Strain Fermentation Research Articles

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

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

The herbicidal activity and mechanism of Talaromyces purpureogenus CY-1 metabolites.

Studies have shown that Talaromyces can produce a large number of secondary metabolites in its metabolic process, many of which have good insecticidal, antibacterial, antitumor, antiviral and other biological activities. In order to explore the herbicidal activity and mechanism of Talaromyces purpureogenus CY-1, we determined the inhibitory effect of the fermentation broth of the CY-1 strain on weeds, identified the major active components, and further investigated the herbicidal mechanism. The results showed that CY-1, with IC50 values of 5.40 g/L and 4.39 g/L, respectively, exhibited good herbicidal activity against Xanthium sibiricum and Amaranthus lividus. Spraying CY-1 strain fermentation broth on redroot pigweed resulted in plant protection efficiencies and fresh weight protection efficiencies of 83.7% and 87%, respectively. The active component identified in the broth was 2-(3-hydroxybenzoyloxy) acrylic acid. Treatment of Amaranthus lividus with 2-(3-hydroxybenzoyloxy) acrylic acid resulted in trends of increasing superoxide dismutase activity, peroxidase activity, respiratory rate, cytochrome oxidase activity, and soluble protein content, followed by a decrease. Peroxidase activity, relative conductance and malondialdehyde content gradually increased, while acetyl lactate synthase and glutamine synthetase initially decreased and gradually returned to normal. The soluble sugar content showed a gradual decrease. The findings of the present study indicate that 2-(3-hydroxybenzoyloxy) acrylic acid can induce oxidative stress in plants, damaging plant cell walls, weakening respiratory activity, inhibiting protein synthesis and sugar metabolism, ultimately leading to weed wilting and death. © 2024 Society of Chemical Industry.

Preparation of safflower fermentation solution and study on its biological activity.

Safflower, a traditional Chinese medicine, is rich in chemical components including flavonoids, polysaccharides, and alkaloids. It exhibits pharmacological effects such as antioxidant, anti-inflammatory, anti-tumor, and anti-thrombosis properties, making it a valuable resource in the medical field. Furthermore, due to its antioxidant and anti-inflammatory effects, safflower is increasingly being utilized in the cosmetics industry. In this study, yeast was employed to ferment safflower, and the optimal fermentation conditions were established through single-factor experiments and response surface methodology. Subsequently, the antioxidant and anti-inflammatory efficacy of the safflower fermentation solution was assessed using both cellular and zebrafish models. Finally, the safety of the safflower fermentation solution was evaluated through a cosmetic eye irritation test. From a total of 20 yeast strains, YF-5 was identified as the dominant strain for safflower fermentation. By optimizing the fermentation conditions, it was established that the optimal parameters for YF-5 fermentation of safflower are as follows: a fermentation temperature of 36.55°C, a material-to-liquid ratio of 1:20.46, a fructose concentration of 6.20%, a fermentation duration of 72 h, and an inoculum volume of 4%. The biological activities of safflower, including its antioxidant and anti-inflammatory properties, were enhanced through yeast fermentation. In HaCaT cell and zebrafish oxidative damage assays, safflower fermentation solution inhibits the production of malondialdehyde (MDA) and increases superoxide dismutase (SOD) activity as well as total antioxidant capacity (T-AOC). In the RAW264.7 cell inflammatory damage assays, a 20% safflower fermentation solution was found to inhibit the release of TNF-α and NO in the inflammatory model, with inhibition rates of 30.94 and 28.86%, respectively. In the zebrafish inflammatory damage assays, the quantity of fluorescent neutral proteins in the 5% safflower fermentation solution was 0.7 times that observed in the dexamethasone (0.1 mg/mL) positive control group, indicating that its anti-inflammatory activity is comparable to that of dexamethasone (0.1 mg/mL). In the chicken embryo chorionic membrane experiment, it was observed that the safflower fermentation solution did not cause significant damage to the blood vessels of the chorionic allantoic membrane (CAM). This finding demonstrates that the safflower fermentation solution possesses a certain degree of safety. Safflower fermentation solution has antioxidant and anti-inflammatory bioactivities, and it has passed cosmetic safety evaluations. It can be used as a new natural cosmetic ingredient added to cosmetic products.

