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1.
Ayora-Talavera T Chappell J Lozoya-Gloria E Loyola-Vargas VM 《Applied biochemistry and biotechnology》2002,97(2):135-145
Catharanthus roseus (L.) G. Don hairy roots harboring hamster 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) (EC 1.1.1.88) cDNA without membrane-binding
domain were evaluated by quantifying the levels of sterols and some indol-alkaloids. Clone 236, with the highest hybridization
signal, had the lowest soluble and microsomal HMGR activity and produced more ajmalicine and catharanthine than the control
but had reduced campesterol concentration. Clone 19, with low hybridization signal, had high soluble HMGR activity and produced
high levels of campesterol and five to seven times more serpentine than the control but a low level of ajmalicine and no accumulation
of catharanthine. These results suggest a possible role for HMGR in indole alkaloid biosynthesis and a possible cosuppression
of both the endogenous and foreign HMGR genes in clone 236. 相似文献
2.
Valduga E Valério A Treichel H Furigo Júnior A Di Luccio M 《Applied biochemistry and biotechnology》2009,157(1):61-69
With the objective of determining the kinetic behavior (growth, substrate, pH, and carotenoid production) and obtain the stoichiometric
parameters of the fermentative process by Sporidiobolus salmonicolor in synthetic and agroindustrial media, fermentations were carried out in shaken flasks at 25°C, 180 rpm, and initial pH of
4.0 for 120 h in the dark, sampling every 6 h. The maximum concentrations of total carotenoids in synthetic (913 μg/L) and
agroindustrial (502 μg/L) media were attained approximately 100 h after the start of the fermentative process. Carotenoid
bioproduction is associated with cell growth and the ratio between carotenoid production and cell growth (Y
P/X) is 176 and 163 μg/g in the synthetic and agroindustrial media, respectively. The pH of the agroindustrial fermentation medium
varied from 4.2 to 8.5 during the fermentation. The specific growth rate (μ
X) for S. salmonicolor in synthetic and agroindustrial media was 0.07 and 0.04 h−1, respectively. The synthetic medium allowed for greater productivity, obtaining maximum cell productivity (P
x) of 0.08 g L−1 h−1 and maximum total carotenoid productivity (P
car) of 14.2 μg L−1 h−1. Knowledge of the kinetics of a fermentative process is of extreme importance when transposing a laboratory experiment to
an industrial scale, as well as making a quantitative comparison between different culture conditions. 相似文献
3.
Mei-Liang Zhou Xue-Mei Zhu Ji-Rong Shao Yan-Min Wu Yi-Xiong Tang 《Applied biochemistry and biotechnology》2012,166(7):1674-1684
Catharanthus roseus (L.) G. Don is a plant species known for its production of a variety of terpenoid indole alkaloids, many of which have pharmacological
activities. Catharanthine can be chemically coupled to the abundant leaf alkaloid vindoline to form the valuable anticancer
drug vinblastine. To study and extract catharanthine and other metabolites from C. roseus, a technique was developed for producing hairy root cultures. In this study, the Agrobacterium rhizogenes A4 was induced in the hairy roots from leaf explants, and the concentration of antibiotics (100 mg/L kanamycin) was elucidated
for selection after transformation. The polymerase chain reaction amplification of rol genes results revealed that transgenic hairy roots contained rol genes from the root induced (Ri)-plasmid. Catharanthine from C. roseus hairy roots was separated and analyzed using high-performance liquid chromatography. Over-expression of CrOrca3 (octadecanoid-responsive Catharanthus AP2/ERF domain), and cytohistochemical staining methods were used to validate transgenic
hairy roots from C. roseus. Hairy root culture of C. roseus is a valuable approach for future efforts in the metabolic engineering of terpenoid indole alkaloids in plants. 相似文献
4.
Optimal design and operation of bioreactors for insect cell culture is facilitated by functional relations providing quantitative
information on cellular metabolite consumption kinetics, as well as on the specific cell growth rates (μG). Initial specific consumption rates of glucose, malate, and oxygen, and associated changes in μG, were measured forSpodoptera frugiperda clone 9 (Sf9) cells grown in batch suspension culture in medium containing 7–35 mM glucose, 0–16 mM malate, and 4–16 mM glutamine.
