• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.3
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• pp.189-193
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• 2001

Critical cell density (CCD), the maximum cell concentration without mutual shading in algal cultures, can be used as a new operating parameter for high-density algal cultures and for the application of the flashing light effect on illuminated algal cultures. CCD is a function of average cell volume and light illumination area. The CCD is thus proposed as an index of estimation of mutual shading in algal cultures. Where cell densities are below the CCD, all the cells in photobioreactors can undergo photosysnthesis at their maximum rate. At cell densities over CCD, mutual shading will occur and some cells in the illumination chamber cannot grow photoautotrophically. When the cell concentration is higher than the CCD, specific oxygen production rates under flashing light were higher than those under continuous light. The CCD was found to be a useful engineering parameter for the application of flashing light, particularly in high-density algal cultures.

• Biotechnology and Bioprocess Engineering:BBE
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• v.4 no.1
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• pp.78-81
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• 1999

Light penetration depth in high-density Chlorella cultures can be successfully estimated by Beer-Lambert's law. The efficiency of light energy absorption algal cultures was so high that algal cells near the illuminating surface shade the cells deep in the culture. To exploit the potential of high-density algal cultures, this mutual shading should be eliminated or minimized. However, providing more light energy will not ease the situation and it will simply drop the overall light utilization efficiency.

• Biotechnology and Bioprocess Engineering:BBE
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• v.8 no.6
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• pp.313-321
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• 2003

This review summarizes the recent advances in high-density algal cultures in the field of algal biotechnology. Photobioreactor engineering for economical and effective utilization of algae and its products has made impressive and promising progress. Bioprocess engineers have expedited the design and the operation of algal cultivation systems. Many of them in use today are open systems due to cost considerations, and closed photobioreactors have recently attracted a considerable attention for the production of valuable biochemicals or for special applications. For high-density cultures, the optimization of environmental factors in the photobioreactors have been explored, including light delivery, CO$_2$and O$_2$gas transfer, medium supply, mixing and temperature. It is expected that further advanced photobioreactor engineering will enable the commercialization of noble algal products within the next decade.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.3
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• pp.186-190
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• 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.

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.817-822
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• 2000

A proper flashing light is expected to enhance microalgal biomass productivity and photosynthetic efficiency. The effect of flashing light on high-density Chlorella kessleri (UTEX 398) cultures was studied using light-emitting diodes. A frequency modulator was designed to flash LEDs, and the device successfully provided wide range of frequencies and various duty cycles of flashing. A relatively high frequencies of 10, 20 and 50 kHz were used in this study. These frequencies have very short flashing time ($2-50{\mu}s$), which corresponded to the time constant of the light reaction of photosynthesis. The specific oxygen production rates of photosynthesis under flashing light were compared with those under an equivalent continuous light in specially designed illumination cuvette. The specific oxygen production rates under flashing light were 5-25% higher than those under the continuous light. A range of cell concentration was discovered, where the benefit of flashing light was maximized. The photosynthetic efficiency was also higher under flashing light with frequencies of over 1 kHz, which was a clear indication of flashing light effect and the degree of mutual shading could by overcome by flashing lights, particularly at high-density algal cultures.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.5
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• pp.352-358
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• 2001

Because algal cells are so efficient at absorbing incoming light energy, providing more light energy to photobioreactors would simply decrease energy conversion efficiency. Furthermore, the algal biomass productivity in photobioreactor is always proportional to the total photosynthetic rate. In order to optimize the productivity of algal photobioreactors (PBRs), the oxygen production rate should be estimated. Based on a simple model of light penetration depth and algal photosynthesis, the oxygen production rate in high-density microalgal cultures could be calculated. The estimated values and profiles of oxygen production rate by this model were found to be in accordance with the experimental data. Optimal parameters for PBR operations were also calculated using the model.

• ALGAE
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• v.33 no.1
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• pp.127-141
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• 2018

