共查询到20条相似文献,搜索用时 15 毫秒
1.
Saurabh Chattopadhyay V. S. Bisaria T. Scheper A. K. Srivastava 《Biotechnology and Bioprocess Engineering》2002,7(6):331-334
Culture conductivity and on-line NADH fluorescence were used to measure cellular growth in plant cell suspension cultures
ofPodophyllum hexandrum. An inverse correlation between dry cell weight and medium conductivity was observed during shake flask cultivation. A linear
relationship between dry cell weight and culture NADH fluorescence was obtained during the exponential phase of batch cultivation
in a bioreactor under the pH stat (pH 6) conditions. It was observed that conductivity measurement were suitable for biomass
characterisation under highly dynamic uncontrolled shake flask cultivation conditions. However, if the acid/alkali feeding
is done for pH control the conductivity measurement could not be applied. On the other hand the NADH fluorescence measurement
allowed online-in situ biomass monitoring of rather heterogenous plant cell suspension cultures in bioreactor even under the
most desirable pH stat conditions. 相似文献
2.
El-Enshasy HA Beshay UI El-Diwany AI Omar HM El-Kholy AG El-Najar R 《Acta microbiologica Polonica》2003,52(3):301-313
The production of rifamycins B and SV using glucose as main C-source by Amycolatopsis mediterranei in batch and fed-batch culture was investigated. Fed-batch culture using glucose as mono feeding substrate either in the form of pulse addition, in case of shake flask, or with constant feeding rate, in bioreactor level, proved to be an alternative production system with a significant increase in both volumetric and specific antibiotic production. The maximal concentrations of about 1146 mg/l and 2500 mg/l of rifamycins B and SV, respectively, was obtained in fed-batch culture in bioreactor level under non-oxygen limitation. On the other hand, the rate of rifamycins production was increased from 6.58 to 12.13 mg/l x h for rifamycin B and from 9.47 to 31.83 mg/l x h for rifamycin SV on the bioprocess transfer and improvement from the conventional batch cultivation in shake flask to fed-batch cultivation in stirred tank bioreactor. 相似文献
3.
Yildirim S Konrad D Calvez S Drider D Prévost H Lacroix C 《Applied microbiology and biotechnology》2007,77(3):525-531
To increase the yield of heterologous production of the class II bacteriocin DvnRV41 with Escherichia coli Origami (DE3) (pLysS/pCR03), induction of bacteriocin gene expression was optimized by varying the inducer isopropyl beta-D-thiogalactopyranoside (IPTG) concentration (0-2 mM), and controlled batch and fed-batch cultures were tested on a 2-L scale. A concentration of 0.5 mM IPTG was found to be optimal for cell growth and bacteriocin production. Shake flask cultivation of E. coli Origami (DE3) (pLysS/pCR03) gave biomass and bacteriocin yields of 1.54 +/- 0.06 g cdw/l and 18 +/- 1 mg DvnRV41/l, respectively. Biomass (2.70 +/- 0.06 and 6.8 +/- 0.6 g cdw/l, respectively) and bacteriocin yields (30 and 74 mg DvnRV41 per liter, respectively) were both increased with batch and fed-batch compared to shake flask cultures. Bacteriocin yields reported in this study are among the highest published for other heterologous expression systems in shake flasks. 相似文献
4.
