Substrate Fermentation Research Articles

Controlling a kisser: fermented products and commercial insects' lures as attractants of kissing bugs.

Triatomines (Hemiptera: Heteroptera: Reduviidae) are hematophagous insects, well-known for their vectorial role in transmitting Trypanosoma cruzi Chagas (Kinetoplastida: Trypanosomatidae) parasites, the etiological agent of Chagas disease. Trapping these insects would limit human-triatomine interaction and, thus, control the disease. In this context, there is a critical need for effective lures to control triatomines. Through double-choice bioassays, we investigated the preference of Triatoma infestans Klug, T. pallidipennis Stal, and Rhodnius prolixus Stal triatomines for: (a) volatiles from fermented products (various fermentation types and substrates) and (b) commercial insect lures. Furthermore, we identified the chemical composition of these volatiles through headspace collection using Solid Phase Micro Extraction coupled with Gas Chromatograph-Mass Spectrometer (HS-SPME-GC-MS). Volatiles from lactic fermentation and certain fermented fruits, along with commercial lures, attracted triatomines, while other products exhibited possible repellent or dislodging properties. These findings hold promise for the control of triatomines and, ultimately, Chagas disease.

Determination of monacolin K and citrinin in red mold dioscorea and dried shrimp products using ultra high-performance liquid chromatography

In this study, red mold dioscorea (RMD) was utilized as a natural colorant to produce dried whiteleg shrimp (Litopenaeus vannamei). Whiteleg shrimp dyed with 50% diluted liquid RMD had a favorable appearance compared to control and undiluted liquid RMD. Dioscorea (Dioscorea hispida Dennst.) or bitter yam flour was used as a substrate in Monascus purpureus TISTR 3615 submerged fermentation to produce RMD. Dioscorea flour concentrations in a range of 10-50 g/L and various nitrogen sources at 4 g/L were investigated. The study showed that 10 g/L of diosorea flour and monosodium glutamate were chosen for further fermentation. Two major secondary metabolites in Monascus fermentation named monacolin K and citrinin, were determined in liquid RMD and dried shrimp products using UHPLC-DAD. Results showed that monacolin K were not detected in liquid RMD and all dried shrimp samples. Although, citrinin at a concentration of 2.06 mg/L was found in liquid RMD. Thus, RMD from the medium containing diosorea flour as a fermentation substrate can be used as colorant in dried shrimp production or other seafood products. However, the citrinin content in pigments remains an issue of concern.

Lactic Acid Production from Distiller’s Dried Grains Dilute Acid Hydrolysates

Lactic acid (LA) is an important chemical with diverse applications in various industries. LA can be produced by the fermentation of different substrates by many microorganisms such as bacteria, fungi, yeasts, and algae. Lactic acid bacteria (LAB) are generally accepted as the main producers of LA. A distinct characteristic of LAB is the complexity of the fermentation media. Distiller’s dried grains with solubles (DDGS), a by-product from bioethanol production, represent a promising substitute for costly sugars in the nutrition media for LA production. In the present paper, the possibility of using dilute acid DDGS hydrolysates as a substrate for LA fermentation was investigated. The influence of different factors (acid concentration, time, pressure, solid-to-liquid ratio) on the reducing sugars (RS) obtained was studied. Additional enzyme hydrolysis was carried out to increase RS content in the hydrolysates. LA production from hydrolysates without and with control of the pH during fermentation was monitored and compared with lactose as a substrate. Inhibition of the process was observed in both substrates in the absence of pH control which was overcome in the case of pH control. A mathematical model based on the Verhulst and Ludeking–Piret equations was proposed and tested, showing very good agreement with experimental data.

Optimisation of the Ethanol Fermentation Process Using Hydrothermal Pretreatment of Cellulose Waste-Effect of Fermentation Pattern and Strain.

