Validation of collection and anaerobic fermentation techniques for measuring prebiotic impact on gut microbiota.

BACKGROUND: Defining the ability of prebiotic dietary carbohydrates to influence the composition and metabolism of the gut microbiota is central to defining their health impact in diverse individuals. Many clinical trials are using indirect methods. This study aimed to validate collection and fermentation methods enabling their use in the context of clinical studies. METHODS AND RESULTS: Parameters tested included stool sample acquisition, storage, and growth conditions. Stool from 3 infants and 3 adults was collected and stored under varying conditions. Samples were cultured anaerobically for two days in the presence of prebiotics, whereupon optical density and pH were measured across time. Whole genome shotgun sequencing and NMR metabolomics were performed. Neither the type of collection vial (standard vial and two different BD anaerobic collection vials) nor cryopreservation (-80 °C or 4 °C) significantly influenced either microbial composition at 16 h of anaerobic culture or the principal components of the metabolome at 8 or 16 h. Metagenomic differences were driven primarily by subject, while metabolomic differences were driven by fermentation sugar (2'-fucosyllactose or dextrose). CONCLUSIONS: These data identified a feasible and valid approach for prebiotic fermentation analysis of individual samples in large clinical studies: collection of stool microbiota using standard vials; cryopreservation prior to testing; and collecting fermentation read-out at 8 and 16 hr. Thus, fermentation analysis can be a valid technique for testing the effects of prebiotics on human fecal microbiota.

Tannic Acid Modulation of Substrate Utilization, Microbial Community, and Metabolic Traits in Sludge Anaerobic Fermentation for Volatile Fatty Acid Promotion.

Anaerobic fermentation is a crucial route to realize effective waste activated sludge (WAS) resource recovery and utilization, while the overall efficiency is commonly restrained by undesirable disruptors (i.e., chemical dewatering agents). This work unveiled the unexpectedly positive effects of biodewatering tannic acid (TA) on the volatile fatty acids (VFAs) biosynthesis during WAS anaerobic fermentation. The total VFAs yield was remarkably increased by 15.6 folds with enriched acetate and butyrate in TA-occurred systems. TA was capable to disintegrate extracellular polymeric substances to promote the overall organics release. However, TA further modulated the soluble proteins structure by hydrogen bonding and hydrophobic interactions, resulting in the decrease of proteins bioavailability and consequential alteration of metabolic substrate feature. These changes reshaped the microbial community and stimulated adaptive regulatory systems in hydrolytic-acidogenic bacteria. The keystone species for carbohydrate metabolism (i.e., Solobacterium and Erysipelotrichaceae) were preferentially enriched. Also, the typical quorum sensing (i.e., enhancing substrate transport) and two-component systems (i.e., sustaining high metabolic activity) were activated to promote the microbial networks connectivity and ecological cooperative behaviors in response to TA stress. Additionally, the metabolic functions responsible for carbohydrate hydrolysis, transmembrane transport, and intracellular metabolism as well as VFA biosynthesis showed increased relative abundance, which maintained high microbial activities for VFAs biosynthesis. This study underscored the advantages of biodewatering TA for WAS treatment in the context of resource recovery and deciphered the interactive mechanisms.

Electrochemical promotion of organic waste fermentation: Research advances and prospects.

The current methods of treating organic waste suffer from limited resource usage and low product value. Research and development of value-added products emerges as an unavoidable trend for future growth. Electro-fermentation (EF) is a technique employed to stimulate cell proliferation, expedite microbial metabolism, and enhance the production of value-added products by administering minute voltages or currents in the fermentation system. This method represents a novel research direction lying at the crossroads of electrochemistry and biology. This article documents the current progress of EF for a range of value-added products, including gaseous fuels, organic acids, and other organics. It also presents novel value-added products, such as 1,3-propanediol, 3-hydroxypropionic acid, succinic acid, acrylic acid, and lysine. The latest research trends suggest a focus on EF for cogeneration of value-added products, studying microbial community structure and electroactive bacteria, exploring electron transfer mechanisms in EF systems, developing effective methods for nutrient recovery of nitrogen and phosphorus, optimizing EF conditions, and utilizing biosensors and artificial neural networks in this area. In this paper, an analysis is conducted on the challenges that currently exist regarding the selection of conductive materials, optimization of electrode materials, and development of bioelectrochemical system (BES) coupling processes in EF systems. The aim is to provide a reference for the development of more efficient, advanced, and value-added EF technologies. Overall, this paper aims to provide references and ideas for the development of more efficient and advanced EF technology.

