Optimization Of Fermentation Conditions Research Articles

Actinomycins produced by endophyte Streptomyces sp. GLL-9 from navel orange plant exhibit high antimicrobial effect against Xanthomonas citri susp. citri and Penicillium italicum.

Citrus canker and citrus blue mold are two severe diseases in citrus plants, which are mainly caused by Xanthomonas citri susp. citri (Xcc) and Penicillium italicum, respectively. The currently widely used pesticides for these two diseases are harmful to human health and the environment. Therefore, searching for novel antimicrobial agents, especially from natural resources, is getting increasing interest. In this study, the crude extract of Streptomyces sp. GLL-9, an endophyte from a navel orange tree, was found to exhibit excellent antimicrobial effects against Xcc and P. italicum. Bioassay-guided isolation led to the discovery of three actinomycins (Acts), actinomycin X2 (Act-X2 ), actinomycin D (ActD), and actinomycin XOβ (Act-XOβ ). The MIC (minimum inhibitory concentration) values of Act-X2 , ActD, and Act-XOβ were 31.25, 62.50, and 62.50 μg mL-1 against Xcc, respectively, while 62.50 (Act-X2 ) and 125.00 μg mL-1 (ActD) against P. italicum, being better or comparable to the positive controls. The highest yield of Acts was obtained by solid-state fermentation with rice containing 1% L-tryptophan as a culture medium, being 6.03, 3.07, and 1.02 mg g-1 , for Act-X2 , ActD, and Act-XOβ , respectively. The ethyl acetate extract of Streptomyces sp. GLL-9 cultivated under the optimal fermentation conditions (EAE-1) can efficiently control these two citrus diseases by excessively producing reactive oxygen species (ROS) in both pathogens, damaging the cell membranes of P. italicum, and inhibiting the growth of Xcc. In addition, Act-X2 , ActD, and EAE-1 displayed broad-spectrum antifungal activity. EAE-1 and Acts produced by Streptomyces sp. GLL-9 have high potential as novel antimicrobial agents against plant pathogens. © 2023 Society of Chemical Industry.

Effective degradation of anthraquinones in Folium Sennae with Monascus fermentation for toxicity reduce and efficacy enhancement

Folium Sennae are widely used around the world, mainly in purging and removal of endogenous active substances, such as anthraquinone and its derivatives. However, the potential toxicity of anthraquinones to the liver, kidney, and intestinal limits the application of Folium Sennae. In this study, we aimed at safe regulation of Folium Sennae to degrade anthraquinones, boosting medicinal properties and reducing toxicity and potency with Monascus fermentation. Monascus strains H1102 for Folium Sennae fermentation were selected as the initial strain which was capable of producing high yields of functional pigment and low yields of hazardous citrinin. The anthraquinone degradation rate reached 41.2%, with 212.2 U mL−1 of the pigment and approximately 0.038 mg L−1 of the citrinin under optimal fermentation conditions followed by response surface streamlining, which met the requirements of reducing toxicity, increasing efficiency of Monascus fermented Folium Sennae. Furthermore, the Monascus/Folium Sennae culture had no observable toxic effect on HK-2 and L-02 cells in vitro and further inhibited cell apoptosis and necrosis. Overall, our results showed that Monascus fermentation could provide an alternative strategy for toxicity reduction of herbal medicines as well as efficacy enhancement.

Mutagenesis enhances gellan gum production by a novel Sphingomonas spp.: upstream optimization, kinetic modeling, and structural and physico-functional evaluation.