Suitable fermentation substrates and fermentation modes can effectively improve the fermentation ethanol yield. In this study, we optimised the hydrothermal pretreatment conditions by orthogonal optimisation using waste tissue paper as substrate. These conditions consisted of 50 min duration in a high-pressure reactor with pure water as solvent at a temperature of 160 °C. The biomass to water ratio was maintained at a constant level. The cellulose content of the pretreated TP was 81.19 ± 4.06%, which was an increase of 21.59% compared to the blank control. The 72 h reducing sugar yield of pretreated TP was 0.61 g sugar/g paper, which was 38.64% higher than that of untreated TP. Subsequently, the pretreated TP was fermented under optimal conditions. The mixed group of Saccharomyces cerevisiae and Candida shehatae (SC) showed a distributed saccharification fermentation pattern, with an ethanol yield of 28.11 g/L in 72 h. On the other hand, the single Saccharomyces cerevisiae (S) exhibited a homobloc saccharification fermentation pattern, with an ethanol yield of 35.15 g/L in 72 h.

Markerless deletion of the putative type I and III restriction-modification systems in the cellulolytic bacterium Clostridium cellulovorans using a codBA-based counterselection technique

Cellulose from lignocellulosic biomass (LB) is of increasing interest for the production of commodity chemicals. However, its use as substrate for fermentations is a challenge due to its structural complexity. In this context, the highly cellulolytic Clostridium cellulovorans has been considered an interesting microorganism for the breakdown of LB. C. cellulovorans does not naturally produce solvents in useful concentrations, but this could be achieved by metabolic engineering. Unfortunately, this is hampered by the lack of tools for genetic engineering. We describe a genetic system that allows strain engineering by the allelic-coupled exchange method. First, the Gram-positive origin of pUB110 was identified as a suitable clostridial 'pseudo-suicide' origin of replication for the construction of deletion vectors. Second, an efficient counterselection strategy based on a codBA cassette and the use of 5-fluorocytosine as the counterselective compound was employed. Third, since the prevention of DNA transfer by host restriction-modification (RM) systems is a critical barrier to genome engineering, deletion plasmids containing flanking regions for the putative type I (Clocel_1114) and III (Clocel_2651) RM systems were constructed and transferred into C. cellulovorans. The restriction-less strains C. cellulovorans ΔClocel_1114 and C. cellulovorans ΔClocel_2651 exhibit high conjugation efficiency and can be easily used for further metabolic engineering.

Effect of microwave-puffed on Auricularia auricula polysaccharide and probiotic fermentation on its biotransformation and quality characteristics during storage period

In this study, probiotics with superior fermentation performance were screened, and the mixed-bacteria fermentation was carried out with Auricularia auricula treated with microwave-puffed process as fermentation substrate, and the changes in nutritional quality under different storage conditions were investigated. The results showed that the acid and bile salt resistance of Lactiplantibacillus plantarum 21,801 and 21,805 reached 95 % and 75 % respectively, and the intestinal adhesion was superior; microwave puffing treatment had the highest retention rate of A. auricula polysaccharides and the lowest loss of polyphenols, and no effect on soluble protein. Mixed bacterial fermentation significantly increased the total polyphenols and total flavonoids of A. auricula (p < 0.05), and the DPPH and ABTS radical scavenging reached 48.31 % and 73.21 % respectively. Furthermore, the viable counts, DPPH radical scavenging, color, and sensory quality of fermented A. auricula remained stable when stored at 4 °C. In contrast, when stored at 25 °C for 7 days, the taste was unfavorable, undesirable odor and spoilage occurred; by 21 days, DPPH clearance rate dropped below 40 % and color changed significantly (△E > 2). In conclusion, the probiotic mixed fermentation and storage conditions had a significant effect on the biometabolic transformation of macromolecules and other substances in A. auricula.

Study on the differences in microscopic oil displacement effects and action mechanisms of different rhamnolipid systems

