Initial Sucrose Concentration Research Articles

Levansucrase production by Zymomonas mobilis : Optimization of process parameters and fructooligosaccharide production

This article investigates the production and optimization of levansucrase using synthetic medium by Zymomonas mobilis NRRL B-14023. Response surface methodology was used to investigate the effects of three different fermentation parameters on levansucrase enzyme production. Maximum levansucrase activity of 13.3 µmol glucose/min was obtained at the optimum levels of process variables (pH 4.90, initial sucrose concentration 159.01 g/L and fermentation temperature 30.3°C). Response surface methodology was found to be useful in optimizing and determining the interactions among process variables. The purity of levansucrase used in enzyme assays were analyzed by SDS-PAGE and a dominant protein band of size around 47 kDa was obtained. The sequence of the produced enzyme was determined by peptide mass fingerprinting. The presence of kestose in the reaction mixture showed that Z. mobilis levansucrase can produce fructooligosaccharides. This study fulfills the lack of mathematical and statistical approaches in optimizing the levansucrase production by Z. mobilis. Practical applications Levansucrases are responsible for the production of valuable fructans; fructooligosaccharides (FOS) and levan. Fructooligosaccharides are sugar polymers three times less sweet than sucrose and naturally present in some foods. FOS have important promising use in the food industry due to their properties such as solubility in water, stability during storage, low calorie value, and prebiotic effects. In this research, the production of microbial levansucrase was optimized for potential industrial applications by response surface methodology. Future studies will focus on conversion of intrinsic sucrose of foods to FOS using levansucrase.

Kinetic Model for Invertase‐Induced Sucrose Hydrolysis: Initial Time Lag

AbstractThe kinetics of sucrose hydrolysis by invertase was studied in order to find a comprehensive model for the reaction pathway and mechanism. First, three common models of Michaelis‐Menten (MM), substrate inhibition (S2), and substrate clusters' inhibition (S3(I)) were investigated. The third model was found to better predict the initial sucrose concentration. Then, the S3(I) model was modified to cover the remaining pathway (S3(II)). Finally, a new comprehensive model (S3(III)) was evaluated, which in addition to what is considered in the two previously mentioned models (S3(I) and S3(II)) also involved the initial time lag. The model predictions showed an excellent agreement with the experimental data. The mean absolute error for the MM model is significantly reduced for the S3(III) model.

Sucrose-enhanced biosynthesis of medicinally important antioxidant secondary metabolites in cell suspension cultures of Artemisia absinthium L.

Natural products are gaining tremendous importance in pharmaceutical industry and attention has been focused on the applications of in vitro technologies to enhance yield and productivity of such products. In this study, we investigated the accumulation of biomass and antioxidant secondary metabolites in response to different carbohydrate sources (sucrose, maltose, fructose and glucose) and sucrose concentrations (1, 3, 5, 7 and 9%). Moreover, the effects of 3% repeated sucrose feeding (day-12, -18 and -24) were also investigated. The results showed the superiority of disaccharides over monosaccharides for maximum biomass and secondary metabolites accumulation. Comparable profiles for maximum biomass were observed in response to sucrose and maltose and initial sucrose concentrations of 3 and 5%. Maximum total phenolic and total flavonoid contents were displayed by cultures treated with sucrose and maltose; however, initial sucrose concentrations of 5 and 7% were optimum for both classes of metabolites, respectively. Following 3% extra sucrose feeding, cultures fed on day-24 (late-log phase) showed higher biomass, total phenolic and total flavonoid contents as compared to control cultures. Highest antioxidant activity was exhibited by maltose-treated cultures. Moreover, sucrose-treated cultures displayed positive correlation of antioxidant activity with total phenolics and total flavonoids production. This work describes the stimulatory role of disaccharides and sucrose feeding strategy for higher accumulation of phenolics and flavonoids, which could be potentially scaled up to bioreactor level for the bulk production of these metabolites in suspension cultures of A. absinthium.

Control strategies for biohydrogen production by immobilized co-culture of Clostridium butyricum and Rhodopseudomonas palustris

ABSTRACTIn this study, we evaluated biohydrogen production of Clostridium butyricum and Rhodopseudomonas palustris by immobilized co-culture. Effects of free cells and immobilized cells, immobilized biomass ratio, sucrose concentration, and initial pH on hydrogen production were investigated. The immobilized co-culture can achieve high cumulative hydrogen volume yield (604 mL) as compared to free co-culture cumulative hydrogen volume (513 mL) while the initial sucrose concentration was 17.8 g/L. The optimum C. butyricum/R. palustris ratio was 1:10, yielding the highest cumulative hydrogen (728 mL). High sucrose concentration (above 35.6 g/L) would inhibit hydrogen production. The optimal initial pH value for hydrogen production of immobilized co-culture was 7.0 (cumulative hydrogen volume 830 mL).

