• Biotechnology and Bioprocess Engineering:BBE
• /
• 제5권3호
• /
• pp.186-190
• /
• 2000

It has been reported that flashing light enhances microalgal biomass productivity and overall photosynthetic efficiency. The algal growth kinetics and oxygen production rates under flashing light with various flashing frequencies (5Hz-37 kHz) were compared with those under equivalent continuous light in photobioreactors. A positive flashing light effect was observed with flashing frequencies over 1kHz. The oxygen production rate under conditions of flashing light was slightly higher than that under continuius ligth. The cells under the hight, particularly at higher cell concentrations. When 37kHz flashing light was applied to an LED-based photobioreactor, the concentration was higher than that obtained under continuous light by about 20%. Flashing light may be a reasonable solution to overcome mutual shading, particularly in high-density algal cultures.

• KSBB Journal
• /
• 제6권1호
• /
• pp.1-7
• /
• 1991

To produce economically important indole alkaloids by cell cultures, we selected protoplastsderived clones (protoclones) of vinca (Catharanthus roseus) for high yields of catharanthine and ajmalicine. Protoplasts were enzymatically isolated from suspension-cultured cells. The highest plating efficiency (1%) was obtained when protoplasts were plated at a density of 1$\times$105 protoplasts/ml in a culture medium solidified with 0.4% Seaplaque agarose. The growth rates of 40 protoclones subcultured on a solid medium varied over a wide range. Protoclone VPC-6, which had the highest growth rate, was observed to produce relatively high yields of catharanthine and ajmalicine when cultured in a liquid medium. Although the original cell line did not produce catharanthine at a detectable level by HPLC, protoclone VPC-10 produced it at a level of 5.9$\mu\textrm{g}$/g fresh weight of cells for 10 days of culture. Under the same conditions, protoclone VPC-15 produced ajmalicine at a level of 133.6$\mu\textrm{g}$/g, of which productivity was improved about ,3 times than that of the original cell line. The results indicate that differences in the growth rate and indole alkaloid yield among the protoclones reflect the somaclonal variation in suspnsion-cultured cells.

• 한국미생물·생명공학회지
• /
• 제24권6호
• /
• pp.717-725
• /
• 1996

We have developed an efficient immobilized cell separator for continuous operation of immobilized fungal cell cultures, and applied this separator to actual fermentation process for the production of cyclosporin A (CyA), a powerful immunosuppressant. In the experiments employing highly viscous polymer (carboxymethyl cellulose) solution, the decantor showed good separating performances at high solution viscosites and fast dilution rates. Air duct and cylindrical separator installed inside the decantor turned out to play key roles for the efficient separation of the immobilized cells. By installing the decantor in an immobilized perfusion reactor system (IPRS), continuous immobilized culture was stably carried out even at high dilution rate for a long period, leading to high productivities of free cells and CyA. Almost no immobilized biomass existed in effuluent stream of the IPRS, demonstrating the effectiveness of the decan- tor system for a long-term continuous fermentation. It was noteworthy that we could obtain these results despite of the unfavorable fermentation conditions, i.e., reduced density of the biosupports caused by overgrowth of cells inside the bead particles and existence of high density of suspended fungal cells (10g/l) in the fermentation broth.

• Journal of Microbiology and Biotechnology
• /
• 제18권7호
• /
• pp.1252-1256
• /
• 2008

Cell recycled culture of succinic acid-producing Anaerobiospirillum succiniciproducens was anaerobically carried out using an internal membrane filter module in order to examine the physiological response of A. succiniciproducens to a high-cell-density environment. The optimal growth of A. succiniciproducens and its enhanced succinic acid productivity were observed under $CO_2$-rich conditions, established by adding $NaHCO_3$ and $Na_2CO_3$, in the cell recycled system. A. succiniciproducens grew up to 6.50 g-DCW/l, the highest cell concentration obtained so far, in cell recycled cultures. The cells did not change their morphology, which is known to be easily changed in unfavorable or stress environments. The maximum productivity of succinic acid was about 3.3 g/l/h, which is 3.3 times higher than those obtained in batch cultures. These results can serve as a guide for designing highly efficient cell recycled systems for succinic acid at a commercial level.

• Journal of Microbiology and Biotechnology
• /
• 제12권3호
• /
• pp.444-448
• /
• 2002

A $CaCO_3$-alginate beads system was developed for high cell density cultivation of Bifidobacterium longum and the cost-effective media were also screened. In batch process with $CaCO_3$, beads, two strains of B. longum showed both the highest viable cells and optical density in TPY medium, resulting in maximum optical density and viable cell counts of 12.40, $2.22{\times}10^10$ cfu/ml for B. longum ATCC 15707 and 13.71, $3.93{\times}10^10$ cfu/ml for B. longum HLC 3742. Released size distribution, according to $CaCO_3$-alginate bead size preparation, was smaller than others. These results were also examined by observing their morphology. The skim milk-based medium was most adequate to cultivate B. longum as the cheapest medium, and $10\%$ skim milk supplemented with $2\%$ glucose and $1\%$ yeast extract was a suitable medium, supporting the growth to $5.57{\times}10^10$ cfu/ml for ATCC 15707 and $6.82{\times}10^9$ cfu/ml for HLC 3742. During the long-term storage at $4^{\circ}C\;and\;-20{\circ}C$, B. longum cultivated with $CaCO_3$ beads had the highest stability. Consequently, $CaCO_3$-alginate beads buffer was found to be useful not only to cultivate B. longum but also to preserve cultures.

