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1.
Ecotoxicol Environ Saf ; 226: 112840, 2021 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-34619473

RESUMEN

Within the framework of biocontrol development, several natural lipopeptides produced by Bacillus subtilis show well-documented anti-microbial properties, especially in orchards. However, the number of studies on their putative insecticidal effects remain low despite the growing interest to develop new strategies of orchards pests' control. The rosy apple aphid Dysaphis plantaginea is the major aphid pest causing great leaf damage to apple trees. In this study, we submitted young adult aphids to topical application of three different families of lipopeptides, Plipastatin (Fengycin), Mycosubtilin (Iturin), and Surfactin, either separately or as a ternary mixture. Their aphicidal effects were investigated at 1, 2.5 and 5 g/L, both at 1 h and 24 h after exposure, and their effects on aphid behavior were studied at the 2.5 g/L concentration at 24 h after exposure. When delivered alone, lipopeptides displayed contrasted effects varying from no aphicidal activity for Mycosubtilin to a mortality induced even at low concentrations by Surfactin. Surprisingly, locomotor activity of the surviving aphids was only affected by the two least lethal treatments, Mycosubtilin and the ternary mix. Their feeding behavior was only impacted by Surfactin, the most lethal treatment, that unexpectedly increased phloem sap ingestion. The results are discussed in the context of lipopeptides applicability for integrated pest management.


Asunto(s)
Áfidos , Bacillus , Insecticidas , Animales , Bacillus subtilis , Insecticidas/toxicidad , Lipopéptidos/farmacología
2.
Amino Acids ; 51(8): 1233-1240, 2019 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-31197572

RESUMEN

The aim of this study was to evaluate the effect of spreading the lipopeptide surfactin, for short time (10/20 s), on dentin wettability. Study groups were surfactin: 2.8; 1.4; 0.7; 0.35; and 0.175 mg/mL and a control group that received no treatment. Dentin discs (4 mm height) were prepared and polished with 600-grit SiC paper. Contact angle determinations were carried out after microbrush spreading of surfactin on dentin specimens for, respectively, 10 and 20 s. Excess liquid was removed, and after 60 s, the specimens were analyzed in a goniometer using the sessile drop method to measure the contact angle. Results were analyzed by two-way ANOVA (concentration × time) and t student, with α = 0.05. Lower contact angles were obtained for surfactin (0.7 mg/mL) spread for 10 s. However, no statistical difference was observed for surfactin (2.8 mg/mL) applied during 20 s. Higher contact angles were observed for surfactin (0.7 mg/mL) spread for 20 s. In conclusion, dentin wettability is dependent on spreading time and surfactin concentration.


Asunto(s)
Dentina/química , Lipopéptidos/administración & dosificación , Péptidos Cíclicos/administración & dosificación , Diente/química , Humectabilidad/efectos de los fármacos , Dentina/efectos de los fármacos , Humanos , Ensayo de Materiales , Propiedades de Superficie
3.
Microb Cell Fact ; 17(1): 121, 2018 Aug 04.
Artículo en Inglés | MEDLINE | ID: mdl-30077177

RESUMEN

BACKGROUND: Lipopeptides are a promising group of surface-active compounds of microbial origin (biosurfactants). These diverse molecules are produced mainly by Bacillus and Pseudomonas strains. Because of their attractive physiochemical and biological properties, biosurfactants are considered to be "green and versatile molecules of the future". The main obstacles in widespread use of biosurfactants are mainly their low yields and high production costs. Pseudofactin (PF) is a lipopeptide produced by Pseudomonas fluorescens BD5. Recently, we identified two analogues, PF1 (C16-Val) and PF2 (C16-Leu), and reported that PF2 has good emulsification and foaming activities, as well as antibacterial, antifungal, anticancer, and antiadhesive properties. Reported production of PF in a mineral salt medium was approximately 10 mg/L. RESULTS: Here, we report successful high-throughput optimization of culture medium and conditions for efficient PF production using P. fluorescens BD5. Compared with production in minimal medium, PF yield increased almost 120-fold, up to 1187 ± 13.0 mg/L. Using Plackett-Burman and central composite design methodologies we identified critical factors that are important for efficient PF production, mainly high glycerol concentration, supplementation with amino acids (leucine or valine) and complex additives (e.g. tryptone), as well as high culture aeration. We also detected the shift in a ratio of produced PF analogues in response to supplementation with different amino acids. Leucine strongly induces PF2 production, while valine addition supports PF1 production. We also reported the identification of two new PF analogues: PF3 (C18-Val) and PF4 (C18-Leu). CONCLUSIONS: Identification of critical culture parameters that are important for lipopeptide production and their high yields can result in reduction of the production costs of these molecules. This may lead to the industrial-scale production of biosurfactants and their widespread use. Moreover, we produced new lipopeptide pure analogues that can be used to investigate the relationship between the structure and biological activity of lipopeptides.