Gellan gum (GG) has gained tremendous attention owing to its diversified applications. However, its high production and hence market cost are still a bottleneck in its widespread utilization. In the present study, high GG producing mutant of Sphingomonas spp. was developed by random mutagenesis using ethyl methylsulphonate (EMS) for industrial fermentation and identified as Sphingomonas trueperi after 16S rRNA and matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) analysis. The fermentation conditions such as pH, temperature, and inoculum ratio were optimized by one factor at a time (OFAT) followed by screening of medium components by the Plackett-Burman statistical design. The most critical nutrients were further optimized by response surface methodology for maximizing GG production. The effect of dissolved oxygen tension in bioreactor on cell growth, substrate consumption, GG production, and batch productivity was elucidated. The highest GG titer (23 ± 2.4g/L) was attained in optimized medium at 10% inoculum (6.45 ± 0.5 log cfu/mL) under controlled fermentation conditions of pH (7), temperature (30°C), agitation (300-600rpm), and aeration (0.5-2.0 SLPM) at 22 ± 2% dissolved oxygen tension in a 10-L bioreactor. Kinetic modeling of optimized batch process revealed that logistic growth model could best explain biomass accumulation, while GG formation and substrate consumption were best explained by Luedeking-Piret and exponential decay model, respectively. Structural and physico-functional features of GG produced by mutant Sphingomonas spp. were characterized by HPLC, FTIR, NMR, DSC, TGA, GPC, SEM, and rheological analysis. The higher productivity (0.51g/L/h) under optimized fermentation conditions suggests potential consideration of mutant and process for commercial utilization.

Systematic optimization of fermentation conditions for in vitro fermentations with fecal inocula.

In vitro fermentation strategies with fecal inocula are considered cost-effective methods to gain mechanistic insights into fecal microbiota community dynamics. However, all in vitro approaches have their limitations due to inherent differences with respect to the in vivo situation mimicked, introducing possible biases into the results obtained. Here, we aimed to systematically optimize in vitro fermentation conditions to minimize drift from the initial inoculum, limit growth of opportunistic colonizers, and maximize the effect of added fiber products (here pectin) when compared to basal medium fermentations. We evaluated the impact of varying starting cell density and medium nutrient concentration on these three outcomes, as well as the effect of inoculation with fresh vs. stored fecal samples. By combining GC-MS metabolite profiling and 16 s rRNA gene-based amplicon sequencing, we established that starting cell densities below 1010 cells/ml opened up growth opportunities for members the Enterobacteriaceae family. This effect was exacerbated when using fecal samples that were stored frozen at -80°C. Overgrowth of Enterobacteriaceae resulted in lowered alpha-diversity and larger community drift, possibly confounding results obtained from fermentations in such conditions. Higher medium nutrient concentrations were identified as an additional factor contributing to inoculum community preservation, although the use of a less nutrient dense medium increased the impact of fiber product addition on the obtained metabolite profiles. Overall, our microbiome observations indicated that starting cell densities of 1010 cells/ml limited opportunities for exponential growth, suppressing in vitro community biases, whilst metabolome incubations should preferably be carried out in a diluted medium to maximize the impact of fermentable substrates.

Screening and optimization of extracellular pectinase produced by Bacillus thuringiensis SH7

Abstract The objective of the current research was to identify and evaluate the possibility of production of pectinase, also known as pectin degrading enzymes, from indigenous bacterial strains. Qualitative screening of isolated bacterial strains showed that among 29 bacterial strains, 5 have maximum enzymatic activity. The highest pectinase producing strains were quantitatively analyzed for enzyme production. SH7 strain was found as highest pectinase producer (0.77 IU/mL) that was further analyzed to molecular level by amplification of 16s rRNA. It was found 100% similar with other reported strains of Bacillus thuringiensis. Medium optimization was performed to optimize fermentation conditions for maximum enzyme yield. An experimental design containing 12 experimental runs was designed by Plackett–Burman design (PBD). Maximum pectinase activity was obtained at 45°C after 24 h when the growth medium was supplemented with 2.5% nitrogen, 5.0% substrate, MgSO4 as metal ion, 1% inoculum size, and pH was adjusted to 6. Factorial regression analysis of the PBD design was performed and the overall design was also found significant in terms of R square value. In PBD, the most significant factors for production were temperature, pH, metal ion concentration, and nitrogen source. Central composite design (CCD) design consisting of 26 experimental runs was employed to optimize these four significant factors. The overall model summary showed maximum pectinase activity (19.2 IU/mL) at 37°C temperature, 0.08 NaCl, 1.7% nitrogen source, and pH 8.4. In CCD, NaCl, nitrogen source, and pH were also reported as significant factors by the Pareto chart, probability plots, and 3D interactions.