Enhanced bioethanol production from Compositae plant stalks: Integrating dry acid pretreatment and fungal mediated biodetoxification

Compositae is an important economic plant with relatively high contents of water-soluble sugars in stalk section. The pretreatment of Compositae plant stalks quickly converts the water-soluble sugars into high titers of furan aldehydes, which inhibits the viability of consequent fermentation strains. This study used the typical Compositae plant stalks of sunflower, Jerusalem artichoke, and stevia as feedstocks for dry acid pretreatment under high solids loading, then a unique solid-state biodetoxification method was applied to degrade these high-titer inhibitors. The results showed that temperature and sulfuric acid concentration were two key parameters for enhancing pretreatment efficiency. Enzymatic hydrolysis yield of the above pretreated stalks reached up to 79.68%, 66.75% and 74.05%, respectively. For pretreated Jerusalem artichoke stalk, it contained 37.36% of cellulose, 2.72% of xylan, 3.04% of glucose, 11.64% of xylose, 20.54% of lignin, and some inhibitors (3.80% of acetate, 0.24% of HMF and 0.23% of furfural). After biodetoxification, 100% of furfural, 40% of HMF, and 36% of acetate were quickly removed by Amorphotheca resinae ZN1, which resulted in 55.57 g/L ethanol production (equivalent to 7.1% by volumetric percentage) with the yield of 55.54% from 73.59 g/L of glucose and 40.69 g/L of xylose by simultaneous saccharification and co-fermentation (SSCF). This study provides a combinational approach of dry acid pretreatment and biodetoxification for Compositae plant stalks biorefinery.

Isolation and identification of epiphytic Pichia kudriavzevii from loquat peels and investigation of its fermentation characteristics for liquor production.

During the process of fruit wine production, yeast plays a crucial role in influencing the taste, flavor, and overall quality of the wine. This study aims to enhance the flavor and quality of loquat wine by isolating strains of Pichia kudriavzevii (P. kudriavzevii) with desirable winemaking characteristics from loquat fruit fermentation broth. A total of 12 strains of P. kudriavzevii were isolated and subjected to morphological and molecular biological identification. Their fermentation performance, ethanol production, ester production, hydrogen sulfide production, killer activity, and tolerance were evaluated. The results revealed that strains Q-2, Q-9, Q-10, Q-12, Q-20, and Q-42 exhibited robust growth and strong tolerance under conditions of 40°C temperature, 12% ethanol concentration, 350g/L glucose concentration, and pH 2.8. Strain Q-42 demonstrated the strongest gas production capacity, killer activity, and good ester and ethanol production. As a highly active fermentation strain with excellent wine making characteristics, P. kudriavzevii Q-42 provides a valuable yeast resource for the industrial production of loquat wine and offers technical support for improving the overall quality of loquat wine.

CRISPRi-mediated metabolic switch enables concurrent aerobic and synthetic anaerobic fermentations in engineered consortium

Replacing petrochemicals with compounds from bio-based manufacturing processes remains an important part of the global effort to move towards a sustainable future. However, achieving economic viability requires both optimized cell factories and innovative processes. Here, we address this challenge by developing a fermentation platform, which enables two concurrent fermentations in one bioreactor. We first construct a xylitol producing Escherichia coli strain in which CRISPRi-mediated gene silencing is used to switch the metabolism from aerobic to anaerobic, even when the bacteria are under oxic conditions. The switch also decouples growth from production, which further increases the yield. The strain produces acetate as a byproduct, which is subsequently metabolized under oxic conditions by a secondary E. coli strain. Through constraint-based metabolic modelling this strain is designed to co-valorize glucose and the excreted acetate to a secondary product. This unique syntrophic consortium concept facilitates the implementation of “two fermentations in one go”, where the concurrent fermentation displays similar titers and productivities as compared to two separate single strain fermentations.