Fungal fermentation is very complex in nature due to its nonlinear relationship with the time, especially in batch culture. Growth and production of carbonyl reductase by Geotrichum candidum NCIM 980 have been studied in a laboratory scale stirred tank bioreactor at different pH (uncontrolled and controlled), agitation, aeration and dissolved oxygen concentration. The yield of the process has been calculated in terms of glucose consumed. Initial studies showed that fermenter grown cells have more than 15 times higher activity than that of the shake flask grown cells. The medium pH was found to have unspecific but significant influence on the enzyme productivity. However, at controlled pH 5.5 the specific enzyme activity was highest (306U/mg). Higher agitation had detrimental effect on the cell mass production. Dissolved oxygen concentration was maintained by automatic control of the agitation speed at an aeration rate of 0.6 volume per volume per minute (vvm). Optimization of glucose concentration yielded 21g/l cell mass with and 9.77x10(3)U carbonyl reductase activity/g glucose. Adaptation of different strategies for glucose feeding in the fermenter broth was helpful in increasing the process yield. Feeding of glucose at a continuous rate after 3h of cultivation yielded 0.97g cell mass/g glucose corresponding to 29.1g/l cell mass. Volumetric oxygen transfer coefficient (K(L)a) increased with the increasing of agitation rate. 相似文献
5.
A Panax notoginseng cell culture was successfully scaled up from shake flask to 1.0-L bubble column reactor and concentric-tube airlift reactor. High-density bioreactor batch cultivation was carried out using a modified MS medium. The maximum cell density in batch cultures reached 20.1, 21.0 and 24.1 g/L in the shake flask, bubble column and airlift reactors, respectively, and their corresponding biomass productivity was 950, 1140 and 1350 mg/(L x d) for each. The productivity of ginseng saponin was 70, 96 and 99 mg/(L x d) in the flask, bubble column and airlift reactors, respectively; and the polysaccharide productivity reached 104, 119 and 151 mg/(L x d) for each. Furthermore, a fed-batch cultivation strategy was developed on the basis of specific oxygen uptake rate (SOUR), i.e., sucrose feeding before a sharp decrease of SOUR, and the highest cell density of 29.7 g/L was successfully achieved in the airlift bioreactor on day 17 with a very high biomass productivity of 1520 mg/(L x d). The concentrations of ginseng saponin and polysaccharide reached about 2.1 and 3.0 g/L, respectively, and their productivity was 106 (saponin) and 158 mg/(L x d) (polysaccharide). This work successfully demonstrated the high-density bioreactor cultivation of P. notoginseng cells in pneumatically agitated bioreactors and the reproduction of the shake flask culture results in bioreactors. The cell density, biomass productivity, production titer and productivity of both ginseng saponin and polysaccharide obtained here were the highest that have been reported on a reactor scale for all the ginseng species. 相似文献
6.
Rozita Omar M. A. Abdullah M. A. Hasan M. Rosfarizan M. Marziah 《Biotechnology and Bioprocess Engineering》2006,11(3):223-229
In this study, we have conducted kinetics and modelling studies ofCentella asiatica cell growth and substrate uptake, in an attempt to evaluate cell growth for a better understanding and control of the process.
In our bioreactor cultivation experiment, we observed a growth rate of 0.18/day, a value only 20% higher than was seen in
the shake flask cultivation trial. However, the observed maximum cell dry weight in the shake flask, 10.5 g/L, was 14% higher
than was achieved in the bioreactor. Ninety seven percentage confidence was achieved via the fitting of three unstructured
growth models; the Monod, Logistic, and Gompertz equations, to the cell growth data. The Monod equation adequately described
cell growth in both cultures. The specific growth rate, however, was not effectively predicted with the Logistic and Gompertz
equations, which resulted in deviations of up to 73 and 393%, respectively. These deviations in the Logistic and Gompertz
models may be attributable to the fact that these models were developed for substrate-independent growth and fungi growth,
respectively. 相似文献
7.
Summary
Lithospermum
erythrorhizon was immobilized in a dual hollow fiber bioreactor (DHFBR) to maintain high cell density and to operate continuously. The cells grew well and its dry biomass density was 325 g/L of the void volume for the cell growth. Volumetric and specific productivities of phenolics were 221 mg/L.day and 0.68 mg/g.dry wt.day, respectively, which are 58 and 2 times of those of shake flask cultures. 相似文献
8.
