Fermentation Broth Research Articles

Research on pear residue dietary fiber and Monascus pigments extracted through liquid fermentation.

Pear residue, a byproduct of pear juice extraction, is rich in soluble sugar, vitamins, minerals, and cellulose. This study utilized Monascus anka in liquid fermentation to extract dietary fiber (DF) from pear residue, and the structural and functional characteristics of the DF were analyzed. Soluble DF (SDF) content was increased from 7.9/100g to 12.6g/100g, with a reduction of average particle size from 532.4 to 383.0nm by fermenting with M. anka. Scanning electron microscopy and infrared spectroscopic analysis revealed more porous and looser structures in Monascus pear residue DF (MPDF). Water-, oil-holding, and swelling capacities of MPDF were also enhanced. UV-visible spectral analysis showed that the yield of yellow pigment in Monascus pear residue fermentation broth (MPFB) was slightly higher than that in the Monascus blank control fermentation broth. The citrinin content in MPFB and M. anka seed broth was 0.90 and 0.98ug/mL, respectively. Therefore, liquid fermentation with M. anka improved the structural and functional properties of MPDF, suggesting its potential as a functional ingredient in food.

Metabolomics combined with intestinal microbiota analysis to reveal the effects of Inonotus hispidus on chronic kidney disease caused by high-protein diet

In recent years, the global incidence of chronic kidney disease caused by popular high-protein diet is on the rise. As a traditional edible and medicinal fungus in Asian countries, Inonotus hispidus has been proved to have a variety of biological activities. In this study, a mouse model of chronic kidney disease was established, and the effect of Inonotus hispidus on chronic kidney disease induced by high protein diet was clarified by the comprehensive strategy of metabonomics and intestinal microbiota analysis. We found that Inonotus hispidus fermentation broth may alleviate renal damage by regulating glycerol phospholipid metabolism and bile acid metabolism, and its fermentation broth can reverse the intestinal microbiota imbalance caused by high-protein diet. In addition, through analysis, it is found that high-protein diet can down-regulate the level of Farnesoid X receptor in the kidney, resulting in lipid accumulation and a series of pathological changes in the kidney. We believe that Inonotus hispidus has the potential to become a new type of safe and effective functional food for the treatment of chronic kidney disease.

Fermentation Analytical Technology (FAT): Monitoring industrial E. coli fermentations using absolute quantitative 1H NMR spectroscopy

BackgroundTo perform fast, reproducible, and absolute quantitative measurements in an automated manner has become of paramount importance when monitoring industrial processes, including fermentations. Due to its numerous advantages – including its inherent quantitative nature – Proton Nuclear Magnetic Resonance (1H NMR) spectroscopy provides an ideal tool for the time-resolved monitoring of fermentations. However, analytical conditions, including non-automated sample preparation and long relaxation times (T1) of some metabolites, can significantly lengthen the experimental time and make implementation in an industrial set up unfeasible. ResultsWe present a high throughput method based on Standard Operating Procedures (SOPs) and 1H NMR, which lays the foundation for what we call Fermentation Analytical Technology (FAT). Our method was developed for the accurate absolute quantification of metabolites produced during Escherichia coli industrial fermentations. The method includes: (1) a stopped flow system for non-invasive sample collection followed by sample quenching, (2) automatic robot-assisted sample preparation, (3) fast 1H NMR measurements, (4) metabolites quantification using multivariate curve resolution (MCR), and (5) metabolites absolute quantitation using a novel correction factor (k) to compensate for the short recycle delay (D1) employed in the 1H NMR measurements. The quantification performance was tested using two sample types: buffer solutions of chemical standards and real fermentation samples. Five metabolites – glucose, acetate, alanine, phenylalanine and betaine – were quantified. Absolute quantitation ranged between 0.64 and 3.40 mM in pure buffer, and 0.71–7.76 mM in real samples. SignificanceThe proposed method is generic and can be straight forward implemented to other types of fermentations, such as lactic acid, ethanol and acetic acid fermentations. It provides a high throughput automated solution for monitoring fermentation processes and for quality control through absolute quantification of key metabolites in fermentation broth. It can be easily implemented in an at-line industrial setting, facilitating the optimization of the manufacturing process towards higher yields and more efficient and sustainable use of resources.

