An extracellular lipase-producing fungus was isolated from oil-rich soil. This fungus belongs to the genus Rhizopus and clades with Rhizopus oryzae. Lipase was purified to homogeneity from this novel fungal source using ammonium sulphate precipitation followed by Q-Sepharose
chromatography. The extracellular lipase was purified 8.6–fold, and enzymatic properties were studied. The molecular mass
of the purified enzyme was estimated to be 17 kD by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and 16.25 kD
by matrix-assisted laser desorption ionization/time-of-flight analysis. The native molecular mass was estimated to be 17.5
kD by gel filtration, indicating the protein to be monomer. The optimum pH and temperature for the enzyme catalysis were 7.0 °C
and 40 °C, respectively. Enzyme was stable in pH range 6.0–7.0 and retains 95–100% activity when incubated at 50 °C for 1 h.
The pI of the purified lipase was 4.2. Enzyme was stable in the organic solvents such as ethanol, hexane and methanol for
2 h. Purified enzyme was used for transesterification of oleic acid in the presence of ethanol for production of oleic acid
ethyl ester with a conversion efficiency of 66% after 24 h at 30 °C. 相似文献
Taxol is the most important member of the clinically useful natural anticancer drug. An endophytic fungus Chaetomella raphigera (strain TAC-15) was isolated from a medicinal plant Terminalia arjuna and screened for its potential in Taxol production. The fungus was identified based on the morphology of the fungal culture
and the characteristics of the spores. This fungus was grown in MID liquid medium and analyzed by chromatographically and
spectrometrically for the presence of Taxol. The amount of Taxol produced by this endophytic fungus was quantified by HPLC
which showed that it produced 79.6 μg/L, and further confirmative analyses were done by using UV, IR, FAB mass spectroscopy,
and NMR spectroscopy. Thus, the fungus can serve as a potential material for fungus engineering to improve the production
of Taxol. 相似文献
Algae in drinking water supplies often bring about impact on the water treatment. In this study, a bipolar pulsed dielectric
barrier discharge system in three-phase discharge plasma reactor was constructed for investigating its ability to control
excessive growth of cyanobacteria, Microcystis aeruginosa. Experimental results show there was almost no change in optical density immediately after the interruption of electrical
discharge, but the decreasing trend of optical density, cell density and chlorophyll-a content was obvious during the incubation period, indicating a significant residual effect of electrical discharge process
on the algal growth inhibition. Scanning electron microscopy investigation of algae revealed surface damage, apparent leakage
of intracellular contents and pores formed after electrical discharge process, which showed that algal cells had no potential
to survive and grow. Compared with the control sample, it was observed that electrical discharge on the algal extracellular
products has almost no residual effect and the growth rate of algae slightly decreased before three days storage. Hydrogen
peroxide was produced by electrical discharge in the μM level and showed a first-order decay. But at such level, the external
addition of hydrogen peroxide alone is not likely to cause the residual effect. These results implicated that the growth inhibition
of M. aeruginosa was mainly caused by electrical discharge process, and it made the algal cells lose ability to survive, demonstrating the
considerable potential of such an alternative process for efficient water purification. 相似文献
In the present frame of work, Macrophomina phaseolina is encapsulated in silica matrices at various concentrations by low temperature sol-gel technique using tetraethylorthosilicate
(TEOS) as precursor. The optical and photophysical properties of these samples have been studied by second harmonics of Nd:YAG
laser at 532 nm. UV-visible absorption spectra of samples have been recorded and it is found that the absorption increases
with increase in concentration of fungus. Further, a decrease in output transmission intensity of the laser has been observed
with increase in fungus concentration. The temporal response of these samples has also been examined. The results show that
the fungus concentration can be measured within ∼15–20 min. This method of optical sensing of fungus in test sample is faster
than other techniques, such as the conventional colorimetric method which takes about 1 h. 相似文献