Effects of Main Nutrient Sources on Improving Monascus Pigments and Saccharifying Power of Monascus purpureus in Submerged Fermentation

Hong Qu (HQ), obtained through fermentation of various grains using Monascus spp., has been widely utilized as the main and characteristic initial saccharification and traditional fermentation starter in the food brewing industry. The quality, color, and flavor of HQ and HQ wine are closely related to the saccharifying power (SP) and Monascus pigments (MPs) of Monascus spp. In this study, to optimize the culture medium in submerged fermentation by M. purpureus G11 for improving SP and MPs, the effects of carbon source, nitrogen source, inorganic salts, and vitamins on SP activity and biosynthesis of MPs were explored through single-factor analysis and response surface Box–Behnken experiments. The results showed that the optimal medium composition was 6.008% rice powder, 1.021% peptone, 0.0049% CuSO4, and 0.052% vitamin B1. Validation experiments performed under the optimized fermentation conditions showed a significant increase in MPs and SP by 14.91% and 36.24%, with maximum MPs and SP reaching 112.61 and 365.12 u/mL, respectively. This study provides a theoretical basis for enhancing MPs and SP in M. purpureus for HQ production, to improve the production efficiency and shorten the production cycle of HQ-related fermentation products.

Isolation and Optimal Fermentation Conditions of Bacillus licheniformis SFD-Y5 for a New Douchi Fibrinolytic Enzyme Producer

Cardiovascular disease (CVD) has become the leading cause of death, and it is critical to develop new functional foods to prevent intravascular thrombosis, the key cause of CVD. Fermented soy-based food is a good choice because of its native fibrinolytic enzyme (FE) activity. In this study, a strain that can produce a new type of fibrinolytic enzyme was selected from Chinese Douchi and identified as Bacillus licheniformis SFD-Y5 by molecular biology experiments and physiological and biochemical experiments. Single factor experiments combined with statistical experiments, including Plackett–Burman experiment, steepest ascent experiment and RSM (Box–Behnken design), were used to optimize the fermentation of FE by B. licheniformis SFD-Y5. The final FE activity was 2434.45 ± 28.49 IU/mL under optimal conditions, which is the highest FE activity produced by wild B. licheniformis so far. Further studies showed that Y5 FE is a serine metalloproteinase with good stability at alkaline pHs (pH 8.0–11.0). The results of our study could lay a foundation for the future production, molecular modification and further application in functional foods of Y5 FE.

Alternative Utilization of Pennisetum purpureum × Pennisetum americanum: Press Cake Conversion to Biobutanol

Conversion of Pennisetum purpureum × Pennisetum americanum (Napier Pak Chong1) press cake into biobutanol using Clostridium beijerinckii TISTR 1461 was proposed as an alternative to combustion in this study. The optimum conditions for biobutanol fermentation were determined using a full factorial design and a central composite design of experiment. The studied factors were initial pHs (5.50–6.50) and sugar concentrations (40–60 g/L), while butanol yield (g/g reducing sugar utilized) was specified as the optimization response. The results showed that the suitable enzyme loading of alkali-pretreated press cake (at 3% w/w NaOH, 10% substrate loading, boiling at 90 °C, with a reaction time of 1 h) was 10 FPU/g biomass, which provided a glucose yield of 345 mg/g pretreated press cake. The optimized pH and reducing sugar concentration were 6.08 and 43 g/L, respectively. At these conditions, the maximum butanol yield from the hydrolysate of NaOH-pretreated press cake was 0.135 g/g reducing sugar utilized (0.30 g/g glucose utilized). Apart from the possibility of generating much less pollution, it was estimated that using the same amount of press cake, butanol production could possibly have a value comparable to that obtained from combustion for electricity production. A new concept for overall Napier Pak Chong1 grass utilization was also presented.

L-Lactic Acid Production via Sustainable Neutralizer-Free Route by Engineering Acid-Tolerant Yeast Pichia kudriavzevii.

