Screening a Strain of Klebsiella sp. O852 and the Optimization of Fermentation Conditions for Trans-Dihydrocarvone Production.
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ABSTRACT:
SUBMITTER: Chen L
PROVIDER: S-EPMC8122266 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:BackgroundHyaluronic acid (HA) has gained significant attention due to its unique physical, chemical, and biological properties, making it widely used in various industries. This study aimed to screen bacterial isolates for HA production, characterize favorable fermentation conditions, and evaluate the inhibitory effect of bacterial HA on cancer cell lines.ResultsA total of 108 bacterial isolates from diverse sources were screened for HA production using HPLC, turbidimetric, and carbazole determination methods. Among the HA-producing isolates, Klebsiella pneumoniae H15 isolated from an animal feces sample, was superior in HA production. The strain was characterized based on its morphological, cultural, and biochemical characteristics. Molecular identification using 16S rDNA sequencing and phylogenetic analysis confirmed its identity. Fermentation conditions, including pH, temperature, time, and agitation rate, were optimized to maximize HA production. The basal medium, comprising sucrose (7.0%) as carbon source and combined yeast extract with peptone (1.25% each) as nitrogen substrate, favored the highest HA production at pH 8.0, for 30 h, at 30 °C, under shaking at 180 rpm. The average maximized HA concentration reached 1.5 g L-1. Furthermore, bacterial HA exhibited a significant inhibitory effect on three cancer cell lines (MCF-7, HepG-2 and HCT), with the lowest concentration ranging from 0.98-3.91 µg mL-1.ConclusionsK. pneumoniae H15, isolated from animal feces demonstrated promising potential for HA production. The most favorable fermentation conditions led to a high HA production. The inhibitory effect of bacterial HA on cancer cell lines highlights its potential therapeutic applications. These findings contribute to a broader understanding and utilization of HA in various industries and therapeutic applications.
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Project description:Aflatoxin B?, a type of highly toxic mycotoxin produced by some species belonging to the Aspergillus genus, such as Aspergillus flavus and Aspergillus parasiticus, is widely distributed in feed matrices. Here, coumarin was used as the sole carbon source to screen microorganism strains that were isolated from types of feed ingredients. Only one isolate (ND-1) was able to degrade aflatoxin B? after screening. ND-1 isolate, identified as a strain of Aspergillus niger using phylogenetic analysis on the basis of 18S rDNA, could remove 26.3% of aflatoxin B? after 48 h of fermentation in nutrient broth (NB). Optimization of fermentation conditions for aflatoxin B? degradation by selected Aspergillus niger was also performed. These results showed that 58.2% of aflatoxin B? was degraded after 24 h of culture under the optimal fermentation conditions. The aflatoxin B? degradation activity of Aspergillus niger supernatant was significantly stronger than cells and cell extracts. Furthermore, effects of temperature, heat treatment, pH, and metal ions on aflatoxin B? degradation by the supernatant were examined. Results indicated that aflatoxin B? degradation of Aspergillus niger is enzymatic and this process occurs in the extracellular environment.
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Project description:The novel exopolysaccharide HZ-7 is produced by Klebsiella sp. H-207, and its fermentation conditions were optimized by response surface methodology (RSM). In this study, the optimized medium consisted of sucrose 31.93 g/L, KNO(3) 2.17 g/L and K(2)HPO(4) 5.47 g/L; while the optimized culture conditions consisted of seed age 13 h, with an inoculum size of 10.6% and incubation temperature of 28.9°C. A maximum HZ-7 yield of about 15.05 g/L was achieved under the optimized conditions using RSM and single-factor experiments. Next the exopolysaccharide HZ-7 was partially purified and characterized. The resulting product showed good properties, such as high concentration of uronic acid (41.67%), low average molecular weight (about 1.94×10(5) Da) and porous surface structure, were very advantageous to biosorption. Therefore HZ-7 was applied to absorb hexavalent chromium (Cr(VI)). The maximum adsorption efficiency (99.2%) which was obtained at an initial pH of 1.0 along with an initial Cr(VI) concentration of 20 mg/L, was not affected by ordinary metal ions and temperature. These data suggest Klebsiella sp. H-207 exopolysaccharide will be promising potential for industrial application.