Rhamnolipids are a class of anionic glycolipid surfactants produced through microbial metabolism. As a widely researched biosurfactant, rhamnolipids possess several advantages over traditional chemical surfactants, including non-toxicity, eco-friendliness, biodegradability, and biocompatibility, particularly in the context of microbial oil recovery applications. This class of surfactants enhances oil recovery by reducing the interfacial tension between oil and water, emulsifying residual oil, and modifying the wettability of rock surfaces. Furthermore, rhamnolipids maintain stability in high-temperature and high-salinity environments. However, rhamnolipids derived from different fermentation substrates exhibit variations in structure, composition, and properties, resulting in distinct displacement effects and mechanisms of action. This study focuses on two types of rhamnolipids: typical rhamnolipid and high-yield rhamnolipid, which are fermented using glycerol and rapeseed oil, respectively. Based on the characteristics of the target heavy oil reservoir, micromodels were designed and manufactured to conduct microfluidic experiments. The results obtained from imaging and video recording were analyzed qualitatively and quantitatively to explore the differences in effects and mechanisms between the two rhamnolipid systems. Results indicate that typical rhamnolipid increased recovery by 4.41% through delayed mechanisms involving wettability modification and residual oil emulsification. Conversely, high-yield rhamnolipid demonstrates an immediate effect by reducing interfacial tension, resulting in a recovery increase in 5.68%. According to the observed experimental phenomena and analytical trends, the conclusions evaluate the production increase, clarify the differences in mechanisms of action, and enhance the microscopic understanding of these surfactants. These findings provide directions for future investigations and serve as a reference for the design of related schemes.

Maximizing microbial activity and synergistic interaction to boost biofuel production from lignocellulosic biomass.

Addressing global environmental challenges and meeting the escalating energy demands stand as two pivotal issues in the current landscape. Lignocellulosic biomass emerges as a promising renewable bio-energy source capable of fulfilling the world's energy requirements on a large scale. One of the most important steps in lowering reliance on fossil fuel and lessening environmental effect is turning lignocellulosic biomass into biofuel. As carbon-neutral substitutes for traditional fuel, biofuel offer a solution to environmental concerns compared to conventional fuel. Effective utilization of lignocellulosic biomass is imperative for sustainable development. Ongoing research focuses on exploring the potential of various microorganisms and their co-interactions to synthesize diverse biofuels from different starting materials, including lignocellulosic biomass. Co-culture techniques demonstrate resilience to nutrient scarcity and environmental fluctuations. By utilising a variety of carbon sources, microbes can enhance their adaptability to environmental stressors and potentially increase productivity through their symbiotic interactions. Furthermore, compared to single organism involvement, co-interactions allow faster execution of multistep processes. Lignocellulosic biomass serves as a primary substrate for pre-treatment, fermentation, and enzymatic hydrolysis processes. This review primarily delves into the pretreatment, enzymatic hydrolysis process and the biochemical pathways involved in converting lignocellulosic biomass into bioenergy.

Impact of coconut water and grape juice blend on physico-chemical and sensory properties of wine

This study focused on crafting a unique wine by blending grape juice with coconut water (CW) in varying ratios (1:2, 2:2, and 2:1), supplemented with crystallized refined table sugar to maintain a 15 oBrix. Fermentation, initiated with the wine yeast S. Cerevisiae VR21, at 0.2 g/100 mL of must, occurred at room temperature. Daily monitoring of physio-chemical parameters [total soluble solids (TSS), pH, and acidity] provided insights into substrate utilization kinetics and fermentation dynamics. The chemical constituent analysis examined the impact of grape juice concentration on the CW wine's chemical properties. During the period of fermentation, the findings indicated significant changes in must constituents; pH was found to be decreased, TSS was decreased and attained steady state however, acidity continuously rose. Subsequently, a sensory analysis using GenStat software version 12.1 evaluated the formulated samples' appearance/color, aroma, taste/body, and overall acceptability. Sensory analysis revealed a preference for formulations with CW and grape juice concentration ratios of 2:2 and 1:2. Particularly, within these ratios, a formulation with a higher proportion of grape juice exhibited superior concentrations of aldehyde, total phenolic content, and antioxidant activity, suggesting enhanced overall quality. This research provides a comprehensive understanding of the chemical and sensory aspects of these distinctive coconut water wine blends.