l-Lactic acid (l-LA) is a platform chemical obtained via microbial fermentation at a near-neutral pH value. Large amounts of neutralizers are required during this process, which increases the production costs in downstream processing as well as environmental burden. To address this challenge, an acid-tolerant yeast Pichia kudriavzevii E1 was isolated and metabolically engineered to produce l-LA without neutralizers. The genome of strain E1 was sequenced and a CRISPR-Cas9 system was developed in this newly isolated strain. Subsequently, the gene encoding pyruvate decarboxylase (pdc) was knocked out to subdue ethanol formation. Furthermore, the l-lactate dehydrogenase gene from Weizmannia coagulans 2-6 and the codon-optimized L-ldhA gene from Bos taurus were introduced into P. kudriavzevii E1 chromosome to redirect the ethanol fermentation pathway to l-LA production. Deletion of the dld(chr3) gene further increased the optical purity of l-LA. After optimizing fermentation conditions, the maximum titer of l-LA in the 5 L fermenter reached 74.57 g/L without any neutralizers, with an optical purity of 100% and a maximum yield of 0.93 g/g glucose. This is the first report of optically pure l-LA production without neutralizers and the engineered acid-tolerant yeast paves the way for the sustainable production of l-LA via a green route.

Efficient Synthesis of Chiral Aryl Alcohol with a Novel Kosakonia radicincitans Isolate in Tween 20/L-carnitine: Lysine-Containing Synergistic Reaction System.

Chiral trifluoromethyl alcohols as vital intermediates are of great interest in fine chemicals and especially in pharmaceutical synthesis. In this work, a novel isolate Kosakonia radicincitans ZJPH202011 was firstly employed as biocatalyst for the synthesis of (R)-1-(4-bromophenyl)-2,2,2-trifluoroethanol ((R)-BPFL) with good enantioselectivity. By optimizing fermentation conditions and bioreduction parameters in aqueous buffer system, the substrate concentration of 1-(4-bromophenyl)-2,2,2-trifluoroethanone (BPFO) was doubled from 10 to 20mM, and the enantiomeric excess (ee) value for (R)-BPFL increased from 88.8 to 96.4%. To improve biocatalytic efficiency by strengthening the mass-transfer rate, natural deep-eutectic solvents, surfactants and cyclodextrins (CDs) were introduced separately in the reaction system as cosolvent. Among them, L-carnitine: lysine (C: Lys, molar ratio 1:2), Tween 20 and γ-CD manifested higher (R)-BPFL yield compared with other same kind of cosolvents. Furthermore, based on the excellent performance of both Tween 20 and C: Lys (1:2) in enhancing BPFO solubility and ameliorating cell permeability, a Tween 20/C: Lys (1:2)-containing integrated reaction system was then established for efficient bioproduction of (R)-BPFL. After optimizing the critical factors involved in BPFO bioreduction in this synergistic reaction system, BPFO loading increased up to 45mM and the yield reached 90.0% within 9h, comparatively only 37.6% yield was acquired in neat aqueous buffer. This is the first report on K. radicincitans cells as new biocatalyst applied in (R)-BPFL preparation, and the developed Tween 20/C: Lys-containing synergistic reaction system has great potential for the synthesis of various chiral alcohols.

Response surface optimization of poly-β-hydroxybutyrate synthesized by Bacillus cereus L17 using acetic acid as carbon source

A strain of Bacillus that can tolerate 10 g/L acetic acid and use the volatile fatty acids produced by the hydrolysis and acidification of activated sludge to produce polyhydroxyalkanoate was screened from the activated sludge of propylene oxide saponification wastewater. The strain was identified by 16S rRNA sequencing and phylogenetic tree analysis and was named Bacillus cereus L17. Various characterization methods showed that the polymer synthesized by strain L17 is poly-β-hydroxybutyrate, which has low crystallinity, good ductility and toughness, high thermal stability and a low polydispersity coefficient. It has wide thermoplastic material operating space as well as industrial and medicinal applications. The optimal fermentation conditions were determined by single factor optimization. Then, Plackett-Burman and Box-Behnken design experiments were carried out according to the single factor optimization results, and the response surface optimization was completed. The final results were: initial pH 6.7, temperature 25 °C, and loading volume 124 mL. The verification experiment showed that the yield of poly-β-hydroxybutyrate after optimization increased by 35.2 % compared to that before optimization.