Ergosterol is an economically important metabolite produced by fungi. Recombinant Saccharomyces cerevisiae YEH56(pHXA42) with increased capacity of ergosterol formation was constructed by combined overexpression of sterol C-24(28)
reductase and sterol acyltransferase in the yeast strain YEH56. The production of ergosterol by this recombinant strain using
cane molasses (CM) as an inexpensive carbon source was investigated. An ergosterol content of 52.6 mg/g was obtained with
6.1 g/l of biomass from CM medium containing 60 g/l of total sugar in 30 h in shake flask. The ergosterol yield was enhanced
through the increasing cell biomass by supplementation of urea to a concentration of 6 g/l in molasses medium. Fermentation
was performed in 5-l bioreactor using the optimized molasses medium. In batch fermentation, the effect of agitation velocity
on ergosterol production was examined. The highest ergosterol yield was obtained at 400 rpm that increased 60.4 mg/l in comparison
with the shake flask culture. In fed-batch fermentation, yeast cells were cultivated, firstly, in the starting medium containing
molasses with 20 g/l of total sugar, 1.68 g/l of phosphate acid, and 6 g/l of urea (pH 5.4) for 5 h, then molasses containing
350 g/l of total sugar was fed exponentially into the bioreactor to keep the ethanol level in the broth below 0.5%. After
40 h of cultivation, the ergosterol yield reached 1,707 mg/l, which was 3.1-fold of that in the batch fermentation. 相似文献
9.
The cell growth and alkaloid production of Catharanthus roseus (L.) G. Don cells cultured in the shake flasks with different volumes and in the stirred tank bioreactor (10 L) were compared. Cell dry weight and alkaloid production showed no significant difference in the small volume scale-up shake flasks. When more broths were added to a certain volume in the shake flask, both cell weight and alkaloid production were decreased. The maximum cell dry weight was similar between the cell cultures in the shake flask and the bioreactor, but the alkaloid production of cells was much less in the bioreactor. Gas regime and shear stress were recognized to be the main factors contributing the important effect on alkaloid production during the scale-up processes. 相似文献
10.
Summary Suspension cultures ofCinchona ledgeriana Moens have been developed which exhibit good growth in shake flasks with dry weight yields of approximately 9.0 g.l–1. Cultures have been scaled up for growth in a 7 l air-lift bioreactor. A typical growth curve in the fermenter is shown with similar growth rates but a reduced biomass levels when compared to shake flasks. The analysis of both flask and bioreactor grown suspension cultures indicated the presence of quinidine and low levels of quinine. 相似文献
11.
Aims: The objective of this study is to optimize the levels of carbon and nitrogen sources of the medium in shake flask experiments and evaluate the effect of pH and dissolved oxygen (DO) on the production of l ‐asparaginase from a newly isolated Serratia marcescens SK‐07 in a batch bioreactor. Methods and Results: Central composite rotatable design (CCRD) was applied to optimize the levels of carbon and nitrogen sources of the medium in shake flask experiments. The optimal levels of l ‐asparagine, glucose, yeast extract and peptone were found to be 4·93, 3·81, 3·65 and 1·47 g l?1, respectively, and maximal l ‐asparaginase production of 25·02 U mg?1 was obtained under these conditions. Among the carbon sources tested, l ‐asparagine was identified to be the most favourable carbon source for enhanced production of l ‐asparaginase. The maximum l ‐asparaginase production of 29·89 U mg?1 was achieved in a batch bioreactor at initial pH of 6·5 (uncontrolled) and DO level of 40% in the culture. Conclusions: We have isolated, screened and identified the potential micro‐organism, S. marcescens, for the production of l ‐asparaginase. An overall 5·55‐fold increase in the production was achieved under optimal levels of carbon and nitrogen sources, DO level and at initial pH of 6·5 (uncontrolled). Significance and Impact of the Study: The experiments illustrate the importance of statistical method for optimization of carbon and nitrogen sources and study the effect of physical process parameters on the production of l ‐asparaginase in shake flask and bioreactor, respectively. This study would be helpful for bioprocess development of bacterial l ‐asparaginase production. 相似文献
12.