The Flour Source’s Role as a Fermentation Substrate Influences the Physical Attributes of Nata de Coco

The production of coconut water in Indonesia is very abundant, but there are problems with its utilization. Discarding unused coconut water often results in an unpleasant odor, as fermentation produces acetic acid pollution. Coconut water has an economic value and is rich in nutrients. One product that can be made from coconut water is nata via fermentation. Making nata requires a source of carbon that can be obtained from flour and is influenced by its type. Thus, this study aims to examine the effect of adding different types of flour as a source of carbon for fermentation in making nata de coco. We use corn, tapioca, rice, and modified cassava flour (mocaf), along with granulated sugar as a sample control. The resulting nata de coco is analyzed for its physical characteristics, including yield, moisture content, and nata thickness. The results showed that nata made using carbon sources from rice flour produced the greatest yield (69.55%), as well as the highest thickness (3.958 mm). Nata, derived from rice flour, also has the highest moisture content (&gt;85%). The study's results indicate that the selection of flour as a carbon source during nata de coco production significantly impacts its yield, thickness, and moisture content

Purified diets containing high levels of soluble fiber and grain-based diets promote similar gastrointestinal morphometry yet distinct microbial communities.

Dietary fibers are essential for maintaining gut health. Insoluble fibers aid in fecal bulking and water retention while soluble fiber is a fermentative substrate for intestinal microbial communities. Grain-based diets (GBDs) are commonly used in preclinical research but the variability in ingredients across batches impedes reproducibility. Purified diets (PDs), which are composed of highly refined ingredients, pose a potential solution but the most widely used low-fat control PDs contain no soluble fiber. This study intended to identify a PD with a combination of fibers that promotes murine gut health and microbial diversity. A PD with optimal fiber composition would aid in the standardization and reproducibility of studies investigating intestinal physiology and the gut microbiota.

Diversity and Anti-Infectious Components of Cultivable Rhizosphere Fungi Derived from Three Species of Astragalus Plants in Northwestern Yunnan, China.

Astragalus, a group of legume plants, has a pronounced rhizosphere effect. Many species of Astragalus with limited resource reserves are distributed in the high-altitude area of northern Yunnan, China. Although some of these plants have high medicinal value, the recognition of them is still at a low level. The aim of this research is to explore the species diversity of cultivable rhizofungi derived from Astragalus acaulis, A. forrestii and A. ernestii growing in a special high-cold environment of northwest Yunnan and discover anti-infective components from these fungi. A total of 93 fungal strains belonging to 38 species in 18 genera were isolated and identified. Antibacterial and antimalarial screening yielded 10 target strains. Among them, the ethyl acetate crude extract of the fermented substrate of the rhizofungus Aspergillus calidoustus AA12 derived from the plant A. acaulis showed broad-spectrum antibacterial activity and the best antimalarial activity. Further chemical investigation led to the first discovery of seven compounds from the species A. calidoustus, including sesterterpine 6-epi-ophiobolin G; three sesquiterpenes, penicisochroman A, pergillin and 7-methyl-2-(1-methylethylethlidene)-furo [3,2-H]isoquinoline-3-one; and three polyketides, trypacidin, 1,2-seco-trypacidin and questin. Among them, the compound 6-epi-ophiobolin G exhibited moderate to strong antibacterial activity against six Gram-positive pathogens with the minimum inhibitory concentration (MIC) ranging from 25 to 6.25 μg/mL and a prominent inhibitory effect on the biofilm of Streptococcus agalactiae at an MIC value of 3.125 μg/mL. This compound also displayed potent antimalarial activity against Plasmodium falciparum strains 3D7 and chloroquine-resistant Dd2 at the half-maximal inhibitory concentration (IC50) values of 3.319 and 4.340 µmol/L at 72 h, respectively. This study contributed to our understanding of the cultivable rhizofungi from characteristic Astragalus plants in special high-cold environments and further increased the library of fungi available for natural anti-infectious product screening.

Optimization of laccase production by Pleurotus pulmonarius through solid substrate fermentation of tender coconut fiber: enhanced laccase production and biomass delignification

Optimization of laccase production by Pleurotus pulmonarius through solid substrate fermentation of tender coconut fiber: enhanced laccase production and biomass delignification

Predicting biohydrogen production from dark fermentation of organic waste biomass using multilayer perceptron artificial neural network (MLP–ANN)

The focus on sustainable energy has increased interest in biohydrogen production through dark fermentation of organic waste biomass, offering dual benefits of energy production and waste management. Optimizing this process is challenging due to complex interactions among substrate composition, microbial consortia, and fermentation parameters. A multilayer perceptron artificial neural network model was developed to predict biohydrogen yield from organic waste. The model, trained on 180 data points from 35 studies, uses inputs, such as substrate type, inoculum type, concentration, pH, and temperature, with hydrogen yield as the output. The multilayer perceptron artificial neural network model achieved high accuracy, with a root mean square error of 0.3838, a mean absolute percentage error of 0.1938, and a coefficient of determination of 0.8381. These results demonstrate the model's effectiveness in predicting biohydrogen production, providing a valuable tool for optimizing the fermentation process and advancing sustainable energy solutions.