An analytical procedure based on matrix solid-phase dispersion (MSPD) and liquid chromatography–mass spectrometry (LC-MS) was developed for determining three microcystins (MCs) in natural water blooms and cyanobacteria strain cultures. The procedure involves sample homogenization with C18, washed with dichloromethane to eliminate interfering compounds, and elution with acidic methanol. Results were compared to those achieved by using an organic solvent standard method. Mean recoveries of MCs with MSPD were 85–92% with intra-day relative standard deviation (RSDs) of 9–19%, whereas organic solvent extraction resulted in recovery rates of 92–105% with intra-day RSDs ranging from 8 to 18%. Limits of quantification (LOQs) were 1 g g–1 dry weight for the MCs either by MSPD or organic solvent extraction. The two analytical methods tested were specific and sensitive to the extraction of MCs and were applied to the detection of MCs in water blooms and culture strains. The concentration of MCs varied from 7 to 3,330 g g–1 of lyophilized cells with MC-LR always showing the highest concentration. MCs levels were higher in culture strains than in water blooms, except for MC-LR, whose concentration in blooms was slightly superior to that determined in culture strains. 相似文献
Single cell-inductively coupled plasma-mass spectrometry (SC-ICP-MS) is an emerging technology. In this work, we have developed a novel SC-ICP-MS method to quantify metal ions in individual cells of a toxic cyanobacterial species, Microcystis aeruginosa (M. aeruginosa), without complicated post-dosing sample preparation, and applied this method to study the treatment effectiveness of copper-based algaecides (cupric sulfate and EarthTec®) on the toxic algae M. aeruginosa. The developed SC-ICP-MS method uses new intrinsic metal element magnesium to determine real transport efficiency and cell concentration. The cell viability and microcystin-LR release by algaecide treatment were studied by flow cytometry and ultra-fast liquid chromatography-tandem mass spectrometry, respectively. The results showed that this novel method was very rapid, highly sensitive (detection limits of intracellular copper and magnesium were 65 ag/cell and 98 ag/cell, respectively), and reproducible (relative standard deviation within 12%). The algaecide effectiveness study further demonstrated that copper in the forms of cupric sulfate and copper-based algaecide EarthTec® successfully diminished M. aeruginosa populations. The higher the copper concentration used to treat the cells, the faster the speeds of copper uptake and cell lysis in the copper concentrations ranged from 0 to 200 μg/L of copper-based algaecide. The cells exhibit obvious heterogeneity in copper uptake. The result suggests that M. aeruginosa cells uptake and cumulate copper followed by cellular lysis and microcystin-LR release. These novel results indicated that though the copper-based algaecides could control this type of harmful algal bloom, further treatment to remove the released algal toxin from the treated water would be needed.
When cells are exposed to high salinity conditions, they develop a mechanism to extrude excess Na+ from cells to maintain the cytoplasmic Na+ concentration. Until now, the ATPase involved in Na+ transport in cyanobacteria has not been characterized. Here, the characterization of ATPase and its role in Na+ transport of alkaliphilic halotolerant Aphanothece halophytica were investigated to understand the survival mechanism of A. halophytica under high salinity conditions. 相似文献
A gene encoding Yarrowia lipolytica lipase LIP2 (YlLIP2) was cloned into a constitutive expression vector pGAPZαA and electrotransformed into the Pichia pastoris X-33 strain. The high-yield clones obtained by high copy and enzyme activity screening were chosen as the host strains for
shaking flask and fermentor culture. The results showed that glucose was the optimum carbon source for YlLIP2 production,
and the maximum hydrolytic activity of recombinant YlLIP2 reached 1,315 U/ml under the flask culture at 28 °C, pH 7.0, for
48 h. The fed-batch fermentation was carried out in 3- and 10-l bioreactors by continuously feeding glucose into the growing
medium for achieving high cell density and YlLIP2 yields. The maximum hydrolytic activity of YlLIP2 and cell density obtained
in the 3-l bioreactor were 10,300 U/ml and 116 g dry cell weight (DCW)/l, respectively. The peak hydrolytic activity of YlLIP2
and cell density were further improved in the 10-l fermentor where the values respectively attained were 13,500 U/ml and 120 g
DCW/l. The total protein concentration in the supernatant reached 3.3 g/l and the cell viability remained approximately 99%
after 80 h of culture. Furthermore, the recombinant YlLIP2 produced in P. pastoris pGAP and pAOX1 systems have similar content of sugar (about 12%) and biochemical characteristics. The above results suggest
that the GAP promoter-derived expression system of P. pastoris is effective for the expression of YlLIP2 by high cell density culture and is probably an alternative to the conventional
AOX1 promoter expression system in large-scale production of industrial lipases. 相似文献
The uranium(VI) accumulation was studied in detail by using the biomass of mangrove endophytic fungus Fusarium sp.#ZZF51 from the South China Sea. The uranium(VI) biosorption process onto the tested fungus powders was optimized at pH 4.0,
