Fermentation Broth Research Articles

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.

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.

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.

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.

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.

Isolation and characterization of Stenotrophomonas pavanii GXUN74707 with efficient flocculation performance and application in wastewater treatment

The identification of microorganisms with excellent flocculants-producing capability and optimization of the fermentation process are necessary for the wide-scale application of bioflocculants. Therefore, we isolated and identified a highly efficient flocculation performance strain of Stenotrophomonas pavanii GXUN74707 from the sludge. The optimal fermentation and flocculation conditions of strain S. pavanii GXUN74707 was in fermentation medium with glucose and urea as the carbon and nitrogen sources, respectively, at pH 7.0 for 36 h, which treatment of kaolin suspension with 0.5 mL of the fermentation broth resulted in a flocculation rate of 99.0%. The bioflocculant synthesized by strain S. pavanii GXUN74707 was found mainly in the supernatant of the fermentation broth. Chemical analysis revealed that the pure bioflocculant consisted of 79.70% carbohydrates and 14.38% proteins. The monosaccharide components of MBF-GXUN74707 are mainly mannose (5.96 μg/mg), galactose (1.86 μg/mg), and glucose (1.73 μg/mg). Infrared spectrometric analysis showed the presence of carboxyl (COO-), hydroxyl (-OH) groups. The SEM images showed clumps of rod-shaped bacteria with adhesion of extracellular products. Furthermore, the strain decolored dye wastewater containing direct black, direct blue, and Congo red by 89.2%, 95.1%, 94.1%, respectively. The chemical oxygen demand (COD) and biological oxygen demand (BOD) removal rates after treatment of aquaculture wastewater with the fermentation broth were 68% and 23%, respectively. This study is the first to report the performance and application of strain Stenotrophomonas pavanii in wastewater flocculation. The results indicate that strain S. pavanii is a good candidate for the production novel bioflocculants and demonstrates its potential industrial practicality in biotechnology processes.

Different wavelengths of LED irradiation promote secondary metabolite production in Pycnoporus sanguineus for antioxidant and immunomodulatory applications.

Pycnoporus sanguineus is a fungus of the phylum Basidiomycota that has many applications in traditional medicine, modern pharmaceuticals, and agricultural industries. Light plays an essential role in the metabolism, growth, and development of fungi. This study evaluated the mycelial growth and antioxidant and anti-inflammatory activities in P. sanguineus fermentation broth (PFB) cultured under different wavelengths of LED irradiation or in the dark. Compared to the dark cultures, the dry weight of mycelia in red- and yellow-light cultures decreased by 37 and 35% and the yields of pigments increased by 30.92 ± 2.18mg and 31.75 ± 3.06mg, respectively. Compared with the dark culture, the DPPH free radical scavenging ability, ABTS+ free radical scavenging capacity, and reducing power of yellow-light cultures increased significantly, and their total phenolic content peaked at 180.0 ± 8.34μg/mL. However, the reducing power in blue-light cultures was significantly reduced, though the total phenol content did not vary with that of dark cultures. In LPS- and IFN-γ-stimulated RAW 264.7 cells, nitrite release was significantly reduced in the red and yellow light-irradiated PFB compared with the dark culture. In the dark, yellow-, and green-light cultures, TNF-α production in the inflamed RAW 264.7 cells was inhibited by 62, 46, and 14%, respectively. With red-, blue-, and white-light irradiation, TNF-α production was significantly enhanced. Based on these results, we propose that by adjusting the wavelength of the light source during culture, one can effectively modulate the growth, development, and metabolism of P. sanguineus.

