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BACKGROUND: Rezafungin is an echinocandin approved in the US and EU to treat candidaemia and/or invasive candidiasis. This post-hoc, pooled analysis of the Phase 2 STRIVE and Phase 3 ReSTORE trials assessed rezafungin versus caspofungin in patients with candidaemia and/or invasive candidiasis (IC) in the intensive care unit (ICU) at randomisation. METHODS: STRIVE and ReSTORE were randomised double-blind trials in adults with systemic signs and mycological confirmation of candidaemia and/or IC in blood or a normally sterile site ≤ 96 h before randomisation. Data were pooled for patients in the ICU at randomisation who received intravenous rezafungin (400 mg loading dose then 200 mg once weekly) or caspofungin (70 mg loading dose then 50 mg once daily) for ≤ 4 weeks. Outcomes were Day 30 all-cause mortality (primary outcome), Day 5 and 14 mycological eradication, time to negative blood culture, mortality attributable to candidaemia/invasive candidiasis, safety, and pharmacokinetics. RESULTS: Of 294 patients in STRIVE/ReSTORE, 113 were in the ICU at randomisation (rezafungin n = 46; caspofungin n = 67). At baseline, ~ 30% of patients in each group had impaired renal function and/or an Acute Physiologic Assessment and Chronic Health Evaluation II score ≥ 20. One patient (in the caspofungin group) was neutropenic at baseline. Day 30 all-cause mortality was 34.8% for rezafungin versus 25.4% for caspofungin. Day 5 and 14 mycological eradication was 78.3% and 71.7% for rezafungin versus 59.7% and 65.7% for caspofungin, respectively. Median time to negative blood culture was 18 (interquartile range, 12.6-43.0) versus 38 (interquartile range, 15.9-211.3) h for rezafungin versus caspofungin (stratified log-rank P = 0.001; nominal, not adjusted for multiplicity). Candidaemia/IC-attributable deaths occurred in two rezafungin patients versus one caspofungin patient. Safety profiles were similar between groups. Overall, 17.4% (rezafungin) versus 29.9% (caspofungin) of patients discontinued due to treatment-emergent adverse events. Rezafungin exposure following the initial 400-mg dose was comparable between patients in the ICU at randomisation (n = 50) and non-ICU patients (n = 117). CONCLUSIONS: Rezafungin was well tolerated and efficacious in critically ill, mainly non-neutropenic patients with candidaemia and/or IC. This analysis provides additional insights into the efficacy and safety of rezafungin in the ICU population.

Rezafungin is a long-acting, intravenously administered echinocandin for the treatment of candidemia and invasive candidiasis (IC). Non-inferiority of rezafungin vs caspofungin for the treatment of adults with candidemia and/or IC was demonstrated in the Phase 3 ReSTORE study based on the primary endpoints of day 14 global cure and 30-day all-cause mortality. Here, an analysis of ReSTORE data evaluating efficacy outcomes by baseline Candida species is described. Susceptibility testing was performed for Candida species using the Clinical and Laboratory Standards Institute reference broth microdilution method. There were 93 patients in the modified intent-to-treat population who received rezafungin; 94 received caspofungin. Baseline Candida species distribution was similar in the two treatment groups; C. albicans (occurring in 41.9% and 42.6% of patients in the rezafungin and caspofungin groups, respectively), C. glabrata (25.8% and 26.6%), and C. tropicalis (21.5% and 18.1%) were the most common pathogens. Rates of global cure and mycological eradication at day 14 and day 30 all-cause mortality by Candida species were comparable in the rezafungin and caspofungin treatment groups and did not appear to be impacted by minimal inhibitory concentration (MIC) values for either rezafungin or caspofungin. Two patients had baseline isolates with non-susceptible MIC values (both in the rezafungin group: one non-susceptible to rezafungin and one to caspofungin, classified as intermediate); both were candidemia-only patients in whom rezafungin treatment was successful based on the day 30 all-cause mortality endpoint. This analysis of ReSTORE demonstrated the efficacy of rezafungin for candidemia and IC in patients infected with a variety of Candida species.

Arbuscular mycorrhizal (AM) fungi (e.g., Rhizophagus species) recruit specific bacterial species in their hyphosphere. However, the chemical interplay and the mutual benefit of this intricate partnership have not been investigated yet, especially as it involves bacteria known as strong producers of antifungal compounds such as Bacillus velezensis. Here, we show that the soil-dwelling B. velezensis migrates along the hyphal network of the AM fungus R. irregularis , forming biofilms and inducing cytoplasmic flow in the AM fungus that contributes to host plant root colonization by the bacterium. During hyphosphere colonization, R. irregularis modulates the biosynthesis of specialized metabolites in B. velezensis to ensure stable coexistence and as a mechanism to ward off mycoparasitic fungi and bacteria. These mutual benefits are extended into a tripartite context via the provision of enhanced protection to the host plant through the induction of systemic resistance. [Display omitted] • B. velezensis colonizes the entire mycelial network more efficiently than roots • Bacillus uses fungal hyphae as highways for soil invasion and to colonize new plant • The lipopeptide surfactin plays key roles in the chemical ecology of the interaction • The microbial partnership enhances the systemic resistance of tomato against Botrytis Anckaert et al. investigate the dynamics of Rhizophagus hyphae colonization by B. velezensis. Reduced production of bacterial toxic metabolites triggered by the fungus leads to an unsuspected stable coexistence. This interaction brings benefits to both microbes and provides positive effects to the host plant, which is more resistant to disease. [ABSTRACT FROM AUTHOR]

Bacillus subtilis is an important probiotic microorganism that secretes a variety of antimicrobial compounds, including lipopeptides, which are a class of small molecule peptides with important application value in the fields of feed additives, food, biopesticides, biofertilizers, medicine and the biological control of plant diseases. In this study, we isolated a novel B. subtilis HF1 strain that is rich in lipopeptide components and homologs, has a strong antagonistic effect on a variety of plant fungi, and is highly efficient in promoting the growth of broilers. The live B. subtilis HF1 and its fermentation broth without cells showed significant inhibitory effects on 20 species of plant fungi. The crude extracts of lipopeptides in the fermentation supernatant of B. subtilis HF1 were obtained by combining acid precipitation and methanol extraction, and the lipopeptide compositions were analyzed by ultrahigh-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS). The results showed that HF1 could produce 11 homologs of surfactin and 13 homologs of fengycin. Among the fengycin homologs, C13-C19 fengycin A and C15-C17 fengycin B were identified; among the surfactin homologs, C11-C17 surfactin A and C13-C16 surfactin B were characterized. C13 fengycin A, C11 surfactin A and C17 surfactin A were reported for the first time, and their functions are worthy of further study. In addition, we found that HF1 fermentation broth with and without live cells could be used as a feed additive to promote the growth of broilers by significantly increasing body weight up to 15.84%. HF1 could be a prospective strain for developing a biocontrol agent for plant fungal diseases and an efficient feed additive for green agriculture. [ABSTRACT FROM AUTHOR]

The study aimed to evaluate the biosurfactants (BSs) production by SM-23 strain of Virgibacillus identified by phenotypical and WGS analysis as Virgibacillus massiliensis. We first demonstrated the lipopeptides production by Virgibacillus massiliensis specie and studied their biochemical and molecular analysis as well as their biological potential. The GC–MS analysis indicated that methyl.2-hyroxydodecanoate was the major fatty acid compound with 33.22%. The maximum BSs production was obtained in LB medium supplemented by 1% olive oil (v/v) at 30 °C and 5% NaCl with 1.92 g/l. The obtained results revealed the significant biosurfactants/bioemulsifier potential compared to triton X100 with E24 of 100%, and an emulsification stability SE of 83%. The lipopeptides types were identified by FTIR analysis. A strong antimicrobial action was observed by the produced lipopeptides by the agar diffusion method against E.coli, K. pneumoniae, S. aureus, Fusarium sp, Alternaria sp, and Phytophtora sp. The complete genome sequencing showed genes involved in the synthesis of multiple compounds identified as amphipathic cyclic lipopeptides such as locillomycin/locillomycin B/locillomycin C and bacillibactin. Our results highlighted significant lipopeptides properties displayed by V. massiliensis that can be exploited to develop a novel strategy in the formulation of natural biocidal and fungicidal agents. [ABSTRACT FROM AUTHOR]