Streptomyces isolate SOM013, a potential agent against microbial resistance and gastric ulcers

The menace of microbial resistance and re-emerging disease is still a problem for healthcare givers globally, and the need for newer sources of potent antibiotics has become paramount. This study investigated the antimicrobial and antiulcer activities of Streptomyces isolate SOM013. Streptomyces isolates were cultivated and purified following standard microbiological protocols. Secondary metabolites were recovered and characterized from Streptomyces isolate SOM013 via broth fermentation and extraction. Varying concentrations (0.5 mg/mL, 0.025 mg/mL and 0.0125 mg/mL) of the SOM013 extract were used for antimicrobial screening against resistant bacteria and medically important fungi (methicillin-resistant Escherichia coli, Oxacillin resistant Helicobacter pylori, Shigella spp, extended broad-spectrum resistant Pseudomonas aeruginosa, Streptococcus spp, Campylobacter spp, Candida albicans, Aspergillus niger, and Aspergillus flavus). The antiulcer activity of the SOM013 was also examined in a methanol-induced gastric ulcer animal model. A total of 23 Streptomyces spp were recovered from the study. Methanolic extract of the SOM013 isolates was more potent across the clinical test microorganisms compared to water extract. The antimicrobial activity was dose dependent, with methanolic extract at 0.05 g/mL displaying the highest zone of inhibition (18.8 ± 0.3 mm) when tested against extended broad-spectrum resistant Pseudomonas aeruginosa. Further, the extract's ulcer index and protection efficacy were significant as the concentration increased (P < 0.01). SOM013 isolate has a moderate antimicrobial and high antiulcer activity worthy of pharmacological exploration.

Inhibition of LPS-Induced Skin Inflammatory Response and Barrier Damage via MAPK/NF-κB Signaling Pathway by Houttuynia cordata Thunb Fermentation Broth.

Houttuynia cordata Thunb is rich in active substances and has excellent antioxidant and anti-inflammatory activity. Scanning electron microscopy and gel permeation chromatography were used to analyze the molecular characteristics of the fermentation broth of Houttuynia cordata Thunb obtained through fermentation with Clavispora lusitaniae (HCT-f). The molecular weight of HCT-f was 2.64265 × 105 Da, and the polydispersity coefficient was 183.10, which were higher than that of unfermented broth of Houttuynia cordata Thunb (HCT). By investigating the active substance content and in vitro antioxidant activity of HCT-f and HCT, the results indicated that HCT-f had a higher active substance content and exhibited a superior scavenging effect on 2,2-diphenyl-1-picrylhydrazyl radicals and hydroxyl radicals, with IC50 values of 11.85% and 9.01%, respectively. Our results showed that HCT-f could effectively alleviate the increase in the secretion of inflammatory factors and apoptotic factors caused by lipopolysaccharide (LPS) stimulation, and had a certain effect on repairing skin barrier damage. HCT-f could exert an anti-inflammatory effect by down-regulating signaling in the MAPK/NF-κB pathway. The results of erythrocyte hemolysis and chicken embryo experiments showed that HCT-f had a high safety profile. Therefore, this study provides a theoretical basis for the application of HCT-f as an effective ingredient in food and cosmetics.

Co-production of 1,3-propanediol and phage phiKpS2 from the glycerol fermentation by Klebsiella pneumoniae

As an alternative to antibiotics in response to antimicrobial-resistant infections, bacteriophages (phages) are garnering renewed interest in recent years. However, the massive preparation of phage is restricted using traditional pathogens as host cells, which incurs additional costs and contamination. In this study, an opportunistic pathogen, Klebsiella pneumoniae used to convert glycerol to 1,3-propanediol (1,3-PDO), was reused to prepare phage after fermentation. The phage infection showed that the fed-batch fermentation broth containing 71.6 g/L 1,3-PDO can be directly used for preparation of phage with a titer of 1 × 108 pfu/mL. Then, the two-step salting-out extraction was adopted to remove most impurities, e.g. acetic acid (93.5%), ethanol (91.5%) and cells (99.4%) at the first step, and obtain 1,3-PDO (56.6%) in the top phase as well as phage (97.4%) in the middle phase at the second step. This integrated process provides a cheap and environment-friendly manner for coproduction of 1,3-PDO and phage.Graphical abstract

