Abundant Enzyme Research Articles

PARROT: Prediction of enzyme abundances using protein-constrained metabolic models.

Protein allocation determines the activity of cellular pathways and affects growth across all organisms. Therefore, different experimental and machine learning approaches have been developed to quantify and predict protein abundance and how they are allocated to different cellular functions, respectively. Yet, despite advances in protein quantification, it remains challenging to predict condition-specific allocation of enzymes in metabolic networks. Here, using protein-constrained metabolic models, we propose a family of constrained-based approaches, termed PARROT, to predict how much of each enzyme is used based on the principle of minimizing the difference between a reference and an alternative growth condition. To this end, PARROT variants model the minimization of enzyme reallocation using four different (combinations of) distance functions. We demonstrate that the PARROT variant that minimizes the Manhattan distance between the enzyme allocation of a reference and an alternative condition outperforms existing approaches based on the parsimonious distribution of fluxes or enzymes for both Escherichia coli and Saccharomyces cerevisiae. Further, we show that the combined minimization of flux and enzyme allocation adjustment leads to inconsistent predictions. Together, our findings indicate that minimization of protein allocation rather than flux redistribution is a governing principle determining steady-state pathway activity for microorganism grown in alternative growth conditions.

Structural insights into the functions of Raf1 and Bsd2 in hexadecameric Rubisco assembly

Hexadecameric form I Rubisco, which consists of eight large (RbcL) and eight small (RbcS) subunits, is the most abundant enzyme on earth. Extensive efforts have been undertaken to engineer an improved Rubisco to speed up its catalytic efficiency and ultimately increase agricultural productivity. However, difficulties with correct folding and assembly in foreign hosts or in vitro have hampered the genetic manipulation of hexadecameric Rubisco. In this study, we reconstituted Synechococcus sp. PCC6301 Rubisco in vitro using the chaperonin system and assembly factors from cyanobacteria and Arabidopsis thaliana (At). Rubisco holoenzyme was produced in the presence of cyanobacterial Rubisco accumulation factor 1 (Raf1) alone or both AtRaf1 and bundle-sheath defective-2 (AtBsd2) from Arabidopsis. RbcL released from GroEL is assembly capable in the presence of ATP, and AtBsd2 functions downstream of AtRaf1. Cryo-EM structures of RbcL8–AtRaf18, RbcL8–AtRaf14–AtBsd28, and RbcL8 revealed that the interactions between RbcL and AtRaf1 are looser than those between prokaryotic RbcL and Raf1, with AtRaf1 tilting 7° farther away from RbcL. AtBsd2 stabilizes the flexible regions of RbcL, including the N and C termini, the 60s loop, and loop 6. Using these data, combined with previous findings, we propose the possible biogenesis pathways of prokaryotic and eukaryotic Rubisco.

Redox mediators stimulated chain elongation process in fluidized cathode electro-fermentation systems for caproate production

Medium chain fatty acids (MCFAs), the secondary products of traditional anaerobic fermentation, can be produced via chain elongation (CE), a process often retarded due to the difficulty during interspecies electron transfer (IET). This study employed redox mediators, neutral red (NR), methyl viologen (MV), and methylene blue (MB) as electron shuttles to expedite the electro-fermentation for caproate production by improving IET. Results showed that MV increased the MCFAs production by promoting acetate to ethanol conversion, leading to the highest MCFAs selectivity of 68.73%. While NR was indicated to improve CE by encouraging H2 production, and the biocathode had the highest electrical activity due to the smallest internal resistance and largest capacitance increase of 96% than the control. A higher proportion of Sutterella, Prevotella, and Hydrogenophaga, linked with the H2 mediated interspecies electron transfer (MIET) during CE process, was observed across redox mediators supplied groups compared to the control. The presence of mediators led to an elevated abundance of key enzymes for enhanced CE process and electron transfer. This study provided the perspective of the stimulated electron transfer for improved MCFAs production in electro-fermentation systems.