Controlled-release systems enhance anti-tumor effects by leveraging local antigen persistence for antigen-presenting cells (APCs) recruitment and T cell engagement. However, constant antigen presentation alone tends to induce dysfunction in tumor-specific CD8+ T cells, neglecting the synergistic effects of co-stimulatory signal. To address this, we developed a soft particle-stabilized emulsion (SPE) to deliver lipopeptides with controlled release profiles by adjusting their hydrophobic chain lengths: C 6 -SPE (fast release), C 10 -SPE (medium release), and C 16 -SPE (slow release). Following administration, C 6 -SPE release antigen rapidly, inducing early antigen presentation, whereas C 16 -SPE's slow-release delays antigen presentation. Both scenarios missed the critical window for coordinating with the expression of CD86, leading to either T cell apoptosis or suboptimal activation. In contrast, C 10 -SPE achieved a spatiotemporally synergetic effect of the MHC-I-peptide complex and co-stimulatory signal (CD86), leading to effective dendritic cell (DC) activation, enhanced T cell activation, and tumor regression in EG7-OVA bearing mice. Additionally, co-delivery of cytosine-phosphate-guanine (CpG) with SPE provided a sustained expression of the CD86 window for DC activation, improving the immune response and producing robust anti-tumor effects with C 6 -SPE comparable to C 10 -SPE. These findings highlight that synchronizing the spatiotemporal dynamics of antigen presentation and APC activation may confer an optimal strategy for enhanced vaccinations. [Display omitted] [ABSTRACT FROM AUTHOR]

Citrus diseases caused by fungal pathogens drastically decreased the yield and quality of citrus fruits, leading to huge economic losses. Given the threats of chemical pesticides on the environment and human health, biocontrol agents have received considerable attention worldwide as ecofriendly and sustainable alternative to chemical fungicides. In the present study, we isolated a Bacillus velezensis strain TZ01 with potent antagonistic effect against three citrus pathogenic fungi: Diaporthe citri, Colletotrichum gloeosporioides and Alternaria alternata. The culture supernatant of this strain exhibited remarkable antifungal activity on potato dextrose agar plates and detached leaves of five citrus varieties. Treatment with TZ01 culture supernatant obviously affected the hyphal morphology and caused nucleic acid leakage. The crude lipopeptides (LPs) extracted from the culture supernatant were found as the major active ingredients, and could maintain the activity under a wide range of temperature and pH and ultraviolet radiation. Furthermore, the type of LPs, produced in vitro, were explored. Whole-genome sequencing of TZ01 revealed secondary metabolite gene clusters encoding synthetases for non-ribosomal peptides and polyketide production, and gene clusters responsible for the synthesis of three important LPs (surfactin, iturin, and fengycin) were identified in the genome. The liquid chromatography-mass spectrometry analysis confirmed the presence of various homologs of surfactin A, bacillomycin D, and fengycin A in the extracted LPs. Taken together, these results contribute to the possible biocontrol mechanisms of B. velezensis strain TZ01, as well as providing a promising new candidate strain as a biological control agent for controlling citrus fungal pathogens. [ABSTRACT FROM AUTHOR]

Introduction: The fungus Fusarium verticillioides significantly threatens maize crops in tropical soils. In light of this, biological control has emerged as a promising strategy to reduce fungicide costs and environmental risks. In this study, we aimed to test the antifungal activity of cell-free supernatant (CFS) from three Bacillus velezensis (CT02, IM14, and LIS05) and one Paenibacillus ottowii (LIS04) against F. verticillioides, thereby contributing to the development of effective biocontrol measures. Methods: The research employed a comprehensive approach. The antifungal activity of the bacterial strains was tested using cell-free supernatant (CFS) from three Bacillus velezensis (CT02, IM14, and LIS05) and one Paenibacillus ottowii (LIS04). The UPLC-MS evaluated the CFS to identify the main bioactive molecules involved in the inhibitory effect on F. verticillioides. Scanning electron microscopy (SEM) was used to assess the impact of CFS on spores and hyphae, and genome sequencing was conducted to identify the genes involved in biological control. These robustmethodologies ensure the reliability and validate our findings. Results: The CFS of the four strains demonstrated significant inhibition of fungal growth. The UPLC-MS analysis revealed the presence of lipopeptides with antifungal activity, including surfactin and fengycins A and B expressed by the three strains of Bacillus velezensis and iturin A expressed by strains LIS05 and IM14. For Paenibacillus ottowii, fusaricidins, ABCDE, and five previously unreported lipopeptides were detected. Scanning electron microscopy (SEM) showed that treatments with CFS led to significant distortion and breakage of the F. verticillioides hyphae, in addition to the formation of cavities in the membrane. Genome mining confirmed the presence of genes coding for the lipopeptides identified by UPLC-MS, including the gene for iturin in CTO2. Genomic sequencing revealed that CT02, IM14, and LIS05 belong to different strains of Bacillus velezensis, and LIS04 belongs to Paenibacillus ottowii, a species recently described. Discussion: The four bacterial strains, including three novel strains identified as Bacillus velezensis and one as the recently described species Paenibacillus ottowii, demonstrate significant potential as biocontrol agents for managing fungal disease. This finding underscores the novelty and potential impact of our research. [ABSTRACT FROM AUTHOR]

This review describes the discovery, structure, activity, engineered constructs, and applications of KR-12, the smallest antibacterial peptide of human cathelicidin LL-37, the production of which can be induced under sunlight or by vitamin D. It is a moonlighting peptide that shows both antimicrobial and immune-regulatory effects. Compared to LL-37, KR-12 is extremely appealing due to its small size, lack of toxicity, and narrow-spectrum antimicrobial activity. Consequently, various KR-12 peptides have been engineered to tune peptide activity and stability via amino acid substitution, end capping, hybridization, conjugation, sidechain stapling, and backbone macrocyclization. We also mention recently discovered peptides KR-8 and RIK-10 that are shorter than KR-12. Nano-formulation provides an avenue to targeted delivery, controlled release, and increased bioavailability. In addition, KR-12 has been covalently immobilized on biomaterials/medical implants to prevent biofilm formation. These constructs with enhanced potency and stability are demonstrated to eradicate drug-resistant pathogens, disrupt preformed biofilms, neutralize endotoxins, and regulate host immune responses. Also highlighted are the safety and efficacy of these peptides in various topical and systemic animal models. Finaly, we summarize the achievements and discuss future developments of KR-12 peptides as cosmetic preservatives, novel antibiotics, anti-inflammatory peptides, and microbiota-restoring agents. [ABSTRACT FROM AUTHOR]

Lipopeptides (LPs) from B. tequilensis EA-CB0015 have antifungal activity against Fusarium species. Specifically, against F. oxysporum f. sp. cubense (Foc), the cause of Fusarium wilt of banana. Cinnamon (CN) extract is also known for its inhibitory activity against the Fusarium genus. The main goal of this research was to determine whether the effect of LPs and CN extract or their combination against a Foc isolate is related to an impairment of mitochondrial function. Our results show that biomass decreased by 74% (p < 0.0001) and 84% (p < 0.0001) when cultures were treated with 128 ppm LPs and 152.5 ppm CN extract, respectively. In parallel, we found a pronounced impairment of the bioenergetic response. That is, the routine oxygen consumption rate diminished by 55% (p = 0.0148) and 38% (p < 0.0001), respectively. Moreover, the ATP-linked respiratory rate decreased by 63% (p = 0.0461) and 44% (p = 0.0005), while the FCCP-simulated respiratory rate by 63% (p = 0.0255) and 45% (p = 0.0002). Therefore, our data suggest that the altered bioenergetic response observed in cultures of Foc treated with LPs or CN is mainly caused by an impairment of the activity of the respiratory complexes. On the other hand, biomass production was reduced by 80% (p < 0.0001) when cultures were treated with a mixture comprising only 10% LPs and 40% CN extract. Furthermore, ATP-linked and FCCP-stimulated respiratory rates decreased by 62% (p = 0.0024) and 68% (p < 0.0001), respectively under the same conditions. A potentially synergistic antifungal effect of cyclic LPs with a CN extract is suggested. [ABSTRACT FROM AUTHOR]

