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1.
Production of lipopeptides fengycin and surfactin in rotating discs bioreactor was studied. The effects of rotation velocity and the addition of agitators between the discs on volumetric oxygen transfer coefficient k L a were firstly studied in model media. Then the production of lipopeptides was also studied at different agitation conditions in the modified bioreactor (with agitators). The effect of agitation on dissolved oxygen, on submerged and immobilized biomass, on lipopeptide concentrations and yields and on the selectivity of the bioreaction was elucidated and discussed. The proposed modified rotating discs bioreactor allowed to obtain high fengycin concentrations (up to 787 mg L?1), but also better selectivity of the bioreaction towards fengycin (up to 88 %) and better yields of fengycin per glucose (up to 62.9 mg g?1), lipopeptides per glucose (up to 71.5 mg g?1), fengycin per biomass (up to 309 mg g?1) and lipopeptides per biomass (up to 396 mg g?1) than those reported in the literature. Highest fengycin production and selectivity were obtained at agitation velocity of 30 min?1. The proposed non-foaming fermentation process could contribute to the scale-up of lipopeptide fermentors and promote the industrial production of fengycin. The proposed bioreactor and bioprocess could be very useful also for the production of other molecules using bioprocesses requiring bubbleless oxygen supply.  相似文献   

2.
Bacillus subtilis fmbj can produce a lipopeptide antimicrobial substance, the main components of which are surfactin and fengycin. In this paper, the sensitivity of Bacillus cereus to antimicrobial lipopeptides from B. subtilis fmbj was observed, and the effect of the microstructure of antimicrobial lipopeptide on spores of B. cereus was investigated. At the same time, the optimization of the inactivation of antimicrobial lipopeptides to spores of B. cereus by a response surface methodology was studied. Results showed that B. cereus had high sensitivity to it, whose minimal inhibitory concentration was 156.25 μg/ml. It could result in the death of spores by destroying the structure of resting spores and sprouting spores, as was observed by transmission electron microscopy. The optimization result indicated that spores of B. cereus could be inactivated by 2 orders of magnitude when the temperature was 29.6°C, the action time was 7.6 h, and the concentration was 3.46 mg·ml−1.  相似文献   

3.
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.  相似文献   

4.
This study evaluates osmolality of a submerged conidia-producing medium in relation to the following spore characteristics: yield, morphology (dimensions and cell wall structure), chemical properties of cell wall surfaces (charge, hydrophobicity, and lectin binding), cytoplasmic polyols and trehalose, and performance (drying stability and pathogenicity). Spore production was increased by the addition of up to 150 g l?1 polyethylene glycol 200 (PEG). Spores from high osmolality medium (HOM spores) containing 100 g l?1 PEG had thin cell walls and dimensions more similar to blastospores than submerged conidia or aerial conidia. However, a faint electron-dense layer separating primary and secondary HOM spores’ cell walls was discernable by transmission electron microscopy as found in aerial and submerged conidia but not found in blastospores. HOM spores also appeared to have an outer rodlet layer, unlike blastospores, although it was thinner than those observed in submerged conidia. HOM spores’ surfaces possessed hydrophobic microsites, which was further evidence of the presence of a rodlet layer. In addition, HOM spores had concentrations of exposed N-acetyl-β-d-glucosaminyl residues intermediate between blastospores and submerged conidia potentially indicating a masking of underlying cell wall by a rodlet layer. All spore types had exposed α-d-mannosyl and/or α-d-glucosyl residues, but lacked oligosaccharides. Similar to blastospores, HOM spores were less anionic than submerged conida. Although HOM spores had thin cell walls, they were more stable to drying than blastospores and submerged conidia. Relative drying stability did not appear to be the result of differences in polyol or trehalose concentrations, since trehalose concentrations were lower in HOM spores than submerged conidia and polyol concentrations were similar between the two spore types. HOM spores had faster germination rates than submerged conidia, similar to blastospores, and they were more pathogenic to Schistocerca americana than submerged conidia and aerial conidia.  相似文献   

5.
Microbial production of two biosurfactants, fengycin and surfactin, by Bacillus subtilis ATCC 21332 in a rotating discs bioreactor was studied. Simultaneous production of these lipopeptides was performed by free and cells immobilized on the surfaces of rotating discs. The aeration applied on surface allowed a non-foaming fermentation process and an important production of lipopeptides for low microbial growth in the culture medium. It was demonstrated that the selectivity of lipopeptides synthesis could be modified varying operating conditions and that the cells immobilization improved greatly fengycin synthesis. The maximal concentration of fengycin and surfactin obtained were 838 mg L?1 and 212 mg L?1, respectively. The development of this bubble-less process could advance the scale-up of the fermenters for production of biosurfactants.  相似文献   