Comparative Proteomic Analysis of Streptomyces aureochromogenes Under Different Carbon Sources and Insights into Polyoxin Production

In this paper, fermentation dynamics process of Streptomyces aureochromogenes was researched. The production of polyoxins in culture medium with moderate concentration sucrose was at a relative high level. And, when the carbon source changed to glucose, the mycelium grew in a better condition, while no poloxins was produced. This phenomenon also happened when increasing the initial concentration of sucrose to three times. To evaluate the regulatory mechanism of polyoxin production, comparative proteomic analysis of S. aureochromogenes was conducted. The result showed that tricarboxylic acid cycle (TCA) cycle, amino metabolism, and fatty acid metabolism were active and glycolysis was repressed. And, the fact that stress and stress-related protein like superoxide dismutase (SOD), Clp protease, and catalase were highly expressed indicated a higher regulator level in synthesis of polyoxins. In addition, enzymes related to transcription or translation processes also revealed a correlation between specific cell growth rate and polyoxin production. This study will be helpful in better understanding the biosynthesis mechanism of polyoxins and metabolism regulation in this process.

Kinetics and model development for enzymatic synthesis of fructo-oligosaccharides using fructosyltransferase.

Experimental investigations were made to synthesize fructo-oligosaccharides (FOS) from sucrose using fructosyltransferase. The influence of various parameters such as temperature (45-55 °C), pH (4-5), initial sucrose concentration (ISC: 300-500 g/L) and enzyme concentration (4-32 U/mL) were varied. A maximum FOS yield of 60% was observed at ISC 500 g/L, pH 4.5 with enzyme activity 32 U/mL and at 55 °C. It was confirmed that 1-kestose (tri-) was the major product of FOS as compared to nystose (tetra-) and fructosylnystose (penta-saccharides). Further, the reaction rate increases with increase in temperature. From separate sets of experiments, it was observed that FOS formation was affected by glucose inhibition. Apart from the increase in the rate of FOS formation with increasing enzyme activity, the final values of FOS yield increase though till 16 U/mL and thereafter attain plateau. A kinetic model was also developed, based on Michaelis-Menten kinetics, and a five-step ten-parameter model, including glucose inhibition, was obtained. Model was solved using COPASI(®) (version 4.8) solver for kinetic parameter estimations followed by time course simulations.

Kinetics of Sucrose Hydrolysis by Immobilized Recombinant Pichia pastoris Cells in a Batch reactors

Sucrose hydrolysis was carried out in a constant-volume batch reactor, using recombinant Pichia pastoris BfrA4X whole cells expressing Thermotoga maritima invertase, entrapped in calcium alginate beads. The kinetics of the enzymatic hydrolysis of sucrose by the biocatalyst was examined at substrate concentrations ranging between 0.03 M and 2.04 M. The reaction rate increases until 0.31 M after which the reaction velocity was constant until 1.16 M, above this concentration, the reaction rate decreases with increasing sucrose concentration. The experimental data obtained with two weight of the biocatalyst were incorporated into two kinetic models to predict the reaction time needed for sucrose hydrolysis. One model was applied for sucrose concentrations bellow 1.16 M while a second one could be used at inhibitory range between 1.46 and 2.04 M with a k value as function of initial sucrose concentration and biocatalyst weight.

Production of fructosyltransferase by Penicillium simplicissimum in batch culture

Seventeen (17) strains of macro-fungi were screened for fructosyltransferase (FTase) production, of which Penicillium simplicissimum was selected for further work. FTase production was carried out in shake flask cultured at 30°C and 250 rpm for 48 h. For optimization of one factor-at-a-time method, effects of initial sucrose concentration, fermentation time, pH, inoculum size and agitation speed were studied on growth and extracellular FTase activity. These fermentation parameters were then optimized using response surface methodology (RSM) coupled with a central composite design (CCD). 20 g/l sucrose concentration, 36 h fermentation time, pH 6, 15 ml inoculum size and 150 rpm agitation speed were the optimum parameters obtained from RSM. The experimental values were in good agreement with predicted values within -0.057 to 0.023 errors. Nevertheless, the biomass and FTase (extracellular and intracellular) activities increased from 4.38 - 12.16 g/l, 92.76 - 99.46 IU/ml and 60.73 - 69.98 IU/ml in the unoptimized parameters to 14.16 g/l, 118.86 IU/ml (extracellular) and 71.98 IU/ml (intracellular) in the parameters optimized by RSM, respectively. Keywords: Fructosyltransferase, Penicillium simplicissimum , response surface methodology, fructooligosaccharides. African Journal of Biotechnology , Vol 13(46) 4294-4307

Kinetics of sucrose conversion to fructo-oligosaccharides using enzyme (invertase) under free condition.