Remediation mechanism of high concentrations of multiple heavy metals in contaminated soil by Sedum alfredii and native microorganisms

Pollution accident of nonferrous metallurgy industry often lead to serious heavy metal pollution of the surrounding soil. Phytoremediation of contaminated soil is an environmental and sustainable technology, and soil native microorganisms in the process of phytoremediation also participate in the remediation of heavy metals. However, the effects of high concentrations of multiple heavy metals (HCMHMs) on plants and native soil microorganisms remain uncertain. Thus, further clarification of the mechanism of phytoremediation of HCMHMs soil by plants and native soil microorganisms is required. Using the plant Sedum alfredii (S. alfredii) to restore HCMHM-contaminated soil, we further explored the mechanism of S. alfredii and native soil microorganisms in the remediation of HCMHM soils. The results showed that (i) S. alfredii can promote heavy metals from non-rhizosphere soil to rhizosphere soil, which is conducive to the effect of plants on heavy metals. In addition, it can also enrich the absorbed heavy metals in its roots and leaves; (ii) native soil bacteria can increase the abundance of signal molecule-synthesizing enzymes, such as trpE, trpG, bjaI, rpfF, ACSL, and yidC, and promote the expression of the pathway that converts serine to cysteine, then synthesize substances to chelate heavy metals. In addition, we speculated that genes such as K19703, K07891, K09711, K19703, K07891, and K09711 in native bacteria may be involved in the stabilization or absorption of heavy metals. The results provide scientific basis for S. alfredii to remediate heavy metals contaminated soils, and confirm the potential of phytoremediation of HCMHM contaminated soil.

Gut microbiota-associated taurine metabolism dysregulation in a mouse model of Parkinson's disease.

PD is recognized as a multisystem disease concerning GI dysfunction and microbiota dysbiosis but still lacks ideal therapies. Recently, aberrant microbiota-derived metabolites are emerging as important participants in PD etiology. However, the alterations of gut microbiota community and serum untargeted metabolite profile have not been fully investigated in a PD mice model. Here, we discover sharply reduced levels of Lactobacillus and taurine in MPTP-treated mice. Moreover, Lactobacillus, Adlercreutzia, and taurine-related metabolites showed the most significant correlation with pathological and GI performance of PD mice. The abundances of microbial transporter and enzymes participating in the degeneration of taurine were disturbed in PD mice. Most importantly, taurine supplement ameliorates MPTP-induced motor deficits, DA neuron loss, and microglial activation. Our data highlight the impaired taurine-based microbiome-metabolism axis during the progression of PD and reveal a novel and previously unrecognized role of genera in modulating taurine metabolism.

Effects of the polyhalogenated carbazoles 3-bromocarbazole and 1,3,6,8-tetrabromocarbazole on soil microbial communities

Soil ecosystems are being more contaminated with polyhalogenated carbazoles (PHCZs), which raising much attention about their impact on soil microorganisms. 3-Bromocarbazole (3-BCZ) and 1,3,6,8-tetrabromocarbazole (1,3,6,8-TBCZ) are two typical PHCZs with high detection rates in the soil environment. However, ecological risk research on these two PHCZs in soil is still lacking. In the present study, after 80 days of exposure, the ecological influence of 3-BCZ and 1,3,6,8-TBCZ was investigated based on 16S rDNA sequencing, ITS sequencing, gene (16S rDNA, ITS, amoA, nifH, narG and cbbL) abundance and soil enzyme activity. The results showed that the bacterial 16S rDNA gene abundance significantly decreased under 3-BCZ and 1,3,6,8-TBCZ exposure after 80 days of incubation. The fungal ITS gene abundance significantly decreased under 1,3,6,8-TBCZ (10 mg/kg) exposure. PHCZs contributed to the alteration of bacteria and fungi community abundance. Bacteria Sphingomonas, RB41 and fungus Mortierella, Cercophora were identified as the most dominant genera. The two PHCZs consistently decreased the relative abundance of Sphingomonas, Lysobacter, Dokdonella, Mortierella and Cercophora etc at 80th day. These keystone taxa are related to the degradation of organic compounds, carbon metabolism, and nitrogen metabolism and may thus have influence on soil ecological functions. Bacterial and fungal functions were estimated using functional annotation of prokaryotic taxa (FAPROTAX) and fungi functional guild (FUNGuild), respectively. The nitrogen and carbon metabolism pathway were affected by 3-BCZ and 1,3,6,8-TBCZ. The soil nitrogen-related functions of aerobic ammonia oxidation were decreased but the soil carbon-related functions of methanol oxidation, fermentation, and hydrocarbon degradation were increased at 80th day. The effects of 3-BCZ and 1,3,6,8-TBCZ on the abundances of the amoA, nifH, narG, and cbbL genes showed a negative trend. These results elucidate the ecological effects of PHCZs and extend our knowledge on the structure and function of soil microorganisms in PHCZ-contaminated ecosystems.