A novel microbial community restructuring strategy for enhanced hydrogen production using multiple pretreatments and CSTR operation.

To achieve rapid enrichment of the targeted hydrogen-producing bacterial population and reconstruction of the microbial community in the biological hydrogen-producing reactor, the activated sludge underwent multiple pretreatments using micro-aeration, alkaline treatment, and heat treatment. The activated sludge obtained from the multiple pretreatments was inoculated into the continuous stirred tank reactor (CSTR) for continuous operations. The community structure alteration and hydrogen-producing capability of the activated sludge were analyzed throughout the operation of the reactor. We found that the primary phyla in the activated sludge population shifted to Proteobacteria, Firmicutes, and Bacteroidetes, which collectively accounted for 96.69% after undergoing several pretreatments. This suggests that the multiple pretreatments facilitated in achieving the selective enrichment of the fermentation hydrogen-producing microorganisms in the activated sludge. The CSTR start-up and continuous operation of the biological hydrogen production reactor resulted in the reactor entering a highly efficient hydrogen production stage at influent COD concentrations of 4000 mg/L and 5000 mg/L, with the highest hydrogen production rate reaching 8.19 L/d and 9.33 L/d, respectively. The main genus present during the efficient hydrogen production stage in the reactor was Ethanoligenens, accounting for up to 33% of the total population. Ethanoligenens exhibited autoaggregation capabilities and a superior capacity for hydrogen production, leading to its prevalence in the reactor and contribution to efficient hydrogen production. During high-efficiency hydrogen production, flora associated with hydrogen production exhibited up to 46.95% total relative abundance. In addition, redundancy analysis (RDA) indicated that effluent pH and COD influenced the distribution of the primary hydrogen-producing bacteria, including Ethanoligenens, Raoultella, and Pectinatus, as well as other low abundant hydrogen-producing bacteria in the activated sludge. The data indicates that the multiple pretreatments and reactor's operation has successfully enriched the hydrogen-producing genera and changed the community structure of microbial hydrogen production.

Tannery sludge valorization through zeolite-assisted anaerobic process for short-chain fatty acids (SCFAs) production.

Tannery sludge, a challenging waste, was utilized as a substrate for the production of Short-Chain Fatty Acids (SCFAs) through a series of six thermophilic Continuous Stirred-Tank Reactor runs. The sludge was subjected to a mild thermal pre-treatment and incorporated zeolites (chabazite in run II, and clinoptilolite in run III) in the acidification process. Results highlighted zeolites' impact on chromium concentration and the SCFAs/CODSOL ratio. Ammonia release remained consistent at around 47 % and 51 % for run I and II, respectively, but surpassed 60% in run III, suggesting limited zeolite effectiveness in NH4 absorption. Chromium release in the liquid fraction, due to thermal pretreatment, reached 335 mg/L. While in tests without zeolite, complete removal proved challenging, in zeolite-amended runs, complete removal was achieved, showcasing the materials' heavy metal absorption capacity. SCFA concentrations reached 20260 mgCOD/L, with acidification efficiency varying; runs I and III had ratios around 0.70 COD/COD, while run II showed substantial improvement (0.92) with chabazite. Anaerobic fermentation-digestion mass balance indicated a 41% reduction in landfill sludge mass, reducing its environmental footprint while yielding valuable byproducts like biogas and SCFAs. These findings underscore zeolites' potential in heavy metal absorption and acidification process enhancement, paving the way for applications with tannery sludge.

A review of volatile fatty acids production from organic wastes: Intensification techniques and separation methods.