Laísa dos Reis Willian Daniel Hahn Schneider Roselei Claudete Fontana Marli Camassola Aldo J. P. Dillon 《Bioenergy Research》2014,7(1):60-67
The effect of pH on the production of cellulases and xylanases by Penicillium echinulatum S1M29 was evaluated in a shake flask and in a bioreactor. To control the pH in a shake flask, a buffer made with citric acid and disodium phosphate was used. The buffer was capable of maintaining the culture pH values for the first 48 h. In the bioreactor, the pH was controlled automatically by the addition of NaOH and H2SO4. In the shake flask, the highest activities of xylanases (18.5 IU/mL) and endoglucanases (8.2 IU/mL), as well as the highest filter paper activity (FPA) (0.9 IU/mL), were obtained at initial pH values of between 6.0 and 7.0. In the bioreactor, the highest activities of these enzymes were obtained in a pH range of 5.5 to 6.5. Different isoforms of the endoglucanases were found in the various cultures depending on the pH. More acidic pH ranges favored the production of β-glucosidases in both the shake flask and the bioreactor. 相似文献
13.
Present investigation involves hairy root cultivation of Azadirachta indica in a modified stirred tank reactor under optimized culture conditions for maximum volumetric productivity of azadirachtin. The selected hairy root line (Az-35) was induced via Agrobacterium rhizogenes LBA 920-mediated transformation of A. indica leaf explants (Coimbatore variety, India). Liquid culture of the hairy roots was developed in a modified Murashige and Skoog medium (MM2). To further enhance the productivity of azadirachtin, selected growth regulators (1.0?mg/l IAA and 0.025?mg/l GA3), permeabilizing agent (0.5?% v/v DNBP), a biotic elicitor (1?% v/v Curvularia (culture filtrate)) and an indirectly linked biosynthetic precursor (50?mg/l cholesterol) were added in the growth medium on 15th day of the hairy root cultivation period in shake flask. Highest azadirachtin production (113?mg/l) was obtained on 25th day of the growth cycle with a biomass of 21?g/l DW. Further, batch cultivation of hairy roots was carried out in a novel liquid-phase bioreactor configuration (modified stirred tank reactor with polyurethane foam as root support) to investigate the possible scale-up of the established A. indica hairy root culture. A biomass production of 15.2?g/l with azadirachtin accumulation in the hairy roots of 6.4?mg/g (97.28?mg/l) could be achieved after 25?days of the batch cultivation period, which was ~27 and ~14?% less biomass and azadirachtin concentration obtained respectively, in shake flasks. An overall volumetric productivity of 3.89?mg/(l?day) of azadirachtin was obtained in the bioreactor. 相似文献
14.
Elias DA Tollaksen SL Kennedy DW Mottaz HM Giometti CS McLean JS Hill EA Pinchuk GE Lipton MS Fredrickson JK Gorby YA 《Archives of microbiology》2008,189(4):313-324
High-throughput analyses that are central to microbial systems biology and ecophysiology research benefit from highly homogeneous
and physiologically well-defined cell cultures. While attention has focused on the technical variation associated with high-throughput
technologies, biological variation introduced as a function of cell cultivation methods has been largely overlooked. This
study evaluated the impact of cultivation methods, controlled batch or continuous culture in bioreactors versus shake flasks,
on the reproducibility of global proteome measurements in Shewanella
oneidensis MR-1. Variability in dissolved oxygen concentration and consumption rate, metabolite profiles, and proteome was greater in
shake flask than controlled batch or chemostat cultures. Proteins indicative of suboxic and anaerobic growth (e.g., fumarate
reductase and decaheme c-type cytochromes) were more abundant in cells from shake flasks compared to bioreactor cultures, a finding consistent with
data demonstrating that “aerobic” flask cultures were O2 deficient due to poor mass transfer kinetics. The work described herein establishes the necessity of controlled cultivation
for ensuring highly reproducible and homogenous microbial cultures. By decreasing cell to cell variability, higher quality
samples will allow for the interpretive accuracy necessary for drawing conclusions relevant to microbial systems biology research. 相似文献
15.