6.
In this study, influence of three critical parameters nitrogen sources, initial pH and metal ions was discussed in the production of antifungal lipopeptides from Bacillus amyloliquefaciens Q-426. The results revealed that lipopeptide biosynthesis might have relations with the population density of strain Q-426 and some special amino acids. Also, the alkali-resistant strain Q-426 could grow well in the presence of Fe2+ ions below 0.8 M l?1 and still maintain the competitive advantage below 0.2 M l?1. Moreover, lipopeptides exhibited significant inhibitory activities against Curvularia lunata (Walk) Boed even at the extreme conditions of temperature, pH and salinity. Finally, biosurfactant properties of lipopeptides mixture were evaluated by use with totally six different methods including bacterial adhesion to hydrocarbons assay, lipase activity, hemolytic activity, emulsification activity, oil displacement test and surface tension measurement. The research suggested that B. amyloliquefaciens Q-426 may have great potential in agricultural and environmental fields.  相似文献   

7.
Abstract

A regeneration protocol from leaf explants of Grindelia robusta Nutt. was developed. The combination of 0.5 mg l?1 IBA plus 0.5 mg l?1 or 1 mg l?1 BA added to Murashige-Skoog (MS) medium resulted in the best callus induction frequency; the combination of 0.4 or 0.9 mg l?1 BA plus 1.2 mg l?1 GA3 resulted in the best shoot regeneration. Rooting was successful on MS medium supplemented with 0.5 mg l?1 IBA. Hardening of G. robusta plants was accomplished in 30 days with 85% survival rate.  相似文献   

8.
To gain a better understanding of the factors influencing spore adhesion in dairy manufacturing plants, casein-modified glass surfaces were prepared and characterized and their effect on the adhesion kinetics of spores from a Geobacillus sp., isolated from a dairy manufacturing plant (DMP) was assessed using a flow chamber. Surfaces were produced by initially silanizing glass using (3-glycidyloxypropyl) trimethoxysilane (GPS) or (3-aminopropyl) triethoxysilane to form epoxy-functionalized (G-GPS) or amino-functionalized glass (G-NH2) substrata. Casein was grafted to the G-GPS directly by its primary amino groups (G-GPS-casein) or to G-NH2 by employing glutaraldehyde as a linking agent (G-NH2-glutar-casein). The surfaces were characterised using streaming potential measurements, contact angle goniometry, infrared spectroscopy and scanning electron microscopy. The attachment rate of spores suspended in 0.1 M KCl at pH 6.8, was highest on the positively charged (+14 mV) G-NH2 surface (333 spores cm?2 s?1) compared to the negatively charged glass (?22 mV), G-GPS (?20 mV) or G-GPS-casein (?21 mV) surfaces (162, 17 or 6 spores cm?2 s?1 respectively). Whilst there was a clear decrease in attachment rate to negatively charged casein-modified surfaces compared to the positively charged amine surface, there was no clear relationship between surface hydrophobicity and spore attachment rate.  相似文献   

9.
A marine Bacillus circulans DMS-2 was able to grow and produce biosurfactant on glucose mineral salts medium (GMSM) with a reduction in the surface tension up to 27 mN m−1. The microorganism produced 1.64 ± 0.1 g l−1 of crude biosurfactant. The lipopeptide nature of the produced biosurfactant was confirmed by primulin and ninhydrin assays using High Performance Thin Layer Chromatography (HPTLC). Preparative thin layer chromatography (TLC) was performed to purify the lipopeptides from the crude biosurfactant. The critical micelle concentrations (CMC) of the crude and purified products were found to be 90 and 40 mg l−1 respectively. Fourier transform infrared spectrophotometer (FTIR) and matrix assisted laser desorption/ionization time of flight (MALDI-ToF) mass spectral analysis revealed the identity of the produced lipopeptides as surfactin (m/z 1,023 Da) and fengycin (m/z 1,495 Da) isoforms. The purified marine lipopeptides displayed a significant antiproliferative activity against the human colon cancer cell lines HCT-15 (IC50 80 μg ml−1) and HT-29 (IC50 120 μg ml−1).  相似文献   