Asunto(s)
Lipopéptidos/metabolismo , Pseudomonas fluorescens/metabolismo , Tensoactivos
4.
Can J Microbiol ; 63(1): 46-60, 2017 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-27912317

RESUMEN

Biosurfactants are important in many areas; however, costs impede large-scale production. This work aimed to develop a global sustainable strategy for the production of biosurfactants by a novel strain of Bacillus amyloliquefaciens. Initially, Bacillus sp. strain 0G was renamed B. amyloliquefaciens subsp. plantarum (syn. Bacillus velezensis) after analysis of the gyrA and gyrB DNA sequences. Growth in modified Landy's medium produced 3 main recoverable metabolites: surfactin, fengycin, and acetoin, which promote plant growth. Cultivation was studied in the presence of renewable carbon (as glycerol) and nitrogen (as arginine) sources. While diverse kinetics of acetoin production were observed in different media, similar yields (6-8 g·L-1) were obtained after 72 h of growth. Glycerol increased surfactin-specific production, while arginine increased the yields of surfactin and fengycin and increased biomass significantly. The specific production of fengycin increased ∼10 times, possibly due to a connecting pathway involving arginine and ornithine. Adding value to crude extracts and biomass, both were shown to be useful, respectively, for the removal of p-xylene from contaminated water and for biodiesel production, yielding ∼70 mg·g-1 cells and glycerol, which could be recycled in novel media. This is the first study considering circular bioeconomy to lower the production costs of biosurfactants by valorisation of both microbial cells and their primary and secondary metabolites.


Asunto(s)
Bacillus amyloliquefaciens/metabolismo , Biocombustibles/análisis , Medios de Cultivo/metabolismo , Contaminantes Químicos del Agua/metabolismo , Xilenos/metabolismo , Bacillus amyloliquefaciens/genética , Bacillus amyloliquefaciens/crecimiento & desarrollo , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Biodegradación Ambiental , Biomasa , Glicerol/metabolismo , Lipopéptidos/metabolismo , Péptidos Cíclicos/metabolismo
5.
Bioprocess Biosyst Eng ; 40(2): 161-180, 2017 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27738757

RESUMEN

Innovations in novel enzyme discoveries impact upon a wide range of industries for which biocatalysis and biotransformations represent a great challenge, i.e., food industry, polymers and chemical industry. Key tools and technologies, such as bioinformatics tools to guide mutant library design, molecular biology tools to create mutants library, microfluidics/microplates, parallel miniscale bioreactors and mass spectrometry technologies to create high-throughput screening methods and experimental design tools for screening and optimization, allow to evolve the discovery, development and implementation of enzymes and whole cells in (bio)processes. These technological innovations are also accompanied by the development and implementation of clean and sustainable integrated processes to meet the growing needs of chemical, pharmaceutical, environmental and biorefinery industries. This review gives an overview of the benefits of high-throughput screening approach from the discovery and engineering of biocatalysts to cell culture for optimizing their production in integrated processes and their extraction/purification.


