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1.
The regenerating activities of chemically modified anionic bacterial polysaccharides by O-sulfonation were investigated using a in vivo model of rat injured muscle regeneration. Glucuronan (GA), a linear homopolysaccharide of -->4)-beta-D-GlcpA-(1--> residues partially acetylated at the C-3 and/or the C-2 position, and glucoglucuronan (GGA), a linear heteropolysaccharide of -->3)-beta-D-GlcpA-(1-->4)-beta-D-Glcp-(1--> residues were sulfated. SO3-DMF sulfatation complex provided polysaccharides with different sulfur contents, however, a depolymerization occurred because we did not use large excess of pyridine to obtain pure modified polysaccharides. A regenerating activity on injured extensor digitorum longus (EDL) muscles on rats was obtained with these two sulfated anionic polymers. The position of sulfate groups on glucoglucuronan (primary or secondary alcohol) seems to have no influence on the biological activity by opposition to the degree of sulfatation both for the glucuronans and the glucoglucuronans. The yield of acetate groups in the glucuronan polymer modulated the specific activity.  相似文献   

2.
The production of pectinase by Aspergillus niger LB-02-SF was focused on a submerged cultivation, before it was evaluated in a solid-state process. This study involved the creation of a defined culture medium and an evaluation of the effects of the addition of the enzyme inducer, citrus pectin, to the medium after the intense biomass growth phase. A culture medium formulated without glucose allowed a reduction of biomass growth and greater pectinase production, facilitated by the control of process parameters such as mixing, pH and oxygen supply. The addition of pectin when a minimum pH of 2.7 was reached at 22 h of cultivation did not affect fungal growth. The maximum biomass concentration was 11.0 g/L at 48 h, a value similar to that observed for the control, in which pectin was included in the medium at the beginning of the process (11.5 g/L, at 41 h). However, this condition favored the production of 14 U/mL pectinase, which was approximately 40% higher than the value observed for the control. These results show that pectinase production by A. niger in a submerged cultivation is strongly affected by the medium composition as well as the delayed addition of pectin to the fermentation broth.  相似文献   

3.
Haloarchaeal strains require high concentrations of NaCl for their growth, with optimum concentrations of 10–30%. They display a wide variety of morphology and physiology including pH range for growth. Many strains grow at neutral to slightly alkaline pH, and some only at alkaline pH. However, no strain has been reported to grow only in acidic pH conditions within the family Halobacteriaceae. In this study, we isolated many halophiles capable of growth in a 20% NaCl medium adjusted to pH 4.5 from 28 commercially available salts. They showed growth at pH 4.0 to 6.5, depending slightly on the magnesium content. The most acidophilic strain MH1-52-1 isolated from an imported solar salt (pH of saturated solution was 9.0) was non-pigmented and extremely halophilic. It was only capable of growing at pH 4.2–4.8 with an optimum at pH 4.4 in a medium with 0.1% magnesium chloride, and at pH 4.0–6.0 (optimum at pH 4.0) in a medium with 5.0% magnesium. The 16S rRNA and DNA-dependent RNA polymerase subunit B' gene sequences demonstrated clearly that the strain MH1-52-1 represents a new genus in the family Halobacteriaceae.  相似文献   

4.
《Process Biochemistry》2010,45(9):1563-1569
A soil-isolated bacterium (strain B4) was identified as a species of Bordetella and deposited with the China General Microbiological Culture Collection (code, CGMCC 2229). The bacterium grew in a mineral medium, on cholesterol as a sole source of carbon and energy. Only one metabolite of cholesterol was accumulated in detectable amounts during the strain growth. It was identified as 4-cholesten-3-one. Cholesterol oxidase (COD) (EC 1.1.3.6), which catalyzes cholesterol into this metabolite, was evidenced from the strain. The conditions of the bacterium growth were optimized for extracellular enzyme production, which then reached around 1700 UL−1 within 24 h culturing. The enzyme was purified from the spent medium of the strain to homogeneity on SDS-PAGE, and characterized. Its molecular mass, as estimated by this technique, was 55 kDa. COD showed an optimum activity at pH 7.0. It was completely stable at pH 5.0 and 4 °C for 48 h, and retained 80% at least of its initial activity at pH 4.0 or at a pH of 6.0–10.0. The optimum temperature for its reaction was 37 °C. The thermal stability of COD was appreciable, as 90% or 80% of its initial activity was recovered after 1 h or 2 h incubation at 50 °C. Ag+ or Hg+ at 1 mM, was inhibitor of COD activity, while Cu2+, at the same concentration, was activator. The COD Km, determined at 37 °C and pH 7.0, was 0.556 mM. The enzyme was stable at pH 7.0 and 37 °C during 24 h mechanical shaking in the presence of 33% (v/v) of either of the solvents, dimethylsulfoxide, ethyl acetate, butanol, chloroform, benzene, xylene or cyclohexane.  相似文献   