High value-added products from organic waste fermentation have garnered increasing concern in modern society. VFAs are short-chain fatty acids, produced as intermediate products during the anaerobic fermentation of organic matter. VFAs can serve as an essential organic carbon source to produce substitutable fuels, microbial fats and oils, and synthetic biodegradable plastics et al. Extracting VFAs from the fermentation broths is a challenging task as the composition of suspensions is rather complex. In this paper, a comprehensive review of methods for VFAs production, extraction and separation are provided. Firstly, the methods to enhance VFAs production and significant operating parameters are briefly reviewed. Secondly, the evaluation and detailed discussion of various VFAs extraction and separation technologies, including membrane separation, complex extraction, and adsorption methods, are presented, highlighting their specific advantages and limitations. Finally, the challenges encountered by different separation technologies and novel approaches to enhance process performance are highlighted, providing theoretical guidance for recycling VFAs from organic wastes efficiently.

Effects of Fe-N co-modified biochar on methanogenesis performance, microbial community, and metabolic pathway during anaerobic co-digestion of alternanthera philoxeroides and cow manure.

The performance of anaerobic digestion (AD) is susceptible to disturbances in feedstock degradation, intermediates accumulation, and methanogenic archaea activity. To improve the methanogenesis performance of the AD system, Fe-N co-modified biochar was prepared under different pyrolysis temperatures (300,500, and 700 °C). Meanwhile, pristine and Fe-modified biochar were also derived from alternanthera philoxeroides (AP). The aim was to compare the effects of Fe-N co-modification, Fe modification, and pristine biochar on the methanogenic performance and explicit the responding mechanism of the microbial community in anaerobic co-digestion (coAD) of AP and cow manure (CM). The highest cumulative methane production was obtained with the addition of Fe-N-BC500 (260.38 mL/gVS), which was 42.37 % higher than the control, while the acetic acid, propionic acid, and butyric acid concentration of Fe-N-BC were increased by 147.58 %, 44.25 %, and 194.06 % compared with the control, respectively. The co-modified biochar enhanced the abundance of Chloroflexi and Methanosarcina in the AD system. Metabolic pathway analysis revealed that the increased methane production was related to the formation and metabolism of volatile fatty acids and that Fe-N-BC500 enhanced the biosynthesis of coenzyme A and the cell activity of microorganisms, accelerating the degradation of propionic acid and enhancing the hydrogenotrophic methanogenesis pathway. Overall, Fe-N co-modified biochar was proved to be an effective promoter for accelerated methane production during AD.

Effects of yeast inoculation methods on caproic acid production and microbial community during anaerobic fermentation of Chinese cabbage waste.

To provide a sufficient supply of electron donors for the synthesis of caproic acid, yeast fermentation was employed to increase ethanol production in the anaerobic fermentation of Chinese cabbage waste (CCW). The results showed that the caproic acid yield of CCW with ethanol pre-fermentation was 7750.3 mg COD/L, accounting for 50.2% of the total volatile fatty acids (TVFAs), which was 32.5% higher than that of the CCW without yeast inoculation. The synchronous fermentation of yeast and seed sludge significantly promoted the growth of butyric acid consuming bacterium Bacteroides, resulting in low yields of butyric acid and caproic acid. With yeast inoculation, substrate competition for the efficient ethanol conversion in the early stage of acidogenic fermentation inhibited the hydrolysis and acidfication. Without yeast inoculation, the rapid accumulation of TVFAs severely inhibited the growth of Bacteroidetes. In the reactor with ethanol pre-fermentation, the key microorganism for caproic acid production, Clostridium_sensu_stricto_12, was selectively enriched.

Recent advancements in wastewater treatment via anaerobic fermentation process: A systematic review.