The study reports the synthesis of fructo-oligosaccharide (FOS) from sucrose using invertase derived from Saccharomyces cerevisiae. The reaction was conducted in a batch mode under free enzyme condition. Fructo-oligosaccharide formation was detected at a high sucrose concentration of over 200g/L. The investigation was extended to study the effect of different parameters such as initial sucrose concentration (ISC), pH, and enzyme concentration. A maximum FOS yield of 10% (dry basis) was observed using 525g/L of ISC, with 6U/mL of the enzyme, and pH 5.5 at 40°C. 1-Kestose was the major product of among different forms of FOS. The FOS yield increased with an increase in sucrose concentration up to 525g/L, beyond which it started to decrease. However, the maximum FOS yield was not affected by the increasing concentration of the enzyme beyond a certain level (2U/mL). Furthermore, the activity of enzyme slightly increased with an increase in the pH up to 6, and thereafter it declined. Addition of glucose decreased the FOS yield because of enzyme inhibition. A five-step, ten-parameter model was developed, for which the simulation was performed in COPASI. The results predicted by the model were consistent with the experimental data.

Characterization of the antifungal activity of Lactobacillus harbinensis K.V9.3.1Np and Lactobacillus rhamnosus K.C8.3.1I in yogurt

Few antifungal protective cultures adapted to fermented dairy products are commercially available because of the numerous constraints linked to their market implementation. Consumer's demand for naturally preserved food products is growing and the utilization of lactic acid bacteria is a promising way to achieve this goal. In this study, using a 25−1 factorial fractional design, we first evaluated the effects of fermentation time, of initial sucrose concentration and of the initial contamination amount of a spoilage yeast, on antifungal activities of single and mixed cultures of Lactobacillus rhamnosus K.C8.3.1I and Lactobacillus harbinensis K.V9.3.1Np in yogurt. L. harbinensis K.V9.3.1Np, the most relevant strain with regard to antifungal activity was then studied to determine its minimal inhibitory inoculation rate, its antifungal stability during storage and its impact on yogurt organoleptic properties. We showed that L. harbinensis K.V9.3.1Np maintained a stable antifungal activity over time, which was not affected by initial sucrose, nor by a reduction of the fermentation time. This inhibitory activity was an all-or-nothing phenomenon. Once L. harbinensis K.V9.3.1Np reached a population of ∼2.5 × 106 cfu/g of yogurt at the time of contamination, total inhibition of the yeast was achieved. We also showed that an inoculation rate of 5 × 106 cfu/ml in milk had no detrimental effect on yogurt organoleptic properties. In conclusion, L. harbinensis K.V9.3.1Np is a promising antifungal bioprotective strain for yogurt preservation.

Organic carbon induced mobilization of iron and manganese in a West Bengal aquifer and the muted response of groundwater arsenic concentrations