5.
The extensive prospects of violacein in the pharmaceutical industry have attracted increasing interest. However, the fermentation levels of violacein are currently inadequate to meet the demands of industrial production. This study was undertaken to develop an efficient process for the production of violacein by recombinant Citrobacter freundii. The effects of dissolved oxygen (DO) and pH on cell growth and violacein production in batch cultures were investigated first. When the DO and pH of the medium were controlled at around 25% and 7.0, respectively, the biomass and concentration of violacein were maximized. Based on the consumption of nutrients in the medium observed during batch culture, a fed-batch fermentation strategy with controlled DO and pH was implemented. By continuously feeding glycerol, NH4Cl, and l-tryptophan at a constant feeding rate of 16 mL h−1, the final concentration of violacein reached 4.13 g L−1, which was 4.09-fold higher than the corresponding batch culture, and the maximal dry cell weight (DCW) and average violacein productivity obtained for the fed-batch culture were 3.34 g DCW L−1 and 82.6 mg L−1 h−1, respectively. To date, this is the first report on the efficient production of violacein by genetically engineered strains in a fermentor.  相似文献   

6.
Extracellular lipase of the yeast Candida rugosa was produced via high cell density fed-batch fermentations using palm oil as the sole source of carbon and energy. Feeding strategies consisted of a pH-stat operation, foaming-dependent control and specific growth rate control in different experiments. Compared to foaming-dependent feeding and the pH-stat operation, the specific growth rate control of feeding proved to be the most successful. At the specific growth rate control set at 0.05 h−1, the final lipase activity in the culture broth was the highest at ∼700 U L−1. This was 2.6-fold higher than the final enzyme activity obtained at a specific growth rate control set at 0.15 h−1. The peak enzyme concentration achieved using the best foaming-dependent control of feeding was around 28% of the peak activity attained using the specific growth rate control of feeding at 0.05 h−1. Similarly, the peak enzyme concentration attained using the pH-stat feeding operation was a mere 9% of the peak activity attained by specific growth rate control of feeding at a set-point of 0.05 h−1. Fed-batch fermentations were performed in a 2 L stirred-tank bioreactor (30 °C, pH 7) with the dissolved oxygen level controlled at 30% of air saturation.  相似文献   

7.
When stationary phase cells of the dimorphic yeast Candida albicans are diluted into fresh medium at pH 4.5 (low pH), they synchronously form ellipsoidal buds, but when diluted into the same medium at pH 6.7 (high pH), they synchronously form elongate mycelia. Using a perfusion chamber to monitor single cells, we have compared the rates of volume growth between budding and mycelium-forming cells. Results are presented which demonstrate that: (1) after release from stationary phase into medium of low or high pH, each original sphere grows in volume to the time of initial evagination, but does not grow subsequently; (2) successive budding on the original mother cell occurs without interruption resulting in continuous volume growth; however, an interruption in volume growth of the initial bud (B1) occurs before it in turn evaginates; and (3) the rate of volume growth of the first bud at low pH is identical to the rate of volume growth of the mycelium at high pH even though the surface to volume ratios are quite different. The last result is unexpected and is therefore considered in relation to cell wall deposition.  相似文献   