This manuscript delves into the realm of wastewater treatment, with a particular emphasis on anaerobic fermentation processes, especially dark, photo, and dark-photo fermentation processes, which have not been covered and overviewed previously in the literature regarding the treatment of wastewater. Moreover, the study conducts a bibliometric analysis for the first time to elucidate the research landscape of anaerobic fermentation utilization in wastewater purification. Furthermore, microorganisms, ranging from microalgae to bacteria and fungi, emphasizing the integration of these agents for enhanced efficiency, are all discussed and compared. Various bioreactors, such as dark and photo fermentation bioreactors, including tubular photo bioreactors, are scrutinized for their design and operational intricacies. The results illustrated that using clostridium pasteurianum CH4 and Rhodopseudomonas palustris WP3-5 in a combined dark-photo fermentation process can treat wastewater to a pH of nearly 7 with over 90% COD removal. Also, integrating Chlorella sp and Activated sludge can potentially treat synthetic wastewater to COD, P, and N percentage removal rates of 99%,86%, and 79%, respectively. Finally, the paper extends to discuss the limitations and future prospects of dark-photo fermentation processes, offering insights into the road ahead for researchers and scientists.

Synergistic effects of peracetic acid and free ammonia pretreatment on anaerobic fermentation of waste activated sludge to promote short-chain fatty acid production for polyhydroxyalkanoate biosynthesis: Mechanisms and optimization.

Peracetic acid (PAA) combined with free ammonia (FA) pretreatment can be utilized to promote anaerobic fermentation (AF) of waste activated sludge (WAS) to produce short-chain fatty acids (SCFAs), and the resulting SCFAs are desirable carbon sources (C-sources) for polyhydroxyalkanoate (PHA) biosynthesis. This work aimed to determine the optimum conditions for PAA + FA pretreatment of sludge AF and the feasibility of using anaerobic fermentation liquor (AFL) for PHA production. To reveal the mechanisms of integrated pretreatment, the impacts of PAA + FA pretreatment on different stages of sludge AF and changes in the microbial community structure were explored. The experimental results showed that the maximum SCFA yield reached 491.35 ± 6.02 mg COD/g VSS on day 5 after pretreatment with 0.1 g PAA/g VSS +70 mg FA/L, which was significantly greater than that resulting from PAA or FA pretreatment alone. The mechanism analysis showed that PAA + FA pretreatment promoted sludge solubilization but strongly inhibited methanogenesis. According to the analysis of the microbial community, PAA + FA pretreatment changed the microbial community structure and promoted the enrichment of bacteria related to hydrolysis and acidification, and Proteiniclasticum, Macellibacteroides and Petrimonas became the dominant hydrolytic and acidifying bacteria. Finally, after alkali treatment, the AFL was utilized for batch-mode PHA production, and a maximum PHA yield of 55.05 wt% was achieved after five operation periods.

Enhancement of medium-chain fatty acids production from sludge anaerobic fermentation liquid under moderate sulfate reduction.

In recent years, there has been a marked increase in the production of excess sludge. Chain-elongation (CE) fermentation presents a promising approach for carbon resource recovery from sludge, enabling the transformation of carbon into medium-chain fatty acids (MCFAs). However, the impact of sulfate, commonly presents in sludge, on the CE process remains largely unexplored. In this study, batch tests for CE process of sludge anaerobic fermentation liquid (SAFL) under different SCOD/SO42- ratios were performed. The moderate sulfate reduction under the optimum SCOD/SO42- of 20:1 enhanced the n-caproate production, giving the maximum n-caproate concentration, selectivity and production rate of 5.49 g COD/L, 21.4% and 4.87 g COD/L/d, respectively. The excessive sulfate reduction under SCOD/SO42- ≤ 5 completely inhibited the CE process, resulting in almost no n-caproate generation. The variations in n-caproate production under different conditions of SCOD/SO42- were all well fitted with the modified Gompertz kinetic model. Alcaligenes and Ruminococcaceae_UCG-014 were the dominant genus-level biomarkers under moderate sulfate reduction (SCOD/SO42- = 20), which enhanced the n-caproate production by increasing the generation of acetyl-CoA and the hydrolysis of difficult biodegradable substances in SAFL. The findings presented in this work elucidate a strategy and provide a theoretical framework for the further enhancement of MCFAs production from excess sludge.