• Journal of Microbiology and Biotechnology
• /
• 제20권11호
• /
• pp.1534-1538
• /
• 2010

Fed-batch cultures of Hansenula polymorpha were studied to develop an efficient biosystem to produce recombinant human serum albumin (HSA). To comply with this purpose, we used a high-purity oxygen-supplying strategy to increase the viable cell density in a bioreactor and enhance the production of target protein. A mutant strain, H. polymorpha GOT7, was utilized in this study as a host strain in both 5-l and 30-l scale fermentors. To supply high-purity oxygen into a bioreactor, nearly 100% high-purity oxygen from a commercial bomb or higher than 93% oxygen available in situ from a pressure swing adsorption (PSA) oxygen generator was employed. Under the optimal fermentation of H. polymorpha with highpurity oxygen, the final cell densities and produced HSA concentrations were 24.6 g/l and 5.1 g/l in the 5-l fermentor, and 24.8 g/l and 4.5 g/l in the 30-l fermentor, respectively. These were about 2-10 times higher than those obtained in air-based fed-batch fermentations. The discrepancies between the 5-l and 30-l fermentors with air supply were presumably due to the higher contribution of surface aeration over submerged aeration in the 5-l fermentor. This study, therefore, proved the positive effect of high-purity oxygen in enhancing viable cell density as well as target recombinant protein production in microbial fermentations.

• Journal of Microbiology and Biotechnology
• /
• 제11권6호
• /
• pp.1024-1030
• /
• 2001

The key factors for high-density Haematococcus pluvialis cultures and conditions for astaxanthin induction were examined to maximize astaxanthin production. Light intensity was found to be the most important factor, and thus experiments were found to be the most important factor, and thus experiments were carried out using different light sources and intensities. A high cell density of over 2.7 g/l was obtained at $75{\mu}E/m^2/s$, whereas a much lower cell concentration (<1.0 g/ 1) was obtained with lower light intensities $(15-30{\mu}E/m^2/s$. A high light intensity and the supplement of 470 nm photons had a more dramatic effect on the final astaxanthin concentration and per cell astaxanthin content. A maximum astaxanthin concentration of 6.5 mg/l was obtained at a light intensity of $160{\mu}E/m^2/s$, whereas only 1.3 and 0.7 mg/l were obtained at 30 and $15{\mu}E/m^2/s$, respectively. A supplement of 470 nm photons enhanced the carotenoid and chlorophyll formation.

• Journal of Microbiology and Biotechnology
• /
• 제19권12호
• /
• pp.1695-1702
• /
• 2009

An animal-component-free and chemically defined fed-batch process for GS-engineered cell lines producing recombinant antibodies has been developed. The fed-batch process relied on supplying sufficient nutrients to match their consumption, simultaneously minimizing the accumulation of by-products (lactate and osmolality). The proportionalities of nutritional consumption were determined by direct analysis. The robust, metabolically responsive feeding strategy was based on the offline measurement of glucose. The fed-batch process was shown to perform equivalently in GS-CHO and GS-NS0 cultures. Compared with batch cultures, the fed-batch technology generated the greater increase in cell yields (5-fold) and final antibody concentrations (4-8-fold). The majority of the increase in final antibody concentration was a function of the increased cell density and the prolonged culture time. This generic and high-yielding fed-batch process would shorten development time, and ensure process stability, thereby facilitating the manufacture of therapeutic antibodies by GS-engineered cell lines.

• Journal of Microbiology and Biotechnology
• /
• 제16권12호
• /
• pp.1890-1896
• /
• 2006

Lumostatic operation for cultivation of Haematococcus pluvialis was assessed to test the scale-up strategy of photobioreactors. Lumostatic operation is a method of maintaining a proper light condition based on the specific light uptake rate ($q_e$), by cells. Lumostatic operations were performed in 0.4-, 2-, 10-, and 30-1 scale bubble column photobioreactors and the results were compared with cultures illuminated with constant light intensity. Significant differences were observed in the maximal cell concentrations obtained from 0.4-, 2-, 10-, and 30-1 scale photobioreactors under constant light intensity, yielding the maximal cell concentrations of $2.8{\times}10^5$, $2.2\times10^5$, $1.5\times10^5$, and $1.1\times10^5$ cells/ml, respectively. The maximal cell concentration in a 0.4-1 photobioreactor under lumostatic operation was $4.3\times10^5$ cells/ml. Furthermore, those in 2-, 10-, and 30-1 scale photobioreactors were about the same as that in the 0.4-1 photobioreactor. The results suggest that lumostatic operation with proper $q_e$ is a good strategy for increasing the cell growth of Haematococcus pluvialis compared with a constant supply of light energy. Therefore, lumostatic operation is not only an efficient way to achieve high cell density cultures with minimal power consumption in microalgal cultures but it is also a perfect parameter for the scale-up of photobioreactors.

• KSBB Journal
• /
• 제13권3호
• /
• pp.268-271
• /
• 1998

Bacteriorhodopsin in the purple membrane (PM) of halobacteria has recently been attracting much attention to be used as a component of molecular electron device and optical computers. In order to increase the productivity of bacteriorhodopsin in high cell density cultures of Halobacterium halobium R1, an internal membrane cell-retention bioreactor system was employed. As a result, the production of cell mass at OD660 of 12 and of bacteriorhodopsin at 125-130 mg/L were obtained using the internal membrane bioreactor system at a dilution rate of 0.066 hr-1. The productivity achieved by the internal membrane system (0.7 mg/L$.$hr) was 3.5-fold higher than that obtained by the corresponding batch cultivations (0.2 mg/L$.$hr).