Effect of Lactobacillus acidophilus Fermentation on the Quality of Glutinous Rice Products

AbstractThe effect of Lactobacillus acidophilus and spontaneous fermentation on the quality characteristics of glutinous rice products are investigated in this study. Optimization of the product quality based on orthogonal experiments on the four fermentation conditions ensure that the product still has good sensory properties when stored during its shelf life at room temperature. The optimal fermentation conditions based on the range analysis are the mass ratio of glutinous rice, corn and rice 8:1:1, L. acidophilus inoculum of 8%, fermentation temperature of 37 °C, and fermentation time of 28 h. Under this condition, rice products’ hardness, viscosity, elasticity, cohesiveness, and taste significantly increase compared to the spontaneously fermented products. The pasting characteristics, water absorption index, thermodynamic properties, short‐range ordered structure, and microstructure of two kinds of fermented rice flours are analyzed. L. acidophilus fermented rice flour is more prone to pasting and ageing, with a higher proportion of short‐chain amylopectin in starch and lower final viscosity and water absorption, with a denser and smoother internal structure. In conclusion, fermentation improves the properties of glutinous rice flour and its products, and L. acidophilus fermentation is more effective than spontaneous fermentation.

Genome Analysis and Optimization of Caproic Acid Production of Clostridium butyricum GD1-1 Isolated from the Pit Mud of Nongxiangxing Baijiu.

Caproic acid is a precursor substance for the synthesis of ethyl caproate, the main flavor substance of nongxiangxing baijiu liquor. In this study, Clostridium butyricum GD1-1, a strain with high caproic acid concentration (3.86 g/l), was isolated from the storage pit mud of nongxiangxing baijiu for sequencing and analysis. The strain's genome was 3,840,048 bp in length with 4,050 open reading frames. In addition, virulence factor annotation analysis showed C. butyricum GD1-1 to be safe at the genetic level. However, the annotation results using the Kyoto Encyclopedia of Genes and Genomes Automatic Annotation Server predicted a deficiency in the strain's synthesis of alanine, methionine, and biotin. These results were confirmed by essential nutrient factor validation experiments. Furthermore, the optimized medium conditions for caproic acid concentration by strain GD1-1 were (g/l): glucose 30, NaCl 5, yeast extract 10, peptone 10, beef paste 10, sodium acetate 11, L-cysteine 0.6, biotin 0.004, starch 2, and 2.0% ethanol. The optimized fermentation conditions for caproic acid production by C. butyricum GD1-1 on a single-factor basis were: 5% inoculum volume, 35°C, pH 7, and 90% loading volume. Under optimal conditions, the caproic acid concentration of strain GD1-1 reached 5.42 g/l, which was 1.40 times higher than the initial concentration. C. butyricum GD1-1 could be further used in caproic acid production, NXXB pit mud strengthening and maintenance, and artificial pit mud preparation.

Preparation of exopolysaccharides by heat-resistant Weizmannia coagulans using anion exchange resins

Weizmannia coagulans with high temperature tolerance and stability was selected for the production of exopolysaccharides (EPS) at the fermentation temperature of 52 °C. The weak basic anion exchange resin was employed in the fermentation for the construction of buffer system to neutralize some lactic acid and form intermolecular hydrogen bonds with hydroxyl groups on capsular polysaccharides of bacteria via tertiary amine groups, resulting in the increased growth space for capsular polysaccharides. In addition, the fermentation at relatively high temperature significantly decreased the adsorption of Ca2+ derived from CaO used for pH adjustment to the resin and sustained the ion exchange capability of the resin available for the formation of intermolecular hydrogen bonds with capsular polysaccharides. The concentration of EPS obtained achieved up to 19.8 g/L, which was increased by approximately 20-fold as compared to the concentration of EPS obtained by conventional fermentation. The conversion rate of carbohydrates to EPS reached up to 35.1%. The EPS produced under the optimized fermentation condition included neutral and acidic heteropolysaccharides with a molecular mass of 3.4 × 104–1.5 × 106 Da. The concentration and molecular mass distribution of EPS exhibited good reproducibility. This study provides a novel method for the industrial production of EPS.

Optimization of Fermentation Conditions and Metabolite Profiling of Grape Juice Fermented with Lactic Acid Bacteria for Improved Flavor and Bioactivity.