Asunto(s)
Enzimas/biosíntesis , Enzimas/química , Enzimas/genética , Ingeniería de Proteínas/métodos , Catálisis
6.
Appl Microbiol Biotechnol ; 98(14): 6255-64, 2014.
Artículo en Inglés | MEDLINE | ID: mdl-24723290

RESUMEN

The use of surfactin and mycosubtilin as an eco-friendly alternative to control lettuce downy mildew caused by the obligate pathogen Bremia lactucae was investigated. Preliminary ecotoxicity evaluations obtained from three different tests revealed the rather low toxicity of these lipopeptides separately or in combination. The EC50 (concentration estimated to cause a 50 % response by the exposed test organisms) was about 100 mg L(-1) in Microtox assays and 6 mg L(-1) in Daphnia magna immobilization tests for mycosubtilin and 125 mg L(-1) and 25 mg L(-1) for surfactin, respectively. The toxicity of the mixture mycosubtilin/surfactin (1:1, w/w) was close to that obtained with mycosubtilin alone. In addition, the very low phytotoxic effect of these lipopeptides has been observed on germination and root growth of garden cress Lepidium sativum L. While a surfactin treatment did not influence the development of B. lactucae on lettuce plantlets, treatment with 100 mg L(-1) of mycosubtilin produced about seven times more healthy plantlets than the control samples, indicating that mycosubtilin strongly reduced the development of B. lactucae. The mixture mycosubtilin/surfactin (50:50 mg L(-1)) gave the same result on B. lactucae development as 100 mg L(-1) of mycosubtilin. The results of ecotoxicity as well as those obtained in biocontrol experiments indicated that the presence of surfactin enhances the biological activities of mycosubtilin. Mycosubtilin and surfactin were thus found to be efficient compounds against lettuce downy mildew, with low toxicity compared to the toxicity values of chemical pesticides. This is the first time that Bacillus lipopeptides have been tested in vivo against an obligate pathogen and that ecotoxic values have been given for surfactin and mycosubtilin.


Asunto(s)
Antifúngicos/farmacología , Lactuca/microbiología , Lipopéptidos/farmacología , Oomicetos/efectos de los fármacos , Péptidos Cíclicos/farmacología , Control Biológico de Vectores/métodos , Enfermedades de las Plantas/microbiología , Antifúngicos/toxicidad , Sinergismo Farmacológico , Lipopéptidos/toxicidad , Lipoproteínas/farmacología , Lipoproteínas/toxicidad , Oomicetos/crecimiento & desarrollo , Péptidos Cíclicos/toxicidad
7.
Microbiol Res ; 280: 127569, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38103466

RESUMEN

Crown gall disease caused by Agrobacterium tumefaciens is considered to be the main bacterial threat of stone fruit plants in Mediterranean countries. In a previous study, Bacillus velezensis strain 32a was isolated from Tunisian rhizosphere soil and revealed high antagonistic potential against A. tumefaciens strains. In order to better characterize the antagonistic activity of this strain against this important plant pathogen, the production of secondary metabolites was analyzed using liquid chromatography coupled with mass spectrometry. The results revealed the production of different compounds identified as surfactins, fengycins, iturins and bacillibactin belonging to the lipopeptide group, three polyketides (macrolactins, oxydifficidin and bacillaenes), bacilysin and its chlorinated derivative; chlorotetaine. The involvement of lipopeptides in this antagonistic activity was ruled out by performing agar and broth dilution tests with pure molecules. Thus, the construction of B. velezensis 32a mutants defective in polyketides and bacilysin biosynthesis and their antagonistic activity was performed and compared to a set of derivative mutants of a comparable strain, B. velezensis GA1. The defective difficidin mutants (△dfnA and △dfnD) were unable to inhibit the growth of A. tumefaciens, indicating the high-level contribution of difficidin in the antagonism process. While the macrolactin deficient mutant (∆mlnA) slightly decreased the activity, suggesting a synergetic effect with difficidin. Remarkably, the mutant △dhbC only deficient in bacillibactin production showed significant reduction in its capacity to inhibit the growth of Agrobacterium.Taken collectively, our results showed the strong synergetic effect of difficidin and macrolactins and the significant implication of siderophore to manage crown gall disease.