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Project description:Efficiency of cellulase-free xylanases is one of the determining factors in paper and pulp industries. Use of microbes which can produce cellulase-free xylanases may help to overcome the current challenges in kraft pulp processing. Isolation and screening of microorganisms from local samples offers a possibility for obtaining the potential microbes for this purpose. This research was therefore aimed to collect, screen, characterize and identify potential cellulase-free xylanase producers. A total of 313 microbial isolates were collected while using selective media (EBAM and XAM) to determine the xylanolytic potential of microbes. Qualitative and quantitative analyses were performed and finally 11 bacterial and 6 fungal strains were selected for characterization and identification. The potential isolates were identified as Bacillus pumilus (388.82 U/mg), Bacillus safensis (385.26 U/mg), Aspergillus flavus (493.33 U/mg) and Aspergillus niger (419.33 U/mg). Optimization of the microbial strains while using agro-industrial waste is suggested.
| S-EPMC4909030 | biostudies-literature
Project description:BackgroundThe production of indole-3-acetic acid (IAA) is an essential tool for rhizobacteria to stimulate and facilitate plant growth. For this, eighty rhizobial bacteria isolated from root nodules of Acacia cyanophylla grown in different regions of Morocco were firstly screened for their ability to produce IAA. Then, IAA production by a combination of isolates and the inoculation effect on the germination of Acacia cyanophylla seeds was studied using the best performing isolates in terms of IAA production. The best IAA producer bacterial isolate (I69) was selected to optimize IAA production using response surface methodology based on the central composite design.ResultsResults showed that the majority of tested isolates were able to produce IAA with a relatively higher concentration of 135 μg/ml for the isolate I69, followed by isolates I22 and I75 with respective concentrations of 116 μg/ml and 105 μg/ml IAA. The IAA production and the seed germination rate were relatively increased by the synergistic effect of I69 and I22. Later, response surface methodology was used to determine optimal operating conditions leading to IAA production optimization. Thus, an incubation temperature of 36 °C, a pH of 6.5, an incubation time of 1 day, and respective tryptophan and NaCl concentrations of 1 g/l and 0.1 g/l were optimal parameters leading to 166 μg/ml IAA which was the maximal produced concentration.ConclusionThe present study highlighted that IAA-producing rhizobacteria could be harnessed to improve plant growth. Furthermore, their production can be easily controlled using response surface methodology, which represents a very useful tool for optimization.
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Project description:Streptomyces species 1-14 isolated from cassava rhizosphere soil were evaluated for their antibacterial efficacy against Fusarium oxysporum f.sp. cubense race 4 (FOC4). Of the 63 strains tested, thirteen exhibited potent antibacterial properties and were further screened against eight fungal pathogens. The strain that showed maximum inhibition against all of the test pathogens was identified by 16S rDNA sequencing as Streptomyces sp. 1-14, was selected for further studies. Through the propagation of Streptomyces sp. 1-14 in soil under simulated conditions, we found that FOC4 did not significantly influence the multiplication and survival of Streptomyces sp. 1-14, while indigenous microorganisms in the soil did significantly influence Streptomyces sp. 1-14 populations. To achieve maximum metabolite production, the growth of Streptomyces 1-14 was optimized through response surface methodology employing Plackett-Burman design, path of steepest ascent determinations and Box-Behnken design. The final optimized fermentation conditions (g/L) included: glucose, 38.877; CaCl2•2H2O, 0.161; temperature, 29.97°C; and inoculation amount, 8.93%. This optimization resulted in an antibacterial activity of 56.13% against FOC4, which was 12.33% higher than that before optimization (43.80%). The results obtained using response surface methodology to optimize the fermentation medium had a significant effect on the production of bioactive metabolites by Streptomyces sp. 1-14. Moreover, during fermentation and storage, pH, light, storage temperature, etc., must be closely monitored to reduce the formation of fermentation products with reduced antibacterial activity. This method is useful for further investigations of the production of anti-FOC4 substances, and could be used to develop bio-control agents to suppress or control banana fusarium wilt.
| S-EPMC6241123 | biostudies-literature
Project description:Effective degradation of ?-carrageenan by isolated Thalassospira sp. fjfst-332 is reported for the first time in this paper. It was identified by 16S rDNA sequencing and morphological observation using Transmission Electron Microscopy (TEM). Based on a Plackett-Burman design for significant variables, Box-Behnken experimental design and response surface methodology were used to optimize the culture conditions. Through statistical optimization, the optimum medium components were determined as follows: 2.0 g/L ?-carrageenan, 1.0 g/L yeast extract, 1.0 g/L FOS, 20.0 g/L NaCl, 2.0 g/L NaNO?, 0.5 g/L MgSO?·7H?O, 0.1 g/L K?HPO?, and 0.1 g/L CaCl?. The highest activity exhibited by Thalassospira sp. fjfst-332 was 267 U/mL, which makes it the most vigorous wild bacterium for ?-carrageenan production. In order to guide scaled-up production, two empirical models-the logistic equation and Luedeking-Piretequation-were proposed to predict the strain growth and enzyme production, respectively. Furthermore, we report the fermentation kinetics and every empirical equation of the coefficients (?, ?, X?, Xm and ?m) for the two models, which could be used to design and optimize industrial processes.