10.
Aims: To compare physical properties of spores that were produced in broth sporulation media at greater than 108 spores ml−1. Methods and Results: Bacillus atrophaeus reproducibly sporulated in nutrient broth (NB) and sporulation salts. Microscopy measurements showed that the spores were 0·68 ± 0·11 μm wide and 1·21 ± 0·18 μm long. Coulter Multisizer (CM3) measurements revealed the spore volumes and volume-equivalent spherical diameters, which were 0·48 ± 0·38 μm3 and 0·97 ± 0·07 μm, respectively. Bacillus cereus reproducibly sporulated in NB, sporulation salts, 200 mmol l−1 glutamate and antifoam. Spores were 0·95 ± 0·11 μm wide and 1·31 ± 0·17 μm long. Spore volumes were 0·78 ± 0·61 μm3 and volume-equivalent spherical diameters were 1·14 ± 0·11 μm. Bacillus atrophaeus spores were hydrophilic and B. cereus spores were hydrophobic. However, spore hydrophobicity was significantly altered after treatment with pH-adjusted bleach. Conclusions: The utility of a CM3 for both quantifying Bacillus spores and measuring spore sizes was demonstrated, although the volume between spore exosporium and spore coat was not measured. This study showed fundamental differences between spores from a Bacillus subtilis- and B. cereus-group species. Significance and Impact of the Study: This is useful for developing standard methods for broth spore production and physical characterization of both living and decontaminated spores.  相似文献   

11.
Abstract

Monitoring of biofilms subjected to different operating conditions was performed using a flow cell system. The system was fed by chlorine-free tap water, with and without added nutrients (0.5 mg l?1 carbon, 0.1 mg l?1 nitrogen and 0.01 mg l?1 phosphorus), and biofilms were grown on polyvinyl chloride (PVC) and stainless steel (SS) coupons, both in laminar and turbulent flow. The parameters analysed were culturable cells, using R2A, and total bacteria, which was assessed using the 4,6-diamino-2-phenylindole (DAPI) staining method. The impact of the different operating conditions in the studied parameters was established using Multivariate Analysis of Variance (MANOVA). From the most relevant to the least relevant factor, the total and culturable bacteria in biofilms increased due to the addition of nutrients to water (F = 20.005; p < 0.001); the use of turbulent (Re = 11000) instead of laminar (Re = 2000) hydrodynamic flows (F = 9.173; p < 0.001); and the use of PVC instead of SS as the support material (F = 2.848; p = 0.060). Interactions between these conditions, namely between surface and flow (F = 8.235; p < 0.001) and also flow and nutrients (F = 5.498; p < 0.05) have also proved to significantly influence biofilm formation. This work highlights the need for a deeper understanding of how the large spectrum of conditions interact and affect biofilm formation potential and accumulation with the final purpose of predicting the total and culturable bacteria attached to real drinking water distribution pipes based on the system characteristics.  相似文献   

12.
Aims: To evaluate the efficacy of chlorine dioxide (ClO2) against seven species of bacterial threat (BT) agents in water. Methods and Results: Two strains of Bacillus anthracis spores, Yersinia pestis, Francisella tularensis, Burkholderia pseudomallei, Burkholderia mallei and Brucella species were each inoculated into a ClO2 solution with an initial concentration of 2·0 (spores only) and 0·25 mg l?1 (all other bacteria) at pH 7 or 8, 5 or 25°C. At 0·25 mg l?1 in potable water, six species were inactivated by at least three orders of magnitude within 10 min. Bacillus anthracis spores required up to 7 h at 5°C for the same inactivation with 2·0 mg l?1 ClO2. Conclusions: Typical ClO2 doses used in water treatment facilities would be effective against all bacteria tested except B. anthracis spores that would require up to 7 h with the largest allowable dose of 2 mg l?1 ClO2. Other water treatment processes may be required in addition to ClO2 disinfection for effective spore removal or inactivation. Significance and Impact of Study: The data obtained from this study provide valuable information for water treatment facilities and public health officials in the event that a potable water supply is contaminated with these BT agents.  相似文献   

13.
The development of natural crop protection products as alternatives to the use of synthetic fungicides is currently popular. The aim of this study is to evaluate the antifungal effects of several essential oils against the fungal pathogens, Botrytis cinerea and Rhizopus stolonifer, under in vitro condition. Four essential oils (fennel, black caraway, peppermint and thyme) were each tested at five concentrations (0, 200, 400, 600 or 800 μl l?1). In vitro results showed that the essential oil of black caraway and fennel had the highest fungicidal effect against B. cinerea and R. stolonifer, respectively. The growth of B. cinerea was completely inhibited by the essential oil of black caraway at 400 μl l?1. Fennel oil perfectly inhibited growth of R. stolonifer fungus colonies at concentration higher than 600 μl L?1 in potato dextrose agar medium. Percentage of spores germination was the lowest in medium of Fennel and black caraway essential oils, and was the highest in Thyme ones. These results show that plant essential oils can have a strong effect on reducing post-harvest decay. These plant essential oils could provide an alternative to synthetic chemicals to control post-harvest phytopathogenic fungi on fruit.  相似文献   