Abstract Background Natural products are important sources for the discovery of new biopesticides to control the worldwide destructive pests Acyrthosiphon pisum Harris. Here, insecticidal substances were discovered and characterized from the secondary metabolites of the bio-control microorganism Bacillus velezensis strain ZLP-101, as informed by whole-genome sequencing and analysis. Results The genome was annotated, revealing the presence of four potentially novel gene clusters and eight known secondary metabolite synthetic gene clusters. Crude extracts, prepared through ammonium sulfate precipitation, were used to evaluate the effects of strain ZLP-101 on Acyrthosiphon pisum Harris aphid pests via exposure experiments. The half lethal concentration (LC50) of the crude extract from strain ZLP-101 against aphids was 411.535 mg/L. Preliminary exploration of the insecticidal mechanism revealed that the crude extract affected aphids to a greater extent through gastric poisoning than through contact. Further, the extracts affected enzymatic activities, causing holes to form in internal organs along with deformation, such that normal physiological activities could not be maintained, eventually leading to death. Isolation and purification of extracellular secondary metabolites were conducted in combination with mass spectrometry analysis to further identify the insecticidal components of the crude extracts. A total of 15 insecticidal active compounds were identified including iturins, fengycins, surfactins, and spergualins. Further insecticidal experimentation revealed that surfactin, iturin, and fengycin all exhibited certain aphidicidal activities, and the three exerted synergistic lethal effects. Conclusions This study improved the available genomic resources for B. velezensis and serves as a foundation for comprehensive studies of the insecticidal mechanism by Bacillus velezensis ZLP-101 in addition to the active components within biological control strains.

Rice sheath blight is a serious disease caused by Rhizoctonia solani that reduces rice yield. Currently, there is a lack of efficient and environmentally friendly control methods. In this study, we found that Bacillus velezensis (B. velezensis) Y6 could significantly inhibit the growth of mycelium in Rhizoctonia solani, and its control efficiency against rice sheath blight was 58.67% (p < 0.01) in a pot experiment. Lipopeptides play an important role in the control of rice sheath blight by B. velezensis Y6, among which iturin and fengycin are essential, and iturin W, a novel lipopeptide in B. velezensis, plays a major role in lipopeptide antagonism to Rhizoctonia solani. In the field, we also found that inoculation with B. velezensis Y6 can increase rice yield (dry weight) by 11.75%. Furthermore, the transcriptome profiling results of the rice roots revealed that there were a total of 1227 differential genes (DEGs) regulated when treated with Y6, of which 468 genes were up-regulated and 971 genes were down-regulated in rice roots compared with the control. Among them, the DEGs were mainly distributed in biological processes (BP) and were mainly enriched in response to stimulus (GO:0050896), response to stress (GO:0006950), and response to abiotic stimulus (GO:0009628). According to the KEGG pathway analysis, there were 338 DEGs classified into 87 KEGG functional pathway categories. Compared with the control, a large number of enriched genes were distributed in phenylpropanoid biosynthesis (map00940), glutathione metabolism (map00480), glycolysis/gluconeogenesis (map00010), and amino sugar and nucleotide sugar metabolism (map00520). In summary, this investigation provides a new perspective for studying the molecular mechanism of B. velezensis in controlling rice sheath blight. [ABSTRACT FROM AUTHOR]

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Lipopeptides are a class of lipid–peptide‐conjugated compounds with differing structural features. This structural diversity is responsible for their diverse range of biological properties, including antimicrobial, antioxidant, and anti‐inflammatory activities. Lipopeptides have been attracting the attention of food scientists due to their potential as food additives and preservatives. This review provides a comprehensive overview of lipopeptides, their production, structural characteristics, and functional properties. First, the classes, chemical features, structure–activity relationships, and sources of lipopeptides are summarized. Then, the gene expression and biosynthesis of lipopeptides in microbial cell factories and strategies to optimize lipopeptide production are discussed. In addition, the main methods of purification and characterization of lipopeptides have been described. Finally, some biological activities of the lipopeptides, especially those relevant to food systems along with their mechanism of action, are critically examined. [ABSTRACT FROM AUTHOR]

Background: Natural products are important sources for the discovery of new biopesticides to control the worldwide destructive pests Acyrthosiphon pisum Harris. Here, insecticidal substances were discovered and characterized from the secondary metabolites of the bio-control microorganism Bacillus velezensis strain ZLP-101, as informed by whole-genome sequencing and analysis. Results: The genome was annotated, revealing the presence of four potentially novel gene clusters and eight known secondary metabolite synthetic gene clusters. Crude extracts, prepared through ammonium sulfate precipitation, were used to evaluate the effects of strain ZLP-101 on Acyrthosiphon pisum Harris aphid pests via exposure experiments. The half lethal concentration (LC50) of the crude extract from strain ZLP-101 against aphids was 411.535 mg/L. Preliminary exploration of the insecticidal mechanism revealed that the crude extract affected aphids to a greater extent through gastric poisoning than through contact. Further, the extracts affected enzymatic activities, causing holes to form in internal organs along with deformation, such that normal physiological activities could not be maintained, eventually leading to death. Isolation and purification of extracellular secondary metabolites were conducted in combination with mass spectrometry analysis to further identify the insecticidal components of the crude extracts. A total of 15 insecticidal active compounds were identified including iturins, fengycins, surfactins, and spergualins. Further insecticidal experimentation revealed that surfactin, iturin, and fengycin all exhibited certain aphidicidal activities, and the three exerted synergistic lethal effects. Conclusions: This study improved the available genomic resources for B. velezensis and serves as a foundation for comprehensive studies of the insecticidal mechanism by Bacillus velezensis ZLP-101 in addition to the active components within biological control strains. [ABSTRACT FROM AUTHOR]

Surfactants, also known as surface-active agents, have emerged as an important class of compounds with a wide range of applications. However, the use of chemical-derived surfactants must be restricted due to their potential adverse impact on the ecosystem and the health of human and other living organisms. In the past few years, there has been a growing inclination towards natural-derived alternatives, particularly microbial surfactants, as substitutes for synthetic or chemical-based counterparts. Microbial biosurfactants are abundantly found in bacterial species, predominantly Bacillus spp. and Pseudomonas spp. The chemical structures of biosurfactants involve the complexation of lipids with carbohydrates (glycolipoproteins and glycolipids), peptides (lipopeptides), and phosphates (phospholipids). Lipopeptides, in particular, have been the subject of extensive research due to their versatile properties, including emulsifying, antimicrobial, anticancer, and anti-inflammatory properties. This review provides an update on research progress in the classification of surfactants. Furthermore, it explores various bacterial biosurfactants and their functionalities, along with their advantages over synthetic surfactants. Finally, the potential applications of these biosurfactants in many industries and insights into future research directions are discussed. [ABSTRACT FROM AUTHOR]

Bacillus velezensis TCS001 is a novel biocontrol bacterium from marine habitat. This study investigated the inhibitory effects of lipopeptides produced by TCS001 on 17 plant pathogens through mycelial growth-rate assay, and the underlying anti-fungal mechanisms were preliminarily explored with Botrytis cinerea. Results showed that the inhibitory rates of TCS001 lipopeptides against 17 plant pathogens ranged from 56.45% to 97.15%. The inhibitory rates to Botryosphaeria dothidea (causing Carya cathayensis canker disease), Macrophoma kawatsukai, Sclerotinia sclerotiorum, Bipolaris sorokiniana, and Botrytis cinerea, were 97.15%, 94.87%, 92.64%, 91.04%, and 89.55%, respectively. Pot test and optical microscope observation demonstrated that lipopeptides could limit B. cinerea growth in vivo and have teratogenic effect on hypha. The fermentation conditions for optimal lipopeptides production of TCS001 were determined by a single-factor experiment combined with response surface methodology. The three key factors that affected lipopeptide production most were the nitrogen source, carbon source, and fermentation temperature. The optimized fermentation conditions were as follows: soluble starch 10.5 g/L, peanut cake powder 18.5 g/L, NaCl 3.0 g/L, liquid volume 32%, inoculum volume 3%, fermentation pH 6.0, fermentation temperature 31 °C, seed age 16 h, rotation speed 164 r/min, fermentation time 48 h. There is a good fit between the predicted values and the actual values of the response surface. The actual yield of lipopeptides increased to 0.653 g/L, which was 1.59 times that of 0.41 g/L before optimization. The results lay a foundation for the subsequent development of TCS001 and provide a theoretical basis. [ABSTRACT FROM AUTHOR]