Asunto(s)
Bacillus , Policétidos , Tumores de Planta , Bacillus/metabolismo , Policétidos/farmacología , Policétidos/metabolismo , Lactonas
8.
Microbiol Res ; 286: 127816, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38964072

RESUMEN

Apple scab, caused by the hemibiotrophic fungus Venturia inaequalis, is currently the most common and damaging disease in apple orchards. Two strains of V. inaequalis (S755 and Rs552) with different sensitivities to azole fungicides and the bacterial metabolite fengycin were compared to determine the mechanisms responsible for these differences. Antifungal activity tests showed that Rs552 had reduced sensitivity to tebuconazole and tetraconazole, as well as to fengycin alone or in a binary mixture with other lipopeptides (iturin A, pumilacidin, lichenysin). S755 was highly sensitive to fengycin, whose activity was close to that of tebuconazole. Unlike fengycin, lipopeptides from the iturin family (mycosubtilin, iturin A) had similar activity on both strains, while those from the surfactin family (lichenysin, pumilacidin) were not active, except in binary mixtures with fengycin. The activity of lipopeptides varies according to their family and structure. Analyses to determine the difference in sensitivity to azoles (which target the CYP51 enzyme involved in the ergosterol biosynthesis pathway) showed that the reduced sensitivity in Rs552 is linked to (i) a constitutive increased expression of the Cyp51A gene caused by insertions in the upstream region and (ii) greater efflux by membrane pumps with the involvement of ABC transporters. Microscopic observations revealed that fengycin, known to interact with plasma membranes, induced morphological and cytological changes in cells from both strains. Sterol and phospholipid analyses showed a higher level of ergosta-7,22-dien-3-ol and a lower level of PI(C16:0/C18:1) in Rs552 compared with S755. These differences could therefore influence the composition of the plasma membrane and explain the differential sensitivity of the strains to fengycin. However, the similar antifungal activities of mycosubtilin and iturin A in the two strains indirectly indicate that sterols are probably not involved in the fengycin resistance mechanism. This leads to the conclusion that different mechanisms are responsible for the difference in susceptibility to azoles or fengycin in the strains studied.


Asunto(s)
Ascomicetos , Azoles , Lipopéptidos , Malus , Enfermedades de las Plantas , Lipopéptidos/farmacología , Malus/microbiología , Enfermedades de las Plantas/microbiología , Ascomicetos/efectos de los fármacos , Ascomicetos/metabolismo , Ascomicetos/genética , Azoles/farmacología , Farmacorresistencia Fúngica/genética , Pruebas de Sensibilidad Microbiana , Antifúngicos/farmacología , Antifúngicos/metabolismo , Fungicidas Industriales/farmacología , Regulación Fúngica de la Expresión Génica/efectos de los fármacos , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo
9.
Soft Matter ; 9(40): 9572-8, 2013 Oct 28.
Artículo en Inglés | MEDLINE | ID: mdl-26029764

RESUMEN

The self-assembly in aqueous solution of three lipopeptides obtained from Bacillus subtilis has been investigated. The lipopeptides surfactin, plipastatin and mycosubtilin contain distinct cyclic peptide headgroups as well as differences in alkyl chain length, branching and chain length distribution. Cryogenic transmission electron microscopy and X-ray scattering reveal that surfactin and plipastatin aggregate into 2 nm-radius spherical micelles, whereas in complete contrast mycosubtilin self-assembles into extended nanotapes based on bilayer ordering of the lipopeptides. Circular dichroism and FTIR spectroscopy indicate the presence of turn structures in the cyclic peptide headgroup. The unexpected distinct mode of self-assembly of mycosubtilin compared to the other two lipopeptides is ascribed to differences in the surfactant packing parameter. This in turn is due to specific features of the conformation of the peptide headgroup and alkyl chain branching.