Effects of four bolete species on ectomycorrhizae formation and development in Pinus thunbergii and Quercus acutissima

BackgroundBolete cultivation is economically and ecologically valuable. Ectomycorrhizae are advantageous for plant development and productivity. This study investigated how boletes affect the formation of Pinus thunbergii and Quercus acutissima ectomycorrhizae using greenhouse-based mycorrhizal experiments, inoculating P. thunbergii and Q. acutissima with four species of boletes (Suillus bovinus, Suillus luteus, Suillus grevillei, and Retiboletus sinensis).ResultsThree months after inoculation, morphological and molecular analyses identified S. bovinus, S. luteus, S. grevillei and R. sinensis ectomycorrhizae formation on the roots of both tree species. The mycorrhizal infection rate ranged from 40 to 55%. The host plant species determined the mycorrhiza morphology, which was independent of the bolete species. Differences in plant growth, photosynthesis, and endogenous hormone secretion primarily correlated with the host plant species. Infection with all four bolete species significantly promoted the host plants’ growth and photosynthesis rates; indole-3-acetic acid, zeatin, and gibberellic acid secretion increased, and the abscisic acid level significantly decreased. Indole-3-acetic acid was also detected in the fermentation broths of all bolete species.ConclusionsInoculation with bolete and subsequent mycorrhizae formation significantly altered the morphology and hormone content in the host seedlings, indicating growth promotion. These findings have practical implications for culturing pine and oak tree species.

Effects of Latilactobacillus delbrueckii fermentation on the bioconversion and antioxidant capacity of phenolic compounds in quinoa sprouts

Polyphenols, serving as a significant active ingredient, exert a crucial role in the antioxidant effect of quinoa. Nevertheless, the preponderance of research has predominantly centered on quinoa grains, with a conspicuous scarcity of studies dedicated to quinoa sprouts. In this study, quinoa sprouts were used as substrate, and Latilactobacillus delbrueckii subsp. bulgaricus was employed as the fermentation agent. The fermentation parameters of quinoa sprouts were meticulously optimized using a combination of single-factor experiments and response surface analysis, successfully leading to the enrichment of polyphenols. A noteworthy elevation in the total polyphenol and flavonoid contents were observed after fermentation, with increments of 46.56% and 57.28%, respectively. Additionally, a statistically significant (P < 0.05) augmentation in antioxidant activity was demonstrated, as measured by 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity, 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) scavenging activity, oxygen radical absorbance capacity (ORAC), and reducing power. To gain further insights, metabolomics analysis was conducted, revealing the identity and quantity of active compounds present in the fermented quinoa sprouts broth. Notably, a marked increase in flavonoids like luteolin and quercetin was evident, whereas a substantial reduction in the levels of certain polyphenols, such as dihydrowogonin and quercetin 3-O-xylosyl-rutinoside, was observed after fermentation. The activities of five key enzymes were quantitatively assayed within the metabolic pathway of polyphenols, revealing significant variations. This comprehensive analysis provides a clearer understanding of the metabolic mechanisms underlying the bioconversion of polyphenols during the fermentation process.

A Novel Bio-Purification Process Employing an Engineered E. coli Strain for Downstream Processing of Lactic Acid Solutions from the Fermentation of Agro-Industrial by-Products

This study aims to integrate a novel bio-purification process employing an engineered E. coli strain in the downstream processing of lactic acid (LA) fermentation broths from low-cost renewable biological feedstocks. Fermentation broth of candy waste and digestate mixture was used as a real biological feedstock. An engineered E. coli strain that selectively catabolize impurities without catabolizing LA was initially adapted on the biological feedstock, followed by shake flask experiments to prove the bio-purification concept. Scale-up and validation in a bench-scale bioreactor followed, before developing a semi-continuous membrane bioreactor (MBR) bio-purification process. The MBR bio-purification was assessed with biological feedstocks which simulated ultrafiltration or nanofiltration permeates. Incomplete removal of impurities and increased fouling was observed in the case of the ultrafiltration permeate. Contrarily, the nanofiltration permeate was successfully treated with MBR bio-purification, since low membrane fouling, 100% maltose and acetic acid removal, and no LA catabolism was achieved. MBR bio-purification as a post-treatment step in the downstream processing of LA was demonstrated as a promising technology for increasing the purity of LA solutions.