The exact circumstances that cause the widespread enrichment of Mn and As in groundwater of the Bengal Delta Plain (BDP) and many other Asian delta areas still remain a matter of debate in the scientific community. We conducted an in situ field experiment in the central BDP region to investigate the influence of organic matter on the mobility of Fe, Mn and As in shallow aquifers. The groundwater at our study site was initially characterized by a circum-neutral pH, low concentrations of O2, NO3− and SO42−, and increased Fe, Mn and As concentrations, reflecting reducing conditions in the aquifer. Since organic matter controls microbially mediated redox processes which are believed to result in the mobilization of Fe, Mn and As from Holocene aquifer sediments, an easily degradable carbon source (sucrose) was introduced into a shallow aquifer via four nested monitoring wells and distributed by circular pumping. Initial sucrose concentrations reached up to 2.55mM in the local groundwater and induced a strong increase in the activity of indigenous microbes that decomposed the sucrose within the following 14days stepwise into intermediate catabolic products (e.g., acetic acid), and finally to CO2/HCO3−. The formation of organic acids was accompanied by a temporary decline in the pH and the redox potential, as well as an increase in the concentration of most major and trace elements in the groundwater by several times. While Mn concentrations rose up to 81.3μM (representing a 7.5 fold increase), Fe (on average 96.7% Fe(II)) concentrations reached a considerable transient maximum of 1390μM, which was 36 times higher than the initial baseline value. The most significant observation of this experiment is that the relative increments of dissolved As (on average 95.8% As(III)) reached between 19 and 49% only, which is in clear contrast to the pronounced mobilization of Fe, Mn and other trace elements. Changes in the groundwater composition during the experiment imply that the mobilization of Fe and Mn was primarily caused by a reductive dissolution of Mn-oxides and Fe-(oxyhydr)oxides, resulting from the stimulation of indigenous bacteria by the addition sucrose. In this context, the release of As can be attributed to the dissolution of Fe-(oxyhydr)oxides, which constitute the principal source of As in the aquifer sediments according to mineralogical and geochemical analyses. In contrast to the pronounced mobilization of Fe, the response of groundwater arsenic concentrations appeared to be muted, as indicated by subsequently declining As to Fe mol ratios that dropped one order in magnitude. The remarkable decoupling of As from Fe mobilization indicates that the aquifer sediments were apparently capable of compensating for the additional release of As. We attribute this As buffer potential to remaining Fe-minerals and potentially newly formed Fe(II)- and mixed Fe(II/III)-mineral phases, which were able to readily immobilize dissolved As. Sequential extraction results of the initial aquifer sediments further support this interpretation, revealing that up to 85% of the total As in the sediments was already present in adsorbed form, with Fe-minerals as principal hosts. Hence, the experimental data implies that a biogeochemically controlled environment of competing As release and retention arose after the addition of sucrose, where Fe-mineral phases played a key role in buffering the release of As. We further conclude that organic carbon limited aquifer systems in the BDP with increased As concentrations in groundwater may exhibit an unexpected buffer potential towards an additional As release, even when vast amounts of easily degradable organic carbon are introduced into the system.

Biohydrogen Production by Immobilized Co-culture of Clostridium Butyricum and Rhodopseudomonas Palustris

In this study, we evaluated biohydrogen production of Clostridium butyricum and Rhodopseudomonas palustris by immobilized co-culture. Effects of the free cells and immobilized cells, immobilized biomass ratio, sucrose concentration and initial pH on hydrogen production were investigated. The immobilized co-culture can achieve high cumulative hydrogen (604ml) as compare to free co-culture cumulative hydrogen while the initial sucrose concentration was 17.8g/l. The optimum C. butyricum/R. palustris ratio was 1/10 yielding the highest cumulative hydrogen (728ml). High sucrose concentration (above 35.6g/l) would inhibit hydrogen production. The optimal initial pH for hydrogen production of immobilized co-culture was 7.0 (cumulative hydrogen 830ml).

Production of fructosyltransferase in mechanically stirred and air-lift bioreactors

AbstractAureobasidium pullulans cells were cultivated in a 3 dm3 stirred bioreactor to optimize the production of fructosyltransferase (FTase). Batch experiments were focused on the influence of the initial sucrose concentration (150–350 g dm−3), sodium nitrate concentration (10–25 g dm−3), and stirring rate (180–540 min−1) on the process. It was found that the FTase specific activity per cell mass was positively influenced by the conditions that impeded the cell growth such as higher sucrose concentrations or lower oxygen transfer rates. Higher content of inorganic nitrogen substrate slightly increased the overall FTase production. The presence of fructose moiety-containing saccharides in the cultivation medium was observed to be indispensable for FTase production. An addition of a concentrated sucrose solution in the 40th cultivation hour therefore essentially resumed the FTase production. Finally, scale-up experiments were performed in 12 dm3 and 100 dm3 mechanically stirred bioreactors and in 20 dm3 and 60 dm3 air-lift bioreactors.