Endomicrobiome of in vitro and natural plants deciphering the endophytes-associated secondary metabolite biosynthesis in Picrorhiza kurrooa, a Himalayan medicinal herb.

Picrorhiza kurrooa an endangered high-altitude Himalayan medicinal herb, is used as a potent hepatoprotective due to the presence of various secondary metabolites, with picrosides being the most bioactive. In-vitro propagation is used as a sustainable strategy for its conservation and cultivation. The in-vitro propagation produces P. kurrooa plants (Tc) in mass, but these plants have reduced secondary metabolites (especially picrosides) compared to plants growing in their natural habitats (Wt). Therefore, considering the well-established role of microbes in secondary metabolite biosynthesis, in this study, endomicrobiome of P. kurroa plants (Tc and Wt) was explored. Using high-throughput DNA-sequencing, the endophytic communities associated with leaves, roots, and rhizomes of Wt and Tc plants were characterized. Diversity analysis revealed a loss of diversity during in-vitro propagation, and the abundant phyla were Proteobacteria, Bacteroidetes, Parcubacteria (OD1), Firmicutes, and Verrucomicrobia. Besides, the presence of distinct genera specific to different parts of Wt plants was also revealed. Quantification of secondary metabolites demonstrated the reduced accumulation of picrosides and intermediates of picroside biosynthesis in the Tc plants compared to Wt plants. Host-secondary metabolite production was positively correlated to microbial community abundance, suggesting a dynamic interplay of host-endomicrobiota interaction. Predictive functional analysis revealed the abundance of enzymes of secondary metabolite biosynthesis (especially MVA/MEP and phenylpropanoid/shikimate pathway involved in picrosides biosynthesis) in the associated-endophytic community with predominance in roots and rhizomes of Wt plants. This investigation provides novel insight into the change in the endomicrobiome of Wt and Tc plants and their correlation to the biosynthesis of secondary metabolites, and that needs to be considered for cultivation practices.IMPORTANCEPicrorhiza kurrooa is a major source of picrosides, potent hepatoprotective molecules. Due to the ever-increasing demands, overexploitation has caused an extensive decline in its population in the wild and placed it in the endangered plants' category. At present plant in-vitro systems are widely used for the sustainable generation of P. kurrooa plants, and also for the conservation of other commercially important, rare, endangered, and threatened plant species. Furthermore, the in-vitro-generated plants had reduced content of therapeutic secondary metabolites compared to their wild counterparts, and the reason behind, not well-explored. Here, we revealed the loss of plant-associated endophytic communities during in-vitro propagation of P. kurrooa plants which also correlated to in-planta secondary metabolite biosynthesis. Therefore, this study emphasized to consider the essential role of plant-associated endophytic communities in in-vitro practices which may be the possible reason for reduced secondary metabolites in in-vitro plants.

Proteomic profile of the germinating seeds reveals enhanced seedling growth in Arabidopsis rpp1a mutant.