8.
Monascus purpureus CCRC31499 produced a protease when it was grown in a medium containing shrimp and crab shell powder (SCSP) of marine wastes. An extracellular protease was purified from the culture supernatant to homology. The protease had a molecular weight of 40,000 and a pI of 7.9. The optimal pH, optimum temperature, pH stability, and thermal stability of the protease were pH 7–9, 40 °C, pH 5–9, and 40 °C, respectively. In addition to protease activity, CCRC31499 also exhibited activity of enhancing vegetable growth in culture supernatant. This is also the first report of isolation of a protease from Monascus species.  相似文献   

9.
《Mycological Research》2006,110(5):612-623
The in vitro growth of Morchella elata was characterized with respect to the effects of a variety of substrates, isolates, developmental status of the parental ascoma, temperature, and pH. Optimal substrates for growth included sucrose, mannose and lactose, but the growth of some isolates was substantially reduced in some composite media. Maltose and potato-dextrose media limited growth and caused changes in colony morphology; mycelial pigmentation was black in the case of maltose, and mycelial margins were plumose in potato-dextrose cultures. Rapid growth was most reliably achieved in a composite medium containing 1:1 sucrose:mannose. Isolates derived from single ascospores shortly after ejection from ascomata varied in ability to grow in the various substrates. This may be related to variable maturity or dormancy; increasing growth rates correlated with pileus length in the parental ascomata, and ascomata that initially produced slower-growing or abortive colonies produced faster-growing colonies after storage at 20 °C for 96 wk. The growth of M. elata derived from recently ejected ascospores was optimal at 16–24 °C or above for a faster-growing isolate, and 20–24 °C or above for a slow-growing isolate. Although neither isolate grew at 8 °C or below in an initial experiment, spawn cultured on puffed wheat at 28 °C produced mycelia that proliferated when transferred to soil media and incubated at 8 °C. Growth of M. elata in liquid cultures adjusted with potassium hydroxide was optimal at pH 7.0, and was relatively sensitive to more acidic or alkaline pH. When calcium carbonate was used to adjust pH, optimal growth shifted to pH 7.7 or above, suggesting that wood ash and other calcium compounds may not only stimulate growth in natural settings, but also alter the optimal pH for proliferation of M. elata. Further studies with other substrate combinations and incubation conditions will be necessary to fully understand the connections between in vitro growth and the ecological behaviour of the fungus.  相似文献   

10.
The levels of pyruvate kinase (PYK), alcohol dehydrogenase (ADH1), phosphoglycerate kinase (PGK1) and phosphoglycerate mutase (GPM1) mRNAs were measured during batch growth and during the yeast-to-hyphal transition in Candida albicans. The four mRNAs behaved in a similar fashion. PYK1, ADH1, PGK1 and GPM1 mRNA levels were shown to increase dramatically during the exponential growth phase of the yeast form, and then to decrease to relatively low levels in the stationary phase. The dimorphic transition was induced using two sets of conditions: (i) an increase in temperature (from 25°C to 37°C) combined with the addition of serum to the medium; and (ii) an increase in temperature (from 25°C to 37°C) and an increase in pH of the growth medium (from pH 4.5 to pH 6.5). Additional cultures were analysed to control for the addition of serum, and for changes in temperature or pH. Immediately following dilution of late-exponential cells into fresh media the levels of all four glycolytic mRNAs decreased rapidly in contrast to the ACT1 mRNA control, the level of which increased under most conditions. The recovery of glycolytic mRNA levels depended on the culture conditions, but there was no direct correlation with the formation of germ tubes, with the addition of serum to the medium, the Increase in culture temperature, the medium pH, or the glucose concentration. This indicates that the changes in glycolytic gene expression that accompany the dimorphic transition in C. albicans reflect the underlying physiological status of the cells during morphogenesis and not alterations to cell shape.  相似文献   