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Project description:Curcumin is a plant secondary metabolite with outstanding therapeutic effects. Therefore, there is a great interest in developing new strategies to produce this high-value compound in a cheaper and environmentally friendly way. Curcumin heterologous production in Escherichia coli using artificial biosynthetic pathways was previously demonstrated using synthetic biology approaches. However, the culturing conditions to produce this compound were not optimized and so far only a two-step fermentation process involving the exchange of culture medium allowed high concentrations of curcumin to be obtained, which limits its production at an industrial scale. In this study, the culturing conditions to produce curcumin were evaluated and optimized. In addition, it was concluded that E. coli BL21 allows higher concentrations of curcumin to be produced than E. coli K-12 strains. Different isopropyl ?-d-thiogalactopyranoside concentrations, time of protein expression induction and substrate type and concentration were also evaluated. The highest curcumin production obtained was 959.3 µM (95.93% of per cent yield), which was 3.1-fold higher than the highest concentration previously reported. This concentration was obtained using a two-stage fermentation with lysogeny broth (LB) and M9. Moreover, terrific broth was also demonstrated to be a very interesting alternative medium to produce curcumin because it also led to high concentrations (817.7 µM). The use of this single fermentation medium represents an advantage at industrial scale and, although the final production is lower than that obtained with the LB-M9 combination, it leads to a significantly higher production of curcumin in the first 24 h of fermentation. This study allowed obtaining the highest concentrations of curcumin reported so far in a heterologous organism and is of interest for all of those working with the heterologous production of curcuminoids, other complex polyphenolic compounds or plant secondary metabolites.
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Project description:Rice-fish coculture (RF) is a small ecosystem in which microorganisms are widely distributed in the fish, water environment, soil, and plants. In order to study the positive effects of microorganisms on common carp and rice in the RF ecosystem, a total of 18 strains with growth-promoting ability were screened from common carp (Cyprinus carpio) gut contents, among which three strains had the ability to produce both DDP-IV inhibitors and IAA. The strain with the strongest combined ability, FYN-22, was identified physiologically, biochemically, and by 16S rRNA, and it was initially identified as Bacillus licheniformis. As the number of metabolites secreted by the strain under natural conditions is not sufficient for production, the FYN-22 fermentation medium formulation was optimized by means of one-factor-at-a-time (OFAT) experiments and response surface methodology (RSM). The results showed that, under the conditions of a soluble starch concentration of 10.961 g/l, yeast concentration of 2.366 g/l, NH4Cl concentration of 1.881 g/l, and FeCl3 concentration of 0.850 g/l, the actual measured number of FYN-22 spores in the fermentation broth was 1.913 × 109 CFU/ml, which was 2.575-fold improvement over the pre-optimization value. The optimized fermentation solution was used for the immersion operation of rice seeds, and, after 14 days of incubation in hydroponic boxes, the FYN-22 strain was found to have a highly significant enhancement of 48.31% (p < 0.01) on the above-ground part of rice, and different degrees of effect on root length, fresh weight, and dry weight (16.73, 17.80, and 21.97%, respectively; p < 0.05). This study may provide new insights into the fermentation process of Bacillus licheniformis FYN-22 and its further utilization in RF systems.
| S-EPMC9802155 | biostudies-literature
Project description:The present study identified the pectinase-producing bacterium isolated from the contaminated broth as Bacillus sp. on 16S rDNA sequence analysis. The bacterium illustrated water-like droplets on the colony grown on the Sabouraud dextrose agar plate. It also exhibited multi-enzymes activities, such as pectinase, polygalacturonase, xylanase, and cellulase by using various agro-wastes as low-cost substrates. The orange peel was observed to be the best substrate among the agro-wastes used for maximum multi-enzymes (pectinase, polygalacturonase, xylanase, and cellulase). However, the bacterium demonstrated its capability to produce different enzymes according to the different substrates/agro-wastes used. The Plackett-Burman design was used to determine the essential influencing factors, while the Box Behnken design response surface methodology was for optimizing cultural conditions. At their optimal conditions (40°C incubation temperature, 24 h of incubation period, 1% w/v orange peel, and 2% v/v inoculum volume), the bacterium exhibited the maximum pectinase (9.49 ± 1.25 U/ml) and xylanase (16.27 ± 0.52 U/ml) activities. Furthermore, the study explored the ability of the bacterium to produce bacterial lipids and observed about 25% bacterial lipid content on a dry weight basis. Therefore, the bacterium is a good candidate for producing important multi-enzymes and subsequent agro-waste degradation controlling the environment, and facilitating waste management. Also, the bacterium can be a potential feedstock in producing renewable biofuel.
| S-EPMC9663924 | biostudies-literature