Isolation, structural characterization and immunomodulatory activity on RAW264.7 cells of a novel exopolysaccharide of Dictyophora rubrovalvata

Fungal polysaccharides have been explored by many for both structural studies and biological activities, but few studies have been done on the extracellular polysaccharides of Dictyophora rubrovalvata, so a new exopolysaccharide was isolated from Dictyophora rubrovalvata and its structure and its immunological activity were investigated. The crude exopolysaccharide (EPS) was purified by DEAE52 cellulose and Sephadex G-200 to obtain a new acidic polysaccharide (DR-EPS). DR-EPS (2.66 × 103 kDa) was consisted mainly of mannose, glucose, galactose and glucuronic acid with a molar ratio of 1: 0.86: 0.20: 0.01. In addition, DR-EPS increased the phagocytic activity of RAW264.7 cells up to 2.67 times of the blank control group. DR-EPS improved intracellular nucleic acid and glycogen metabolism as observed by AO and PAS staining. DR-EPS(40 μg/mL) promoted NO production up to 30.66 μmol, enhanced acid phosphatase (ACP) and superoxide dismutase (SOD) activities, with activity maxima of 660 U/gprot and 96.27 U/mgprot, respectively, and DR-EPS (160 μg / mL) significantly increased the lysozyme content as 2.73 times of the control group. The good immunological activity of extracellular polysaccharides of Dictyophora rubrovalvata provides directions for the use of fermentation broths.

Diversity and Functionality of Bacteria Associated with Different Tissues of Spider Heteropoda venatoria Revealed through Integration of High-Throughput Sequencing and Culturomics Approaches

Spiders host a diverse range of bacteria in their guts and other tissues, which have been found to play a significant role in their fitness. This study aimed to investigate the community diversity and functional characteristics of spider-associated bacteria in four tissues of Heteropoda venatoria using HTS of the 16S rRNA gene and culturomics technologies, as well as the functional verification of the isolated strains. The results of HTS showed that the spider-associated bacteria in different tissues belonged to 34 phyla, 72 classes, 170 orders, 277 families, and 458 genera. Bacillus was found to be the most abundant bacteria in the venom gland, silk gland, and ovary, while Stenotrophomonas, Acinetobacter, and Sphingomonas were dominant in the gut microbiota. Based on the amplicon sequencing results, 21 distinct cultivation conditions were developed using culturomics to isolate bacteria from the ovary, gut, venom gland, and silk gland. A total of 119 bacterial strains, representing 4 phyla and 25 genera, with Bacillus and Serratia as the dominant genera, were isolated. Five strains exhibited high efficiency in degrading pesticides in the in vitro experiments. Out of the 119 isolates, 28 exhibited antibacterial activity against at least one of the tested bacterial strains, including the pathogenic bacteria Staphylococcus aureus, Acinetobacter baumanii, and Enterococcus faecalis. The study also identified three strains, GL312, PL211, and PL316, which exhibited significant cytotoxicity against MGC-803. The crude extract from the fermentation broth of strain PL316 was found to effectively induce apoptosis in MGC-803 cells. Overall, this study offers a comprehensive understanding of the bacterial community structure associated with H. venatoria. It also provides valuable insights into discovering novel antitumor natural products for gastric cancer and xenobiotic-degrading bacteria of spiders.

Recovery of organic acids from lactic acid-enriched fermentation broth via salting-out assisted solvent extraction

The organic acids (OAs) market has been expanding due to their versatility and wide range of uses in several industrial applications, thus prompting a transition from chemical to biological approaches to generate such high-value products. Nevertheless, downstream processes to separate OAs from the fermentation broth represent, to date, the main cost in biological OAs manufacturing. With the goal of limiting this issue, the present study investigated OAs separation from a lactic acid (LA)-rich fermentation broth through physical, i.e., using ethyl acetate (EA), and physico-chemical, i.e., using tributyl phosphate (TBP), solvent extraction. In addition, the salting-out effect of different salts on OAs recovery was investigated. Among the investigated conditions, a combination of pure TBP and 40% ammonium sulfate ensured the highest LA extraction efficiency of 65%. Besides, mixing TBP with EA (50:50) enabled to maintain the same LA extraction efficiency while reducing the process costs, limiting the toxicity of the chemicals involved, and obtaining an extracted OAs mix with lower co-metabolites, i.e. volatile fatty acids, compared to using pure TBP. Overall, solvent extraction assisted by salting-out agents were shown to be a promising method to separate OAs from a fermentation broth in liquid form, especially for fermentation processes operated at low pH.