Ribosomal phosphoprotein P1 (RPP1) is an integral component of the P-protein stalk in the 60S subunit of eukaryotic ribosomes and is required for the efficient elongation of translation. Previously, Arabidopsis RPP1A was revealed to be involved in the regulation of seed size and seed storage protein accumulation. In this work, the seedling growth analysis shows that the knockout mutation of Arabidopsis RPP1A significantly promoted seedling growth, particularly in the shoots. The label-free quantitative proteomic analysis demonstrated that a total of 593 proteins were differentially accumulated between the germinating seeds of the wild-type Col-0 and rpp1a mutant. And these proteins were significantly enriched in the intracellular transport, nitrogen compound transport, protein transport, and organophosphate metabolic process. The abundance of proteins involved in the RNA and protein processing processes, including ncRNA processing and protein folding, were significantly increased in the rpp1a mutant. Mutation in RPP1A highlighted the effects on the ribosome, energy metabolism, and nitrogen metabolism. The abundance of enzymes involved in glycolysis and pyruvate mechanism was decreased in the germinating seeds of the rpp1a mutant. Whereas the processes of amino acid biosynthesis, protein processing in endoplasmic reticulum, and biosynthesis of cofactors were enhanced in the germinating seeds of the rpp1a mutant. Taken together, the lack of RPP1A triggered changes in other ribosomal proteins, and the higher amino acid contents in the seedlings of the rpp1a mutant probably contributed to enhanced biosynthesis, processing, and transport of proteins, resulting in accelerated growth. Our results show the novel role of a P-protein and shed new light on the regulatory mechanism of seedling growth.

Enhanced biological wastewater treatment supplemented with anaerobic fermentation liquid of primary sludge

The wastewater treatment performance in an inverted A2/O reactor supplemented with fermentation liquid of primary sludge was explored comparing to commercial carbon sources sodium acetate and glucose. Similar COD removal rate was observed with the effluent COD stably reaching the discharge standard for those 3 carbon sources. However, the fermentation liquid distributed more carbon source in the anaerobic zone. Fermentation liquid and sodium acetate tests achieved better nitrogen removal rate than glucose test. The fermentation liquid test showed the best biological phosphorus removal performance with the effluent phosphorus barely reaching the discharge standard. The microbial community characterization revealed that the fermentation liquid test was dominated by phylum Proteobacter in all the anoxic, anaerobic and aerobic zones. Denitrifying phosphorus accumulating organisms (PAOs) (i.e., genera Dechloromonas and unclassified_f__Rhodocyclaceae) were selectively enriched with high abundances (over 20%), which resulted in improved phosphorus removal efficiency. Moreover, the predicted abundances of enzymes involved in nitrogen and phosphorus removal were also enhanced by the fermentation liquid.

Sorghum grain germination as a route to improve kafirin digestibility: Biochemical and label free proteomics insights.

Kafirin, the sorghum grain storage protein presents lower digestibility compared to its cereals counterparts. Germination has been proposed as an adequate bioprocessing method to improve seed protein digestibility. Here, germination was rationalized so as to evenly sample germinated seeds and the dynamic changes of the proteome and several biochemical markers was connected for the first time with the in vitro protein digestibility of germinated seeds. Free sulfhydryl groups increased during germination and in vitro protein digestibility enhanced. The dynamic in abundance of several enzymes out of which 3 cysteine proteases were found to coincide with appearance of aqueous soluble peptides derived from kafirin at boot time of their degradation. The study provides deep information about the molecular events occurring during sorghum seed germination and reveals potential biomarkers of the kafirin proteolysis. It points a way to improve sorghum nutritional value through controlled germination.

Uncovering Microbiome Adaptations in a Full-Scale Biogas Plant: Insights from MAG-Centric Metagenomics and Metaproteomics.