Asunto(s)
Bacillus subtilis , Ácidos Grasos/química , Membrana Dobles de Lípidos/química , Lipopéptidos/química , Micelas , Oligopéptidos/química , Péptidos Cíclicos/química , Microscopía por Crioelectrón , Lipoproteínas/química , Microscopía Electrónica de Transmisión , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos X
10.
Pest Manag Sci ; 78(3): 929-937, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34719104

RESUMEN

BACKGROUND: Lipopeptides extracted from the Bacillus genus are emerging biopesticides, especially in protecting crops against phytopathogens. Among the three main families of lipopeptides, surfactins have been identified as having insecticidal properties against several insect orders. However, the sublethal effects of these promising biopesticides on insect pests and their natural enemies remain largely unknown. The aim of this study was to evaluate the effects of surfactins topically applied on black bean aphid Aphis fabae mortality. First, the effects of surfactins on aphid mortality were determined by delivering increasing concentrations to adults and nymphs. Second, the sublethal effects of surfactins on locomotor activity and feeding behavior of surviving aphids were evaluated using the electropenetrography method. Finally, the effect of host exposure to surfactins on host selection behavior by Aphidius matricariae parasitoid females was analyzed. RESULTS: Four surfactins concentrations were studied (0.5, 1, 2.5 and 5 g L-1 ). There was concentration-dependent mortality in response to surfactins at 24 h after treatment. Surfactins impacted aphid behavior when delivered at 1 g L-1 by inducing a greater locomotor activity and a reduction in feeding activity. By contrast, at the third trophic level, exposure of aphid hosts to surfactins did not affect behaviors leading to host recognition and acceptance by parasitoid females. CONCLUSION: This study highlighted the consequences of aphid exposure to surfactins in the context of bottom-up regulation. Although surfactins could directly impact aphid behavior, they had no apparent consequences on the host selection behavior exhibited by parasitoid wasps.


Asunto(s)
Áfidos , Bacillus , Avispas , Animales , Femenino , Interacciones Huésped-Parásitos , Lipopéptidos/farmacología
11.
Front Microbiol ; 13: 914713, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-35794911

RESUMEN

This study investigated the antiradical and antioxidant potential of the three families of lipopeptides (i.e., surfactin, mycosubtilin, and plipastatin/fengycin) produced by Bacillus subtilis strains. The antiradical/antioxidant activities of highly purified lipopeptides were studied in acellular models using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide anion ( O 2 . - ), hydrogen peroxide, (H2O2) and hydroxyl radical (HO.). At a lipopeptide concentration of 500 mg.L-1, the maximum inhibition of DPPH reached 22.88% (obtained for plipastatin). Moreover, the scavenging effects of O 2 . - , H2O2, and HO. at the highest concentration tested (250 mg.L-1) were found to be 6, 21, and 3% for surfactin, 19, 9, and 15% for mycosubtilin, 21, 18, and 59% for plipastatin, 21, 31, and 61% for the mixture of surfactin/plipastatin, and 13, 16, and 15% for the mixture of surfactin/mycosubtilin, respectively. These results showed that plipastatin was the best candidate due to its antioxidant activities.

12.
Metabolites ; 12(2)2022 Jan 24.
Artículo en Inglés | MEDLINE | ID: mdl-35208182

RESUMEN

Mycosubtilin belongs to the family of lipopeptides. Different isoforms with various antifungal activities can be obtained according to the length and the isomery of the fatty acid. In this work, the activities of the mycosubtilin isoforms were first studied against the pathogen Aspergillus niger, revealing the high activity of the anteiso-C17 isoform. Modification of the mycosubtilin isoform patterns during cultures of the natural strain Bacillus subtilis ATCC 6633 was then investigated through amino acid feeding experiments. In parallel, single-gene knockouts and single-gene overexpression, leading to the overproduction of the anteiso-C15 fatty acid chains, were predicted using informatics tools which provide logical reasoning with formal models of reaction networks. In this way, it was in silico predicted that the single overexpression of the ilvA gene as well as the single knockout of the codY gene may lead to the overproduction of anteiso-C15 fatty acid chains. For the first time, it has been demonstrated that overexpression of ilvA helps to enhance the furniture of odd anteiso fatty acids leading to a favored mycosubtilin anteiso-C17 production pattern (+41%). Alternatively, a knock-out codY mutant led to a higher furniture of even iso fatty acids, leading to a favored mycosubtilin iso-C16 production pattern (+180%). These results showed that increased selective synthesis of particular isoforms of mycosubtilin through metabolic engineering is feasible, disclosing the interest of these approaches for future development of lipopeptide-producing strains.