The initial specific glucose consumption rate (q
G
) could be described by a modified Michaelis-Menten equation treating malate as a “competitive” inhibitorK
1 = 6.5 mM) and glutamine as a “noncompetitive” inhibitorK
I
= 14 mM) ofq
G
, with aK
m
of 7.1 mM for glucose. All three carbon sources were found to increase μG in a saturable manner, and a modified Monod equation was employed to describe this relationship (μGmax = 0.047 h-1). The initial specific oxygen consumption rate (qO2) in Sf9 cells could be related to μG by the maintenance energy model, and it was calculated that, under typical culture conditions, about 15–20% of the cellular
energy demand comes from functions not related to growth. Fitted parameters in mathematical expression for μg: K4, Monod constant for glucose (mM); K5, modified Monod constant for malate (mM); K6, Monod constant for glutamine (mM); mo2, specific consumption rate of oxygen by the cells under zero-growth conditions (nmol/cell/h); qF, initial specific fumarate production rate (nmol/cell/ h);q
G
, initial specific glucose consumption rate (nmol/cell/h); qGmax, maximum initial specific glucose consumption rate (nmol/cell/h);q
M
, initial specific malate consumption rate (nmol/cell/h); qo2, initial specific oxygen consumption rate (nmol/cell/h); Yo2, cell yield on oxygen (cells/nmol); μ, initial specific cell growth rate (h-1); μg, initial specific cell growth rate (h-1); μGmax, maximum initial specific cell growth rate (h-1). 相似文献
5.
Silva Daniel Pereira Pessoa Adalberto Roberto Inês-Conceição Vitolo Michele 《Applied biochemistry and biotechnology》2001,91(1-9):605-613
A batch culture of Saccharomyces cerevisiae for the production of hexokinase was carried out in a 5-L fermentor containing 3 L of culture medium, which was in oculated
with cell suspension (about 0.7 g/L), and left ferm entingat 35°C and pH 4.0. The aeration and agitation were adjusted to
attain k
La values of 15, 60, 135, and 230 h−1. The highest hexokinase productivity (754.6 U/[L h]) and substrate-cell conversion yield (0.21 g/g) occurred for a k
La of 60 h−1. Moreover, the formation of hexokinase and cell growth are coupled events, which is in accordance with the constitutive character
of this enzyme. Hexokinase formation for k
La>60 h−1 was not enhanced probably owing to saturation of the respiratory pathway by oxygen. 相似文献
6.
L. A. Sáenz-Carbonell I. E. Maldonado-Mendoza O. Moreno-Valenzula R. Clau-Uitz M. López-Meyer C. Oropeza V. M. Loyola-Vargas 《Applied biochemistry and biotechnology》1993,38(3):257-267
The release of alkaloids from root culturesDatura stramonium andCatharanthus roseus and thiophenes from root cultures ofTagetes patula was found to increase when the pH of the culture media (ranging from 4.8 to 7.0) was reduced to 3.5. The extent of the effect
was different in each type of culture. Increases ranged from 4- to 20-fold, which in some cases accounted for 75% of the total
secondary metabolite pool produced per flask. When the release of individual metabolites was measured, even larger increases,
were observed (nearly 400-fold for ajmalicine). Increased release of alkaloids fromC. roseus roots were also observed in cultures growing in a 14-L fermentor, when the medium pH was reduced. Reduction of the pH of
the media did not affect growth of the root cultures in subsequent subcultures. The importance of this treatment as a stategy
to improve the recovery of secondary metabolites from producing cultures is discussed. 相似文献
7.
Microbial lipid is a promising source of oil to produce biofuel if it can be generated from lignocellulosic materials. Mortierella isabellina is a filamentous fungal species featuring high content of oil in its cell biomass. In this work, M. isabellina was studied for lipid production from corn stover. The experimental results showed that M. isabellina could grow on different kinds of carbon sources including xylose and acetate, and the lipid content reached to 35 % at C/N ratio of 20. With dilution, M. isabellina could endure inhibition effects by dilute acid pretreatment of corn stover (0.3 g/L furfural, 1.2 g/L HMF, and 1 g/L 4-hydroxybenozic acid) and the strain formed pellets in the cell cultivations. An integrated process was developed combining the dilute acid pretreatment, cellulase hydrolysis, and cell cultivation for M. isabellina to convert corn stover to oil containing fungal biomass. With 7.5 % pretreated biomass solid loading ratio, the final lipid yield from sugar in pretreated biomass was 40 % and the final lipid concentration of the culture reached to 6.46 g/L. 相似文献
8.