Preparation and flavor characteristics of plant-based meat flavoring by mixed fermentation of multiple strains

In this study, plant-based meat flavoring was prepared by mixed fermentation and thermal reaction. Based on the changes of amino acid nitrogen, reducing sugar, and total acid during the fermentation process and combined with sensory evaluation, the optimal fermentation conditions were determined as 4 % total inoculation amount of Monascus purpureus, Actinomucor elegans, and Bacillus subtilis with an inoculation ratio of 1:1:1, and 36 h of fermentation time. The odor characterization of meat flavoring was identified by solid-phase microextraction–gas chromatography–mass spectrometry (SPME–GC–MS), aroma extract dilution analysis (AEDA), quantitative measurements, odor activity value (OAV) and aroma recombination and omission experiments. 64 volatile compounds and 28 odorants were identified. The compounds with FD ≥ 27 were precisely quantified by five-point standard calibration curve and OAVs were calculated, and those with OAV ≥ 1 were subjected to aroma recombination and omission experiments. Finally, furfuryl mercaptan, difurfuryl disulfide, 4-methyl-5-ethylthiazole, 1-octanal, and 2-acetylthiophene were determined to be the key aroma-active compounds. These compounds have been recognized as key aroma-active compounds or detected in other meats or meat flavorings. In this study, we prepared plant-based meat flavoring through mixed fermentation, and provide new ideas for the development of plant-based thermal reaction meat flavoring process.

Co-fermentation of hot-pressed rapeseed meal with multiple strains and cellulase: Evaluating changes in protein quality and metabolite profiles

Hot-pressed rapeseed meal (RSM) is rich in resources, but high fiber content and low protein utilization result from high temperature processing. This study aims to obtain a high-quality protein source through the fermentation of hot-pressed RSM. We used Plackett-Burman and Box-Behnken designs to optimize parameters for solid state fermentation with Bacillus subtilis strain 5, Saccharomyces cerevisiae, and Aspergillus niger. Under the optimum conditions, the synergistic fermentation with multiple strains and cellulase significantly reduced anti-nutritional factors in hot-pressed RSM, while also increasing protein solubility, digestibility and extraction rate by 466.80%, 45.56% and 816.44%, respectively. 77 enzymes secreted by fermentation strains were identified in fermented hot-pressed RSM, which were mainly involved in the metabolism of proteins, peptides and amino acids (28.6%), energy (28.5%), carbohydrates (13.0%) and lipids (5.2%). Non-targeted metabonomics identified 1245 differential metabolites, with the top five being peptides and amino acids, lipids, heterocyclic compounds, alkaloids, and organic acids. These differential metabolites enhanced the nutritional, functional and flavor properties of fermented hot-pressed RSM. Among them, 702 differential metabolites were enriched into metabolic pathways, with the largest number of pathways associated with amino acid metabolism. These results hold important implications for understanding the quality of fermented hot pressed RSM.

Effects of fermentation conditions (salt concentration, temperature, and pH) on Lactobacillus strains for induction of interleukin-12 in the exposed murine splenocytes

The present work aimed to assess the stresses of temperature, salt concentration, and pH on Lactobacillus gasseri 54C, Lactiplantibacillus plantarum ATCC 14917, and 11SH in fermented culture medium and their effects on interleukin-12 (IL-12) induction in murine splenocytes. First, the strains were fermented under different microbial stress conditions at salt concentrations (2, 4, and 6% w/v), temperatures (30 and 42°C), and pH levels (4, 5, and 6), as well as their combined conditions. Then, they were heat-killed, lyophilized, and exposed to murine splenocytes. The IL-12 induction was measured using the immunoassay method. It was indicated that the induction of IL-12 depended on the culture conditions, as it was increased by the fermentation strains under microbial stresses with the higher temperature (42°C), lower salt concentration (%2), and lower pH level (4). It seems that the bacterial cells' integrity is essential to stimulate the induction of IL-12.