The current focus on renewable energy in global policy highlights the importance of methane production from biomass through anaerobic digestion (AD). To improve biomass digestion while ensuring overall process stability, microbiome-based management strategies become more important. In this study, metagenomes and metaproteomes were used for metagenomically assembled genome (MAG)-centric analyses to investigate a full-scale biogas plant consisting of three differentially operated digesters. Microbial communities were analyzed regarding their taxonomic composition, functional potential, as well as functions expressed on the proteome level. Different abundances of genes and enzymes related to the biogas process could be mostly attributed to different process parameters. Individual MAGs exhibiting different abundances in the digesters were studied in detail, and their roles in the hydrolysis, acidogenesis and acetogenesis steps of anaerobic digestion could be assigned. Methanoculleus thermohydrogenotrophicum was an active hydrogenotrophic methanogen in all three digesters, whereas Methanothermobacter wolfeii was more prevalent at higher process temperatures. Further analysis focused on MAGs, which were abundant in all digesters, indicating their potential to ensure biogas process stability. The most prevalent MAG belonged to the class Limnochordia; this MAG was ubiquitous in all three digesters and exhibited activity in numerous pathways related to different steps of AD.

Degradation of polypropylene by fungi Coniochaeta hoffmannii and Pleurostoma richardsiae

The urgent need for better disposal and recycling of plastics has motivated a search for microbes with the ability to degrade synthetic polymers. While microbes capable of metabolizing polyurethane and polyethylene terephthalate have been discovered and even leveraged in enzymatic recycling approaches, microbial degradation of additive-free polypropylene (PP) remains elusive. Here we report the isolation and characterization of two fungal strains with the potential to degrade pure PP. Twenty-seven fungal strains, many isolated from hydrocarbon contaminated sites, were screened for degradation of commercially used textile plastic. Of the candidate strains, two identified as Coniochaeta hoffmannii and Pleurostoma richardsiae were found to colonize the plastic fibers using scanning electron microscopy (SEM). Further experiments probing degradation of pure PP films were performed using C. hoffmannii and P. richardsiae and analyzed using SEM, Raman spectroscopy and Fourier transform infrared spectroscopy with attenuated total reflectance (FTIR-ATR). The results showed that the selected fungi were active against pure PP, with distinct differences in the bonds targeted and the degree to which each was altered. Whole genome and transcriptome sequencing was conducted for both strains and the abundance of carbohydrate active enzymes, GC content, and codon usage bias were analyzed in predicted proteomes for each. Enzymatic assays were conducted to assess each strain’s ability to degrade naturally occurring compounds as well as synthetic polymers. These investigations revealed potential adaptations to hydrocarbon-rich environments and provide a foundation for further investigation of PP degrading activity in C. hoffmannii and P. richardsiae.

The OsTIL1 lipocalin protects cell membranes from reactive oxygen species damage and maintains the 18:3-containing glycerolipid biosynthesis under cold stress in rice.

Lipocalins constitute a conserved protein family that binds to and transports a variety of lipids while fatty acid desaturases (FADs) are required for maintaining the cell membrane fluidity under cold stress. Nevertheless, it remains unclear whether plant lipocalins promote FADs for the cell membrane integrity under cold stress. Here, we identified the role of OsTIL1 lipocalin in FADs-mediated glycerolipid remodeling under cold stress. Overexpression and CRISPR/Cas9 mediated gene edition experiments demonstrated that OsTIL1 positively regulated cold stress tolerance by protecting the cell membrane integrity from reactive oxygen species damage and enhancing the activities of peroxidase and ascorbate peroxidase, which was confirmed by combined cold stress with a membrane rigidifier dimethyl sulfoxide or a H2 O2 scavenger dimethyl thiourea. OsTIL1 overexpression induced higher 18:3 content, and higher 18:3/18:2 and (18:2 + 18:3)/18:1 ratios than the wild type under cold stress whereas the gene edition mutant showed the opposite. Furthermore, the lipidomic analysis showed that OsTIL1 overexpression led to higher contents of 18:3-mediated glycerolipids, including galactolipids (monoglactosyldiacylglycerol and digalactosyldiacylglycerol) and phospholipids (phosphatidyl glycerol, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl serine and phosphatidyl inositol) under cold stress. RNA-seq and enzyme linked immunosorbent assay analyses indicated that OsTIL1 overexpression enhanced the transcription and enzyme abundance of four ω-3 FADs (OsFAD3-1/3-2, 7, and 8) under cold stress. These results reveal an important role of OsTIL1 in maintaining the cell membrane integrity from oxidative damage under cold stress, providing a good candidate gene for improving cold tolerance in rice.