Purification, characterisation and visualisation of soybean protein hydrolysis by aspergillopepsin I from mangrove Aspergillus tubingensis

In order to obtain marine-derived enzymes suitable for protein hydrolysis in acid brewing system, seven species of fungi were obtained from mangrove soil. One of the strains, Aspergillus tubingensis, was determined to have a high acid protease production capacity. Its fermentation broth was purified and the obtained protease was identified as aspergillopepsin I (ATAP) with a molecular weight of 41205 Da. Homology modelling shows that ATAP contains three active sites, each containing to a Zn atom and amino acid residues. The enzyme had the highest relative activity at pH 3.5 and 35–40 °C, and exhibited pH and thermal stability at pH 3.0–3.5 and 20–50 °C. ATAP showed certain salt tolerance. K+ and Zn2+ promoted the enzyme activity, while Mg2+, Cu2+, Co2+, and Fe2+ slightly inhibited the enzyme activity. The results of inhibitors suggest that ATAP belongs to the class of aspartic proteases containing metal atoms. Furthermore, the results of enzymatic hydrolysis of defatted soybean powder showed that the contents of soluble protein, amino nitrogen, free amino acids and the proportion of small molecular peptides (<3 kDa) in the hydrolysate increased significantly. In addition, molecular docking of ATAP and aspergillopepsin I from Aspergillus oryzae 3.042 with soybean globulins (7S and 11S), respectively, showed better hydrolysis of soybean globulins by the former, especially for 7S globulin. Visual analysis indicated the interaction forces between enzymes and soybean globulins were dominated by hydrogen bond and hydrophobic interaction. In conclusion, ATAP may be a candidate as an enzyme preparation for the hydrolysis of soybean proteins.

Optimization of the fermentation media and growth conditions of Bacillus velezensis BHZ-29 using a Plackett-Burman design experiment combined with response surface methodology.

Bacillus velezensis occurs extensively in the soil environment. It produces a range of antimicrobial compounds that play an important role in the field of biological control. However, during the actual application process it is often affected by factors such as the medium formulation and fermentation conditions, and therefore biocontrol measures often do not achieve their expected outcomes. In this study, the B. velezensis BHZ-29 strain was used as the research object. The carbon and nitrogen sources, and inorganic salts that affect the number of viable bacteria and antibacterial potency of B. velezensis BHZ-29, were screened by a single factor test. A Plackett-Burman design experiment was conducted to determine the significant factors affecting the number of viable bacteria and antibacterial potency, and a Box-Behnken design experiment was used to obtain the optimal growth of B. velezensis BHZ-29. The medium formula that produced the highest number of viable bacteria and most antibacterial substances was determined. The initial pH, temperature, amount of inoculant, liquid volume, shaking speed, and culture time were determined by a single factor test. The factors that had a significant influence on the number of viable bacteria of B. velezensis BHZ-29 were selected by an orthogonal test. A Box-Behnken design experiment was conducted to obtain the optimal fermentation conditions, and highest number of viable bacteria and antibacterial titer. Molasses, peptone, and magnesium sulfate had significant effects on the viable count and antibacterial titer of B. velezensis BHZ-29. The viable count of B. velezensis BHZ-29 increased from 7.83 × 109 to 2.17 × 1010 CFU/mL, and the antibacterial titer increased from 111.67 to 153.13 mm/mL when the optimal media were used. The optimal fermentation conditions for B. velezensis BHZ-29 were as follows: temperature 25.57°C, pH 7.23, culture time 95.90 h, rotation speed 160 rpm, amount of inoculant 2%, and liquid volume 100 ml. After the optimization of fermentation conditions, the number of viable bacteria increased to 3.39 × 1010 CFU/mL, and the bacteriostatic titer increased to 158.85 mm/ml.The plant height and leaf number of cotton plants treated with BHZ-29 fermentation broth were higher than those of cotton inoculated with Verticillium dahliae. The number of bacteria was 1.15 × 107 CFU/g, and the number of fungi was 1.60 × 105 spores/g. The disease index of the cotton seedlings treated with the optimized fermentation broth was 2.2, and a control effect of 93.8% was achieved. B. velezensis BHZ-29 could reduce the disease index of cotton Verticillium wilt and had a controlling effect on the disease. The best effect was achieved in the treatment group with an inoculation concentration of 2 × 108 CFU/ml, the disease index was 14.50, and a control effect of 84.18% was achieved. The fermentation process parameters of the number of viable bacteria and antibacterial titer by strain B. velezensis BHZ-29 were optimized to lay a foundation for the practical production and application of strain B. velezensis BHZ-29 in agriculture.