Effect of bacterial fermentation on the ability of myofibrillar proteins to bind esters and its potential mechanism: Based on protein metabolism and structural changes

The effect of fermentation strains (Lactiplantibacillus plantarum CQ01107 and Staphylococcus simulans CD207) on the binding properties of porcine myofibrillar proteins (MPs) to esters was investigated from two perspectives: metabolism degree and structural alterations. Results demonstrated that S. simulans could reduce the particle size and α-helix content of MPs, while simultaneously increasing the absolute zeta potential and active sulfhydryl content. This process decreased protein aggregation and facilitated the unfolding of MPs, thereby enhancing their binding to esters. Conversely, L. plantarum showed limited promotion, which might be related to its robust acid production and protein hydrolysis capacities. In addition, ethyl octanoate, with a longer carbon chain length, was found to have the highest binding capacity to MPs (28.38 %–41.59 %). Molecular docking results further revealed that the binding of the four esters to MPs was spontaneous, with ethyl octanoate exhibiting the lowest binding energy to MPs (−5.635 kcal/mol). The primary forces involved in the binding of the four selected esters to MPs were hydrophobic interactions, hydrogen bonding, and van der Waals forces. These findings can provide new insights into the mechanisms by which fermentation strains influence flavor formation in fermented foods.

Utilizing a high-throughput visualization screening technology to develop a genetically encoded biosensor for monitoring 5-aminolevulinic acid production in engineered Escherichia coli

5-Aminolevulinic acid (5-ALA) is a non-protein amino acid widely used in agriculture, animal husbandry and medicine. Currently, microbial cell factories are a promising production pathway, but the lack of high-throughput fermentation strain screening tools often hinders the exploration of engineering strategies to increase cell factory yields. Here, mutant AC103-3H was screened from libraries of saturating mutants after response-specific engineering of the transcription factor AsnC of L-asparagine (Asn). Based on mutant AC103-3H, a whole-cell biosensor EAC103-3H with a specific response to 5-ALA was constructed, which has a linear dynamic detection range of 1–12 mM and a detection limit of 0.094 mM, and can be used for in situ screening of potential high-producing 5-ALA strains. With its support, overexpression of the C5 pathway genes using promoter engineering assistance resulted in a 4.78-fold enhancement of 5-ALA production in the engineered E. coli. This study provides an efficient strain screening tool for exploring approaches to improve the 5-ALA productivity of engineered strains.

Pathway and Production Differences in Branched-Chain Hydroxy Acids as Bioactive Metabolites in Limosilactobacillus fermentum, Ligilactobacillus salivarius, and Latilactobacillus sakei.

Branched-chain hydroxy acids (BCHAs) as bioactive metabolites of Lactobacillaceae include 2-hydroxy isovaleric acid (HIVA), 2-hydroxy isocaproic acid (HICA), and 2-hydroxy-3-methyl isovaleric acid (HMVA). Combining targeted and untargeted metabolomics, this study elucidates differences in extracellular BCHA production in Limosilactobacillus fermentum, Ligilactobacillus salivarius, and Latilactobacillus sakei alongside comparing comprehensive metabolic changes. Through targeted metabolomics, BCHA production among 38 strains exhibited strain specificity, except for L. sakei, which showed significantly lower BCHA production. Explaining the lower production in L. sakei, which lacks the branched-chain amino acid (BCAA)-utilizing pathway, comparison of BCHA production by precursor reaction revealed that the pathway utilizing BCAAs is more dominant than the pathway utilizing pyruvate. Expanding upon the targeted approach, untargeted metabolomics revealed the effects of the reaction compound on other metabolic pathways besides BCHAs. Metabolism alterations induced by BCAA reactions varied among species. Significant differences were observed in glycine, serine, and threonine metabolism, pyruvate metabolism, butanoate metabolism, and galactose metabolism (p < 0.05). These results emphasize the importance of the synergy between fermentation strains and substrates in influencing nutritional components of fermented foods. By uncovering novel aspects of BCAA metabolism pathways, this study could inform the selection of fermentation strains and support the targeted production of BCHAs.