13.
Microorganisms ; 10(2)2022 Jan 19.
Artículo en Inglés | MEDLINE | ID: mdl-35208667

RESUMEN

The exploration of certain microbial resources such as beneficial endophytic microorganisms is considered a promising strategy for the discovery of new antimicrobial compounds for the pharmaceutical industries and agriculture. Thirty-one endophytic bacterial strains affiliated with Bacillus, Janthinobacterium, Yokenella, Enterobacter, Pseudomonas, Serratia, and Microbacterium were previously isolated from vetiver (Chrysopogon zizanioides (L.) Roberty) roots. These endophytes showed antifungal activity against Fusarium graminearum and could be a source of antimicrobial metabolites. In this study, in particular, using high-throughput screening, we analyzed their antagonistic activities and those of their cell-free culture supernatants against three species of Fusarium plant pathogens, a bacterial strain of Escherichia coli, and a yeast strain of Saccharomyces cerevisiae, as well as their capacity to produce lipopeptides. The results showed that the culture supernatants of four strains close to B. subtilis species exhibited antimicrobial activities against Fusarium species and E. coli. Using mass spectrometry analyses, we identified two groups of lipopeptides (surfactins and plipastatins) in their culture supernatants. Whole-genome sequencing confirmed that these bacteria possess NRPS gene clusters for surfactin and plipastatin. In vitro tests confirmed the inhibitory effect of plipastatin alone or in combination with surfactin against the three Fusarium species.

14.
Front Plant Sci ; 13: 1074447, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-36777540

RESUMEN

Plant immunity induction with natural biocontrol compounds is a valuable and promising ecofriendly tool that fits with sustainable agriculture and healthy food. Despite the agroeconomic significance of wheat, the mechanisms underlying its induced defense responses remain obscure. We reveal here, using combined transcriptomic, metabolomic and cytologic approach, that the lipopeptide mycosubtilin from the beneficial bacterium Bacillus subtilis, protects wheat against Zymoseptoria tritici through a dual mode of action (direct and indirect) and that the indirect one relies mainly on the priming rather than on the elicitation of plant defense-related mechanisms. Indeed, the molecule primes the expression of 80 genes associated with sixteen functional groups during the early stages of infection, as well as the accumulation of several flavonoids during the period preceding the fungal switch to the necrotrophic phase. Moreover, genes involved in abscisic acid (ABA) biosynthesis and ABA-associated signaling pathways are regulated, suggesting a role of this phytohormone in the indirect activity of mycosubtilin. The priming-based bioactivity of mycosubtilin against a biotic stress could result from an interaction of the molecule with leaf cell plasma membranes that may mimic an abiotic stress stimulus in wheat leaves. This study provides new insights into induced immunity in wheat and opens new perspectives for the use of mycosubtilin as a biocontrol compound against Z. tritici.