11.
《Reproductive biology》2014,14(2):122-127
The objective of this study was to evaluate the influence of epidermal growth factor (EGF) and insulin like growth factor-I (IGF-1) on the in vitro maturation of cat oocytes recovered from follicular and luteal stage ovaries. Oocytes from follicular (n = 580) and luteal (n = 209) stages were harvested and divided into four groups, which were cultured in FSH-mediated maturation medium supplemented with: (1) EGF alone (25 ng/mL); (2) IGF-1 alone (100 ng/mL); (3) EGF + IGF-1 (25 ng/mL EGF + 100 ng/mL IGF-I); or (4) no growth factor (control). The proportion of follicular stage oocytes reaching the metaphase II stage was significantly higher than that of oocytes obtained at the luteal stage in both control and study groups (p < 0.001). The percentages of oocytes reaching the metaphase II stage during the follicular period were 62.6% in control; 70.9% in EGF; 72.8% in IGF-1, and 78.1% in EGF + IGF-1 groups, whereas the respective values for gametes collected from luteal stage ovaries were 12.5%, 17.5%, 12.5%, and 16.9%. Additionally, the differences between the study and control groups were significant in the case of follicular stage oocytes. Finally, supplementing the maturation medium with EGF and/or IGF-1 significantly enhanced the meiotic maturation of oocytes recovered from follicular stage ovaries. The present study also demonstrated that the combination of EGF and IGF-I provides an additional or synergic effect on meiotic maturation of oocytes recovered from the follicular stage.  相似文献   

12.
Release of Staphylococcus aureus enterotoxin B (SEB) into the culture medium was initiated during the mid-log phase of growth. A medium consisting of 4% N-Z Amine A (Sheffield), 0.2% dextrose, and 1% yeast extract supported maximum production of SEB. Although pH of the medium during cultivation did not significantly affect the growth curve of the organism, the time required for detectable excretion was affected, as was the final yield. Optimal conditions for SEB production were achieved with pH control at 7.0; alkaline control (pH 8.0) produced only minimal amounts of toxin, whereas acid control (pH 6.0) resulted in 50% reduction in yield. Slightly less SEB was produced when there was no extrinsic pH control, and cultures were buffered only by media constituents and by-products of growth. With pH control at 7.0, deletion of 0.2% dextrose from the medium resulted in 40% reduction in the 8-h yield. There was also a delay in production during early stages of fermentation.  相似文献   

13.
AIMS: To study the effect of sugars and sugar mixtures on the growth kinetics of Oenococcus oeni NCIMB 11648 in batch culture with the aim of producing a high cell productivity system for starter cultures. METHODS AND RESULTS: The growth of O. oeni was investigated on single sugars (glucose, fructose or sucrose) and their mixtures (glucose-fructose, glucose-sucrose or fructose-sucrose). Better growth was obtained on sugar mixtures compared with growth on a single sugar. The production system of O. oeni biomass was investigated in batch culture with or without pH control with respect to kinetics, specific growth rate and biomass yield. The effect of pH and substrate concentration on fermentation balances and ATP yield were determined. The optimal growth of O. oeni was achieved on the glucose-fructose mixture (9 g l(-1), 1 : 1) at pH 4.5 and 25 degrees C with pH control, with highest cell volumetric productivity (7.9 mg cell l(-1) h(-1)), biomass yield (0.041 g cell g(-1) sugar) and specific growth rate (0.066 h(-1)). CONCLUSIONS: The limitations to the growth of O. oeni were pH and inhibition by end product resulting in poor utilization of the medium with low cell yields. The cell productivity of the system can be improved by the appropriate use of mixed sugar growth medium. SIGNIFICANCE AND IMPACT OF THE STUDY: This study uniquely showed that appropriate sugar mixtures with the correct environmental conditions can significantly improve the productivity of O. oeni cultures.  相似文献   