Innovative Lactic Acid Production Techniques Driving Advances in Silage Fermentation

Lactic acid (LA) plays a crucial role in the silage process, which occurs through LA fermentation. Consequently, there is a strong correlation between lactic acid production and the efficiency of the silage. However, traditional methods face challenges like long fermentation times, low acid production, and unstable quality, limiting agricultural preservation. This paper aims to explore innovations in lactic acid production technologies and show how these technologies have driven the development of silage fermentation for agricultural conservation. First, the important role of LA in agricultural preservation and the limitations of traditional silage techniques are presented. Next, advancements in LA production methods are thoroughly examined, covering the selection of microbial strains and the substitution of fermentation substrates. Following this, new technologies for silage fermentation are explored, drawing from innovations in LA production. These include the selection of LA strains, optimization of fermentation conditions, and improvements in fermentation techniques. These innovations have proven effective in increasing LA production, improving feed quality, extending shelf life, and providing new solutions to enhance agricultural production and sustainability.

Potential and characteristics of bio-H2 production from the garden waste of ornamental plant Malus prunifolia fruit: Comparison between ethanol-type and butyrate-type fermentations

Edible fruit waste has been studied and considered to be the ideal substrates for low-cost bio-H2 production. However, the garden waste of ornamental plant fruits, which has similar components to edible fruit, is rarely studied. In this study, the potential and characteristics of fermentative bio-H2 production from the campus ornamental plant fruit of Malus prunifolia (MP) in both ethanol- and butyrate-types were investigated. Ethanoligenens harbinense B49 and Clostridium neuense G1 were employed as the inoculums for the ethanol- and butyrate-type fermentations, respectively. The results indicated that the fruit of MP is rich in fructose and sucrose, both of strains B49 and G1 could grow well with MP powder as the carbon source for fermentation. However, a higher H2 yield of 1.77 mol-H2/mol-hexose was obtained in the butyrate-type fermentation with strain G1, which was 1.4 times of that with strain B49. C. neuense G1 was more suitable for the H2 fermentation with MP as substrate, and a H2 yield of 0.55 mol-H2/mol-hexose by strain B49 was observed in the test of buffer system exchanging from ethanol-type to the butyrate-type. In summary, garden wastes such as fruits of ornamental plants should be given more attention as low-cost fermentation substrate candidates for bio-H2 production.

Evaluation of a phenolic-tolerant Wickerhamomyces anomalus strain for efficient ethanol production based on omics analysis

Lignocellulosic biomass is a highly economically viable substrate for ethanol fermentation, but the presence of phenolic compounds (PCs) in lignocellulosic hydrolysates severely hinders the growth of ethanologenic strains. This study aims to investigate the ethanol fermentation capabilities and PC tolerance of the newly isolated Wickerhamomyces anomalus CAP5. The strain demonstrated remarkable ethanol production, achieving 87.1 g/L with a yield of 0.47 g/g, exceeding 90 % of the theoretical maximum. To elucidate the mechanisms responsible for this high ethanol yield, comprehensive genome and transcriptome analyses were performed. These analyses identified key metabolic pathways, including acetate reabsorption and an incomplete glycerol synthesis pathway, as crucial factors contributing to the strain's superior performance. Moreover, W. anomalus CAP5 demonstrated notable resistance to PCs, particularly phenolic acids. In fermenting with coffee husk hydrolysate (CHH), which is rich in PCs, W. anomalus CAP5 achieved noteworthy ethanol production of 60.5 g/L, marking a record for ethanol production from CHH. Hence, W. anomalus CAP5 emerges as a promising strain for effective ethanol production from lignocellulosic hydrolysates enriched with PCs.

Biosynthesis of pterostilbene in Escherichia coli from resveratrol on macroporous adsorption resin using a two-step substrate addition strategy.