In a scenario of increasing alarm about food waste due to rapid urbanization, population growth and lifestyle changes, this study aims to explore the valorization of waste from the retail sector as potential substrates for the biotechnological production of biosurfactants. With a perspective of increasingly contributing to the realization of the circular bioeconomy, a vegetable juice, derived from unsold fruits and vegetables, as a carbon source was used to produce lipopeptides such as surfactin and fengycin. The results from the shake flask cultivations revealed that different concentrations of vegetable juice could effectively serve as carbon sources and that the fed-batch bioreactor cultivation strategy allowed the yields of lipopeptides to be significantly increased. In particular, the product/substrate yield of 0.09 g/g for surfactin and 0.85 mg/g for fengycin was obtained with maximum concentrations of 2.77 g/L and 27.53 mg/L after 16 h, respectively. To conclude, this study provides the successful fed-batch cultivation of B. subtilis using waste product as the carbon source to produce secondary metabolites. Therefore, the consumption of agricultural product wastes might be a promising source for producing valuable metabolites which have promising application potential to be used in several fields of biological controls of fungal diseases. [ABSTRACT FROM AUTHOR]

Botrytis cinerea is the causal agent of gray mold disease affecting Paeoniaceae plants. The present study focuses on the potential of Bacillus amyloliquefaciens Mdgb15 as a promising biological control agent toward B. cinerea. The strain Mdgb15 showed the highest inhibition rate (55.56%) among all the strains tested against B. cinerea growth and was effective in controlling tree peony gray mold in detached leaves. A conventional PCR with three specific primer pairs detected lipopetides substance synthesis genes of Mdgb15. The results revealed that Mdgb15 had srfAA, fenA and ituD genes, which resulted in the production of surfactin, fengycin and iturin. An accurate qRT-PCR protocol with specifically designed primers was developed to characterize the expression of lipopeptide synthesis gene of Mdgb15. The qRT-PCR analysis revealed that the expression of the ituD and fenA genes in Mdgb15 was significantly up-regulated during the coculture with B. cinerea. Subsequently, the lipopeptides were extracted by acid precipitation of culture filtrate of Mdgb15 and were separated by Sephadex Gel chromatography. Three separated fractions exhibited strong antifungal activity against B. cinerea, and were identified as bacillomycin D by MALDI-TOF-MS analysis. This study has demonstrated that B. amyloliquefaciens Mdgb15 is a future potential new biocontrol agent for controlling tree peony gray mold disease. It suggests that the antifungal mechanisms of Mdgb15 may be at least partially associated with its ability to secret bacillomycin D. [ABSTRACT FROM AUTHOR]

Peptide-drug conjugates (PDCs) have recently gained significant attention for the targeted delivery of anticancer therapeutics, mainly due to their cost-effective and chemically defined production and lower antigenicity compared to ADCs, among other benefits. In this study, we designed and synthesized novel PDCs by conjugating new thiol-functionalized tubulysin analogs (tubugis) to bombesin, a peptide ligand with a relevant role in cancer research. Two tubulysin analogs bearing ready-for-conjugation thiol groups were prepared by an on-resin multicomponent peptide synthesis strategy and subsequently tested for their stand-alone in vitro anti-proliferative activity against human cancer cells, which resulted in IC50 values in the nanomolar range. In addition, various fluorescently labeled [K5]-bombesin(6–14) peptides, non-lipidated and lipidated with fatty acid chains of variable length, were also synthesized using the versatile multicomponent chemistry. These bombesin derivatives were tested for their gastrin-related peptide receptor (GRPR)-mediated internalization into cancer cells using flow cytometry, proving that the lipid tail (especially C14) enhances the cell internalization. Using the tubugi toxins and bombesin peptides, three different bombesin-tubugi conjugates were synthesized with different cleavage propensity and lipophilicity. Preliminary in vitro experiments revealed that, depending on the linker and the presence of a lipid tail, these novel PDCs possess good to potent anticancer activity and moderate selectivity for GRPR-overexpressing cancer cells.

Citrus melanose, caused by Diaporthe citri, is one of the most prevalent and important fungal diseases of citrus crops globally. However, the overuse of chemical fungicides for disease control has an adverse impact on citrus production. In contrast, biological control agents (BCAs) are environmentally friendly and have become essential tools for plant disease control. In the present study, Bacillus subtilis M23, a strain isolated from healthy citrus leaves, significantly suppressed the mycelial growth of D. citri, the causal agent of citrus melanose, in vitro. The fermentation broth of B. subtilis M23 also exhibited good antifungal activity against D. citri on citrus plants in greenhouse and field experiments. Lipopeptides (LPs) produced by M23 exhibited excellent antifungal activity against D. citri, with a 50 % effective concentration (EC50) of approximately 1.00 and 0.28 μg/mL for inhibition of mycelial growth and conidial germination, respectively. LP biosynthesis genes were identified in the M23 genome using a PCR assay. Notably, LP extracts significantly reduced the formation of pycnidia and ATP biosynthesis in D. citri. Comparative transcriptome analysis revealed significant differences in the expression of genes associated with fungal vacuole, oxidoreductase activity and endopeptidase activity in D. citri treated with B. subtilis M23. Both RNA-seq and RT-qPCR analysis provided similar results on the expression of selected genes. Collectively, our data provided convincing data supporting the biocontrol potential of B. subtilis M23 for the management of citrus melanose.

Beneficial endophytic bacteria can suppress the development of insect pests through direct antagonism, with the help of metabolites, or indirectly by the induction of systemic resistance through the regulation of hormonal signaling pathways. Lipopeptides are bacterial metabolites that exhibit direct antagonistic activity against many organisms, including insects. Also, lipopeptides are able to trigger induced systemic resistance (ISR) in plants against harmful organisms, but the physiological mechanisms of their action are just beginning to be studied. In this work, we studied ten strains of bacteria isolated from the tissues of wheat and potatoes. Sequencing of the 16S rRNA gene showed that all isolates belong to the genus Bacillus and to two species, B. subtilis and B. velezensis. The genes for lipopeptide synthetase – surfactin synthetase (Bs_srf ), iturin synthetase (Bs_ituA, Bs_ituB) and fengycin synthetase (Bs_fenD) – were identified in all bacterial isolates using PCR. All strains had high aphicidal activity against the Greenbug aphid (Schizaphis graminum Rond.) due to the synthesis of lipopeptides, which was proven using lipopeptiderich fractions (LRFs) isolated from the strains. Endophytic lipopeptide-synthesizing strains of Bacillus spp. indirectly affected the viability of aphids, the endurance of plants against aphids and triggered ISR in plants, which manifested itself in the regulation of oxidative metabolism and the accumulation of transcripts of the Pr1, Pr2, Pr3, Pr6 and Pr9 genes due to the synthesis of lipopeptides, which was proven using LRF isolated from three strains: B. subtilis 26D, B. subtilis 11VM, and B. thuringiensis B-6066. We have for the first time demonstrated the aphicidal effect of fengycin and the ability of the fengycin-synthesizing strains and isolates, B. subtilis Ttl2, Bacillus sp. Stl7 and B. thuringiensis B-6066, to regulate components of the pro-/antioxidant system of aphid-infested plants. In addition, this work is the first to demonstrate an elicitor role of fengycin in triggering a systemic resistance to S. graminum in wheat plants. We have discovered new promising strains and isolates of endophytes of the genus Bacillus, which may be included in the composition of new biocontrol agents against aphids. One of the criteria for searching for new bacteria active against phloem-feeding insects can be the presence of lipopeptide synthetase genes in the bacterial genome.