Reactive liquid-liquid extraction of lactic acid from microfiltered sweet sorghum silage press juice in an agitated extraction column using a hydrophobic natural deep eutectic solvent as modifier

Downstream processing of carboxylic acids from fermentation broths is cost-intensive and environmentally harmful, which makes methods such as reactive liquid-liquid extraction attractive separation technologies. To enhance the sustainability of liquid-liquid extraction processes, research focuses on sustainable solvents like hydrophobic natural deep eutectic solvents. However, the application of hydrophobic natural deep eutectic solvents in agitated extraction columns for continuous processing has never been investigated. This is essential to facilitate their use in industrial processes, promoting cleaner production. In this study, the application of a thymol-menthol-based deep eutectic solvent as a modifier (with the extractant tri-n-octylamine) in an agitated extraction column is evaluated for reactive liquid-liquid extraction of lactic acid from microfiltered sweet sorghum silage press juice. In addition, three-stage cross-current extractions were performed. The thymol-menthol-based deep eutectic solvent showed efficient phase separation, stable operation, and chemical stability in the agitated extraction column. In three-step cross-current operation, the extraction efficiency for lactic acid was increased from 37.0 ± 0.4 % without feed pH adjustment to 76.2 ± 0.9 % with continuously adjusting the pH value to 2.05 ± 0.02. The thymol-menthol-based deep eutectic solvent shows great potential for application in agitated extraction columns. This is highly promising because natural deep eutectic solvents can enhance the sustainability of extraction processes and reduce the reliance on fossil-based solvents.

Chicken manure-derived biochar enhanced the potential of Comamonas testosteroni ZG2 to remediate Cd contaminated soil.

The combined remediation of Cd-contaminated soil using biochar and microorganisms has a good application value. In this study, the effect of chicken manure-derived biochar on CdCO3 precipitation induced by Comamonas testosteroni ZG2 was investigated. The results showed that biochar could be used as the carrier of strain ZG2, enhance the resistance of strain ZG2 to Cd, and reduce the toxicity of Cd to bacterial cells. Cd adsorbed by biochar could be induced by strain ZG2 to form CdCO3 precipitation. Strain ZG2 could also induce CdCO3 precipitation when biochar was added during precipitation formation and fermentation broth formation. The CdCO3 precipitation could enter the pores of the biochar and attach to the surface of the biochar. The single and combined effects of strain ZG2 and biochar could realize the remediation of Cd-contaminated soil to a certain extent. The overall effect was in the order of strain ZG2 with biochar > biochar > strain ZG2. The combination of strain ZG2 and biochar reduced soil available Cd by 48.2%, the aboveground biomass of pakchoi increased by 72.1%, and the aboveground Cd content decreased by 73.3%. At the same time, it promoted the growth and development of the root system and improved the microbial community structure of the rhizosphere soil. The results indicated that chicken manure-derived biochar could enhance the stability of CdCO3 precipitation induced by strain ZG2, and strain ZG2 combined with biochar could achieve a more stable remediation effect on Cd-contaminated soil.

Modulation effect of sulfated polysaccharide from Sargassum fusiforme on gut microbiota and their metabolites in vitro fermentation.

The present study demonstrated the digestion behavior and fermentation characteristics of a sulfated polysaccharide from Sargassum fusiforme (SFSP) in the simulated digestion tract environment. The results showed that the molecular weight of two components in SFSP could not be changed by simulated digestion, and no free monosaccharide was produced. This indicates that most of SFSP can reach the colon as prototypes. During the fermentation with human intestinal flora in vitro, the higher-molecular-weight component of SFSP was utilized, the total sugar content decreased by 16%, the reducing sugar content increased, and the galactose content in monosaccharide composition decreased relatively. This indicates that SFSP can be selectively utilized by human intestinal flora. At the same time, SFSP also changed the structure of intestinal flora. Compared with the blank group, SFSP significantly increased the abundance of Bacteroidetes and decreased the abundance of Firmicutes. At the genus level, the abundances of Bacteroides and Megamonas increased, while the abundances of Shigella, Klebsiella, and Collinsella decreased. Moreover, the concentrations of total short-chain fatty acids (SCFAs), acetic, propionic and n-butyric acids significantly increased compared to the blank group. SFSP could down-regulate the contents of trimethylamine, piperidone and secondary bile acid in fermentation broth. The contents of nicotinic acid, pantothenic acid and other organic acids were increased. Therefore, SFSP shows significant potential to regulate gut microbiota and promote human health.