Improved anaerobic sludge fermentation mediated by a tryptophan-degrading consortium: Effectiveness assessment and mechanism deciphering.

The hydrolysis of extracellular polymeric substances (EPS) represents a critical bottleneck in the anaerobic fermentation of waste activated sludge (WAS), while tryptophan is identified as an underestimated constituent of EPS. Herein, we harnessed a tryptophan-degrading microbial consortium (TDC) to enhance the hydrolysis efficiency of WAS. At TDC dosages of 5%, 10%, and 20%, a notable increase in SCOD was observed by factors of 1.13, 1.39, and 1.88, respectively. The introduction of TDC improved both the yield and quality of short chain fatty acids (SCFAs), the maximum SCFA yield increased from 590.6 to 1820.2, 1957.9 and 2194.9 mg COD/L, whilst the acetate ratio within SCFAs was raised from 34.1% to 61.2-70.9%. Furthermore, as TDC dosage increased, the relative activity of protease exhibited significant increments, reaching 116.3%, 168.0%, and 266.1%, respectively. This enhancement facilitated WAS solubilization and the release of organic substances from bound EPS into soluble EPS. Microbial analysis identified Tetrasphaera and Soehngenia as key participants in WAS solubilization and the breakdown of protein fraction. Metabolic analysis revealed that TDC triggered the secretion of enzymes associated with amino acid metabolism and fatty acid biosynthesis, thereby fostering the decomposition of proteins and production of SCFAs.

The Autocatalytic Chemical Reaction of a Soluble Biopolymer Derived from Municipal Biowaste.

The paper discusses the perspectives of further implementation of the autocatalytic properties of a soluble biopolymer (SBP) derived from municipal biowastes for the realisation of a biorefinery producing value-added bio-products for consumer use. The reaction of an SBP and water is reported to cause the depolymerisation and oxidation of the pristine SBP organic matter with the formation of carboxyl-functionalised polymers having lower molecular weight and CO2. These findings demonstrate the oxidation of the SBP via water, which could only occur through the production of O and OH radicals catalysed by the SBP. According to the adopted experimental plan, the anaerobic digestate supplied by an Italian municipal biowaste treatment plant was hydrolysed in pH 13 water at 60 °C. The dry product was re-dissolved in plain water at pH 10 and used as a control against the same solution with hydrogen peroxide at 0.1-3 H2O2 moles per SBP carbon mole added. The control and test solutions were kept at room temperature, in the dark or in a climatic chamber under irradiation with simulated solar light, until the pH of the solutions remained constant. Afterwards, the solutions were processed to recover and analyse the crude soluble products. The present work reports the results obtained for the control solution and for the test solutions treated in the presence and absence of H2O2, with and without pH control, in the dark and under irradiation with simulated solar light.

Carbon dioxide and trace oxygen concentrations impact growth and product formation of the gut bacterium Phocaeicola vulgatus.

BACKGROUND: The promising yet barely investigated anaerobic species Phocaeicola vulgatus (formerly Bacteroides vulgatus) plays a vital role for human gut health and effectively produces organic acids. Among them is succinate, a building block for high-value-added chemicals. Cultivating anaerobic bacteria is challenging, and a detailed understanding of P. vulgatus growth and metabolism is required to improve succinate production. One significant aspect is the influence of different gas concentrations. CO2 is required for the growth of P. vulgatus. However, it is a greenhouse gas that should not be wasted. Another highly interesting aspect is the sensitivity of P. vulgatus towards O2. In this work, the effects of varying concentrations of both gases were studied in the in-house developed Respiratory Activity MOnitoring System (RAMOS), which provides online monitoring of CO2, O2, and pressure under gassed conditions. The RAMOS was combined with a gas mixing system to test CO2 and O2 concentrations in a range of 0.25-15.0 vol% and 0.0-2.5 vol%, respectively. RESULTS: Changing the CO2 concentration in the gas supply revealed a CO2 optimum of 3.0 vol% for total organic acid production and 15.0 vol% for succinate production. It was demonstrated that the organic acid composition changed depending on the CO2 concentration. Furthermore, unrestricted growth of P. vulgatus up to an O2 concentration of 0.7 vol% in the gas supply was proven. The viability decreased rapidly at concentrations larger than or equal to 1.3 vol% O2. CONCLUSIONS: The study showed that P. vulgatus requires little CO2, has a distinct O2 tolerance and is therefore well suited for industrial applications.