Enhanced nitrogen removal from micro-polluted water by Fe (II)/Fe (III) cycling under intermittent aeration

The potential of iron-carbon materials (FeC) as inorganic electron donors for aerobic denitrification of micro-polluted source water was explored. For this purpose, a raw water denitrification experiment was conducted using FeC under intermittent aeration condition. Then, magnetite was added as a biological carrier to construct another system, FeC and magnetite intermittent aeration reaction system, (FCMI). The findings revealed that FCMI notably removed the pollutants with the removal efficiencies of nitrate, total nitrogen, total phosphorus, and chemical oxygen demand reaching 95.63%, 88.62%, 90.48%, and 61.46%, respectively. Cycling between aerobic and anaerobic environments facilitated the formation of Fe (II)/Fe (III) cycle within FeC materials. Microbial profiling revealed that Proteobacteria and Actinobacteria comprised 97.14% of the total abundance in FCMI. Iron-reducing bacteria and aerobic denitrifying bacteria were found to thrive on the surface of magnetite materials. The abundance of enzymes related to denitrification under intermittent aeration exceeded those under continuous aeration levels by 8–19%. This study verifies the ability of iron electrons to cycle under intermittent aeration condition and proposes a novel strategy for in-situ denitrification of micro-polluted source water.

Differential metabolic reprogramming in developing soybean embryos in response to nutritional conditions and abscisic acid.

Seed storage compound deposition is influenced by both maternal and filial tissues. Within this framework, we analyzed strategies that operate during the development and filling of soybean embryos, using in vitro culture systems combined with metabolomics and proteomics approaches. The carbon:nitrogen ratio (C:N) of the maternal supply and the hormone abscisic acid (ABA) are specific and interacting signals inducing differential metabolic reprogrammings linked to changes in the accumulation of storage macromolecules like proteins or oils. Differences in the abundance of sugars, amino acids, enzymes, transporters, transcription factors, and proteins involved in signaling were detected. Embryos adapted to the nutritional status by enhancing the metabolism of both carbon and nitrogen under lower C:N ratio condition or only carbon under higher C:N ratio condition. ABA turned off multiple pathways especially in high availability of amino acids, prioritizing the storage compounds biosynthesis. Common responses induced by ABA involved increased sucrose uptake (to increase the sink force) and oleosin (oil body structural component) accumulation. In turn, ABA differentially promoted protein degradation under lower nitrogen supply in order to sustain the metabolic demands. Further, the operation of a citrate shuttle was suggested by transcript quantification and enzymatic activity measurements. The results obtained are useful to help define biotechnological tools and technological approaches to improve oil and protein yields, with direct impact on human and animal nutrition as well as in green chemistry.

Trimethylation profile of histones H3 lysine 4 and 9 in late preantral and early antral caprine follicles grown in vivo versus in vitro in the presence of anethole.