Antioxidant Peptides Derived from Cheese Products via Single and Mixed Lactobacillus Strain Fermentation.

Probiotics are used in cheese fermentation to endow the product with unique functional properties, such as enhanced flavor and aroma development through proteolysis and lipolysis. In this study, two probiotic Lactobacillus strains, Lactobacillus plantarum A3 and Lactobacillus reuteri WQY-1, were selected to develop new probiotic cheeses in the form of single- and mixed-strain starters. The results demonstrated that the L. plantarum A3 single-strain group and the L. plantarum A3/L. reuteri WQY-1 mixed fermentation group exhibited superior product performance, particularly the release of functional hydrolysates during cheese ripening. Furthermore, Label-free quantitative proteomic analysis revealed 26 unique antioxidant peptides in the L. plantarum A3 single-strain group and 53 in the L. plantarum A3/L. reuteri WQY-1 mixed fermentation group. Among these, CMENSAEPEQSLACQCL (β-lactoglobulin), CMENSAEPEQSLVCQCL (β-lactoglobulin), and IQYVLSR (κ-casein) have been found to possess potential antioxidant properties both in vitro and in vivo. This confirmed that milk-derived protein peptides in cheese products exhibit potential antioxidant functions through the hydrolysis of probiotic strains.

Exploring the Diversity and Potential Use of Flower-Derived Lactic Acid Bacteria in Plant-Based Fermentation: Insights into Exo-Cellular Polysaccharide Production.

Isolation of new plant-derived lactic acid bacteria (LAB) is highly prioritized in developing novel starter cultures for plant-based fermentation. This study explores the diversity of LAB in Danish flowers and their potential use for plant-based food fermentation. A total of 46 flower samples under 34 genera were collected for LAB isolation. By introducing an enrichment step, a total of 61 LAB strains were isolated and identified using MALDI-TOF and 16S rRNA sequencing. These strains represent 24 species across 9 genera, predominantly Leuconostoc mesenteroides, Fructobacillus fructosus, Apilactobacillus ozensis, and Apilactobacillus kunkeei. Phenotypic screening for exo-cellular polysaccharide production revealed that 40 strains exhibited sliminess or ropiness on sucrose-containing agar plates. HPLC analysis confirmed that all isolates produced exo-cellular polysaccharides containing glucose, fructose, or galactose as sugar monomers. Therefore, the strains were glucan, fructan, and galactan producers. The suitability of these strains for plant-based fermentation was characterized by using almond, oat, and soy milk. The results showed successful acidification in all three types of plant-based matrices but only observed texture development in soy by Leuconostoc, Weissella, Lactococcus, Apilactobacillus, and Fructobacillus. The findings highlight the potential of flower-derived LAB strains for texture development in soy-based dairy alternatives.

Outer Membrane Vesicle Production by Escherichia coli Enhances Its Defense against Phage Infection.

Several studies have investigated the multifunctional characteristics of outer membrane vesicles (OMVs), but research on their role in mediating phage-bacteria interactions is limited. Employing Escherichia coli as a model, we engineered a mutant strain overproducing OMVs for protective experiments against phage infections. The addition of exogenous OMVs proved highly effective in safeguarding the bacterial host against various phages, mitigating predatory threats. Screening for phage-resistant strains and adsorption experiments revealed that inhibiting phage adsorption is a crucial pathway through which OMVs protect against phage predation. Although OMVs conferred tolerance to the phage-sensitive strains (those easily infected by phages), they could not restore the phage-resistant strains (those that effectively resist phage infection) to a sensitive phenotype. This study provides valuable insights for the future development of novel biotechnological approaches aimed at utilizing OMVs to protect fermentative strains and reduce the risk of phage contamination.