Fabrication of polyvinyl alcohol/vanillin/Ti3C2Tx/ZnONPs composite film with excellent antimicrobial properties for pepper preservation

Novel food packaging materials with antimicrobial properties must be developed to ensure that fresh agricultural products are free from microbial contamination during the process from “farm” to “table”. Herein, we synthesized an antimicrobial composite packaging film by employing a green in situ synthesis method. The method involves the in situ reduction of zinc oxide nanoparticles (ZnONPs) on the substrate Ti3C2Tx by Lactobacillus plantarum fermentation broth, which was then blended with polyvinyl alcohol and vanillin to obtain the polyvinyl alcohol/vanillin/Ti3C2Tx/ZnONPs (PVA/VAN/T-Z) composite film. The incorporation of crosslinker VAN and antimicrobial material Ti3C2Tx/ZnONPs did not affect the transparency and color of the film, but improved its mechanical strength, hydrophobicity, and antimicrobial ability of the film. Antimicrobial experiments demonstrated that the PVA/VAN/T-Z composite film exhibited significant antimicrobial activity against bacteria (Gram-negative E. coli and Gram-positive S. aureus) and fungi isolated from peppers. Furthermore, pepper preservation experiments demonstrated that the PVA/VAN/T-Z composite film exhibited commendable antimicrobial and preservation performance compared to commercially available film. As a novel antimicrobial packaging material, the multifunctional PVA/VAN/T-Z composite film has a broad application prospect in food preservation.

Pseudomonas protegens controlled brown rot (Monilinia fructicola) on peach fruit in postharvest by suppressing fungal development and inducing plant defense-related genes

Brown rot, caused by Monilinia fructicola (Wint.) Honey is one of the world's most economically important stone fruit diseases, incurring significant losses during pre-harvest and post-harvest phases. This study demonstrated the inhibitory effect of an endophyte bacterium Pseudomonas protegens (strain QNF1), isolated from Chinese leek (Allium tuberosum) roots. The in vitro experiment revealed that P. protegens significantly suppressed M. fructicola mycelial growth by 67.11 % and 95.53 % in the potato dextrose agar (PDA) and potato dextrose broth (PDB) media compared to the control. P. protegens cell-free supernatants (CFS) also restrained M. fructicola mycelial growth by 97.95 %. In addition, the in vivo experiment displayed P. protegens reduced brown rot of peach fruit inoculated with M. fructicola plug and dipped into M. fructicola fermentation broth by 81.57 % and 85.58 %, respectively. Moreover, P. protegens improved the peach fruit quality to a certain extent. In the early stage, P. protegens significantly induced defense-related genes, including superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO), chitinase (CHI), and beta-1,3-glucanases (GLU). Through gas chromatography-mass spectrometry and liquid chromatography-MS analysis, 21 and 19 principal metabolites were identified in P. protegens, respectively. Among them, pyroglutamic acid, lysine, valine, tryptophan, tyrosine, and methionine were discovered by both analysis methods. The study revealed that P. protegens reduced the brown rot of peach fruit by suppressing fungal development and inducing plant defense systems. The main findings of the study proved that P. protegens represents a promising alternative for controlling the brown rot of peach fruit and other stone fruits during storage.