The influence of Lactobacillus plantarum fermentation in selenium-enriched Brassica napus L.: changes in the nutritional constituents, bioactivities and bioaccessibility

Selenium (Se)-enriched Brassica napus L. is a valuable organic Se supplement. In this study, the fermentation broth enriched with organic Se (FFS) was prepared using Lactobacillus plantarum to ferment the substrate of Se-enriched Brassica napus L. Significant increases were observed after fermentation in total sugars, reducing sugars, soluble proteins, total phenolic content (TPC), and total flavonoid content (TFC). The organic Se was retained at a concentration of 54.75 ​mg/g in the freeze-dried sample. Principal component analysis and cluster analysis showed good separation between the FFS and unfermented (FS) groups. Fragrant 2-ethyloxetane had the highest content among all volatiles, while sinapine had the highest content among all phenolic compounds. The fermentation process showed remarkable improvement in the abundance and concentration of volatile compounds and phenolic contents, making FFS exhibit strong antioxidant activity and inhibitory capacity against α-glucosidase activity. The bioaccessibility of phenolic compounds was significantly greater in FFS compared to FS. ADMET analysis revealed that the majority of phenolic compounds contained in FFS did not exhibit mutagenicity toxicity, hepatotoxicity, skin sensitization, or blood-brain barrier penetration, indicating a favorable level of biosafety. Overall, our study provides a new insight into the further utilization of Se-enriched Brassica napus L. in foods.

A new Schizophyllum commune strain as a potential biocontrol agent against blueberry root rot.

In recent years, blueberry root rot has been caused mainly by Fusarium commune, and there is an urgent need for a green and efficient method to control this disease. To date, research on Schizophyllum commune has focused on antioxidant mechanisms, reactive dye degradation, etc., but the mechanism underlying the inhibition of pathogenic microorganisms is still unclear. Here, the control effects of S. commune on F. commune and blueberry root rot were studied using adversarial culture, tissue culture, and greenhouse pot experiments. The results showed that S. commune can dissolve insoluble phosphorus and secrete various extracellular hydrolases. The results of hyphal confrontation and fermentation broth antagonism experiments showed that S. commune had a significant inhibitory effect on F. commune, with inhibition rates of 70.30% and 22.86%, respectively. Microscopy results showed distortion of F. commune hyphae, indicating that S. commune is strongly parasitic. S. commune had a significant growth-promoting effect on blueberry tissue-cultured seedlings. After inoculation with S. commune, inoculation with the pathogenic fungus, or inoculation at a later time, the strain significantly reduced the root rot disease index in the potted blueberry seedlings, with relative control effects of 79.14% and 62.57%, respectively. In addition, S. commune G18 significantly increased the antioxidant enzyme contents in the aboveground and underground parts of potted blueberry seedlings. We can conclude that S. commune is a potential biocontrol agent that can be used to effectively control blueberry root rot caused by F. commune in the field.

Fabrication of Ti3C2Tx/ZnONPs composite with high antibacterial and antibiofilm activity

Microbial contamination poses a serious threat to human health and socioeconomics, and is a major problem for global public health. Herein, we fabricated Ti3C2Tx/ZnONPs composite by using Lactobacillus plantarum fermentation broth to reduce ZnONPs in situ on few-layer Ti3C2Tx MXene after etching and delamination treatment. Antibacterial experiments showed that the Ti3C2Tx/ZnONPs composite exhibited significant antibacterial and antibiofilm activity against both Gram-negative E. coli and Gram-positive S. aureus. Moreover, the exploration of the antibacterial mechanisms demonstrated that Ti3C2Tx/ZnONPs could inhibit and kill bacteria by disrupting bacterial cell wall/membrane, causing leakage of intracellular contents, interfering with metabolism and inducing oxidative stress. The synergistic effect of multiple antibacterial mechanisms can effectively prevent the evolution of resistance. Overall, as a novel inorganic antibacterial material, Ti3C2Tx/ZnONPs composite holds a promising application in the antimicrobial field.