This study assessed the histones methylation profile (H3K4me3 and H3K9me3) in late preantral (PA)and early antral (EA)caprine follicles grown in vivo and in vitro, and the anethole effect during in vitro culture of PA follicles. Uncultured in vivo-grown follicles (PA,n = 64; EA, n = 73) were used as controls to assess the methylation profile and genes' expression related to apoptosis cascade (BAX, proapoptotic; BCL2, antiapoptotic), steroidogenesis (CYP17, CYP19A1), and demethylation (KDM1AX1, KDM1AX2, KDM3A). The isolated PA follicles (n = 174) were cultured in vitro for 6 days in α-MEM+ in either absence (control) or presence of anethole. After culture, EA follicles were evaluated for methylation, mRNA abundance, and morphometry. Follicle diameter increased after culture, regardless of treatment. The methylation profile and the mRNA abundance were similar between in vivo-grown PA and EA follicles. Anethole treatment led to higher H3K4me3 fluorescence intensity in EA follicles. The mRNA abundances of BAX, CYP17, and CYP19A1 were higher, and BCL2 and KDM3A were lower in in vitro-grown EA follicles than in vivo-grown follicles. In conclusion, in vitro follicle culture affected H3K4me3 fluorescence intensity, mRNA abundance of apoptotic genes, and steroidogenic and demethylase enzymes compared with in vivo-grown follicles.

Sodium butyrate promotes gastrointestinal development of preweaning bull calves via inhibiting inflammation, balancing nutrient metabolism, and optimizing microbial community functions.

Butyrate promotes the growth and gastrointestinal development of calves. But, the mechanisms behind its effects on signaling pathways of the gastrointestinal tract and rumen microbiome is unclear. This study aimed to reveal transcriptomic pathways of gastrointestinal epithelium and microbial community in response to butyrate supplementation in calves fed a high fiber starter. Fourteen Holstein bull calves (39.9±3.7kg, 14d of age) were assigned to 2 groups (sodium butyrate group, SB; control group, Ctrl). The SB group received 0.5% SB supplementation. At d 51, the calves were slaughtered to obtain samples for analysis of the transcriptome of the rumen and jejunum epithelium as well as ruminal microbial metagenome. Sodium butyrate supplementation resulted in a higher performance in average daily gain and development of jejunum and rumen papillae. In both the rumen and jejunum epithelium, SB down-regulated pathways related to inflammation including NF-κB (PPKCB, CXCL8, CXCL12), interleukin-17 (IL17A, IL17B, MMP9), and chemokine (CXCL12, CCL4, CCL8) and up-regulated immune pathways including the intestinal immune network for immunoglobulin A (IgA) production (CD28). Meanwhile, in the jejunum epithelium, SB regulated pathways related to nutritional metabolism including nitrogen metabolism (CA1, CA2, CA3), synthesis and degradation of ketone bodies (HMGCS2, BDH1, LOC100295719), fat digestion and absorption (PLA2G2F, APOA1, APOA4), and the PPAR signaling pathway (FABP4, FABP6, CYP4A11). The metagenome showed that SB greatly increased the relative abundance of Bacillus subtilis and Eubacterium limosum, activated ruminal microbial carbohydrate metabolism pathways and increased the abundance of carbohydrate hydrolysis enzymes. In conclusion, butyrate exhibited promoting effects on growth and gastrointestinal development by inhibiting inflammation, enhancing immunity and energy harvesting, and activating microbial carbohydrate metabolism. These findings provide new insights into the potential mechanisms behind the beneficial effects of butyrate in calf nutrition.

Dosage effect of micron zero-valent iron during thermophilic anaerobic digestion of waste activated sludge: Performance and functional community