Two molecularly imprinted polymers (MIP) for catharanthine and vindoline have been synthesized in order to specifically extract these natural indole alkaloids from Catharanthus roseus by solid-phase extraction (SPE). Each MIP was prepared by thermal polymerisation using catharanthine (or vindoline) as template, methacrylic acid (or itaconic acid) as functional monomer, ethylene glycol dimethacrylate (EDMA) as cross-linking agent and acetonitrile (or acetone) as porogenic solvent.For catharanthine-MIP, a SPE protocol (ACN–AcOH 99/1 washing and MeOH–AcOH 90/10 elution) allows a good MIP/NIP selectivity (imprinting factor 12.6). The specificity of catharanthine-MIP versus related bisindole alkaloids was assessed by cross-reactivity study. The catharanthine-MIP specifically retained catharanthine and its N-oxide analogue but displayed a weak cross-reactivity for other Vinca alkaloids (vinorelbine, vincristine, vinblastine, vindoline, vinflunine). It appears that the catharanthine-like unit of these molecules are hardly trapped in catharanthine cavities located in the MIP, probably due to the sterical hindrance of the vindoline moiety. Finally, the MIP-SPE applied to C. roseus extract enabled quantitative recovery of catharanthine (101%) and the total removal of vindoline. Its capacity was determined and was equal to 2.43 μmol g−1.Vindoline is a weaker base than catharanthine, so the vindoline-MIP was achieved with a strong acidic monomer (itaconic acid) to increase vindoline–monomer interactions and a modified washing solvent (ACN–HCOOH 99/1) to reduce non-specific interactions. The influence of the amount of HCOOH (protic modifier) percolated during the washing step upon the elution yield and the imprinting factor for vindoline was investigated. This preliminary optimisation of the washing step, and in particular the number of moles of acid percolated, seems useful to emphasize the use of MIP in conditions of high selectivity or high yield. A compromise was obtained with an imprinting factor equal to 7.6 and an elution recovery of 33%. However MIP-vindoline failed to achieve a specific extraction of vindoline since catharanthine was also extracted probably because of strong non-specific interactions occurring between catharanthine and the sorbent. 相似文献
9.
The effect of arsenic compounds on indole alkaloid production by cell suspension cultures of Catharanthus roseus was investigated. The analysis of indole alkaloids was achieved by using thermospray liquid chromatography-mass spectrometry (LC MS) which facilitated the rapid screening of alkaloid composition in cultures treated with different arsenicals at different times in their growth cycle. Treatment with dimethylarsinate (DMA), a non-selective herbicide, has a drastic inhibitory effect on alkaloid production although it is the least toxic arsenical to growth. Tryptamine, an early precursor in the biosynthesis of indole alkaloids, accumulates in cells treated with DMA, indicating that the initial step of condensation of tryptamine with secologanin is inhibited. Treatment with DMA during the early stationary phase of culture growth enhances the accumulation of some alkaloids, although some, such as catharanthine, are suppressed. The arsenicals arsenate and methylarsonate (MMA) have an inhibitory effect on alkaloid production when applied during the early growth stages. In contrast to MMA and DMA, arsenate has a stimulatory effect on catharanthine production when introduced to the culture during its early stationary phase. Thus the changes in the pattern of alkaloid accumulation on addition of arsenicals are dependent on the arsenic species and its concentration, as well as the time of application. This variable response indicates that each arsenical has a distinct mode of action on the secondary metabolic pathways of C. roseus. 相似文献
10.
