes and medication usage. Several microbial taxa differed between patients with PA and controls, particularly for Center 1, e.g., Proteobacteria levels were increased, whereas butyrate-producing genera, such as Roseburia and Faecalibacterium, were decreased. Most measured microbial metabolites were lower in patients with PA, and butyrate was particularly depleted in patients from Center 1. Furthermore, microbiota profile of these patients showed the lowest compositional and functional diversity, and lowest stability over 3 months. As the first study to map the gut microbiota of patients with PA, this work represents an important step forward for developing new therapeutic strategies to further improve PA clinical status. New dietary strategies should consider microbial propionate production as well as butyrate production and microbiota stability.

Microaerobic metabolism of lactate and propionate enhances vitamin B12 production in Propionibacterium freudenreichii

BackgroundPropionibacterium freudenreichii is used in biotechnological applications to produce vitamin B12. Although cultured mainly in anaerobic conditions, microaerobic conditions can greatly enhance biomass formation in P. freudenreichii. Since B12 yields may be coupled to biomass formation, microaerobic conditions show great potential for increasing B12 yields in P. freudenreichii.ResultsHere we show biomass formation increases 2.7 times for P. freudenreichii grown in microaerobic conditions on lactate versus anaerobic conditions (1.87 g/L vs 0.70 g/L). Consumption of lactate in microaerobic conditions resulted first in production of pyruvate, propionate and acetate. When lactate was depleted, pyruvate and propionate were oxidised with a concomitant sixfold increase in the B12 titer compared to anaerobic conditions, showing potential for propionate and pyruvate as carbon sources for B12 production. Consequently, a fed-batch reactor with anaerobically precultured lactate-grown cells was fed propionate in microaerobic conditions resulting in biomass increase and production of B12. Vitamin yields increased from 0.3 mu g B12 per mmol lactate in anaerobic conditions to 2.4 mu g B12 per mmol lactate and 8.4 mu g B12 per mmol propionate in microaerobic conditions. Yield per cell dry weight (CDW) increased from 41 mu g per g CDW in anaerobic conditions on lactate to 92 mu g per g CDW on lactate and 184 mu g per g CDW on propionate in microaerobic conditions.ConclusionsHere we have shown both B12 yield per substrate and per CDW were highest on cells oxidising propionate in microaerobic conditions, showing the potential of propionate for biotechnological production of vitamin B12 by P. freudenreichii.

Production of propionate using metabolically engineered strains of Clostridium saccharoperbutylacetonicum

The carboxylic acid propionate is a valuable platform chemical with applications in various fields. The biological production of this acid has become of great interest as it can be considered a sustainable alternative to petrochemical synthesis. In this work, Clostridium saccharoperbutylacetonicum was metabolically engineered to produce propionate via the acrylate pathway. In total, the established synthetic pathway comprised eight genes encoding the enzymes catalyzing the conversion of pyruvate to propionate. These included the propionate CoA-transferase, the lactoyl-CoA dehydratase, and the acryloyl-CoA reductase from Anaerotignum neopropionicum as well as a D-lactate dehydrogenase from Leuconostoc mesenteroides subsp. mesenteroides. Due to difficulties in assembling all genes on one plasmid under the control of standard promoters, the PtcdB-tcdR promoter system from Clostridium difficile was integrated into a two-plasmid system carrying the acrylate pathway genes. Several promoters were analyzed for their activity in C. saccharoperbutylacetonicum using the fluorescence-activating and absorption-shifting tag (FAST) as a fluorescent reporter to identify suitable candidates to drive tcdR expression. After selecting the lactose-inducible PbgaL promoter, engineered C. saccharoperbutylacetonicum strains produced 0.7 mM propionate upon induction of gene expression. The low productivity was suspected to be a consequence of a metabolic imbalance leading to acryloyl-CoA accumulation in the cells. To even out the proposed imbalance, the propionate-synthesis operons were rearranged, thereby increasing the propionate concentration by almost four-fold. This study is the first one to report recombinant propionate production using a clostridial host strain that has opened a new path towards bio-based propionate to be improved further in subsequent work.Key points• Determination of promoter activities in C. saccharoperbutylacetonicum using FAST.• Implementation of propionate production in C. saccharoperbutylacetonicum.• Elevation of propionate production by 375% to a concentration of 3 mM.