14.
Aim: To investigate the viability, surface characteristics and ability of spores of a Geobacillus sp. isolated from a milk powder production line to adhere to stainless steel surfaces before and after a caustic (NaOH) wash used in clean‐in‐place regimes. Methods and Results: Exposing sessile spores to 1% NaOH at 65°C for 30 min decreased spore viability by two orders of magnitude. The zeta potential of the caustic treated spores decreased from ?20 to ?32 mV and they became more hydrophobic. Transmission electron microscopy revealed that caustic treated spores contained breaks in their spore coat. Under flow conditions, caustic treated spores suspended in 0·1 mol l?1 KCl were shown to attach to stainless steel in significantly greater numbers (4·6 log10 CFU cm?2) than untreated spores (3·6 log10 CFU cm?2). Conclusions: This research suggests that spores surviving a caustic wash will have a greater propensity to attach to stainless steel surfaces. Significance of Study: The practice of recycling caustic wash solutions may increase the risk of contaminating dairy processing surfaces with spores.  相似文献   

15.
Atrazine-degrading microorganisms designated EAA-3 and EAA-4, belonging to the genus Nocardioides, were obtained from an agricultural soil in Nigeria. The degradation kinetics of the two strains revealed total disappearance of 25 mg l?1 of atrazine in less than 72 h of incubation at the rate of 0.42 mg l?1 h?1 and 0.35 mg l?1 h?1, respectively. Screening for atrazine catabolic genes in these organisms revealed the presence of trzN, atzB, and atzC. Other genes, specifically atzA, atzD, and trzD, were not detected. Potential intermediates of atrazine catabolic route such as hydroxyatrazine, desethylatrazine, and desisopropylatrazine were utilized as sources of carbon and energy, while desisopropyl desethyl-2-hydroxyatrazine and desisopropyl-2-hydroxyatrazine were attacked but in the presence of glucose. A soil microcosm study showed that degradation was faster in microcosms contaminated with 13 mg of atrazine per g?1 of soil compared with 480 mg g?1 of soil. In the former, degradation was 10% higher in the inoculated soil than the non-inoculated control (natural attenuation) over the 28-day study period. Corresponding value obtained for the latter was nearly 70% higher. This study has demonstrated that the bacterial strains isolated enhanced atrazine degradation and the catabolic activities of these strains were not affected with increasing soil atrazine concentration.  相似文献   

16.
《Process Biochemistry》2010,45(11):1795-1799
Microbial production and isolation of biosurfactants was studied. The production of lipopeptides surfactin and fengycin was performed by free and immobilized aerobic cells of Bacillus subtilis ATCC 21332. After preliminary tests with 5 polymer materials, the particles of polypropylene foamed with powder activated carbon (PPch) were selected for lipopeptides production for their thermal and mechanical stability and for the high colonizing effect. To avoid foaming during biosurfactant production, biofilm grown on solid floating support was aerated by air injected over the surface of cultural medium. The synthesis of both lipopeptides and especially of the fengycin was greatly enhanced by the immobilization. The relationship between support wettability, colonization of the cells, and lipopeptide production was discussed. Extraction behaviour of the lipopeptides into alkanes was studied. The distribution ratio of surfactin was found to be higher than this of fengycin at the same conditions and the n-heptane was more efficient solvent for both lipopeptides. Kinetics of surfactin recovery from fermentation broth applying batch pertraction in a rotating discs contactor was studied. Lipopeptide was successfully extracted (more than 75% in the first hour) using n-heptane as liquid membrane and a 0.2 mol L−1 phosphate buffer solution (pH  7.3) as receiving solution. However, the stripping of the organic liquid and surfactin accumulation into the receiving phase were less efficient.  相似文献   