Mewis I Smetanska IM Müller CT Ulrichs C 《Applied biochemistry and biotechnology》2011,164(2):148-161
Cell cultures established from plants represent an attractive alternative to whole plants for effective production of bioactive
secondary metabolites. Cell culture from Vitis vinifera L. cv. Gamay Fréaux accumulated high amounts of hydroxycinnamic acid derivatives and anthocyanins. Two new compounds were
identified: 3-O-glucosylresveratrol, a stilbene derivative, abundant in cell suspension culture, and a hydroxyphenol, 4-(3,5-dihydroxyphenyl)-phenol,
abundant in callus culture. The major anthocyanin monoglucosides present in cell suspension culture were cyanidin 3-O-glucoside and peonidin 3-O-glucoside, and the major cinnamoyl derivatives were cyanidin 3-O-p-coumaryl glucoside and peonidin 3-O-p-coumaryl glucoside. Three minor anthocyanin compounds were found in V. vinifera cell culture: delphinidin 3-O-glucoside, petunidin 3-O-glucoside, and delphinidin 3-O-p-coumaryl glucoside. Anthocyanin levels of cell suspension cultures increased significantly—about eight fold—after 4-day cultivation
in new medium. Salicylic acid at a concentration of 50 μM did not enhance anthocyanin accumulation in cell suspension culture,
and similar levels of jasmonic acid significantly reduced the anthocyanin content. 相似文献
11.
A fed-batch culture system with constant feeding (glucose 80 g L−1, 0.25 ml min−1) was used to study the influence of glucose on cell dry weight and exopolysaccharides production from submerged Tremella fuciformis spores in a 5-L stirred-tank bioreactor. The results showed that high levels of cell mass (9.80 g L−1) and exopolysaccharides production (3.12 g L−1) in fed-batch fermentation were obtained after 1 h of feeding, where the specific growth rate (μ) and exopolysaccharides yield on substrate consumed (YP/S) were 0.267 d−1 and 0.14 g g−1. Unlike batch fermentation, maximal cell mass and exopolysaccharides production merely reached 7.11 and 2.08 g L−1; the specific growth rate (μ) and exopolysaccharides yield on substrate consumed (YP/S) were 0.194 d−1 and 0.093 g g−1, respectively. It is concluded that the synthesis of exopolysaccharides can be promoted effectively when feeding glucose
at a late exponential phase. 相似文献
12.
James P. Kutney Lewis S. L. Choi Pawel Kolodziejczyk Stephen K. Sleigh Kenneth L. Stuart Brian R. Worth Wolfgang G. W. Kurz Kenneth B. Chatson Friedrich Constabel 《Helvetica chimica acta》1981,64(6):1837-1842
Catharanthus roseus cells suspended in production medium showed the presence of four Aspidosperma-type alkaloids, however, no vindoline. Cells grown in media with the pH adjusted to 7.0 produced 3.3 mg total alkaloids/g dry weight. At pH 5.0, 1.7 mg of alkaloid/g dry weight was produced. (S)-Adenosyl-methionine did not stimulate the production of vindoline. When added to cell suspensions for 21 days, vindoline and catharanthine were degraded to non-alkaloidal substances, not dimerized to bisindole alkaloids. 相似文献
13.
Chattopadhyay S Srivastava AK Bisaria VS 《Applied biochemistry and biotechnology》2002,102(1-6):381-393
The root explants of the germinated seedlings of Podophyllum hexandrum were grown in MS medium supplemented with indole acetic acid (IAA) (2 mg/L) and activated charcoal (0.5%), and healthy callus
culture was obtained after incubation for 3 wk at 20°C. The cultivation of plant cells in shake flask was associated with
problems such as clumping of cells and browning of media, which were solved by the addition of pectinase and polyvinylpyrrolidone.
The effect of major media components and carbon source was studied on the growth and podophyllotoxin production in suspension
culture. It was found that glucose was a better carbon source than sucrose and that NH4
+:NO3
− ratio (total nitrogen concentration of 60 mM) and PO4
3− did not have much effect on the growth and product formation. The relative effect of culture parameters (inoculum level,
pH, IAA, glucose, NH4
+:NO3
− ratio, and PO4
3−) on the overall growth and product response of the plant cell suspension culture was further investigated by Plackett-Burman
design. This indicated that inoculum level, glucose, IAA, and pH had significant effects on growth and production of podophyllotoxin.