Developing a High-Umami, Low-Salt Soy Sauce through Accelerated Moromi Fermentation with Corynebacterium and Lactiplantibacillus Strains.

The traditional fermentation process of soy sauce employs a hyperhaline model and has a long fermentation period. A hyperhaline model can improve fermentation speed, but easily leads to the contamination of miscellaneous bacteria and fermentation failure. In this study, after the conventional koji and moromi fermentation, the fermentation broth was pasteurized and diluted, and then inoculated with three selected microorganisms including Corynebacterium glutamicum, Corynebacterium ammoniagenes, and Lactiplantibacillus plantarum for secondary fermentation. During this ten-day fermentation, the pH, free amino acids, organic acids, nucleotide acids, fatty acids, and volatile compounds were analyzed. The fermentation group inoculated with C. glutamicum accumulated the high content of amino acid nitrogen of 0.92 g/100 mL and glutamic acid of 509.4 mg/100 mL. The C. ammoniagenes group and L. plantarum group were rich in nucleotide and organic acid, respectively. The fermentation group inoculated with three microorganisms exhibited the best sensory attributes, showing the potential to develop a suitable fermentation method. The brewing speed of the proposed process in this study was faster than that of the traditional method, and the umami substances could be significantly accumulated in this low-salt fermented model (7% w/v NaCl). This study provides a reference for the low-salt and rapid fermentation of seasoning.

Water-soluble biopolymers calcium polymalate derived from fermentation broth of Aureobasidium pullulans markedly alleviates osteoporosis and fatigue

Osteoporosis is a prevalent condition characterized by bone loss and decreased skeletal strength, resulting in an elevated risk of fractures. Calcium plays a crucial role in preventing and managing osteoporosis. However, traditional calcium supplements have limited bioavailability, poor solubility, and adverse effects. In this study, we isolated a natural soluble biopolymer, calcium polymalate (PMACa), from the fermentation broth of the fungus Aureobasidium pullulans, to investigate its potential as an anti-osteoporosis therapeutic agent. Characterization revealed that linear PMA-Ca chains juxtaposed to form a porous, rod-like state, in the presence of Ca2+. In vivo mouse models demonstrated that PMA-Ca significantly promoted the conversion of serum calcium into bone calcium, and stimulated bone growth and osteogenesis. Additionally, PMA-Ca alleviated exercise fatigue in mice by facilitating the removal of essential metabolites, such as serum lactate (BLA) and blood urea nitrogen (BUN), from their bloodstream. In vitro studies further showed that PMA-Ca strengthened osteoblast cell activity, proliferation, and mineralization. And PMA-Ca upregulated the expression of some genes involved in osteoblast differentiation, indicating a potential correlation between bone formation and PMACa. These findings indicate that soluble PMA-Ca has the potential to be a novel biopolymer-based calcium supplement with sustainable production sourced from the fermentation industry.

Bacillus velezensis strain NA16 shows high poultry feather-degrading efficiency, protease and amino acid production

Isolated Bacillus velezensis strain NA16, which produces proteases, amino acids and the transcription levels of different keratinolytic enzymes and disulfide reductase genes in whole gene sequencing, was evaluated during feather degradation. The result shows under optimum fermentation conditions, chicken feather fermentation showed total amino acid concentration of 7599 mg/L, degradation efficiency of 99.3% at 72 h, and protease activity of 1058 U/mL and keratinase activity of 288 U/mL at 48 h. Goose feather fermentation showed total amino acid concentration of 4918 mg/L (96 h), and degradation efficiency was 98.9% at 120 h. Chicken feather fermentation broth at 72 h showed high levels of 17 amino acids, particularly phenylalanine (1050 ± 1.90 mg/L), valine (960 ± 1.04 mg/L), and glutamic (950 ± 3.00 mg/L). Scanning electron microscopy and Fourier transform infrared analysis revealed the essential role of peptide bond cleavage in structural changes and degradation of feathers. Protein purification and zymographic analyses revealed a key role in feather degradation of the 39-kDa protein encoded by gene1031, identified as an S8 family serine peptidase. Whole genome sequencing of NA16 revealed 26 metalloproteinase genes and 22 serine protease genes. Among the proteins, S8 family serine peptidase (gene1031, gene1428) and S9 family peptidase (gene3132) were shown by transcription analysis to play major roles in chicken feather degradation. These findings revealed the transcription levels of different families of keratinolytic enzymes in the degradation of feather keratin by microorganisms, and suggested potential applications of NA16 in feather waste management and amino acid production.