15.
Microorganisms ; 10(9)2022 Sep 09.
Artículo en Inglés | MEDLINE | ID: mdl-36144412

RESUMEN

Apple scab is an important disease conventionally controlled by chemical fungicides, which should be replaced by more environmentally friendly alternatives. One of these alternatives could be the use of lipopeptides produced by Bacillus subtilis. The objective of this work is to study the action of the three families of lipopeptides and different mixtures of them in vitro and in vivo against Venturia inaequalis. Firstly, the antifungal activity of mycosubtilin/surfactin and fengycin/surfactin mixtures was determined in vitro by measuring the median inhibitory concentration. Then, the best lipopeptide mixture ratio was produced using Design of Experiment (DoE) to optimize the composition of the culture medium. Finally, the lipopeptides mixtures efficiency against V. inaequalis was assessed in orchards as well as the evaluation of the persistence of lipopeptides on apple. In vitro tests show that the use of fengycin or mycosubtilin alone is as effective as a mixture, with the 50-50% fengycin/surfactin mixture being the most effective. Optimization of culture medium for the production of fengycin/surfactin mixture shows that the best composition is glycerol coupled with glutamic acid. Finally, lipopeptides showed in vivo antifungal efficiency against V. inaequalis regardless of the mixture used with a 70% reduction in the incidence of scab for both mixtures (fengycin/surfactin or mycosubtilin/surfactin). The reproducibility of the results over the two trial campaigns was significantly better with the mycosubtilin/surfactin mixture. The use of B. subtilis lipopeptides to control this disease is very promising.

16.
Front Bioeng Biotechnol ; 9: 678469, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34124025

RESUMEN

In this work, an original culture process in bioreactor named overflowing continuous culture (O-CC) was developed to produce and recover continuously mycosubtilin, a lipopeptide antifungal biosurfactant of major interest. The lipopeptide production was first investigated in shake conical flasks in different culture media [ammonium citrate sucrose (ACS), Difco sporulation medium (DSM), and Landy], followed by a pH condition optimization using 3-(N-morpholino)propanesulfonic acid (MOPS) and 2-(N-morpholino)ethanesulfonic acid (MES) buffered media. A simple theoretical modeling of the biomass evolution combined with an experimental setup was then proposed for O-CC processed in stirred tank reactor at laboratory scale. Seven O-CC experiments were done in modified Landy medium at the optimized pH 6.5 by applying dilution rates comprised between 0.05 and 0.1 h-1. The O-CC allowed the continuous recovery of the mycosubtilin contained in the foam overflowing out of the reactor, achieving a remarkable in situ product removal superior to 99%. The biomass concentration in the overflowing foam was found to be twofold lower than the biomass concentration in the reactor, relating advantageously this process to a continuous one with biomass feedback. To evaluate its performances regarding the type of lipopeptide produced, the O-CC process was tested with strain BBG116, a mycosubtilin constitutive overproducing strain that also produces surfactin, and strain BBG125, its derivative strain obtained by deleting surfactin synthetase operon. At a dilution rate of 0.1 h-1, specific productivity of 1.18 mg of mycosubtilin⋅g-1(DW)⋅h-1 was reached. Compared with other previously described bioprocesses using almost similar culture conditions and strains, the O-CC one allowed an increase of the mycosubtilin production rate by 2.06-fold.

17.
Front Bioeng Biotechnol ; 9: 815337, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-35004661

RESUMEN

Lipopeptides produced by Bacillus subtilis display many activities (surfactant, antimicrobial, and antitumoral), which make them interesting compounds with a wide range of applications. During the past years, several processes have been developed to enable their production and purification with suitable yield and purity. The already implemented processes mainly end with a critical drying step, which is currently achieved by freeze-drying. In this study, the possibility to replace this freeze-drying step with a spray-drying one, more suited to industrial applications, was analyzed. After evaluating their thermal resistance, we have developed a spray-drying methodology applicable for the three lipopeptides families produced by B. subtilis, i.e., surfactin, mycosubtilin (iturin family), and plipastatin (fengycin family). For each lipopeptide, the spray-drying procedure was applied at three steps of the purification process by ultrafiltration (supernatant, diafiltered solution, and pre-purified fraction). The analysis of the activities of each spray-dried lipopeptide showed that this drying method is not decreasing its antimicrobial and biosurfactant properties. The methodology developed in this study enabled for the first time the spray-drying of surfactin, without adjuvants' addition and regardless of the purification step considered. In the case of fengycin and mycosubtilin, only diafiltered solution and purified fraction could be successfully spray-dried without the addition of adjuvant. Maltodextrin addition was also investigated as the solution for the direct drying of supernatant. As expected, the performances of the spray-drying step and the purity of the powder obtained are highly related to the purification step at which the product was dried. Interestingly, the impact of mycosubtilin concentration on spray-drying yield was also evidenced.