Pterostilbene (PST), a 3',5'-O-methylated derivative of resveratrol (RSV), is a potent natural antioxidant produced by some plants in trace amounts as defense compound. It exhibits various health-promoting activities, such as anticancer, antiviral, and antimicrobial effects. Large-scale biosynthesis of PST is crucial due to the challenges associated with extracting it from plants. This study aims to develop an efficient method for PST production using an engineered Escherichia coli strain by feeding RSV as a precursor. We introduced a two-step substrate addition strategy combined with immobilized RSV (IMRSV) on macroporous adsorption resin (MAR) to enhance PST production. Five MARs were selected for RSV immobilization, and the substrate addition strategy and fermentation parameters for PST synthesis were optimized. A maximum PST concentration of 403 ± 9 mg/L was achieved, representing a 239% increase over the control, which in a one-step addition of free RSV. The PST titer reached 395 ± 24 mg/L in a 3-L bioreactor. In conclusion, the combination of a two-step substrate addition system and IMRSV is a promising approach for the economical and industrial-scale production of PST.

Sustainable enhancement of biogas production from a cold-region municipal wastewater anaerobic digestion process using optimized sludge-derived and commercial biochar additives

Anaerobic digestion (AD) of municipal wastewater sludges produces valuable solid digestate and biogas. Biogas is a source of clean energy and enhancement of its production has been of recent interest for increased electricity generation, among other products. The objective of this study was the development of a novel municipal sludge-derived biochar and its application in a municipal wastewater AD system to increase the biogas production rate. Thickened waste-activated sludge (TWAS) samples were collected from the cold-region municipal wastewater treatment plant and used to synthesize biochar applied in the simulation of AD processes using laboratory-scale reactors. The TWAS-derived biochar was synthesized using the commonly used furnace pyrolysis (sludge-based biochar, SBC), and more novel microwave pyrolysis including phosphoric acid as a microwave activator (activated sludge-based biochar, ASBC). The microwave pyrolysis conditions were optimized using a computational fluid dynamics (CFD) technique. In addition, various commercially available carbon-based additives were assessed for their impacts on the AD process including activated carbon, wood-derived biochar, and forest residue-derived biochar. Results showed that the ASBC increased the cumulative methane production by 50% (333 mL/g VS) versus the control sample (221 mL/g VS) after 30 d. The ASBC showed higher surface area, electrical conductivity, and metal contents versus the other biochars which boosted the AD microbial community growth leading to higher organic matter conversion into biogas. In the ASBC-amended digesters, the bacterial phylum Bacteroidota, which contains a major genus of the dgA-11-gut-group, exhibited a synergy between organic substrate fermentation and volatile fatty acid production, resulting in enhanced biogas production. The TWAS biochar demonstrated promising performance in enhancing the AD process fostering energy and resource self-sufficiency at municipal wastewater treatment plants. This smart sludge management aligns well with sustainable waste management practices and clean energy production strategies, especially considering that the biochar was sourced from a readily available continuous waste-product stream.

Production of Magnesium Dilactate through Lactic Acid Fermentation with Magnesium Carbonate.

Magnesium dilactate is increasingly sought after for its applications in the pharmaceutical, food, and dietary supplement industries due to its essential role in various physiological processes. This study explores a sustainable method for synthesizing magnesium dilactate through lactic acid fermentation using tomato juice, coupling the neutralization of lactic acid with hydrated magnesium carbonate hydroxide. Utilizing the lactic acid bacteria Lactobacillus paracasei and Lactobacillus plantarum, fermentation was optimized in a 50% diluted MRS medium supplemented with glucose and tomato juice supplemented with glucose, yielding a maximum lactate concentration of 107 g/L. Notably, fermentation in diluted media proved more effective than in undiluted tomato juice, highlighting the inhibitory effects of certain organic compounds and the physical nature of the original tomato juice. Post-fermentation, magnesium lactate was crystallized, achieving high recovery rates of up to 95.9%. Characterization of the product through X-ray diffraction and scanning electron microscopy confirmed its crystalline purity. This research underscores the viability of tomato juice as a fermentation substrate, promoting the valorization of agricultural by-products while providing an eco-friendly alternative to traditional chemical synthesis methods for magnesium dilactate production.