Abstract Background Barely Hordeum vulgare L. is considered one of the most important cereal crops with economic and industrial importance in the world, but its productivity is affected by climate change and abiotic stresses. One of the most recent and important microbiological promising aspects is the use of associated microorganisms, especially the endophytic bacteria producers for non-ribosomal peptides which play an important role in promoting plant growth, productivity, and tolerance to biotic and abiotic stresses. This work aims to identify vertically transferred or inherited endophytic bacterial communities in barely seeds, detect the presence of non-ribosomal peptides from these isolated endophytic strains and study their effect on protein patterns as a response to salinity stress. Results From two different tolerant (Giza 126) and sensitive (Giza 123) barely seeds cultivars, six different endophytic bacterial strains were isolated and identified using 16S rRNA. Bacterial strains belonged to Bacillus, Paenibacillus, Staphylococcus and Acinetobacter genera. Three of them have been isolated from both sensitive and tolerant barely cultivar (Uncultured Staphylococcus, Acinetobacter and Priestia endophytica or Bacillus endophyticus), while the other three endophytes have been isolated uniquely from the tolerant barely cultivar (Paenibacillus glucanolyticus, Bacillus cereus and Bacillus sp.). Non-ribosomal peptide synthetases genes NRPs of two lipopeptide families; surfactins and kurstakins have been detected using both bioinformatic analysis and degenerate primers. On the other hand, fragments similar to NRPs genes might be considered new NRPS molecules in Paenibacillus glucanolyticus, Acinetobacter and Priestia endophytica which have been detected using degenerate primers and required whole genome sequencing. The effect of soaking barely seeds exposed to 2.5% NaCl using SDS-PAGE electrophoresis revealed the presence of 24 bands, 10 of them were monomorphic with 41.5%, and 14 were polymorphic with 58.5% polymorphism. Conclusion The overnight soaking and co-cultivation of isolated endophytic strains with barely seeds before planting proved their capability in conferring salt stress tolerance to barely seedlings which appeared in protein patterns. We could consider these barely seeds endophytic among the PGPR strains promising to improve plant growth during abiotic stresses.

Isonitrile lipopeptides (INLPs) are known to be related to the virulence of pathogenic mycobacteria by mediating metal transport, but their biosynthesis remains obscure. In this work, we use in vitro biochemical assays, site-directed mutagenesis, chemical synthesis, and spectroscopy techniques to scrutinize the activity of core enzymes required for INLP biosynthesis in mycobacteria. Compared to environmental Streptomyces, pathogenic Mycobacterium employ a similar chemical logic and enzymatic machinery in INLP biosynthesis, differing mainly in the fatty-acyl chain length, which is controlled by multiple enzymes in the pathway. Our in-depth study on the non-heme iron(II) and α-ketoglutarate-dependent dioxygenase for isonitrile generation, including Rv0097 from Mycobacterium tuberculosis (Mtb), demonstrates that it recognizes a free-standing small molecule substrate, different from the recent hypothesis that a carrier protein is required for Rv0097 in Mtb. A key residue in Rv0097 is further identified to dictate the varied fatty-acyl chain length specificity between Streptomyces and Mycobacterium.

Fire blight, a devastating disease caused by Erwinia amylovora, poses a significant threat to pear and apple trees in Xinjiang province, China. In an effort to combat this pathogen, we isolated 10 bacteria from various components of apple and crabapple trees and conducted screenings to assess their ability to inhibit E. amylovora in vitro. Through biochemical tests and partial 16S rRNA gene sequencing, we identified two promising strains, Priestia megaterium strain H1 and Bacillus subtilis strain I2. These strains were then evaluated for their efficacy in biocontrol under controlled laboratory conditions, focusing on immature fruits and leaves. Remarkably, all selected antagonists exhibited the capability to reduce the severity of the disease on both fruit and leaves. P. megaterium strain H1 and B. subtilis strain I2 exhibited significant reductions in disease incidence on both immature fruits and leaves compared to the control. Specifically, on immature fruits, they achieved reductions of 53.39% and 44.76%, respectively, while on leaves, they demonstrated reductions of 59.55% and 55.53%, respectively. Furthermore, during the study, we detected the presence of lipopeptides, including surfactin, iturins, bacillomycin D, and fengycins, in the methanol extract obtained from these two antagonistic bacteria using thin-layer chromatography (TLC). Based on the results obtained, B. subtilis strain I2 and P. megaterium strain H1 exhibit considerable potential for controlling fire blight. However, further evaluation of their efficacy under natural field conditions is essential to validate their practicality as a biocontrol method. [ABSTRACT FROM AUTHOR]

As lead molecules, cyclic lipopeptides with antibacterial, antifungal, and antiviral properties have garnered a lot of attention in recent years. Because of their potential, cyclic lipopeptides have earned recognition as a significant class of antimicrobial compounds with applications in pharmacology and biotechnology. These lipopeptides, often with biosurfactant properties, are amphiphilic, consisting of a hydrophilic moiety, like a carboxyl group, peptide backbone, or carbohydrates, and a hydrophobic moiety, mostly a fatty acid. Besides, several lipopeptides also have cationic groups that play an important role in biological activities. Antimicrobial lipopeptides can be considered as possible substitutes for antibiotics that are conventional to address the current drug-resistant issues as pharmaceutical industries modify the parent antibiotic molecules to render them more effective against antibiotic-resistant bacteria and fungi, leading to the development of more resistant microbial strains. Bacillus species produce lipopeptides, which are secondary metabolites that are amphiphilic and are typically synthesized by non-ribosomal peptide synthetases (NRPSs). They have been identified as potential biocontrol agents as they exhibit a broad spectrum of antimicrobial activity. A further benefit of lipopeptides is that they can be produced and purified biotechnologically or biochemically in a sustainable manner using readily available, affordable, renewable sources without harming the environment. In this review, we discuss the biochemical and functional characterization of antifungal lipopeptides, as well as their various modes of action, method of production and purification (in brief), and potential applications as novel antibiotic agents. [ABSTRACT FROM AUTHOR]

Background: Barely Hordeum vulgare L. is considered one of the most important cereal crops with economic and industrial importance in the world, but its productivity is affected by climate change and abiotic stresses. One of the most recent and important microbiological promising aspects is the use of associated microorganisms, especially the endophytic bacteria producers for non-ribosomal peptides which play an important role in promoting plant growth, productivity, and tolerance to biotic and abiotic stresses. This work aims to identify vertically transferred or inherited endophytic bacterial communities in barely seeds, detect the presence of non-ribosomal peptides from these isolated endophytic strains and study their effect on protein patterns as a response to salinity stress. Results: From two different tolerant (Giza 126) and sensitive (Giza 123) barely seeds cultivars, six different endophytic bacterial strains were isolated and identified using 16S rRNA. Bacterial strains belonged to Bacillus, Paenibacillus, Staphylococcus and Acinetobacter genera. Three of them have been isolated from both sensitive and tolerant barely cultivar (Uncultured Staphylococcus, Acinetobacter and Priestia endophytica or Bacillus endophyticus), while the other three endophytes have been isolated uniquely from the tolerant barely cultivar (Paenibacillus glucanolyticus, Bacillus cereus and Bacillus sp.). Non-ribosomal peptide synthetases genes NRPs of two lipopeptide families; surfactins and kurstakins have been detected using both bioinformatic analysis and degenerate primers. On the other hand, fragments similar to NRPs genes might be considered new NRPS molecules in Paenibacillus glucanolyticus, Acinetobacter and Priestia endophytica which have been detected using degenerate primers and required whole genome sequencing. The effect of soaking barely seeds exposed to 2.5% NaCl using SDS-PAGE electrophoresis revealed the presence of 24 bands, 10 of them were monomorphic with 41.5%, and 14 were polymorphic with 58.5% polymorphism. Conclusion: The overnight soaking and co-cultivation of isolated endophytic strains with barely seeds before planting proved their capability in conferring salt stress tolerance to barely seedlings which appeared in protein patterns. We could consider these barely seeds endophytic among the PGPR strains promising to improve plant growth during abiotic stresses. [ABSTRACT FROM AUTHOR]