To identify the exact optimum concentrations of these parameters on culture growth and podophyllotoxin production, central
composite design experiments were formulated. The overall response equations with respect to growth and podophyllotoxin production
as a function of these culture parameters were developed and used to determine the optimum concentrations of these parameters,
which were pH 6.0, 1.25 mg/L of IAA, 72 g/L of glucose, and inoculum level of 8 g/L. 相似文献
14.
Gregory M. Colores Petra M. Radehaus Steven K. Schmidt 《Applied biochemistry and biotechnology》1995,54(1-3):271-275
ASphingomonas species that mineralizes high concentrations of pentachlorophenol (PCP) was isolated from a PCP-contaminated EPA Superfund
site. This bacterium, identified asSphingomonas sp. strain RA2, is able to degrade PCP at concentrations of up to 300 μg/mL in liquid culture. This organism was tested for
its ability to degrade high concentrations of PCP in a soil that did not contain organisms capable of degrading high concentrations
of PCP. When inoculated into contaminated soil,Sphingomonas sp. RA2 mineralized PCP at concentrations of 300, 600, 900, and 1200 μg PCP/g of soil, but was unable to mineralize 1500
μg PCP/g of soil. Only very minimal loss of PCP was seen in uninoculated soils. The results of this study demonstrate thatSphingomonas sp. RA2 may be a useful organism for remediation of sites contaminated with high concentrations of PCP. 相似文献
15.
《Magnetic resonance in chemistry : MRC》2003,41(8):633-635
A new diterpenoid, 12β,13α‐dihydroxytriptonide, was obtained from the transformed culture of triptonide by Catharanthus roseus cell suspension cultures. The complete 1H and 13C NMR assignments of the compound were carried out by using DEPT, COSY, HSQC, g‐HMBC and NOESY techniques. Copyright © 2003 John Wiley & Sons, Ltd. 相似文献
16.
The damped glycolytic oscillation phenomenon occurring in starved cells of the yeast Saccharomyces cerevisiae (NBRC 0565) was characterization for application to a toxicity bioassay. S. cerevisiae was grown under semi-anaerobic conditions. The transient oscillations were observed photometrically as the time course of
the fluorescent intensity of reduced pyridine nucleotide resulting from instantaneous addition of glucose to a cell suspension.
In this study, simple and reproducible conditions inducing damped oscillations were obtained by modifying a literature method.
For estimation of the wave shapes of the damped oscillations we used six indexes. To investigate the total reproducibility
as the averaged relative standard deviation (RSDav) for the six indexes obtained from the wave shapes, the damped oscillations were induced under the optimum conditions and
the RSDav values were calculated as 14% in a buffer cell suspension (n = 62) and 22% in a water cell suspension (n = 78). Finally, the effects of glucose concentration on the six indexes were examined, and all the indexes changed when the
glucose concentration was changed. Excellent correlations were obtained between the index of oscillation-state time and the
concentration of glucose in a buffer cell suspension (r = 0.9985, 0.5–250 mmol L−1, 10 points) and in a water cell suspension (r = 0.9989, 2.5 μmol L−1–250 mmol L−1, 12 points), respectively.
Figure Characterization of damped glycolytic oscillation, (a) typical shape, and (b) its estimation
Electronic supplementary material The online version of this article (doi:)contains supplementary material, which is available to authorized users. 相似文献
17.
In order to continue our previous studies concerning Geranium pyrenaicum Burm. (Geraniaceae), we have performed spectrophotometric determinations and a HPLC study of some polyphenols. We have analyzed
the dried Geranii pyrenaici herba (harvested from Cluj-Napoca, district of Cluj, Romania). We have established the content in flavonoids (0.316%), phenolic
acids (0.099%), tannins (5.295%), and anthocyanins (12.030 mg/100 g vegetal product). We have identified and measured by HPLC
the following compounds: hyperoside (21.61 μg/100 mg), ellagic acid (1810.44 μg/100 mg), isoquercitrine (11.197 μg/100 mg),
and caftaric acid (76.83 μg/100 mg). We have also analyzed by HPLC a hydrolyzed sample of the same drug in which we have identified
and measured: ellagic acid (4139.33 μg/100 mg), quercetol (29.65 μg/100 mg), kaempherol (41.48 μg/100 mg), and caftaric acid
(20.721 μg/100 mg).