A new mechanism and kinetic model for the enzymatic synthesis of short-chain fructooligosaccharides from sucrose

A kinetic model based on a ping-pong mechanism was developed under the steady-state hypothesis to account for the short-chain fructooligosaccharides (sc-FOS) synthesis using the commercial cellulolytic enzyme preparation, Rohapect CM. This new mechanism takes into account the interactions between the enzyme species and potential substrates (sucrose and sc-FOS) as a single complex reaction, allowing a better understanding of the reaction kinetics.The initial reaction rate laws appropriately describe the kinetic profiles of the examined substrates. Whereas sucrose exhibited Michaelis–Menten behavior with substrate inhibition, 1-kestose and nystose followed Michaelis–Menten and sigmoid enzyme kinetics. In addition, the enzyme was competitively inhibited by glucose and exhibited significant hydrolytic activity in the presence of nystose.The overall model was simultaneously fitted to experimental data from three initial sucrose concentrations (0.5, 1.5 and 2.1M) using a multi-response regression with kinetic parameters that have biochemical relevance and are independent of the enzyme concentration. According to the model, sucrose acts almost exclusively as a fructosyl donor substrate. The mathematical development described herein is expected to be suitable for modeling similar enzymatic reaction systems.

Characterization of hyperbranched glycopolymers produced in vitro using enzymes

Asymmetrical flow field flow fractionation (AF4) has proven to be a very powerful and quantitative method for the determination of the macromolecular structure of high molar mass branched biopolymers, when coupled with multi-angle laser light scattering (MALLS). This work describes a detailed investigation of the macromolecular structure of native glycogens and hyperbranched α-glucans (HBPs), with average molar mass ranging from 2 × 10(6) to 4.3 × 10(7)gmol(-1), which are not well fractionated by means of classical size-exclusion chromatography. HBPs were enzymatically produced from sucrose by the tandem action of an amylosucrase and a branching enzyme mimicking in vitro the elongation and branching steps involved in glycogen biosynthesis. Size and molar mass distributions were studied by AF4, coupled with online quasi-elastic light scattering (QELS) and transmission electron microscopy. AF4-MALLS-QELS has shown a remarkable agreement between hydrodynamic radii obtained by online QELS and by AF4 theory in normal mode with constant cross flow. Molar mass, size, and dispersity were shown to significantly increase with initial sucrose concentration, and to decrease when the branching enzyme activity increases. Several populations with different size range were observed: the amount of small size molecules decreasing with increasing sucrose concentration. The spherical and dense global conformation thus highlighted was partly similar to native glycogens. A more detailed study of HBPs synthesized from low and high initial sucrose concentrations was performed using complementary enzymatic hydrolysis of external chains and chromatography. It emphasized a more homogeneous branching pattern than native glycogens with a denser core and shorter external chains.

A new strategy to enhance gellan production by two-stage culture in Sphingomonas paucimobilis

The effects of different initial sucrose concentrations and temperatures on gellan biosynthesis by Sphingomonas paucimobilis ATCC 31461 were investigated. Lower sucrose concentrations and higher temperatures were favorable for cell growth. Higher sucrose concentrations and lower temperatures promoted gellan production but retarded cell growth. Based on these results, a two-stage culture strategy was developed to improve gellan production. During the first 24h, S. paucimobilis was cultured in a pulse fed-batch mode with an initial sucrose concentration 10g/L. Ten grams per liter of sucrose were added at 12h and 24h, and the temperature was controlled at 33°C. Batch culture was performed, and the temperature was reduced to 28°C to achieve a high gellan accumulation. The two-stage culture strategy achieved the highest gellan production (22.61g/L) at 60h that was 35.71% higher than the result of the best conventional batch operation (16.66g/L). Meanwhile, high gellan yield was related to high UDPG-pyrophosphorylase activity and glucosyltransferase activity.

Centrifugal freeze concentration

Assisted techniques that improve the efficiency of freeze concentration in one-step configurations are important in achieving commercial viability. An alternative reported in this study is the use of the centrifugation for separating the concentrated solution from the ice matrix. By applying centrifugation, high values of solute had recovered, reaching approximately 0.73 kg of sucrose obtained per 1 kg of initial sucrose at 1600 RCF of centrifugation speed, independent of initial concentration of sucrose (5 to 20 wt.%) and freezing procedure (radial or unidirectional freezing). The performance of centrifugal freeze concentration may be attributed to ice matrix that acted as a porous solid through which the concentrated solution percolates through drainage channels between ice crystals improved by the centrifugal force. The freeze concentration allows producing food concentrates with high quality as compared to evaporation and membrane technology. In this work, centrifugation has been applied as an assisted technique to improve the efficiency of freeze concentration in one-step configuration, obtaining promissory results.