Aims Stem rot caused by Fusarium concentricum is a new disease of Paris polyphylla reported by our research group. The present study investigates the growth inhibitory and apoptotic effects of Bacillus velezensis FJAT-54560 lipopeptide against F. concentricum. Methods and Results HPLC preparation and LC-MS analysis results show that the crude lipopeptides secreted by Bacillus velezensis FJAT-54560 isolated from Jasminum sambac consist of C14–17 iturin A, C14 fengycin B, C16 fengycin A/A2, C18 fengycin A, C20 fengycin B2, C21 fengycin A2, C22–23 fengycin A, C12–16 surfactin A, and C15 surfactin A derivatives. The mass ratios (g/g) of iturin, fengycin, and surfactin in lipopeptides are 2.40, 67.51, and 30.08%, respectively. Through inhibition zone and inhibition rate experiments, we found that crude lipopeptides and purified fengycin exhibit strong antifungal activity against F. concentricum , including accumulation of reactive oxygen species, loss of mitochondrial membrane potential, DNA fragmentation, Ca2+ accumulation, chromatin condensation, and phosphatidylserine externalization. Transcriptomic analysis indicates that crude lipopeptide-induced apoptosis in F. concentricum cells may be mediated by apoptosis-inducing factors and apoptosis mediators and can serve as a metacaspase-independent model. Conclusion Lipopeptides from Bacillus velezensis FJAT-54560 can control the pathogenic fungus F. concentricum by inducing apoptosis. [ABSTRACT FROM AUTHOR]

Pathogenic bacteria contaminations and related diseases in food industries is an urgent issue to solve. The present study aimed to explore natural food biopreservatives from microorganisms. Using dilution-plate method, a strain BBW1542 with antimicrobial activities against various foodborne pathogenic bacteria was isolated from the seabed silt of Beibu Gulf, which was identified as Bacillus subtilis by the morphological observation and 16S rDNA sequences. The antimicrobial substances of B. subtilis BBW1542 exhibited an excellent stability under cool/heat treatment, UV irradiation, acid/alkali treatment, and protease hydrolysis. The genome sequencing analysis and antiSMASH prediction indicated that B. subtilis BBW1542 contained the gene cluster encoding lipopeptides and bacteriocin subtilosin A. MALDI–TOF-MS analysis showed that the lipopeptides from B. subtilis BBW1542 contained C14 and C15 surfactin homologues, together with fengycin homologues of C18 fengycin A/C16 fengycin B and C19 fengycin A/C17 fengycin B. In silico analysis showed that an eight-gene (sboA-albABCDEFG) operon was involved in the biosynthesis of subtilosin A in B. subtilis BBW1542, and the encoded subtilosin A presented an evident closed-loop structure containing 35 amino acids with a molecular weight of 3425.94 Da. Overall, the antagonistic B. subtilis BBW1542 displayed significant resource value and offered a promising alternative in development of food biopreservation. [ABSTRACT FROM AUTHOR]

Microbial interactions, which regulate the dynamics of eco- and agrosystems, can be harnessed to enhance antagonism against phytopathogenic fungi in agriculture. This study tests the hypothesis that plant growth-promoting rhizobacteria (PGPR) can also be potential biological control agents (BCAs). Antifungal activity assays against potentially phytopathogenic fungi were caried out using cultures and cell-free filtrates of nine PGPR strains previously isolated from agricultural soils. Cultures of Bacillus sp. BS36 inhibited the growth of Alternaria sp. AF12 and Fusarium sp. AF68 by 74 and 65%, respectively. Cell-free filtrates of the same strain also inhibited the growth of both fungi by 54 and 14%, respectively. Furthermore, the co-cultivation of Bacillus sp. BS36 with Pseudomonas sp. BS95 and the target fungi improved their antifungal activity. A subsequent metabolomic analysis using Fourier-transform ion cyclotron resonance mass spectrometry (FTICR-MS) identified fengycin- and surfactin-like lipopeptides (LPs) in the Bacillus sp. BS36 cell-free filtrates, which could explain their antifungal activity. The co-production of multiple families of LPs by Bacillus sp. BS36 is an interesting feature with potential practical applications. These results highlight the potential of the PGPR strain Bacillus sp. BS36 to work as a BCA and the need for more integrative approaches to develop biocontrol tools more accessible and adoptable by farmers. [ABSTRACT FROM AUTHOR]

A root bacterium identified as Bacillus velezensis CMML21–47 with strong biocontrol activity against two major sweet potato fungal diseases caused by Fusarium oxysporum and Ceratocystis fimbriata was isolated in this study. In vitro bioassays and field assays were conducted to test the biocontrol activity and confirmed its strong antagonistic activity. The complete genome of B. velezensis CMML21–47 was sequenced, and multiple gene clusters related to the synthesis of antibiotics by antiSMASH analysis were detected. Selective ultra–high performance liquid chromatography with quadrupole time–of–flight mass spectrometry (UPLC–Q–TOF–MS) was performed and detected lipopeptides, including fengycin, surfactin, and bacillomycin. Lipopeptide fractions were separated, tested against the pathogens, and showed strong growth inhibition. Accordingly, CMML21-47 is an attractive antifungal agent, with high potential to control the sweet potato pathogens causing wilt and black rot diseases.

IntroductionThe fungus Fusarium verticillioides significantly threatens maize crops in tropical soils. In light of this, biological control has emerged as a promising strategy to reduce fungicide costs and environmental risks. In this study, we aimed to test the antifungal activity of cell-free supernatant (CFS) from three Bacillus velezensis (CT02, IM14, and LIS05) and one Paenibacillus ottowii (LIS04) against F. verticillioides, thereby contributing to the development of effective biocontrol measures.MethodsThe research employed a comprehensive approach. The antifungal activity of the bacterial strains was tested using cell-free supernatant (CFS) from three Bacillus velezensis (CT02, IM14, and LIS05) and one Paenibacillus ottowii (LIS04). The UPLC-MS evaluated the CFS to identify the main bioactive molecules involved in the inhibitory effect on F. verticillioides. Scanning electron microscopy (SEM) was used to assess the impact of CFS on spores and hyphae, and genome sequencing was conducted to identify the genes involved in biological control. These robust methodologies ensure the reliability and validate our findings.ResultsThe CFS of the four strains demonstrated significant inhibition of fungal growth. The UPLC-MS analysis revealed the presence of lipopeptides with antifungal activity, including surfactin and fengycins A and B expressed by the three strains of Bacillus velezensis and iturin A expressed by strains LIS05 and IM14. For Paenibacillus ottowii, fusaricidins, ABCDE, and five previously unreported lipopeptides were detected. Scanning electron microscopy (SEM) showed that treatments with CFS led to significant distortion and breakage of the F. verticillioides hyphae, in addition to the formation of cavities in the membrane. Genome mining confirmed the presence of genes coding for the lipopeptides identified by UPLC-MS, including the gene for iturin in CTO2. Genomic sequencing revealed that CT02, IM14, and LIS05 belong to different strains of Bacillus velezensis, and LIS04 belongs to Paenibacillus ottowii, a species recently described.DiscussionThe four bacterial strains, including three novel strains identified as Bacillus velezensis and one as the recently described species Paenibacillus ottowii, demonstrate significant potential as biocontrol agents for managing fungal disease. This finding underscores the novelty and potential impact of our research.