17.
Aims: Our goal was to find a novel, biosurfactant‐producing bacterium from Pacific Ocean deep‐sea sediments. Methods and Results: An oil‐degrading biosurfactant‐producing bacterium TW53 was obtained from deep‐sea sediment, and was identified through 16S rDNA analysis as belonging to the genus Rhodococcus. It lowered the surface tension of its culture to 34·4 mN m?1. Thin layer chromatography (TLC) showed that the crude biosurfactants of TW53 were composed of lipopeptides and free fatty acids (FA). The lipopeptides were purified with column chromatography and then hydrolysed with 6 mol l?1 HCl. Gas chromatography‐mass spectrometry analysis showed that the hydrolyte in the hydrophobic fraction contained five kinds of FA with chain lengths of C14–C19, and C16H32O2 was a major component making up 59·18% of the total. However, 3‐hydroxyl FA was not found, although it is usually found in lipopeptides. Silica gel TLC revealed that the hydrolyte in the hydrophilic fraction was composed of five kinds of amino acids; consistently, ESI‐Q‐TOF‐MS analysis confirmed the composition results and provided their sequence tentatively as Ala‐Ile‐Asp‐Met‐Pro. Furthermore, the yield and CMC (critical micelle concentrations) of purified lipopeptides were examined. The purified product reduced the surface tension of water to 30·7 mN m?1 with a CMC value of 23·7 mg l?1. These results suggest that Rhodococcus sp. TW53 produces a novel lipopeptide that we have named rhodofactin. Conclusion: The deep‐sea isolate Rhodococcus sp. TW53 was the first reported lipopeptide‐producing bacterium of this genus. The lipopeptides had novel chemical compositions. Significance and Impact of the Study: Rhodococcus sp. TW53 has potential in the exploration of new biosurfactants and could be used in bioremediation of marine oil pollution.  相似文献   

18.
The potential of arsenic-resistant bacteria in association with Pteris vittata to reduce the level of arsenic from soil was studied. The physicochemical characteristics of contaminated paddy soil were analyzed, and 3 bacterial isolates amongst 11 were screened and were selected for further study. These three isolates were characterized by 16S rDNA sequencing and identified as Bacillus altitudinis Strain SS8 (KJ432582), Bacillus megaterium Strain SS9 (KJ432583) and Lysinibacillus sp. Strain SS11 (KJ432584). Of these, Lysinibacillus sp. Strain SS11 displayed arsenic tolerance of 3256 mg L?1 for arsenate and 1136 mg L?1 for arsenite. Additionally, it showed bioaccumulation capacity of 23.43 mg L?1 for arsenate and 5.65 mg L?1 for arsenite. It also showed resistance to other heavy metals, especially towards iron, copper and chromium. It was also observed that Pteris vittata was able to take up more arsenic and iron from soil in the presence of these bacterial strains than in their absence, leading to contaminant-free soil. Thus, this system appears to be an effective bioremediating process to remove arsenic from contaminated soil.  相似文献   

19.
Abstract

Biology, tolerance, and metal (Pb and Cr) accumulating ability of Gomphrena celosoides were studied under hydroponic conditions. The seedlings were raised in Hoagland’s solution containing different concentrations of Pb (0, 500, 1000, 1500, 2000, 3000, 4000, and 5000?mg l?1) and Cr (0, 50, 100, 150, 200, 300, and 400?mg l?1). Biomass and metal accumulation in different plant parts were determined at seven (7) and fourteen (14) days after stress. Antioxidant enzyme activities, protein, and proline contents were estimated in stressed and unstressed plants. Gomphrena celosoides was able to tolerate Pb and Cr concentrations up to 4000 and 100?mg l?1, respectively in hydroponic solution. Metal accumulation was concentration and duration dependent with the highest Pb (21,127.90 and 117,985.29?mg kg?1) and Cr (3130.85 and 2428.90?mg kg?1) in shoot and root, respectively found in the plants exposed to 5000?mg l?1 Pb and 400?mg l?1 Cr for 14?days. Proline, antioxidant enzyme activities, and protein contents were the highest in plant exposed to higher Pb and Cr concentrations for 7 and 14?days. Gomphrena celosoides could be considered as Pb and Cr accumulator with proline and increase in antioxidant enzyme activities being the tolerance mechanisms.  相似文献   

20.
《Process Biochemistry》2014,49(10):1699-1707
This study reports the potential of a marine bacterium, Bacillus mojavensis A21, to produce lipopeptide biosurfactants. The crude lipopeptide mixture was found to be very effective in reducing surface tension to 31 mN m−1. PCR experiments using degenerate primers revealed the presence of nonribosomal peptide synthetases genes implied in the biosyntheses of fengycin and surfactin. Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) performed on whole cells of B. mojavensis A21 confirmed the presence of lipopeptides identified as members of surfactin and fengycin families. Further, a detailed analysis performed by MALDI-TOF-TOF revealed the presence of pumilacidin compounds. The crude lipopeptide mixture was tested for its inhibitory activity against Gram-positive and Gram-negative bacteria, and fungal strains. It was found to display significant antimicrobial activity. Strain A21 lipopeptide mixture was insensitive to proteolytic enzymes, stable between pH 3.0 and 11.0, and resistant to high temperature. Production of lipopeptides is a characteristic of several Bacillus species, but to our knowledge this is the first report involving identification of pumilacidin, surfactin and fengycin isoforms in a B. mojavensis strain.  相似文献   

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