In Vitro Synthesis of Hyperbranched α-Glucans Using a Biomimetic Enzymatic Toolbox

Glycogen biosynthesis requires the coordinated action of elongating and branching enzymes, of which the synergetic action is still not clearly understood. We have designed an experimental plan to develop and fully exploit a biomimetic system reproducing in vitro the activities involved in the formation of α(1,4) and α(1,6) glycosidic linkages during glycogen biosynthesis. This method is based on the use of two bacterial transglucosidases, the amylosucrase from Neisseria polysaccharea and the branching enzyme from Rhodothermus obamensis . The α-glucans synthesized from sucrose, a low cost agroresource, by the tandem action of the two enzymes, have been characterized by using complementary enzymatic, chromatographic, and imaging techniques. In a single step, linear and branched α-glucans were obtained, whose proportions, morphology, molar mass, and branching degree depended on both the initial sucrose concentration and the ratio between elongating and branching enzymes. In particular, spherical hyperbranched α-glucans with a controlled mean diameter (ranging from 10 to 150 nm), branching degree (from 10 to 13%), and weight-average molar mass (3.7 × 10(6) to 4.4 × 10(7) g.mol(-1)) were synthesized. Despite their structure, which is similar to that of natural glycogens, the mechanisms involved in their in vitro synthesis appeared to be different from those involved in the biosynthesis of native hyperbranched α-glucans.

SAXS Conformational Tracking of Amylose Synthesized by Amylosucrases

Amylose, a linear polymer of α(1,4)-linked glucosyl units and a major constituent of starch granules, can also be enzymatically synthesized in vitro from sucrose by bacterial amylosucrases. Depending on the initial sucrose concentration and the enzyme used, amylose oligomers (or polymers) are formed and self-associate during synthesis into various semicrystalline morphologies. This work describes for the first time a synchrotron SAXS study of the structure in solution of two amylosucrases, namely, NpAS and the thermostable DgAS, under conditions of polymer synthesis and, simultaneously, the amylose conformation. The structure in solution of both amylosucrases during the reaction was shown to be similar to the known crystallographic structures. The conformation of amylose produced at an early stage consists of a mixture of wormlike chains and double helical cylindrical structures. In the case of NpAS, in a second stage, individual double helices pack into clusters before crystallizing and precipitating. Amylose produced by DgAS never self-associates in such clusters due to the higher temperature used for amylose synthesis. All the dimensions determined for wormlike chains and cylindrical conformations at different times of NpAS synthesis are in very good agreement with structural features usually observed on gels of amylose extracted from starch. This provides new insights in understanding the mechanisms of amylose gelation.

Scale-up of adventitious root cultures of Echinacea angustifolia in a pilot-scale bioreactor for the production of biomass and caffeic acid derivatives

In an attempt to scale-up of adventitious root cultures of Echinacea angustifolia for the production of biomass and caffeic acid derivatives, i.e. echinacoside, chlorogenic acid, cichoric acid, caftaric acid, and cynarin, the effects of Murashige and Skoog (MS) medium dilutions, and initial sucrose concentrations were investigated in a 5-L airlift bioreactor. In addition, the kinetics of adventitious root growth and accumulation of secondary metabolites were also studied. The greatest root dry weight (6.50 g L−l) and accumulation of total phenolics [22.06 mg g−1 DW (dry weight)], total flavonoids (5.77 mg g−1 DW) and total caffeic acid derivatives (10.63 mg g−1 DW) were obtained at quarter-strength MS medium. Of the various gradients of sucrose tested, 5 % sucrose supplementation was regarded as an optimal concentration for enhancing productivity of biomass and bioactive compounds. Neither higher salt strength (3/4–2 MS) nor sucrose concentrations (7 and 9 %) showed promotive effect on root growth and metabolite production. The kinetic studies revealed that 4 weeks of culture period is the optimal time to achieve highest productivity of metabolites. Based on these results, a large-scale (20 L) and a pilot-scale (500 L) adventitious root culture system was established. In the pilot-scale bioreactor, adventitious roots were elicitor-treated with 100 μM methyl jasmonate (MJ) on day 28. After 1 week of elicitation, 1.75 kg dry root biomass was harvested containing 60.41 mg g−1 DW of total phenolics, 16.45 mg g−1 DW of total flavonoids, and 33.44 mg g−1 DW of total caffeic acid derivatives. Among the caffeic acid derivatives, the accumulation of echinacoside (the major bioactive compound) in MJ-treated adventitious roots grown in the 500-L bioreactor was the highest (12.3 mg g−1 DW), which is approximately threefold more than the non-MJ-treated roots cultured in 5- and 20-L bioreactors.