Potato is a major crop whose yield and quality are impacted by the pathogenic fungus Rhizoctonia solani. Here, a control approach based on the use of endophytic bacillus spores and bacterial consortia was assessed as an innovative solution to improve tuber productivity and health. Endophytic potato bacteria extracted from several organs (leaves, stems and roots) evaluated for their antagonistic activity against R. solani by the double culture technique and their ability to rot potato slices. Additionally, the ability of Bacillus strains to generate hydrolytic enzymes and produce lipopeptides (using ESI-MS/MS mass spectrometry) was assessed. Then, the effectiveness of selected bacterial strains, either individually or in consortium (after carrying out compatibility tests) in the suppression of R. solani was evaluated in the greenhouse. Six endophytes not affecting potato tubers and having an antagonistic effect against R. solani were identified: B. halotolerans SpS5, B. velezensis KnL15, B. aryabhattai FaR1, B. amyloliquefaciens LiR9, B. haynesii ReR10 which produce several types and combinations of lipopeptides belonging to different families (iturins, fengycins and surfactins) as well as hydrolytic enzymes (chitinase, protease and cellulase). The vegetative and sporulating strain SpS5, as well as the 5 other Bacillus and their Sps5-based consortia, were found to be very effective in increasing the yield of healthy, high-quality tubers and reducing disease severity. Based on the results mentioned above, a microbial fungicide from the spores of (i) B. halotolerans SpS5 for which we sequenced the whole genome or (ii) bacterial consortia could be developed to combat black scurf potato and other fungal infections.

Active films based on different biopolymers (pectin, Brea gum, carboxymethylcellulose, and composite starch-gelatin) were prepared by casting. Lipopeptides synthesized by bacteria of the Bacillus genus were added as antimicrobial active agent. Antibacterial activity was evaluated and quantified by direct contact technique against different strains of Listeria monocytogenes, Escherichia coli, and Salmonella enterica sp. Incorporation of lipopeptides did not cause significant changes in mechanical and barrier properties of films. Antibacterial effect of functionalized films depended on the nature of the polymeric matrix and the pathogen studied. Functionalized pectin films inhibited the growth of all strains studied, with non-functionalized pectin films also showing inhibition of L. monocytogenes. Functionalized starch-gelatin films only inhibited the growth of L. monocytogenes. The other matrices did not show adequate performance for this type of application. The results suggested that pectin and starch-gelatin composite can be used as effective lipopeptide carriers to produce antimicrobial active films for food preservation. [ABSTRACT FROM AUTHOR]

[Display omitted] We evaluated the use of lipopeptides capable to bind to nucleic acids towards plasmid DNA (pDNA) delivery. The investigations were particularly focused on arising retinal pigment epithelial cells (ARPE-19) as motivated by the considerable number of ocular disorders linked to gene aberrations. The lipopeptides comprised the artificial oligoamino acid succinyl - tetraethylene pentamine (Stp) as well as incorporated lysines, histidines, cysteines, fatty acids, and tyrosine trimers. Regardless of the structural differences, the lipopeptides demonstrated to efficiently condense pDNA at nitrogen-to-phosphate molar ratio (N/P) ≥ 6. Spheric nanoparticles were observed by cryo-TEM and dynamic light scattering determined hydrodynamic sizes ranging from 50 to 130 nm. The biological assays evidenced highly efficient pDNA delivery with a lower degree of cytotoxicity compared to the well-known transfecting agent linear polyethylenimine (LPEI). Although more efficient than LPEI, cysteine-containing carriers were demonstrated to be less efficient than the other counterparts possibly due to exceeding polyplex stabilization via disulfide cross links, which could hamper pDNA unpacking at the target site. Therefore, clearly a balance between complex stability and cargo release should be taken into account to optimize the transfection efficiency of the non-viral vectors. The gene transfer activity in ARPE-19 cells suggests the applicability of this kind of carrier for ocular treatments based on retinal gene delivery. [ABSTRACT FROM AUTHOR]

[Display omitted] Lipopeptides synthesized from the Nucleolar Targeting Peptide (NrTP6) with one, two or four dodecanoic fatty acid (FA) chains, display large head to tail volumes, which together with the number of lipid chains per molecule, impacts their self-assembly behavior. In phosphate buffer (PB), peptide to peptide interactions are triggered by the presence of phosphate ions that act as ionic crosslinkers, affecting the organization of the lipid assemblies. The NrTP6 lipopeptides were synthesized by the solid phase peptide synthesis technique. The critical micellar concentration (CMC) of the lipopeptides was determined in water and PB by pyrene fluorescence. The size and morphology of lipopeptide assemblies were characterized by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Circular dichroism (CD) was used to study the secondary structures of the lipopeptide assemblies. For NrTP6 lipopeptides with two and four lipid chains, CMCs in water are larger than in PB. TEM images of the lipopeptide assemblies show different morphologies including fibers, rods, and spheres depending on the number of lipid chains, concentration and whether they are assembled in water or PB. CD spectroscopy shows that the peptide conformation, either random or beta, correlates with the morphology of the assemblies. [ABSTRACT FROM AUTHOR]

Biohybrid catalysts that operate in aqueous media are intriguing for systems chemistry. In this paper, we investigate whether control over the self‐assembly of biohybrid catalysts can tune their properties. As a model, we use the catalytic activity of functional hybrid molecules consisting of a catalytic H‐dPro‐Pro‐Glu tripeptide, derivatized with fatty acid and nucleobase moieties. This combination of simple biological components merged the catalytic properties of the peptide with the self‐assembly of the lipid, and the structural ordering of the nucleobases. The biomolecule hybrids self‐assemble in aqueous media into fibrillar assemblies and catalyze the reaction between butanal and nitrostyrene. The interactions between the nucleobases enhanced the order of the supramolecular structures and affected their catalytic activity and stereoselectivity. The results point to the significant control and ordering that nucleobases can provide in the self‐assembly of biologically inspired supramolecular catalysts. [ABSTRACT FROM AUTHOR]

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Alternaria alternata wreaks havoc on fruit and vegetable production globally, threatening food security by causing black leaf spot disease. Bacillus subtilis, a natural inhabitant of soil, is a promising biological control agent for the management of A. alternata. In the present study, the antagonistic potential of B. subtilis NBAIR-BSWG1 was initially confirmed against A. alternata through a dual culture technique with 43.03% inhibition of mycelial growth. Subsequently, we extracted the cell-free extract from the NBAIR-BSWG1 pure culture and assessed its impact on A. alternata through the poison food technique and found mycelial growth inhibition of 85.82%. Identification of secondary metabolites biosynthetic genes using specific PCR markers showed the presence of surfactin genes (sfp, srf AA) with an amplicon size of 675 bp and 201 bp, respectively. Amplification of fengycin (fenB) and iturin (ituD) at 670 bp and 423 bp respectively, by using a specific PCR primer confirms the contribution of fengycin and iturin for the antagonistic potential of NBAIR-BSWG1. This study identifies NBAIR-BSWG1 as an effective bacterial biocontrol agent for control of A. alternata, unlocks the genetic basis of antifungal activity NBAIRBSWG1, depicts molecular mechanisms involved in biological suppression of A. alternata by NBAIR-BSWG1 paving the way for the development of bioformulations for management of A. alternata. [ABSTRACT FROM AUTHOR]

ABSTRACTBiological control is a more sustainable and environmentally friendly alternative to chemical fungicides for controlling Fusarium spp. infestations. In this work, Bacillus siamensis Sh420 isolated from wheat rhizosphere showed a high antifungal activity against Fusarium graminearum as a secure substitute for fungicides. Sh420 was identified as B. siamensis using phenotypic evaluation and 16S rDNA gene sequence analysis. An in vitro antagonistic study showed that Sh420’s lipopeptide (LP) extract exhibited strong antifungal properties and effectively combated F. graminearum. Meanwhile, lipopeptides have the ability to decrease ergosterol content, which has an impact on the overall structure and stability of the plasma membrane. The PCR-based screening revealed the presence of antifungal LP biosynthetic genes in this strain’s genomic DNA. In the crude LP extract of Sh420, we were able to discover several LPs such as bacillomycin, iturins, fengycin, and surfactins using ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry. Microscopic investigations (fluorescent/transmission electron microscopy) revealed deformities and alterations in the morphology of the phytopathogen upon interaction with LPs. Sh420 LPs have been shown in grape tests to be effective against F. graminearum infection and to stimulate antioxidant activity in fruits by avoiding rust and gray lesions. The overall findings of this study highlight the potential of Sh420 lipopeptides as an effective biological control agent against F. graminearum infestations.IMPORTANCEThis study addresses the potential of lipopeptide (LP) extracts obtained from the strain identified as Bacillus siamensis Sh420. This Sh420 isolate acts as a crucial player in providing a sustainable and environmentally friendly alternative to chemical fungicides for suppressing Fusarium graminearum phytopathogen. Moreover, these LPs can reduce ergosterol content in the phytopathogen influencing the overall structure and stability of its plasma membrane. PCR screening provided confirmation regarding the existence of genes responsible for biosynthesizing antifungal LPs in the genomic DNA of Sh420. Several antibiotic lipopeptide compounds were identified from this bacterial crude extract using ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry. Microscopic investigations revealed deformities and alterations in the morphology of F. graminearum upon interaction with LPs. Furthermore, studies on fruit demonstrated the efficacy of Sh420 LPs in mitigating F. graminearum infection and stimulating antioxidant activity in fruits, preventing rust and gray lesions.