Metagenomic characterization of biomethane transformation by lipid-catalyzed anaerobic fermentation of lignite.

The present study aims at using lipid in a novel way to improve the efficiency of methane production from lignite anaerobic digestion. The obtained results showed an increase by 3.13 times of the cumulative biomethane content of lignite anaerobic fermentation, when 1.8 g lipid was added. The gene expression of functional metabolic enzymes was also found to be enhanced during the anaerobic fermentation. Moreover, the enzymes related to fatty acid degradation such as long-chain Acyl-CoA synthetase and Acyl-CoA dehydrogenase were increased by 1.72 and 10.48 times, respectively, which consequently, accelerated the conversion of fatty acid. Furthermore, the addition of lipid enhanced the carbon dioxide trophic and acetic acid trophic metabolic pathways. Hence, the addition of lipids was argued to promote the production of methane from lignite anaerobic fermentation, which provided a new insight for the conversion and utilization of lipid waste.

Anaerobic fermentation of organic solid waste: Recent updates in substrates, products, and the process with multiple products co-production.

The effective conversion and recycling of organic solid waste contribute to the resolution of widespread issues such as global environmental pollution, energy scarcity and resource depletion. The anaerobic fermentation technology provides for the effective treatment of organic solid waste and the generation of various products. The analysis, which is based on bibliometrics, concentrates on the valorisation of affordable and easily accessible raw materials with high organic matter content as well as the production of clean energy substances and high value-added platform products. The processing and application status of fermentation raw materials such as waste activated sludge, food waste, microalgae and crude glycerol are investigated. To analyse the status of the preparation and engineering applications of the products, the fermentation products biohydrogen, VFAs, biogas, ethanol, succinic acid, lactic acid, and butanol are employed as representatives. Simultaneously, the anaerobic biorefinery process with multiple product co-production is sorted out. Product co-production can reduce waste discharge, enhance resource recovery efficiency, and serve as a model for improving anaerobic fermentation economics.

Distinct effects of typical sludge pretreatment approaches on the antibiotic resistance genes variations, associated bacterial community dynamics and metabolic activities during anaerobic fermentation process.

Anaerobic fermentation is effective for waste activated sludge (WAS) disposal to realize resource generation and pollutants reduction, and various pretreatments were commonly applied to improve the performance. This work mainly investigated the effects of typical WAS pretreatment approaches on the antibiotic resistance genes (ARGs, as emerging contaminants) removal during anaerobic fermentation processes and unveiled the underlying mechanisms. The results indicated that all the pretreatment strategies exhibited evident effects on the overall ARGs removal with the order of Fe2+ activated persulfate (PS/Fe2+) > pH 10 > Ultrasonication > Heat, and showed selective removal tendency for the specific ARGs (namely easily removed (aadA1 and sul1) and persistent ARGs). Mechanistic analysis demonstrated that the pretreatments disrupted the extracellular polymeric substances (EPS) and rose the cell membrane permeability (particularly for PS/Fe2+ and Heat). Then the increased ARGs release benefitted the subsequent reduction of mobile genetic elements (MGEs) and extracellular ARGs (especially for PS/Fe2+ and pH10), resulting the ARGs attenuation. Pretreatments significantly shifted the microbial community structure and the abundances of potential ARGs hosts (i.e., Sulfuritalea, and Denitratisoma). Also, the different pretreatments exhibited distinct effects on the microbial metabolic traits related with ARGs proliferation (i.e., ABC transporters, two-component system and bacterial secretion systems), which also contributed to the ARGs attenuations during WAS fermentation. The partial least-squares path modeling (PLS-PM) analysis indicated that the bacterial community (total effects = 0.968) was key factor determining ARGs fates.