Low efficiency in microalgal biomass production was largely attributed to the low density of algal cell cultures. Though mutations that reduced the level of chlorophyll or pigment content increased efficiency of photon usage and thus the cell-culture density under high-illumination growth conditions (e.g., >$500{\mu}mol\;photon\;m^{-2}\;s^{-1}$), it was unclear whether algae could increase cell-culture density under low-illumination conditions (e.g., ${\sim}50{\mu}mol\;photon\;m^{-2}\;s^{-1}$). To address this question, we performed forward genetic screening in Chlamydomonas reinhardtii. A pool of >1,000 insertional mutants was constructed and subjected to continuous subcultures in shaking flasks under low-illumination conditions. Complexity of restriction fragment length polymorphism (RFLP) pattern in cultures indicated the degree of heterogeneity of mutant populations. We showed that the levels of RFLP complexity decreased when cycles of subculture increased, suggesting that cultures were gradually populated by high cell-culture density (HCD) strains. Analysis of the 3 isolated HCD mutants after 30 cycles of subcultures confirmed that their maximal biomass production was 50-100% higher than that of wild type under low-illumination. Furthermore, levels of chlorophyll content in HCD mutant strains were similar to that of wild type. Inverse polymerase chain reaction analysis identified the locus of insertion in two of three HCD strains. Molecular and transcriptomic analyses suggested that two HCD mutants were a result of the gain-of-function phenotype, both linking to the abnormality of mitochondrial functions. Taken together, our results demonstrate that HCD strains can be obtained through continuous subcultures under low illumination conditions.

• KSBB Journal
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• v.14 no.2
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• pp.160-166
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• 1999

High cell density cultivation of microalga Dunaliella bardawil using nitrogen fed-batch cultures was studied in batch flask. Optimum environmental conditions include concentrated nutrients except NaCl and carbon sources, carbon sources, pH, light, agitation, nitrate and phosphate ions. Cell growth, consumption rates of nitrate and phosphate ions were monitored. Optimal conditions for higher cell density were found to be(in the range tested): 5 times concentrated media(1 times-10 times concentrated media) pH 8.0 (7.0-9.0) white light(blue and red light) 15mM of nitrate (0.94-15mM) 250mM $NaHCO_3$ and $CO_2$ gas. However, the addition of phosphate ions did not enhance the algal maximum cell density and specific growth rate. Nitrate was found to be effective for the cell growth. The maximum cell density of fed-batch culture using nitrate ions in $8.955{\times}106$cells/ml after 189hr incubation.

• ALGAE
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• v.35 no.3
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• pp.277-292
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• 2020

Omega-3 fatty acids, including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), are polyunsaturated fatty acids beneficial to human health. A limited number of microalgae have been used for commercial omega-3 production, which necessitates the identification of new microalgae with high omega-3 contents. We explored the fatty acid composition and EPA and DHA contents of the mixotrophic dinoflagellate Gymnodinium smaydae fed with the optimal algal prey species Heterocapsa rotundata. Cells of G. smaydae were found to be rich in omega-3 fatty acids. In particular, the DHA content of G. smaydae was 21 mg g^-1 dry weight, accounting for 43% of the total fatty acid content. The percentage of DHA in the total fatty acid content of G. smaydae was the highest among the reported microalgae except for Crypthecodinium cohnii. Moreover, to determine if the prey supply interval affected the growth rate of G. smaydae and its fatty acid content, three different prey supply intervals (daily, once every 2 d, and once for 4 d) were tested. Daily prey supply yielded the highest total fatty acid and DHA contents in G. smaydae. Furthermore, we successfully produced high-density G. smaydae cultures semi-continuously for 43 d with daily prey supply. During the semi-continuous cultivation period, the highest density of G. smaydae was 57,000 cells mL^-1, with an average growth rate of 0.7 d^-1. Taken together, the percentage of EPA and DHA in the total fatty acid content was maintained in the range of 54.2-56.9%. The results of this study support G. smaydae as a promising microalgal candidate for commercial DHA production and demonstrate that daily supply of prey can efficiently produce high-density G. smaydae cultures for more than a month.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.3
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• pp.194-199
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• 2001

Chlorella kessleri cultivated in artificial wastewater using diurnal illumination of 12h light/12h dark (L/D) cycles. The inoculum density was 10(sup)5 cells/mL and the irradiance in light cycle was 45$\mu$mol㎡s(sup)-1 at the culture surface. As a control culture, another set of flasks was cultivated under continuous illumination. Regardless of the illumination scheme, the total organic carbon (TOC) and chemical oxygen demand (COD) was reduced below 20% of the initial concentration within a day. However, cell concentration under the L/D lighting scheme was lower tan that under the continuous illuminating scheme. Thus the specific removal rate of organic carbon under L/D cycles was higher than that under continuous illumination. This result suggested that C. kessleri grew chemoorganotrophically in the dark periods. After 3 days, nitrate was reduced to 136.5 and 154.1mg NO$_3$-N/L from 168.1mg NO$_3$-N/L under continuous illumination and under diurnal cycles, respectively. These results indicate nitrate removal efficiency under continuous light was better than that under diurnal cycles. High-density algal cultures using optimized photobioreactors with diurnal cycles will save energy and improve organic carbon sources removal.