adsorption time 60 min, and uranium(VI) initial concentration 50 mg L−1 with 61.89% of removal efficiency. According to Fourier transform infrared spectra for the tested fungus before and after
loaded with uranium(VI), the results showed that both of hydroxyl and carboxyl groups acted as the important roles in the
adsorption process. In addition, the experimental data were analyzed by using parameter and kinetic models, and it was obtained
that the Langmuir isotherm model and the pseudo-second-order kinetic model provided better correlation with the experimental
data for adsorption of uranium(VI). 相似文献
Two new secoanthraquinone derivatives, wentiquinones A ( 1 ) and B ( 2 ), together with the eight known compounds 3 – 10 , were isolated from the culture extracts of Aspergillus wentii EN‐48, an endophytic fungus derived from an unidentified marine brown algal species of the genus Sargassum. The structures of these compounds were elucidated on the basis of extensive spectroscopic analysis. 相似文献
Rising global temperatures have been suggested to favor cyanobacteria over eukaryotic algae, but UV-B fluxes are also predicted to remain high and may interact with temperature to affect algal growth. To understand the interactive effects of temperature and UV-B radiation, cultures of Microcystis aeruginosa and Anabaena circinalis were grown at either 25 or 30°C and then exposed to an acute irradiance of UV-B (1.4 W m−2). Both species showed differences in growth rates at both temperature regimes. The growth rates of M. aeruginosa (0.41 ± 0.02 day−1) and A. circinalis (0.38 ± 0.01 day−1) were higher at 25 and 30°C, respectively. Rates of damage (k) and repair (r) were calculated from the kinetics of change in effective quantum yield, Fv'/Fm'. Analysis of the estimates of r and k shows that M. aeruginosa exhibited relatively high values for both parameters, compared to A. circinalis, at both growth temperatures. In both species, repair rates were higher at 30°C than at 25°C but in A. circinalis damage was also greater at the higher temperature. In contrast, M. aeruginosa showed a lower damage rate at the higher temperature. For both species, the ratio of r:k was higher at the higher temperature. However, the percent inhibition of effective quantum yield by UV-B was greater in A. circinalis than in M. aeruginosa as the r:k was lower A. circinalis. Therefore, it could be concluded that temperature may influence growth and bloom formation of cyanobacteria and that different species may respond differently to UV-B and temperature interactions. 相似文献
One of the compounds suggested to be responsible for the cyanobacterial dominance over competing green algae is identified. Evidence is provided on the molecular, chemical level that the iron chelator anachelin from the cyanobacterium Anabaena cylindrica promotes both the growth of cyanobacteria and reduces the growth of competing chlorophytes. These results illustrate a molecular strategy of addressing two challenges (nutrient availability and algal competition) by one molecule. Such strategies could be implied in harmful algal blooms in marine and freshwater environments. 相似文献
The objective of this research was to develop large-scale technologies to produce oil-rich algal biomass from wastewater.
The experiments were conducted using Erlenmeyer flasks and biocoil photobioreactor. Chlamydomonas reinhardtii was grown in artificial media and wastewaters taken from three different stages of the treatment process, namely, influent,
effluent, and centrate. Each of wastewaters contained different levels of nutrients. The specific growth rate of C. reinhardtii in different cultures was monitored over a period of 10 days. The biomass yield of microalgae and associated nitrogen and
phosphorous removal were evaluated. Effects of CO2 and pH on the growth were also studied. The level of nutrients greatly influenced algae growth. High levels of nutrients
seem to inhibit algae growth in the beginning, but provided sustained growth to a high degree. The studies have shown that
the optimal pH for C. reinhardtii is in the range of 7.5. An injection of air and a moderate amount of CO2 promoted algae growth. However, too much CO2 inhibited algae growth due to a significant decrease in pH. The experimental results showed that algal dry biomass yield
reached a maximum of 2.0 g L−1 day−1 in the biocoil. The oil content of microalgae of C. reinhardtii was 25.25% (w/w) in dry biomass weight. In the biocoil, 55.8 mg nitrogen and 17.4 mg phosphorus per liter per day were effectively removed
from the centrate wastewater. Ferric chloride was found to be an effective flocculent that helps the algae settle for easy
harvest and separation from the culture media. 相似文献
Since the recombinant thyroid-stimulating hormone (rhTSH) is secreted by stably transfected Chinese hamster ovary (CHO-hTSH)
cells, a bioprocess consisting of immobilizing the cells on a substrate allowing their multiplication is very suitable for