Utilization of cotton stalk waste for sustainable isopropanol production via hydrolysis and coculture fermentation

The present investigation focused on isopropanol production from lignocellulosic cotton stalk biomass (CSB) using conventional pretreatment, enzymatic hydrolysis and fermentation methods. In comparison to alkaline (NaOH) pretreatment, dilute sulphuric acid (H2SO4) pretreatment showed higher cellulose exposure (448.5 mg/g). Further, ultrasono assisted acid and alkali pretreatment was performed for maximum exposure of cellulose and it was found 616.9 and 586.15 mg/g respectively. Chemical pretreated CSB was additionally exposed to independent enzymatic hydrolysis using commercial enzymes (Celluclast and Viscozymes) following Response Surface Methodology (RSM), which revealed a maximal production of glucose and xylose yield (544.6 mg/g and 41.8 mg/g). Pretreated and enzymatic hydrolyzed cotton stalk biomass at various conditions were analysed using SEM, FTIR, and XRD to determine the structural and functional changes. Further, a co-culture strategy was employed on pretreated and hydrolyzed CSB using two fermented yeast strains (Saccharomyces cerevisiae and Pichia pastoris) for isopropanol production. HR-MS analysis revealed that the maximum concentration of isopropanol (126.228 mM) was produced in 2:1 proportionate ratio of two fermented yeasts with 20 g/L of substrate loadings at 72 h of incubation time. These results indicate that the production of isopropanol (7.46 g/L) from CSB with different parametric conditions is an encouraging step and can be exploited further for various industrial applications.

Preparation, flavor component and antioxidant activity analysis of Lactiplantibacillus plantarum NCU137 fermented rice protein

Rice protein is the main component in the byproducts of rice processing such as rice dregs and rice bran. This study aims to develop lactic acid bacteria-fermented rice protein and analyze the changes of component and bioactivity. Lactiplantibacillus plantarum NCU137 was selected as the optimum fermentation strain from sixty lactic acid bacteria strains using fermentation performance and sensory score as the criteria. NCU137 was then applied to rice protein fermentation and the process parameters were optimized by single factor and orthogonal experiments. NCU137 fermentation significantly increased the acidity, antioxidant activity, and peptide content of rice protein, and eight volatile flavor compounds were identified as characteristic aroma components as determined by SPME-GC-MS. Of them, the OAV value of 1-hexanol was up to 50.94, contributing the most to the flavor of fermented rice protein. Furthermore, the antioxidant activity and mechanism of rice protein antioxidant peptide (RPAP) on H2O2-induced Caco-2 cell damage were investigated by CCK-8 method and fluorescence microscopy. RPAP showed protective effect on cell damage in a dose-dependent manner, accompanying by reduction of oxidative components such as ROS and MDA, and increasement of antioxidant components such as SOD, CAT, and GSH-PX. The protective effect might lie in the regulation of gene transcription in Nrf2/HO-1 and apoptosis Bax/Bcl pathway, as validated by RT-qPCR assay. Shortly, the rice protein fermented by NCU137 displayed significant improvement of flavor, nutritional components, and antioxidant bioactivity. The fermentation technology provides alternative strategy for processing rice protein in rice dregs and rice bran into high-valued products.

Comparison of Pediococcus pentosaceus YC64-fermented soybean meal and raw soybean meal in diets for Nile tilapia: Growth performance, feed utilization, and intestine health