Voltage-gated sodium channel (VGSC) activators promote neurite outgrowth by augmenting intracellular Na+ concentration ([Na+]i) and upregulating N-methyl-d-aspartate receptor (NMDAR) function. NMDAR activation stimulates calcium (Ca2+) influx and increases brain-derived neurotrophic factor (BDNF) release and activation of tropomyosin receptor kinase B (TrkB) signaling. The BDNF-TrkB pathway has been implicated in activity-dependent neuronal development. We have previously shown that antillatoxin (ATX), a novel lipopeptide isolated from the cyanobacterium Moorea producens, is a VGSC activator that produces an elevation of [Na+]i. Here we address the effect of ATX on the synthesis and release of BDNF and determine the signaling mechanisms by which ATX enhances neurite outgrowth in immature cerebrocortical neurons. ATX treatment produced a concentration-dependent release of BDNF. Acute treatment with ATX also resulted in increased synthesis of BDNF. ATX stimulation of neurite outgrowth was prevented by pretreatment with a TrkB inhibitor or transfection with a dominant-negative Trk-B. The ATX activation of TrkB and Akt was blocked by both a NMDAR antagonist (MK-801) and a VGSC blocker (tetrodotoxin). These results suggest that VGSC activators such as the structurally novel ATX may represent a new pharmacological strategy to promote neuronal plasticity through a NMDAR-BDNF-TrkB-dependent mechanism.

Heavy metal pollution damages the ecosystems and presents a major problem for public health. Thus, an urgent need was developed to decrease the high levels of heavy metals in the soil and aquatic environments. With this aim, numerous physicochemical strategies were developed. However, they are money-consuming, require the use of energy and chemical additives and can release secondary compounds that can pollute and cause great damage to the environment. Then, biological methods based on the investigation of bacteria, fungi and plants along with their derived secondary active metabolites became the best alternatives. Using plant capacities, different phytoremediation strategies were developed such as phytoextraction, phytovolatilization, rhizofiltration and phytostabilization. Regarding bioremediation, bacterial biosorption of heavy metals, biolixiviation and lagooning offer great potential for their environmental cleaning. Additionally, the use of secondary active metabolites, such as biosurfactants, is well-studied. Generally, they are a class of structurally very varied molecules commonly synthesized by many microorganisms with amphiphilic character. Owing to their anionic charge, they have the capacity to sequestrate heavy metals permitting their elimination. Glycolipids and lipopeptides are among the most recognized biosurfactants with interesting heavy metal chelating properties. HIGHLIGHTS Studied heavy metal pollution and toxicology.; Bioremediation and phytoremediation technologies for heavy metal depollution.; Use of biosurfactants as heavy metal chelators.;

Abstract Natural rhizosphere bacteria has the potential to act as an alternative of chemical pesticides for sustainable agriculture. In the current study, tobacco rhizosphere Bacillus velezensis D exhibited great antibacterial effect against Ralstonia solanacearum, and significantly enhanced the tobacco resistance against bacterial wilt in pot experiments. Then Bacillus velezensis D was labeled with gfp marker and found to stably colonize in tobacco root, the colonization density of strain D in root still remained 5.33 × 104 CFU/gat 30 days post-inoculation. Subsequently, field trials for two years (2021–2022) showed the control effects of the strain D on the tobacco bacterial wilt were 12.26% and 36.37%, respectively, indicating the application of B. velezensis D could improve plant resistance to R.solanacearum. In order to further study the antibacterial activities of strain D, effects of the crude extracts on the swimming ability, cell viability and the morphology of R. solanacearum were analyzed. The results showed that the crude extracts reduced the motility of R. solanacearum, and caused cell wall rupture and cell death. Furthermore, MALDI-TOF-MS and HPLC-QTOF-MS analysis indicated that lipopeptides (fengycin and iturin) and polyketides (bacillaene) were detected in the crude extracts of strain D. Based on these findings, we speculated that Bacillus velezensis D firstly colonized in tobacco root, then produced antibacterial substances at ecological sites to exert antagonistic effects, inhibiting motility traits of R. solanacearum and damaging the cell well. Hence, Bacillus velezensis D could be used as a potential biological control agents against tobacco bacterial wilt.

This review describes the discovery, structure, activity, engineered constructs, and applications of KR-12, the smallest antibacterial peptide of human cathelicidin LL-37, the production of which can be induced under sunlight or by vitamin D. It is a moonlighting peptide that shows both antimicrobial and immune-regulatory effects. Compared to LL-37, KR-12 is extremely appealing due to its small size, lack of toxicity, and narrow-spectrum antimicrobial activity. Consequently, various KR-12 peptides have been engineered to tune peptide activity and stability via amino acid substitution, end capping, hybridization, conjugation, sidechain stapling, and backbone macrocyclization. We also mention recently discovered peptides KR-8 and RIK-10 that are shorter than KR-12. Nano-formulation provides an avenue to targeted delivery, controlled release, and increased bioavailability. In addition, KR-12 has been covalently immobilized on biomaterials/medical implants to prevent biofilm formation. These constructs with enhanced potency and stability are demonstrated to eradicate drug-resistant pathogens, disrupt preformed biofilms, neutralize endotoxins, and regulate host immune responses. Also highlighted are the safety and efficacy of these peptides in various topical and systemic animal models. Finaly, we summarize the achievements and discuss future developments of KR-12 peptides as cosmetic preservatives, novel antibiotics, anti-inflammatory peptides, and microbiota-restoring agents.

Rice sheath blight is a serious disease caused by Rhizoctonia solani that reduces rice yield. Currently, there is a lack of efficient and environmentally friendly control methods. In this study, we found that Bacillus velezensis (B. velezensis) Y6 could significantly inhibit the growth of mycelium in Rhizoctonia solani, and its control efficiency against rice sheath blight was 58.67% (p < 0.01) in a pot experiment. Lipopeptides play an important role in the control of rice sheath blight by B. velezensis Y6, among which iturin and fengycin are essential, and iturin W, a novel lipopeptide in B. velezensis, plays a major role in lipopeptide antagonism to Rhizoctonia solani. In the field, we also found that inoculation with B. velezensis Y6 can increase rice yield (dry weight) by 11.75%. Furthermore, the transcriptome profiling results of the rice roots revealed that there were a total of 1227 differential genes (DEGs) regulated when treated with Y6, of which 468 genes were up-regulated and 971 genes were down-regulated in rice roots compared with the control. Among them, the DEGs were mainly distributed in biological processes (BP) and were mainly enriched in response to stimulus (GO:0050896), response to stress (GO:0006950), and response to abiotic stimulus (GO:0009628). According to the KEGG pathway analysis, there were 338 DEGs classified into 87 KEGG functional pathway categories. Compared with the control, a large number of enriched genes were distributed in phenylpropanoid biosynthesis (map00940), glutathione metabolism (map00480), glycolysis/gluconeogenesis (map00010), and amino sugar and nucleotide sugar metabolism (map00520). In summary, this investigation provides a new perspective for studying the molecular mechanism of B. velezensis in controlling rice sheath blight.