18.
Appl Microbiol Biotechnol ; 87(2): 499-507, 2010 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-20221757

RESUMEN

Surfactin and fengycin are lipopeptide biosurfactants produced by Bacillus subtilis. This work describes for the first time the use of bubbleless bioreactors for the production of these lipopeptides by B. subtilis ATCC 21332 with aeration by a hollow fiber membrane air-liquid contactor to prevent foam formation. Three different configurations were tested: external aeration module made from either polyethersulfone (reactor BB1) or polypropylene (reactor BB2) and a submerged module in polypropylene (reactor BB3). Bacterial growth, glucose consumption, lipopeptide production, and oxygen uptake rate were monitored during the culture in the bioreactors. For all the tested membranes, the bioreactors were of satisfactory bacterial growth and lipopeptide production. In the three configurations, surfactin production related to the culture volume was in the same range: 242, 230, and 188 mg l(-1) for BB1, BB2, and BB3, respectively. Interestingly, high differences were observed for fengycin production: 47 mg l(-1) for BB1, 207 mg l(-1) for BB2, and 393 mg l(-1) for BB3. A significant proportion of surfactin was adsorbed on the membranes and reduced the volumetric oxygen mass transfer coefficient. The degree of adsorption depended on both the material and the structure of the membrane and was higher with the submerged polypropylene membrane.


Asunto(s)
Bacillus subtilis/metabolismo , Reactores Biológicos/microbiología , Microbiología Industrial/instrumentación , Lipopéptidos/metabolismo , Péptidos Cíclicos/metabolismo , Tensoactivos/metabolismo , Bacillus subtilis/química , Bacillus subtilis/crecimiento & desarrollo , Biomasa , Microbiología Industrial/métodos , Cinética , Lipopéptidos/química , Péptidos Cíclicos/química , Tensoactivos/química
19.
Microorganisms ; 8(5)2020 May 07.
Artículo en Inglés | MEDLINE | ID: mdl-32392736

RESUMEN

Biofilm bioreactors are promising systems for continuous biosurfactant production since they provide process stability through cell immobilization and avoid foam formation. In this work, a two-compartment biofilm bioreactor was designed consisting of a stirred tank reactor and a trickle-bed reactor containing a structured metal packing for biofilm formation. A strong and poor biofilm forming B. subtilis 168 strain due to restored exopolysaccharides (EPS) production or not were cultivated in the system to study the growth behavior of the planktonic and biofilm population for the establishment of a growth model. A high dilution rate was used in order to promote biofilm formation on the packing and wash out unwanted planktonic cells. Biofilm development kinetics on the packing were assessed through a total organic carbon mass balance. The EPS+ strain showed a significantly improved performance in terms of adhesion capacity and surfactin production. The mean surfactin productivity of the EPS+ strain was about 37% higher during the continuous cultivation compared to the EPS- strain. The substrate consumption together with the planktonic cell and biofilm development were properly predicted by the model (α = 0.05). The results show the efficiency of the biofilm bioreactor for continuous surfactin production using an EPS producing strain.

20.
Front Bioeng Biotechnol ; 8: 585815, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33102467

RESUMEN

Production of bioactive peptides (BAPs) by Lactobacillus species is a cost-effective approach compared to the use of purified enzymes. In this study, proteolytic Lactobacillus helveticus strains were used for milk fermentation to produce BAPs capable of inhibiting angiotensin converting enzyme (ACE). Fermented milks were produced in bioreactors using batch mode, and the resulting products showed significant ACE-inhibitory activities. However, the benefits of fermentation in terms of peptide composition and ACE-inhibitory activity were noticeably reduced when the samples (fermented milks and non-fermented controls) were subject to simulated gastrointestinal digestion (GID). Introducing an ultrafiltration step after fermentation allowed to prevent this effect of GID and restored the effect of fermentation. Furthermore, an integrated continuous process for peptide production was developed which led to a 3 fold increased peptide productivity compared to batch production. Using a membrane bioreactor allowed to generate and purify in a single step, an active ingredient for ACE inhibition.

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