__________
Published in Khimiya Prirodnykh Soedinenii, No. 4, pp. 322–324, July–August, 2005. 相似文献
18.
Minakata K Nozawa H Gonmori K Yamagishi I Suzuki M Hasegawa K Watanabe K Suzuki O 《Analytical and bioanalytical chemistry》2011,400(7):1945-1951
An electrospray ionization tandem mass spectrometric (ESI-MS-MS) method has been developed for the determination of cyanide
(CN–) in blood. Five microliters of blood was hemolyzed with 50 μL of water, then 5 μL of 1 M tetramethylammonium hydroxide solution
was added to raise the pH of the hemolysate and to liberate CN– from methemoglobin. CN– was then reacted with NaAuCl4 to produce dicyanogold, Au(CN)2–, that was extracted with 75 μL of methyl isobutyl ketone. Ten microliters of the extract was injected directly into an ESI-MS-MS
instrument and quantification of CN– was performed by selected reaction monitoring of the product ion CN– at m/z 26, derived from the precursor ion Au(CN)2– at m/z 249. CN– could be measured in the quantification range of 2.60 to 260 μg/L with the limit of detection at 0.56 μg/L in blood. This
method was applied to the analysis of clinical samples and the concentrations of CN– in the blood were as follows: 7.13 ± 2.41 μg/L for six healthy non-smokers, 3.08 ± 1.12 μg/L for six CO gas victims, 730 ± 867 μg
for 21 house fire victims, and 3,030 ± 97 μg/L for a victim who ingested NaCN. The increase of CN– in the blood of a victim who ingested NaN3 was confirmed using MS-MS for the first time, and the concentrations of CN– in the blood, gastric content and urine were 78.5 ± 5.5, 11.8 ± 0.5, and 11.4 ± 0.8 μg/L, respectively. 相似文献
19.
Martin Adam Petr Dobiáš Petra Pavlíková Karel Ventura 《Central European Journal of Chemistry》2009,7(3):303-311
The analytical microextraction methods of gas chromatography coupled with flame ionisation detector (GC-FID) for determination
of selected essential oils in herbs were proposed. Two microextraction methods for the isolation of essential oils from plants
such as Lavandula spica L., Melissa officinalis L., Mentha piperita L. and Salvia officinalis L. were used. The methods of solid-phase and single-drop microextractions, were optimised and compared. The obtained LOD
values for all studied essential oils were found to be within 2.5–20.5 μg for SDME and 57.0–139.8 μg for SPME method per 100
g of dried sample leaves. The appropriate LOQ values were then 8.4–68.4 μg for SDME and 189.8–466.1 μg for SPME of target
analytes per 100 g of dried sample leaves.
相似文献
20.
Marcelo Donadel Malesuik Simone Gonçalves Cardoso Martin Steppe 《Chromatographia》2008,67(1-2):131-136
A reversed-phase liquid chromatographic (LC) method was developed for the assay of nitazoxanide (NTZ) in solid dosage formulations.
An isocratic LC separation was performed on a Phenomenex Synergi Fusion C18 column (250 mm × 4.6 mm, i.d., 4 μm particle size) using a mobile phase of 0.1% o-phosphoric acid solution, pH 6.0: acetonitrile (45:55, v/v) at a flow rate of 1.0 mL min−1. Detection was achieved with a photodiode array detector at 240 nm. The detector response for NTZ was linear over the concentration
range from 2 to 100 μg mL−1 (r = 0.9999). The specificity and stability-indicating capability of the method were proved using stress conditions. The RSD
values for intra-day precision were less than 1.0% for tablets and powder for oral suspension. The RSD values for inter-day
precision were 0.6 and 0.7% for tablets and powder for oral suspension. The accuracy was 100.4% (RSD = 1.8%) for tablets and
100.9% (RSD = 0.3%) for powder for oral suspension. The limits of quantitation and detection were 0.4 and 0.1 μg mL−1. There was no interference of the excipients on the determination of the active pharmaceutical ingredient. The proposed method
was precise, accurate, specific, and sensitive. It can be applied to the quantitative determination of drug in tablets and
powder for oral suspension. 相似文献