Assessing effect of Trichoderma asperellum T16 on management of Bursaphelenchus xylophilus

Pine wilt disease caused by Bursaphelenchus xylophilus is an extremely serious issue worldwide. Trichoderma asperellum is used as a biocontrol agent due to its efficient inhibition of various pathogens. Three solvents, methanol, petroleum ether, and acetone, were used for extracting the fermentation broth of T. asperellum. All three extracts displayed a significant difference in nematocidal efficacy of more than 70% at 72 hours. Water-soluble natural products or polyphenolic compounds exhibited the greatest nematocidal ability after 24 hours of treatment with methanol. However, lipophilic substances exhibit long-lasting nematocidal ability. During this process, the antioxidant enzyme activities of T. asperellum significantly increased, including those of catalase, superoxide dismutase, and peroxidase. The interaction between T. asperellum and B. xylophilus induces the synthesis of glycoside hydrolase family 18 (chitinases) and peptidase S8, potential subtilisin, which dissolve the body wall of the nematodes. Fatty acid metabolism was significantly increased, and a defensive response against substances secreted by the nematode was produced. Antioxidant reaction mechanisms and detoxification metabolism were enriched and included a large number of upregulated DEGs, including GST, CYP450, LysM, and RTA1. The dual and toxic mechanisms exhibited by T. asperellum during its response to B. xylophilus involve multiple pathways and genes that are collectively regulated. This is the first study to investigate the effects of T. asperellum on B. xylophilus. This study provides a theoretical basis for the potential use of T. asperellum as a biocontrol agent against B. xylophilus.

Integrative Genomics and Bioactivity-Guided Isolation of Novel Antimicrobial Compounds from Streptomyces sp. KN37 in Agricultural Applications.

Actinomycetes have long been recognized as an important source of antibacterial natural products. In recent years, actinomycetes in extreme environments have become one of the main research directions. Streptomyces sp. KN37 was isolated from the cold region of Kanas in Xinjiang. It demonstrated potent antimicrobial activity, but the primary active compounds remained unclear. Therefore, we aimed to combine genomics with traditional isolation methods to obtain bioactive compounds from the strain KN37. Whole-genome sequencing and KEGG enrichment analysis indicated that KN37 possesses the potential for synthesizing secondary metabolites, and 41 biosynthetic gene clusters were predicted, some of which showed high similarity to known gene clusters responsible for the biosynthesis of antimicrobial antibiotics. The traditional isolation methods and activity-guided fractionation were employed to isolate and purify seven compounds with strong bioactivity from the fermentation broth of the strain KN37. These compounds were identified as 4-(Diethylamino)salicylaldehyde (1), 4-Nitrosodiphenylamine (2), N-(2,4-Dimethylphenyl)formamide (3), 4-Nitrocatechol (4), Methylsuccinic acid (5), Phenyllactic acid (6) and 5,6-Dimethylbenzimidazole (7). Moreover, 4-(Diethylamino)salicylaldehyde exhibited the most potent inhibitory effect against Rhizoctonia solani, with an EC50 value of 14.487 mg/L, while 4-Nitrosodiphenylamine showed great antibacterial activity against Erwinia amylovora, with an EC50 value of 5.715 mg/L. This study successfully isolated several highly active antimicrobial compounds from the metabolites of the strain KN37, which could contribute as scaffolds for subsequent chemical synthesis. On the other hand, the newly predicted antibiotic-like substances have not yet been isolated, but they still hold significant research value. They are instructive in the study of active natural product biosynthetic pathways, activation of silent gene clusters, and engineering bacteria construction.