14.
The design of serum-free media for suspension culture of genetically engineered Chinese hamster ovary (CHO) cells using general commercial media as a basis was investigated. Subcultivation using a commercial serum-free medium containing insulin-like growth factor (IGF)-1 with or without FCS necessitated additives other than IGF-1 to compensate for the lack of FCS and improve cell growth. Suspension culture with media containing several combinations of growth factors suggested the effectiveness of addition of both IGF-1 and the lipid signaling molecule lysophosphatidic acid (LPA) for promoting cell growth. Subcultivation of CHO cells in suspension culture using the commercial serum-free medium EX-CELL™302, which contained an IGF-1 analog, supplemented with LPA resulted in gradually increasing specific growth rate comparable to the serum-containing medium and in almost the same high antibody production regardless of the number of generations. The culture with EX-CELL™302 supplemented with LPA in a jar fermentor with pH control at 6.9 showed an apparently higher cell growth rate than the cultures without pH control and with pH control at 6.8. The cell growth in the medium supplemented with aurintricarboxylic acid (ATA), which was much cheaper than IGF-1, in combination with LPA was synergistically promoted similarly to that in the medium supplemented with IGF-1 and LPA. In conclusion, the serum-free medium designed on the basis of general commercial media could support the growth of CHO cells and antibody production comparable to serum-containing medium in suspension culture. Moreover, the possibility of cost reduction by the substitution of IGF-1 with ATA was also shown.  相似文献   

15.
The biological role of cyanobacteria secondary metabolites is relatively unknown although several possible hypotheses have been discussed. In the following study the effect of cylindrospermopsin (CYN) and metabolites of non-CYN producing Cylindrospermopsis raciborskii strain on growth, alkaline phosphatase (ALP) activity and microcystin-LR (MC-LR) production in Microcystis aeruginosa was evaluated. Higher concentrations of CYN (10 and 50 μg L−1) induced toxicity effects demonstrated by significant growth inhibition and M. aeruginosa cell necrosis. Lower concentrations of CYN (1 and 5 μg L−1) slightly decreased growth rates but significantly up-regulated ALP activity. Moreover, under all studied CYN concentrations MC-LR production strongly decreased. Spent C. raciborskii medium mimicked the CYN action by inducing strong inhibition of M. aeruginosa growth and MC-LR production and through up-regulation of ALP activity. On the other hand, spent M. aeruginosa medium did not affect C. raciborskii growth and no alterations in ALP activity were observed. Co-culturing of these two species resulted in an increase of C. raciborskii contribution at the expense of M. aeruginosa. From the results we conclude that CYN can be involved in interspecific competition in cyanobacteria and that non-CYN producing C. raciborskii strains may produce a hitherto unknown bioactive compound(s) which can mimic CYN action.  相似文献   

16.
The catalytic center (CC) of water oxidation in photosystem II passes through four stepwise increased oxidized states (S(0)-S(4)) before O(2) evolution takes place from 2H(2)O in the S(4) --> S(0) transition. The pattern of the release of the four protons from the CC cannot be followed directly in the medium, because proton release from unknown amino acid residues also takes place. However, pH-independent net charge oscillations of 0:0:1:1 in S(0):S(1):S(2):S(3) have been considered as an intrinsic indicator for the H(+) release from the CC. The net charges have been proposed to be created as the charge difference between electron abstraction and H(+) release from the CC. Then the H(+) release from the CC is 1:0:1:2 for the S(0) --> S(1) --> S(2) --> S(3) --> S(0) transition. Strong support for this conclusion is given in this work with the analysis of the pH-dependent pattern of H(+) release in the medium measured directly by a glass electrode between pH 5.5 and 7.2. Improved and crystallizable photosystem II core complexes from the cyanobacterium Synechococcus elongatus were used as material. The pattern can be explained by protons released from the CC with a stoichiometry of 1:0:1:2 and protons from an amino acid group (pK approximately 5.7) that is deprotonated and reprotonated through electrostatic interaction with the oscillating net charges 0:0:1:1 in S(0):S(1):S(2):S(3). Possible water derivatives that circulate through the S states have been named.  相似文献   

17.
The presence of Cs and Sr in culture medium of Azolla filiculoides caused about 27.4% and 46.3% inhibition of biomass growth, respectively, in comparison to A. filiculoides control weight which had not metals. Biosorption batch experiments were conducted to determine the Cs and Sr binding ability of native biomass and chemically modified biosorbents derived from Azolla namely ferrocyanide Azolla sorbents type 1 and type 2 (FAS1 and FAS2) and hydrogen peroxide Azolla sorbent (HAS). The best Cs and Sr removal results were obtained when A. filiculoides was treated by 2 M MgCl2 and 30 ml H2O2 8 mM at pH 7 for 12 h and it was then washed by NaOH solution at pH 10.5 for 6 h. Pretreatment of Azolla have been suggested to modify the surface characteristics which could improve biosorption process. The binding of Cs and Sr on the cell wall of Azolla was studied with micro-PIXE and FT-IR.  相似文献   