Suspension cells of Taxus chinensis were cultivated in both shake flasks and bioreactors. The production of taxuyunnanine C (TC) was greatly reduced when the cell cultures were transferred from shake flasks to bioreactors. Oxygen supply, shear stress and stripping-off of gaseous metabolites were considered as potential factors affecting the taxane accumulation in bioreactors. The effects of oxygen supply on the cell growth and metabolism were investigated in a stirred tank bioreactor by altering its oxygen transfer rate (OTR). It was found that both the pattern and amount of TC accumulation were not much changed within the range of OTR as investigated. Comparative studies on the cell cultivation in low shear and high shear generating bioreactors suggest that the decrease of TC formation in bioreactors was not due to the different shear environments in different cultivation vessels. An incorporation of 2% CO(2) in the inlet air was beneficial for the cell growth, but did not improve the TC production in bioreactors. Furthermore, the effects of different levels of ethylene addition into the inlet air on the cell growth and TC production were investigated in a bubble column reactor. The average cell growth rate increased from 0.146 to 0.204 d(-1) as the ethylene concentration was raised from 0 to 50 ppm, and both the content and production of TC were also greatly improved by ethylene addition. At an ethylene concentration of 18 ppm, the highest TC content and volumetric production in the reactor reached 13.28 mg/(g DW) and 163.7 mg/L, respectively, which were almost the same as those in shake flasks. Compared with the control reactor (bubble column without ethylene supplementation), the maximum TC content was increased by 82% and the total production of TC was doubled. The results indicate that ethylene is a key factor in scaling up the process of the suspension cultures of T. chinensis from a shake flask to a bioreactor. 相似文献
16.
The effect of antioxidants on the production of an important cyclohexadepsipeptide congener destruxin E (dtx E) was investigated using the entomopathogenic fungus Metarhizium anisopliae F061. In shaker flask cultivations, 0.015% of menadione-enhanced dtx E production of 220.4 mg/l compared to the control cultivation 90.2 mg/l, which was illustrated by stimulation of dtx E biosynthesis through two electron reduction DT-diaphorase processes in cultivation of M. anisopliae. In 5-l stirred-tank bioreactor cultivation with menadione addition and of control pH 4.0, a yield of 454.6 mg/l of dtx E was obtained after 7 days, and was nearly 30 and 15-fold higher than that from no pH control, and controlled pH 2.0 cultivations, respectively. Further cultivation in a 20-l airlift bioreactor, at pH 4.0, dtx E obtained on the 9th day was 406.0 mg/l, which was much higher than the standard cultivation of no pH control yield 203.3 mg/l on the 11th day. Thus, the present study provides useful information for enhancing dtx E production in cultivation. 相似文献
17.
Serrano-Carreón L Balderas-Ruíz K Galindo E Rito-Palomares M 《Applied microbiology and biotechnology》2002,58(2):170-174
The final concentration of 6-pentyl-a-pyrone (6PP) produced in cultures of Trichoderma spp. is limited by the fact that inhibition of biomass growth occurs at 6PP concentrations as low as 100 mg/l. The aim of this work was to evaluate liquid-liquid extractive fermentation systems as an alternative to overcome the toxicity problems and to increase the production of 6PP by this fungus. Two alkanes (n-decane and n-hexadecane) and two dicarboxylic esters (dibutyl phthalate and dioctyl phthalate) were evaluated in shake flask cultures. The highest 6PP production (173 ppm) was achieved when n-hexadecane was used, being 3.5-fold the maximum 6PP concentration of a culture without the solvent. Cultivation of Trichoderma harzianum in a 10-1 bioreactor with n-hexadecane yielded 6PP production ninefold higher than that from control cultures. However, 6PP production in the bioreactor (83 ppm) was lower than in shake flasks. Differences in the power drawn to the fluid at each scale could account for such behavior. Even in the presence of the solvent, 6PP content decreased after reaching its maximal concentration. 相似文献
18.