Effects of Marquandomyces Marquandii SGSF043 on Maize Growth Promotion and Soil Enzyme Activity

In order to further clarify the growth-promoting effect of the non-core Metarhizium sp. Marquandomyces marquandii on plants, M. marquandii SGSF043, which was obtained via pre-screening in the laboratory, was selected as a test strain and the seed soaking method was adopted. The effects of a fermentation broth obtained from this strain on the seed germination, seedling growth, and rhizosphere soil enzyme activity of maize were studied. The results were as follows: In seed germination tests, M. marquandii SGSF043 fermentation liquid had a certain inhibitory effect on corn seed germination, and the germination rate was only 15%. When the fermentation solution was diluted 10 times, the germination rate reached 97%. After the germination test, the growth of maize plumules was promoted in the groups treated with 10-times and 1000-times dilutions. In the field community experiment, based on the comprehensive evaluation of seedling biomass indicators, the solution diluted 100 times had the best growth-promoting effect. The aboveground fresh weight was increased by 127.13% compared with the control group. The results show that M. marquandii SGSF043 has the potential to promote the growth of maize and improve the soil environment, which provides a theoretical basis for the research on and the application of M. marquandii in farmland.

New Analogues of Amides and Quinolinones Produced by Streptomyces sp. YIM S01983.

Streptomide (1), a new amide analogue, streptomynone (2), a new quinolinone, and ten known compounds including three aliphatic acids (3-5), two amides (6-7), four cyclic dipeptides (8-11), and an adenosine (12) were isolated from the fermentation broth of Streptomyces sp. YIM S01983 isolated from a sediment sample collected in Bendong Village, Huadong Town, Chuxiong, China. Their structures were determined by analysis of the 1D/2D-NMR and HR-ESI-MS spectra. Compound 12 presented weak antimicrobial activities against Candida albicans and Aligenes faecalis (MIC=64 μg/mL). Compounds 7 and 12 showed weak cytotoxic activity against MHCC97H.

Mineral Acid Co-Extraction in Reactive Extraction of Lactic Acid Using a Thymol-Menthol Deep Eutectic Solvent as a Green Modifier.

Carboxylic acids can be isolated from fermentation broths using reactive liquid-liquid extraction, offering an alternative to the environmentally harmful state-of-the-art process of precipitating calcium lactate. To enhance the sustainability of liquid-liquid extraction processes, greener solvents, such as natural deep eutectic solvents, are investigated. However, fermentation broths often exhibit pH values unsuitable for carboxylic acid extraction, which can be adjusted using mineral acids, though mineral acids may be co-extracted. In this study, we systematically examine the co-extraction of hydrochloric, nitric, sulfuric, and phosphoric acid during extraction and back-extraction of lactic acid. The solvent phase consisted of tri-n-octylamine, trioctylphosphine oxide, or tributyl phosphate diluted in a thymol-menthol deep eutectic solvent. The back-extraction was conducted using a diluent swing with p-cymene as the antisolvent and water as the receiving phase. Tri-n-octylamine showed the highest efficiency for lactic acid (up to 29.8%) but also the highest co-extraction of mineral acids (up to 50.9%). In contrast, trioctylphosphine oxide exhibited a lower but more selective lactic acid extraction (5.94%) with low mineral acids co-extraction (0.135%). Overall, the highest co-extraction was observed for phosphoric acid and the lowest for nitric acid. In conclusion, the selected solvent phase composition and mineral acid influence the co-extraction and, thus, final product purity. The successful application of the natural deep eutectic solvent as the modifier enhances the sustainability of liquid-liquid extraction processes.

Separation of ochratoxins by centrifugal partition chromatography

The present research work was dedicated to developing an efficient method based on liquid-liquid chromatography (centrifugal partition chromatography, CPC) applicable to routine purifications of ochratoxins (OT) from the liquid culture of the strain A. albertensis SZMC 2107. The crude extract contained numerous components in addition to OTA (90.1 %,) and OTB (1.1 %,) according to HPLC examinations. For the separation of OTs by CPC, several tertiary systems based on acetonitrile, acetone, and short-chain alcohols were examined to find the most applicable biphasic system. The hexane/i-propanol/water 35:15:50 system supplemented with 0.1 % acetic acid was found to be the most efficient for use in CPC separation. Using liquid–liquid instrumental separation, the two OTs, namely OTA (2.23 mg) and OTB (0.031 mg), were successfully isolated with 96.3 % and–72.8 % purity, respectively, from 1 L ferment broth. The identities and purities of the purified components were confirmed and the performance parameters of each separation step and the whole procedure were determined. The developed method could be used effectively to purify OTs for analytical or toxicological applications.