18.
The successful use of hay extract medium for growth of Verticillium marquandii in the presence of toxic black slag, resulted in selective and efficient Zn2+ recovery. An alkaline black waste slag originating from short rotary furnace, contained Zn2+ and Pb2+ (20% and 3.5%, respectively) in a form of compounds only slightly soluble in water. After 90 h of the culturing in bioreactor on hay extract medium with starch (50 g l–1) and black waste (5 g l–1) supplements, 80% of zinc and 0.5% of lead were bound by the mycelium. The pH value of the culture medium played a crucial role for limitation of the waste inhibitory effect on mycelium growth and for heavy metal recovery by the fungus.  相似文献   

19.
《Process Biochemistry》2007,42(5):847-855
The psychrotrophic bacterium Pseudomonas fluorescens BM07 was induced to excrete an extracellular biopolymer when cells were grown aerobically at 10 °C and its secretion was inhibited at 30 °C. The biopolymer was easily torn apart from the cells by using a shear force under centrifugation (8700 × g, 30 min) and collected as a well-separated mucoid layer in centrifuge tube. The production of the biopolymer was affected by factors such as the types of carbon and nitrogen sources, temperature, and pH. The best production of 2.5 g/l was obtained when the cells were grown on M1 medium containing 70 mM sucrose and 0.2% (w/v) Casamino Acids. In Kings B enriched medium a maximum biopolymer production of up to 3.4 g/l and growth rate of 2.1 g/l, were achieved using 1:1 ratio of C/N. Addition of NaCl and ethanol to the medium led to a decrease in biopolymer production and growth rate of BM07 strain. FT-IR spectroscopy demonstrated the presence of carboxyl, amine, hydroxyl and methoxyl functional groups in the biopolymer. BM07 biopolymer showed high ion binding capacity with particular preference to uptake cadmium and mercury (∼45 and 70%, respectively). The percentage removal of cobalt, zinc, nickel and copper cations were between 20 and 30%. Overall ion uptake by BM07 biopolymer showed a definite preference for larger over smaller cations (Hg > Cd > Ni > Zn > Cu > Co).  相似文献   

20.
This paper analyzes the feasibility of the autotrophic production of vegetative cells of Haematococcus pluvialis under conditions resembling outdoors. The experimental design simulates in laboratory with artificial light an outdoors circadian cycle similar to solar illumination. The influence of the irradiance and nutrient concentration on the growth rate and carotenoids accumulation in batch cultures is studied. The cultures were not photoinhibited even under the maximum irradiance-level tested (2500 μE m−2 s−1). Growth was kept nutrient-limited by using nutrients concentration below the standard inorganic medium (10 mM nitrate). When no nutrient-limitation occurs, the growth rate and biomass productivity measured 0.57 day−1 and 0.28 g L−1 day−1, respectively, were similar to the maximum values reported, regardless of the nutritional regime: autotrophic, mixotrophic or heterotrophic. On the other hand, carotenogenesis was only observed under nutrient-limiting conditions when the medium strength was reduced to 0.2- or 0.3-fold of the standard medium. On the other hand, carotenogenesis ceased under severe nutrient deprivation (i.e. nutrient strength of 0.1-fold of the standard medium). The growth rate and the carotenoids accumulation rate were demonstrated to be a function of the average irradiance inside the culture, and of the nutrient content of the medium. A mathematical model for the observed behaviour is proposed. This model was adequate to fit all the experimental data obtained. The values determined for the characteristics parameters are in agreement with those found by other authors. Therefore, the proposed model can be a useful tool for the design and management of Haematococcus cultures, and could allow improving the yield of this production process.  相似文献   

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