Cyclotides are a family of backbone‐cyclized cystine‐knot‐containing macrocyclic peptides from plants that possess extremely interesting biological activities. Suspension cultures of Oldenlandia affinis, a model plant containing cyclotides, were scaled‐up from shake flask to photobioreactor operation in order to produce these plant peptides under controlled conditions. Cell growth was highly dependent on inoculation culture; cell density as well as culture age had an effect on the growth rates and thus affected the kalata B1 productivity of the bioprocess. In a 25 l scale bioreactor the maximum doubling time was about 1.12 days compared to 2.24 days in shake flasks. The accumulation of kalata B1 of 0.09 mg g?1 DW and 0.07–0.10 mg g?1 DW respectively, however, was on a similar level during the corresponding stationary growth phases in both bioreactor and flask processes. An adjustment of cell culture growth via culture preparation and inoculum density to high cyclotide accumulation results in an estimated output during the most productive retardation phase of about 21 mg kalata B1 per day in the 25 l system. This makes the biotechnological cyclotide synthesis under GMP conditions a competitive production tool compared to field cultivation, chemical, and recombinant synthesis in drug discovery for structure analysis and bioactivity assays. 相似文献
19.
Growth of Aeropyrum pernix, the first reported aerobic neutrophilic hyperthermophilic archaeon, was investigated under different cultivation parameters. Different sources of seawater, pH, and the cultivation methods were tested with the aim to improve the biomass production. A 1-L glass flask fitted with a condenser and air diffuser was used as a bioreactor. The optimum conditions for maximizing A. pernix biomass were obtained when Na2S2O3.5H2O (1 g/L) with added marine broth 2216 at pH 7.0 (20 mmol HEPES buffer/L) was used as a growing medium in a 1-L flask. The biomass production was 0.45 g dry cell mass/L in 40 h under the optimum conditions, which is more than the 0.42 g dry cell mass/L in 60 h previously obtained. 相似文献
20.
Model‐based high cell density cultivation of Rhodospirillum rubrum under respiratory dark conditions
The potential of facultative photosynthetic bacteria as producers of photosynthetic pigments, vitamins, coenzymes and other valuable products has been recognized for decades. However, mass cultivation under photosynthetic conditions is generally inefficient due to the inevitable limitation of light supply when cell densities become very high. The previous development of a new cultivation process for maximal expression of photosynthetic genes under semi‐aerobic dark conditions in common bioreactors offers a new perspective for utilizing the facultative photosynthetic bacterium Rhodospirillum rubrum for large‐scale applications. Based on this cultivation system, the present study aimed in determining the maximal achievable cell density of R. rubrum in a bioreactor, thereby providing a major milestone on the way to industrial bioprocesses. As a starting point, we focus on aerobic growth due to higher growth rates and more facile process control under this condition, with the option to extend the process by an anaerobic production phase. Process design and optimization were supported by an unstructured computational process model, based on mixed‐substrate kinetics. Key parameters for growth and process control were determined in shake‐flask experiments or estimated by simulation studies. For fed‐batch cultivation, a computer‐controlled exponential feed algorithm in combination with a pH‐stat element was implemented. As a result, a maximal cell density of 59 g cell dry weight (CDW) L?1 was obtained, representing so far not attainable cell densities for photosynthetic bacteria. The applied exponential fed‐batch methodology therefore enters a range which is commonly employed for industrial applications with microbial cells. The biochemical analysis of high cell density cultures revealed metabolic imbalances, such as the accumulation and excretion of tetrapyrrole intermediates of the bacteriochlorophyll biosynthetic pathway. Biotechnol. Bioeng. 2010. 105: 729–739. © 2009 Wiley Periodicals, Inc. 相似文献