Fermentation is a promising approach for improving the utilization of soybean meal (SM) by aquaculture fish, and the strain is the core of microbial fermentation processes. Herein, Pediococcus pentosaceus YC64, an autochthonous probiotic isolated from the intestine of Nile tilapia (Oreochromis niloticus), was applied to the fermentation of raw SM. In an eight-week feeding trial, tilapia juveniles were divided into two treatments fed on diets with 0 % (Control) and 100 % of SM protein replaced by fermented SM (FSM). Results showed that solid-state fermentation improved SM quality indicated by increased acid-soluble protein contents. Fish fed the FSM diet had significantly higher weight gain, feed efficiency, and protein efficiency compared to those fed the SM diet. Furthermore, whole-body protein content significantly increased in the FSM-fed fish, accompanied with the increased amino acid transport and reduced protein catabolism. Texture profile analysis indicated that fish fed the FSM diet had reduced muscle hardness compared with the control, which was associated with changes in collagen deposition and myofiber development. Moreover, feeding an FSM diet enhanced goblet cell proliferation and stimulated mucin secretion, ultimately leading to improved intestinal tight junctions and reduced inflammatory response. Besides, the FSM diet feeding altered intestinal microbiota community, characterized by increased abundance of Pediococcus. Overall, P. pentosaceus YC64 exhibits promise as a fermentation strain in the production of FSM. The FSM performed better than SM in improving the growth performance, feed utilization, and intestine health of Nile tilapia.

Molluscicidal activity and biochemical impacts of borrelidins against an aquatic invasive snail Pomacea canaliculata for crop protection

The invasive golden apple snail Pomacea canaliculata is one of the devastating threats to aquatic ecosystems and wetland agriculture worldwide. Macrolides from microbes display various advantages over other compounds in controlling snails. However, emergence of antibiotic-resistant phenotypes against certain macrolides in the field appeals for exploring more effectively molluscicidal macrolides. Here, two borrelidins, borrelidin BN1 and BN2, from the extract of a Streptomyces strain fermentation were evaluated for molluscicidal potential against P. canaliculata using both immersion and contact bioassay methods. Borrelidin BN1 (borrelidin A) presented a significant molluscicidal activity comparable to the chemical pesticide metaldehyde, and had a much lower median lethal concentration value (LC50, 522.984 μg·ml−1) than avermectin B1 at 72 h of contact-killing treatment. Snail growth was inhibited by borrelidin BN1 more than by metaldehyde at sublethal concentrations, consistent with responses of key biochemical parameters. Exposure to borrelidin BN1 decreased the activity of acetylcholinesterase (AChE), glutathione S-transferase (GST), aspartate aminotransferase (AST), alanine aminotransferase (ALT) as well as the levels of energy reserves and sex steroids in snail tissues, while increased the activity of superoxide dismutase (SOD), catalase (CAT), lactate dehydrogenase (LDH) and the level of lipid peroxidation (LPO). Further application assay confirmed that borrelidin BN1 protected crop plant Zizania latifolia from P. canaliculata damage via suppressing snail population density. These findings suggest great potential of borrelidin BN1 as a molluscicide. Additionally, its higher activity than the stereoisomeric borrelidin BN2 (borrelidin F) implied better molluscicidal borrelidins could be acquired through structural optimization.

Screening and fermentation of high-yield glycolic acid strains

Glycolic acid is an important chemical product widely used in various fields, including cosmetics, detergents, textiles, and more. Currently, microbial production of glycolic acid has disadvantages such as poor genetic stability, low yield, and high cost. Additionally, whole-cell catalytic production of glycolic acid typically requires the addition of relatively expensive sorbitol as a carbon source, which limits its industrial production. To develop an industrially applicable method for glycolic acid production, this study used ethylene glycol as a substrate to screen the glycolic acid-producing strains through whole-cell catalysis, obtaining a Rhodotorula sp. capable of producing glycolic acid. The strain was then subjected to UV mutagenesis and high throughput screening, and the positive mutant strain RMGly-20 was obtained. After optimization in shake flasks, the glycolic acid titer of RMGly-20 reached 17.8 g/L, a 10.1-fold increase compared to the original strain. Using glucose as the carbon source and employing a fed-batch culture in a 5 L fermenter, strain RMGly-20 produced 61.1 g/L of the glycolic acid. This achievement marks the preliminary breeding of a genetically stable glycolic acid-producing strain using a cheap carbon source, providing a new host for the biosynthesis of glycolic acid and promoting further progress toward industrial production.