rhTSH recovering from supernatants at relative high degree of purity. In addition, such a system has also the advantage of
easily allowing delicate manipulations of culture medium replacement. In the present study, we show the development of a laboratory
scale bioprocess protocol of CHO-hTSH cell cultures on cytodex microcarriers (MCs) in a 1 L bioreactor, for the preparation
of rhTSH batches in view of structure/function studies. CHO-hTSH cells were cultivated on a fetal bovine serum supplemented
medium during cell growth phase. For rhTSH synthesis phase, 75% of supernatant was replaced by animal protein-free medium
every 24 h. Cell cultures were monitored for agitation (rpm), temperature (°C), dissolved oxygen (% DO), pH, cell concentration,
MCs coverage, glucose consumption, lactate production, and rhTSH expression. The results indicate that the amount of MCs in
the culture and the cell concentration at the beginning of rhTSH synthesis phase were crucial parameters for improving the
final rhTSH production. By cultivating the CHO-hTSH cells with an initial cell seeding of four cells/MC on 4 g/L of MCs with
a repeated fed batch mode of operation at 40 rpm, 37 °C, 20% DO, and pH 7.2 and starting the rhTSH synthesis phase with 3 × 106 cells/mL, we were able to supply the cultures with enough glucose, to maintain low levels of lactate, and to provide high
percent (∼80%) of fully covered MCs for a long period (5 days) and attain a high cell concentration (∼9 × 105 cells/mL). The novelty of the present study is represented by the establishment of cell culture conditions allowing us to
produce ∼1.6 mg/L of rhTSH in an already suitable degree of purity. Batches of produced rhTSH were purified and showed biological
activity. 相似文献
Bi-doped nano-crystalline TiO2 (Bi–TiO2) has been synthesized by sonocrystallization at low temperature. The Bi–TiO2 materials have narrower bandgaps than pristine TiO2, which endow them with significant visible light absorption. Accordingly, these materials had enhanced photocatalytic activity
in the degradation of organic dye pollutants and the cyanotoxin microcystin-LR (MC-LR) under visible irradiation. It was found
that degradation of MC-LR is rather efficient. After irradiation with visible light for 12 h the original MC-LR was removed
completely, and 78% of the organic carbon was mineralized into CO2 after irradiation for 20 h. The hydroxyl radical (·OH) is the major active species responsible for the degradation reaction.
Identified intermediates primarily originate from attack of ·OH radicals on the double bonds between C4 and C5 (C6 and C7)
of Adda and the ethylenic bond of Mdha in MC-LR. Some peptide bonds are also broken with longer irradiation time. 相似文献
To obtain a high level expression of phytase with favorable characteristics, a codon-optimized phytase gene from Citrobacter freundii was synthesized and transferred into Pichia pastoris. Small-scale expression experiments and activity assays were used to screen positive colonies. After purified by Ni2+–NTA agarose affinity column, the characterizations of the recombinant phytase were determined. The recombinant phytase (r-phyC)
had two distinct pH optima at 2.5 and 4.5 and an optimal temperature at 50 °C. It retained more than 80% activity after being
incubated under various buffer (pH 1.5–8.0) at 37 °C for 1 h. The specific activity, Km, and Vmax values of r-phyC for sodium phytate were 2,072 ± 18 U mg−1, 0.52 ± 0.04 mM, and 2,380 ± 84 U mg−1 min−1, respectively. The enzyme activity was significantly improved by 1 mM of K+, Ca2+, and Mg2+. These characteristics contribute to its potential application in feed industry. 相似文献
A surface plasmon resonance (SPR) biosensor for the detection of microcystins (MCs) in drinking water has been developed.
Several assay formats have been evaluated. The selected format is based on a competitive inhibition assay, in which microcystin-LR
(MCLR) has been covalently immobilized onto the surface of an SPR chip functionalized with a self-assembled monolayer. The
influence of several factors affecting sensor performance, such as the nature and concentration of the antibody, the composition
of the carrier buffer, and the blocking and regeneration solutions, has been evaluated. The optimized SPR biosensor provides
an IC50 0.67 ± 0.09 μg L−1, a detection limit of 73 ± 8 ng L−1, and a dynamic range from 0.2 to 2.0 μg L−1 for MCLR. Cross-reactivity to other related MCs, such as microcystin-RR (88%) and microcystin-YR (94%), has also been measured.
The SPR biosensor can perform four simultaneous determinations in 60 min, and each SPR chip can be reused for at least 40
assay–regeneration cycles without significant binding capacity loss. The biosensor has been successfully applied to the direct
analysis of MCLR in drinking water samples, below the provisional guideline value of 1 μg L−1 established by the World Health Organization for drinking water. 相似文献