The role of hydrochloric acid pretreated activated carbon in chain elongation of D-lactate to caproate: Adsorption and facilitation.

Medium chain fatty acids (MCFA) generation is attracting growing interest due to fossil fuel depletion. To promote the production of MCFA, especially caproate, hydrochloric acid pretreated activated carbon (AC) was introduced into chain elongation fermentation. In this study, the role of pretreated AC on caproate production was investigated using lactate and butyrate as electron donor and electron acceptor, respectively. The results showed that AC did not improve the chain elongation reaction at beginning but promoted the caproate production at later stage. The addition of 15 g/L AC facilitated reactor reaching the peak of caproate concentration (78.92 mM), caproate electron efficiency (63.13%), and butyrate utilization rate (51.88%). The adsorption experiment revealed a positive correlation between the adsorption capacity of pretreated AC and the concentration as well as the carbon chain length of carboxylic acids. Moreover, the adsorption of undissociated caproate by pretreated AC contributed to a mitigated toxicity towards microorganisms, thereby facilitating the production of MCFA. Microbial community analysis revealed an increasing enrichment of key functional chain elongation bacteria, including Eubacterium, Megasphaera, Caproiciproducens, and Pseudoramibacter, but a suppression on acrylate pathway microorganism Veillonella, as the dosage of pretreated AC increasing. The findings of this study demonstrated the substantial impact of the adsorption effect of acid-pretreated AC on promoting caproate production, which would aid to the development of more efficient caproate production process.

Regulation of Cat8 in energy metabolic balance and glucose tolerance in Saccharomyces cerevisiae.

Cat8 is a C6 zinc cluster transcription activator in yeast. It is generally recognized that the transcription of CAT8 is inhibited and that Cat8 is inactive in the presence of high concentrations of glucose. However, our recent study found that constitutively overexpressed Cat8 played a regulatory role in Saccharomyces cerevisiae in the presence of 20 g/L glucose. To explore the regulatory network of Cat8 at high glucose concentrations, CAT8 was both overexpressed and deleted in this study. Cell growth and glucose consumption in different media were significantly accelerated by the deletion of CAT8, while the lag period was greatly shortened. RNA-seq and genetic modification showed that the deletion of CAT8 changed the type of energy metabolism in yeast cells. Many genes related to the mitochondrial respiratory chain were downregulated, resulting in a reduction in aerobic respiration and the tricarboxylic acid cycle. Meanwhile, both the energy supply of anaerobic ethanol fermentation and the Crabtree effect of S. cerevisiae were enhanced by the deletion of CAT8. CAT8 knockout cells show a higher sugar uptake rate, a higher cell growth rate, and higher tolerance to glucose than the wild-type strain YS58. This study expands the understanding of the regulatory network of Cat8 and provides guidance for modulating yeast cell growth. KEY POINTS: • The deletion of CAT8 promoted cell growth of S. cerevisiae. • Transcriptome analysis revealed the regulation network of Cat8 under 1% glucose condition. • CAT8 deletion increases the glucose tolerance of cells by enhancing the Crabtree effect.

Enhanced anaerobic hydrogen production from cotton straws assisted by copper molybdate.

Hydrogen production from dark fermentation has potential application due to its environmental friendliness, low production cost, and sustainability. However, there is still an obstacle to improving the efficiency of bioH2 production to meet the requirements in practical applications. In this research, copper molybdates are synthesized under different pH conditions as additives to study their different influence processes during anaerobic hydrogen production from cotton straws with the pure cultural system. A series of results indicate that CuMoO4 with appropriate experimental conditions has the highest H2 yield at 191.3 mL/g straws at 37 °C, which is 236% higher than the control group. It can be shown that O. ethanolica 8KG-4 has an obvious accompanying with high stability and low cytotoxicity for this clean energy production system as well as the improvement of metabolic pathway. These results extend new thinking of obtaining higher H2 yield as a biofuel in future production.