To enrich the flavor compounds and retain the content of polyphenolics in grape juice (GJ) under long-term storage, Lactiplantibacillus plantarum, Lactobacillus acidophilus, Lacticaseibacillus casei, and Lacticaseibacillus paracasei, were screened and the optimal fermentation conditions were determined as fermentation temperature of 41.2 °C for 24 h with an initial LAB density of 8.5 × 106 CFU/mL. Surprisingly, the retention rates of TPC still remained at 50% after storage for 45 days at 4 °C. Moreover, 251 different metabolites were identified, include 23 polyphenolics, 11 saccharides, and 9 organic acids. Most importantly, the total content of polyphenolics reserved was 92.65% at the end of fermentation. Among them, ephedrannin A content significantly decreased; however, 2',6'-Di-O-acetylononin gradually increased with the fermentation time, which resulted in FGJ maintaining excellent bioactivity. Meanwhile, organic acid content (palmitoylethanolamide, tetraacetylethylenediamine) increased with saccharides (linamarin) decreasing, which leads to FGJ having a unique taste. Furthermore, a total of 85 Volatile organic compounds (VOCs) were identified, mainly including esters, aldehydes, and alcohols. Interestingly, key VOCs could be formed by carboxylic acids and derivatives, and fatty acyls via complex metabolic pathways.

Optimized fermentation conditions for dragon fruit yogurt

Flavored yogurt which includes fruit yogurt is more popular with consumers, the addition of fruit to yogurt improves the flavor making the yogurt product more appealing to consumers. Dragon fruit has been used in yogurt production to enhance the nutritional and physiological characteristics of final product. However, limited information exists on optimizing the processing parameters of dragon fruit yogurt production. This study, therefore, adopted a central composite design to optimize the processing parameters of dragon fruit yogurt production and evaluated the statistical relation between the independent factors (dragon fruit puree, milk powder, corn starch, and fermentation time) and their responses (viscosity, syneresis, pH and titratable acidity). Results indicated that milk powder and fermentation time were the principal factors that affect the various responses. Based on the surface response models, the optimum formulation conditions for dragon fruit yogurt were dragon fruit puree (31.42 g), corn starch (7.0 g), milk powder (49.02 g), and fermentation time (6.0 hr). This produced a measured response of pH (4.77 ± 0.12), viscosity (30.33 ± 9.29 cP), syneresis (25.24 ± 5.36 %), and titratable acidity (1.05 ± 0.03 %). The optimum fermentation conditions had a favorable effect on the response values, suggesting that dragon fruit yogurt production could benefit from the optimized formula.

Effect of sourdough on the quality of whole wheat fresh noodles fermented with exopolysaccharide lactic acid bacteria

In this study, the impact of exopolysaccharides (EPS)-positive strain Weissella cibaria (W. cibaria) fermented sourdough on the quality of whole wheat fresh noodles (WWNs) and its improvement mechanisms were studied. The optimal fermentation conditions were found to be 30% sucrose content, fermented at 25 °C for 12 h, which yielded the highest EPS, 28.06 g/kg, in the W. cibaria fermented sourdough with sucrose (DW+). During storage, the sourdough reduced polyphenol oxidase activities and delayed the browning rate of noodles. The DW+ increased the hardness by 11.98% from 2184.99 to 2446.83 g, and the adhesiveness increased by 19.60%, i.e., from 72.01 to 86.13 g∙s of the noodles. The EPS mitigated acidification of sourdough, prevented the disaggregation of glutenin macropolymers (GMP), and increased sourdough elastic modulus. In addition, scanning electron microscope and confocal laser scanning microscopy of noodles containing EPS sourdough also demonstrated the uniform distribution of gluten proteins. The starch granules were also closely embedded in the gluten network. Thus, the present work indicated that the EPS produced sourdough delayed browning and improved the WWNs texture, indicating its potential to enhance the quality of whole grain noodles.

Exploring the Effects of Solid-State Fermentation on Polyphenols in Acanthopanax senticosus Based on Response Surface Methodology and Nontargeted Metabolomics Techniques

