• Journal of Environmental Health Sciences
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• v.29 no.3
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• pp.72-78
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• 2003

In the process of producing chitosan from crustacean shell, the use of excessive acid and alkli is causing the problems of environmental pollution and of production cost. In this study, one way to solve these problems is to cultivate fungi, then, to extract chitosan from the cell wall. By means of flask incubation and batch cultivation, the optimum cultivation conditions for mass production of continuous cultivation was found. Four strains used for the production of fungal chitosan were Gongronella butleri IF08080, Absidia coerulea IF05301, Rhizopus delemar IF04775, Mucor tuberculisporus IF09256. In flask incubation to select strain of producing much chitosan by means of experiment of the effect of initial pH, Absidia coerulea IFO 5301 had highest yield in FCs, 258.1 $\pm$ 47.3 mg/200 $m\ell$l at pH 6.5. In flask incubation under the optimum cultivation condition, temperature 27$^{\circ}C$, culture time 6days, glucose 2%, peptone 1%, (NH$_4$)$_2$ SO$_4$ 0.5%, $K_2$HPO$_4$ 0.1 %, Nacl 0.1 %, MgSO$_4$ㆍ7$H_2O$ 0.05%, CaCl$_2$ㆍ2$H_2O$ 0.01 %, the yield of DCW brought the highest yields. In batch bioreactor, the optimum cultivation condition was that cell suspended solution was 70 $m\ell$, aeration rate 0.5 l/min, agitation rate 800 rpm, culture time 36 hr. In continuous bioreactor, the optimum substrate flow rate was 4 ι/day.

• Journal of Plant Biotechnology
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• v.37 no.1
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• pp.110-114
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• 2010

Highest increase of biomass was observed when tissue-cultured potato (Solanum tuberosum L. cv. Chubaek) shoots were cultured in a liquid medium containing 1/3 MS solution in a 18 L bioreactor, as compared to 1/4 and 1/2 MS solution. The medium containing 1/4 MS solution showed higher increase of shoot biomass than one containing 1/2 MS solution. Potato microtubers were formed when the medium was exchanged with the medium for microtuber formation and incubated under dark condition. The microtubers were observed first at some axillary buds one week after incubation under dark condition and then at most of the axillary buds by the end of 3 weeks. The 1.5 MS liquid medium and $20^{\circ}C$ were optimal conditions. By the end of 6 weeks, more 1,000 microtubers were formed in the 18 L bioreactor. Then, greened microtubers were harvested after one week culture under light condition.

• Journal of Hydrogen and New Energy
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• v.15 no.2
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• pp.166-174
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• 2004

Purple non-sulfur bacteria, Rhodobacter sphaeroides KD131 grew to reach the maximum cell concentration in 45 hrs of incubation in the synthetic media containing (NH4)2SO4, L-aspartic acid and succinic acid as the carbon and nitrogen sources, respectively, at 30oC under 8 klux irradiance using halogen lamp. The strain produced hydrogen from the middle of the logarithmic growth phase and continued until the cell growth leveled out. The strain grew and produced hydrogen under the irradiance of 3-30 klux, but cell growth was inhibited over 100 klux. In addition, anaerobic/light culture condition was better than the aerobic/dark on the hydrogen production. Among various photo-bioreactors examined, the flat-vertical reactor manufactured using clear acrylic plastic material showed the best hydrogen production rate at the given culture condition.

• Journal of Plant Biotechnology
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• v.33 no.2
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• pp.117-122
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• 2006

Comparative studies on medium supply in bioreactors (raft, immersion and ebb and flood) have revealed that multiplication and growth of Alocasia Amazonica were greatest in the raft system, while lowest in ebb and flood system. In the raft system, the basal part of the shoots was continuously in contact with medium, which enabled a constant uptake of nutrients as well as aeration to the explants. The number and the size of leaf stomata were higher in the raft system compared with immersion and ebb&flood system. In the immersion system, plantlets were deformed and epidermal cells in leaves were irregular with a large intercellular space. The results suggested that the medium supply should be controlled properly to maintain normal and healthy plantlets during liquid cultures in bioreactors Which affects morphology and physiology Of the plantlets.

• KSBB Journal
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• v.31 no.4
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• pp.284-290
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• 2016

In this research, recombinant human cytotoxic T-lymphocyte antigen 4-immunoglobulin (hCTLA4Ig) was produced by transgenic rice cells. RAmy3D promoter was used for overcome the limitation of low expression level in transgenic plant cells, and the secretion of target protein was accomplished by signal peptide. However, the RAmy3D promoter system which can be induced only by sugar starvation causes the decrease of cell viability. As a result, cell death promotes the release of protease which degrades the target proteins. The protein stability and productivity can be significantly influenced by proteolysis activity. Therefore, development of new strategies are necessary for the in situ recovery of target proteins from cell culture media. In this study, in situ recovery was performed by various strategies. Direct addition of Protein A resin with nylon bag leads to cell death by increased shear stress and decrease in production of hCTLA4Ig by protease. Medium exchange through modified flask could recover hCTLA4Ig with high cell viability and low protease activity, on the other hand, the productivity was lower than that of control. When in situ recovery was conducted at day 7 after induction in air-lift bioreactor, 1.94-fold of hCTLA4Ig could be recovered compared to control culture without in situ recovery. Consequently, in situ recovery of hCTLA4Ig from transgenic rice cell culture could enhance productivity significantly and prevent degradation of target proteins effectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.3
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• pp.163-170
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• 2002