This work examined the performance and microbial traits in a thermophilic anaerobic digestion (TAD) of waste activated sludge that was impacted by micron zero valent iron (mZVI). Results showed that methane production was promoted by 0.8, 11.9, and 12.0 times, respectively, when mZVI was at dosages of 25, 100, and 250 mg/g total solid (TS). Also, the consumption of volatile fatty acids was increased by mZVI at higher dosages (100 and 250 mg/g TS). Furthermore, 16S rRNA sequencing demonstrated that microbial community stabilized after day 18 regardless of the dosage of mZVI, and that different dosages of mZVI induced different shifts in the functional community of the archaea rather than the bacteria involved in TAD. As a result, mZVI at 100 mg/g TS could increase the relative abundance of archaeal genera Methanothermobacter the most, increasing by 22.8% at the end of TAD compared to CK. Besides, redundancy analysis revealed that the physicochemical properties explained 79.65% and 89.10% of the variations of bacterial and archaeal abundance, respectively. Also, the findings of the correlation analysis revealed that total dissolved iron, ferrous iron, pH, and ammonium nitrogen, may be the key divers of altering functional communities, particularly archaea. Moreover, mZVI at 100 and 250 mg/g TS boosted the metabolic pathways of environmental information processing (ABC transporters) in bacteria and carbon metabolism and methane metabolism for archaea, as well as relative abundances of enzymes and their activities involved in various methanogenic pathways. This study provides new perspectives on the application of mZVI in solid wastes treatments.

Insight into biofilm formation of wastewater treatment processes: Nitrogen removal performance and biological mechanisms

Biofilm formation affects biological nitrogen (N) removal, and a sequencing batch biofilm reactor (SBBR) was set up to evaluate the changes in N removal and microbial characteristics during biofilm formation. The results indicated that the average effluent concentration of chemical oxygen demand (COD), ammonia nitrogen (NH4+-N) and total nitrogen (TN) in the SBBR were 27.48, 1.41, and 13.52 mg L−1, respectively after biofilm formation. Furthermore, this process increased microbial richness, but reduced microbial diversity. Patescibacteria, Proteobacteria, and Bacteroides were the dominant phyla that did not change after biofilm formation. After biofilm formation, Firmicutes was eliminated while Spirochaetes involved in the interspecies relationship. Biofilm increased the nitrification and denitrification relating coding genes abundance (hao, narG, narZ, nxrA, narH, narY, nxrB, napA, napB, norB, norC and nosZ), and enhanced the processes of N respiration and denitrification, carbohydrate metabolism, amino acid metabolism and membrane transport. Meanwhile, correlation analysis between genera and transcriptome reflected that Zooglea, Micropruina, Aeromonas and Tessaracoccus played essential roles in biofilm formation and N removal. The key enzyme abundance of EC:1.7.99.1, EC:1.7.2.4, and EC:1.1.1.42 of N and tricarboxylic acid (TCA) cycle increased after biofilm formation. This study can reveal the effect of biofilm formation on biological N removal and provide a theoretical foundation for the application of biofilm process.

Discrepant diversity patterns and function of bacterial and fungal communities on an earthquake-prone mountain gradient in Northwest Sichuan, China.

Patterns of microbial diversity on elevational gradients have been extensively studied, but little is known about those patterns during the restoration of earthquake-fractured alpine ecosystems. In this study, soil properties, soil enzyme activities, abundance and diversity of soil bacterial and fungal communities at four positions along a 2.6-km elevational gradient in the Snow Treasure Summit National Nature Reserve, located in Pingwu County, Southwest China. Although there were no significant changes in the soil chemical environment, bacterial and fungal communities were significantly different at different elevations. The overall fungal community presented an N-shaped diversity pattern with increasing elevation, while bacterial diversity decreased significantly with elevation. Changes in microbial diversity were associated with soil phosphorus, plant litter, and variations in dominant microbial taxa. Differences in enzyme activities among elevations were regulated by microbial communities, with changes in catalase and acid phosphatase activities mainly controlled by Acidobacteria and Planctomycetaceae bacteria, respectively (catalase: p < 0.001; acid phosphatase: p < 0.01), and those in β-glucosidase, sucrase, and urease activities mainly controlled by fungi. The β-glucosidase and sucrase were both positively correlated with Herpotrichiellaceae, and urease was positively correlated with Sebacinaceae (p < 0.05). These findings contribute to the conservation and management of mountain ecosystems in the face of changing environmental conditions. Further research can delve into the specific interactions between microbial communities, soil properties, and vegetation to gain deeper insights into the intricate ecological dynamics within earthquake-prone mountain ecosystems.