In order to optimize the process of solid-state fermentation and understand the changes of active ingredients during fermentation of A.senticosus, we used response surface methodology to optimize the solid-state fermentation process of A.senticosus and analyzed the effect of fermentation on the active components of A.senticosus by nontargeted metabolomics techniques. The effects of fermentation conditions on polyphenol content were studied in terms of fermentation time, fermentation temperature, water content, and the inoculum level through Plackett–Burman design and response surface experiments. The optimized fermentation conditions were a fermentation time of 60 h, a fermentation temperature of 37°C, a water content of 55%, and an inoculum level of 12.5%, and the polyphenol content reached 10.25 ± 0.05 mg·g−1, which was 40% higher than that before fermentation. Furthermore, the metabolites and metabolic pathways of A. senticosus were analyzed before and after fermentation using nontargeted metabolomics techniques, and differences in metabolites were investigated based on the analysis of the principal components. The results showed that A.senticosus contained 57 phenolics, and a total of 12 polyphenolic metabolites were significantly different before and after fermentation (including phenols and isoflavonoids), among which the content of caffeic acid and its derivatives with antioxidant capacity was significantly increased, which may improve the antioxidant capacity of fermented A.senticosus. Therefore, solid-state fermentation opens new avenues for the clinical development and application of A. senticosus.

Mitigation of N2O emissions via enhanced denitrification in a biological landfill leachate treatment using external carbon from fermented sludge

The effects of an external carbon source (C-source) on the mitigation of N2O gas (N2O(g)) emissions from landfill leachate were investigated via enhanced denitrification using anaerobically fermented sewage sludge. Anaerobic fermentation of sewage sludge was conducted under thermophilic conditions with progressively increasing organic loading rates (OLR). Optimal conditions for fermentation were determined based on the efficiency of hydrolysis and the concentrations of sCOD and volatile fatty acids (VFAs) as follows: at an OLR of 40.48 ± 0.77 g COD/L·d with 1.5 days of solid retention time (SRT), 14.68 ± 0.59% of efficiency of hydrolysis, 14.42 ± 0.30 g sCOD/L and 7.85 ± 0.18 g COD/L of VFAs. Analysis of the microbial community in the anaerobic fermentation reactor revealed that degradation of sewage sludge might be potentially affected by proteolytic microorganisms producing VFAs from proteinaceous materials. Sludge-fermentate (SF) retrieved from the anaerobic fermentation reactor was used as the external C-source for denitrification testing. The specific nitrate removal rate (KNR) of the SF-added condition was 7.54 mg NO3−N/g VSS·hr, which was 5.42 and 2.43 times higher than that of raw landfill leachate (LL) and a methanol-added condition, respectively. In the N2O(g) emission test, the liquid phase N2O (N2O–N(l)) of 20.15 mg N/L was emitted as N2O(g) of 19.64 ppmv under only LL-added condition. On the other hand, SF led to the specific N2O(l) reduction rate (KN2O) of 6.70 mg N/g VSS hr, resulting in mitigation of 1.72 times the N2O(g) emission compared to under the only-LL-added condition. The present study revealed that N2O(g) emissions from biological landfill leachate treatment plants can be attenuated by simultaneous reduction of NO3−N and N2O(l) during enhanced denitrification via a stable supply of an external C-source retrieved from anaerobically fermented organic waste.

Optimization of fermentation conditions as a metabolic strategy for the high-yield and high-selectivity bio-based 2,3-butanediol production

Optimization of culture conditions can be used as an alternative metabolic control strategy to genetic engineering to improve bio-based 2,3-butanediol (2,3-BDO) production, especially its yield and optical purity. In this study, a first-order model was determined to predict the 2,3-BDO yield, which reached about 90% in batch cultures at 32° C, pH = 5, and kLa ∼ 5.0 ± 2.7 h−1, using a new and safe Paenibacillus peoriae NRRL BD-62. No acetoin accumulation and an absolute levo-2,3-BDO optical purity were also verified. About 39.4 g/L of 2,3-BDO, with a yield of 0.43 g/g, and a levo-:meso-2,3-BDO ratio of 1.9:1, was obtained at the expense of acetoin, lactic acid, and ethanol in fed-batch fermentation at kLa adjusted within its optimized range (∼7.5 h−1) to overcome operational limitations using high initial glucose concentrations. The low pH contributed to the carbon flux shift toward the levo-isomer synthesis, possibly due to the preferential butanediol dehydrogenase activation, while the highly oxygen-limited environment favored the complete acetoin conversion, enhancing the 2,3-BDO yield. These results provide significant contributions to the high-yield, acetoin-free, and safe bio-based 2,3-BDO production of particular interest in large-scale fermentations.