This report describes the use of a transtubular bioreactor to study the relative effects of diffusion versus perfusion of medium on antibody production by a hybridoma cell line. The study was performed with a high-density cell culture maintained in a serum-free, low-protein medium for 77 days. It was determined that the reactor possessed a macro-mixing pattern residence time distribution similar to a continuous stirred tank reactor (CSTR), However, due to the arrangement of the medium lines in the reactor, the flow patterns for nutrient distribution consist of largely independent medium path lengths ranging from short to long. When operated with cyclic, reversing, transtubular medium flow, some regions of the reactor (with short residence times) are more accessible to medium than others (with long residence times). From this standpoint, the reactor can be divided into three regions: a captive volume, which consists of medium primarily delivered via diffusion; a lapped volume, which provides nutrients through unilateral convection; and a swept volume, which operates through bilateral convection. The relative sizes of these three volumes were modified experimentally by changing the period over which the direction of medium flow was reversed from 15 min (larger captive volume) to 9 h (larger swept volume). The results suggest that antibody concentration increases as the size of the diffusion-limited (captive) volume is increased to a maximum at around 30 min with a sharp decrease thereafter. As reflected by changes in measured consumption of glucose and production of lactate, no significant difference in cellular metabolism occurred as the reactor was moved between these different states. These results indicate that the mode of operation of the transtubular bioreactor may influence antibody productivity under serum-free, low-protein conditions with minimal effects on cellular metabolism.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.443-449
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• 1999

Bacillus subtilis BK-17 which produces a novel protease with fibrinolytic activity was isolated from soybean paste. Bioreactor production of the enzyme was studied in order to optimize fermentation conditions such as medium concentration, pH, agitation speed, and temperature. Under most cultural conditions, enzyme production initially began when the cell growth stopped. The onset of the enzyme production was indicated by rapid increase in both dissolved oxygen (DO) and pH. Two- to three-times more concentrated medium than the flask optimum medium yielded higher enzyme production in the bioreactor fermentation. When the medium pH was controlled constant, pH 6.5 exhibited the highest activity in the range of 6.0 to 7.5, but the activity was similar to the case when the pH was initially adjusted to 7.5 and subsequently maintained within a relatively wide range of 6.4 to 7.8. Agitation speed did not affect the enzyme production with an exception of DO reaching zero. Fermentation time was reduced when temperature increased within the range of $25^{\circ}C$ to$37^{\circ}C$. However, the highest activity, along with the slow decrease of the enzymatic activity after reaching the maximum value, was observed at $25^{\circ}C$. By shifting the temperature from $37^{\circ}C$ to $25^{\circ}C$immediately after DO reached the minimum level, the high enzyme production of 1,100 U/ml along with the short fermentation period of 13 h could be obtained.

• Korean Journal of Plant Tissue Culture
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• v.21 no.2
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• pp.77-84
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• 1994

To assess the feasibility of anthocyanin production in cell cultures of Euphorbia splendens Bojer the role of sucrose in pigment production was investigated and pilot scale cultures were attempted to establish mass production system. And also, several instrumental analyses were conducted to identify the pigment extracted from cultured rolls. Anthocyanin production was promoted prominently with concenetrations of sucrose ranging from 3% to 9% while cell growth was maximized at 3% of sucrose . This suggested that high osmolarity of sucrose enhance pigment production. When cells were cultured in two types of bioreactor better cell growth was achieved with draft-type air lift bioreactor than impeller type bioreactor and the pigment productivity was reached to 2.2 mg/L/day. The major pigment extracted from cultured cells was characterized as cyanidin-3-glucoside.

• Journal of Microbiology and Biotechnology
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• v.27 no.7
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• pp.1281-1287
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• 2017

Bone morphogenetic protein-4 (BMP-4) is considered to have therapeutic potential for various diseases, including cancers; however, the high expression of biologically active recombinant human BMP-4 (rhBMP-4) needed for its manufacture for therapeutic purposes has yet to be established. In the current study, we established a recombinant Chinese hamster ovary (rCHO) cell line overexpressing rhBMP-4 as well as a production process using 7.5-l bioreactor (5 L working volume). The expression of the mature rhBMP-4 was significantly enhanced by recombinant furin expression. The combination of a chemically defined medium and a nutrient supplement solution for high expression of rhBMP-4 was selected and used for bioreactor cultures. The 11-day fed-batch cultures of the established rhBMP-4-expressing rCHO cells in the 7.5-L bioreactor produced approximately 32 mg/l of rhBMP-4. The mature rhBMP-4 was purified to homogeneity from the culture supernatant using a two-step chromatographic procedure, resulting in a recovery rate of approximately 55% and a protein purity greater than 95%. The N-terminal amino acid sequences and N-linked glycosylation of the purified rhBMP-4 were confirmed by N-terminal sequencing and de-N-glycosylation analysis, respectively. The mature purified rhBMP-4 has been proved to be functionally active, with an effective dose concentration of $EC_{50}$ of 2.93 ng/ml.

• Biotechnology and Bioprocess Engineering:BBE
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• v.7 no.3
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• pp.138-149
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• 2002

Plant cell culture provides a viable alternative over whole plant cultivation for the production of secondary metabolites. In order to successfully cultivate the plant cells at large scale, several engineering parameters such as, cell aggregation, mixing, aeration, and shear sensitivity are taken into account for selection of a suitable bioreactor. The media ingredients, their concentrations and the environmental factors are optimized for maximal synthesis of a desired metabolite. Increased productivity in a bioreactor can be achieved by selection of a proper cultivation strategy (batch, fed-batch, two-stage etc.), feeding of metabolic precursors and extraction of intracellular metabolites. Proper understanding and rigorous analysis of these parameters would pave the way towards the successful commercialization of plant cell bioprocesses.