• Journal of Life Science
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• v.14 no.2
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• pp.235-238
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• 2004

The possibility for developing multistage culture system to cultivate larvae by computer simulation with basic experiments was investigated. This culture system was composed of 3 stages. At the 1st stage, Chlorella sp. were cultivated and at 2nd stage Chlorella sp. were supplied to rotifer (Brachionus plicatilis), and rotifer were supplied to larvae at the 3rd stage. In this study, Chlorella sp. were cultivated by batch culture to search for the possibility of continuous feeding to rotifer at 2 nd stage. The maximum specific growth rate ($\mu$$_{max}$) of Chlorella sp. at the logarithmic phase was 0.56 [1/day]. Rotifer was cultivated by fed-batch culture at the feeding rate of Chlorella sp., $10^3$, $10^4$, $10^{5}$ and $10^{6}$ [cells/rotiferㆍh] to search for the relation between the feeding rate of Chlorella sp. and the growth rate of rotifer. As the results, the minimum feeding rate of Chlorella sp. was 2.8 ${\times}$ $10^4$ [cells/rotiferㆍday] in the multistage culture system, then the change of rotifer concentration at 2nd stage was simulated by computer. The required amount of rotifer for the growth of larvae was also increased as the growth of larvae. On the 9th day of the culture, the rotifer uptake rate of larvae was 250 [cells/rotiferㆍday]. Based on these basic experiments and results, It was suggested that the possibility of multistage culture system to cultivate larvae with continuous feeding of Chlorella sp. and rotifer.r.

• KSBB Journal
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• v.13 no.2
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• pp.214-219
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• 1998

Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

• KSBB Journal
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• v.10 no.4
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• pp.401-405
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• 1995

An open pond type photobioreactor for mass cultivation of S. platensis was designed and the growth parameters from different cultivation processes were compared. 0.30(1/day) of specific growth rate and 1.69(g/$\ell$) of maximum cell density were obtained from batch cultivation. In fed-batch cultivation, specific growth rate and maximum cell density were estimated as 0.22(1/day) and 1.75(g/$\ell$), respectively. Maximum biomass productively from continuous cultivation was obtained as 0.44 (g/$\ell$/day). It proves that an outdoors-mass cultivation of S. platensis considering optimal environmental condition is economically feasible. In addition, the biomass productivity was studied in two different mixing systems such as agitation and air sparging methods. The biomass productivity by an agitation method was better than that in an air sparging method.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.451-456
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• 2004

The effects of variation in composition of the medium on the conversion of Gluconacetobacter hanseii PJK cells producing cellulose ($Cel^+$) to non-cellulose producing ($Cel^-$) mutants and the production of bacterial cellulose (BC) in an agitated culture were investigated. The impeller speed greater than 500 rpm was required to decrease the population of $Cel^-$ mutants to minimum in a basal medium containing $1.5\%$ ethanol because the optimum impeller speed to minimize the population of $Cel^-$ mutants increased with the concentration of ethanol added to a basal medium. Ethanol fed-batch culture could not increase the BC production in an agitated culture unlike that of a shaking culture. The amount of BC produced in a basal medium containing $1\%$ ethanol was $39\%$ more than that of the same medium with $0.27\%\;Na_{2}HPO_4$. Increase in the concentration of acetic acid in a basal medium decreased the BC production. The pH control of the culture broth increased the cell mass in the batch culture and improved the production yield of water-soluble polysaccharide (WSPS), but did not affect the production of BC.

• Membrane Journal
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• v.12 no.1
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• pp.21-27
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• 2002

The formation of soluble microbial products(SMP) and molecular weight distribution on loading rate were observed in batch-type culture medium, which phenol was fed as a substrate. The molecular weight destribution was obtained by using 30K, 100K dalton and $0.45{\mu}$ membrane filters. When the phenol concentrationas a substrate was 120, 230 and 440 mg/L , the specific substrate utilization rate(q) showed 0.639, 1.281 and 1.744 mgTOC/mgMLSS/day, respectively. The endogenous biomass decay rate constant($K_d$) at each substrate concentration was 0.00536, 0.0661 and 0.0749($day^1$), respectively. The $SMP_e$ product rate constant($k_{SMP}_ e$) showed 0.006, 0.0058 and 0.0057($day^1$), respectively. The initial influent substrate during the course of time degraded and produced $SMP_s$. The $SMP_s$ was converted to the $SMP_{nd}$ and endogenous phase converted to the $SMP_e$ ingredients. The molecula weight distribution on loading rate was converted to a higher MW during the course of time.

• Journal of Life Science
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• v.18 no.7
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• pp.980-985
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• 2008

This study was carried out to develope an alcoholic drink by fermentation of onion extract using Saccharomyces cerevisiae. The optimal conditions for ethanol production were obtained by standing culture at $25^{\circ}C$ for 5 days with 5% inoculum volume. At the results by flask culture, the growth curve of used S. cerevisiae reached to the stantionary phase at 48 hr and the death phase at 90 hr, whereas ethanol production reached maximum at 114 hr. Under the above conditions, a large scale production was carried out. A standing culture in 5 l fermenter showed the similar results to its flask culture, but progressed 24 hr rapidly more than that of the flask culture. A fed-batch culture was performed by addition of the onionic medium supplemented with 10% (v/v) sucrose after 72 hr from the fermenting start. The fed-batch culture could prevent S. cerevisiae from entering into the death phase and maintain constant level of alcohol production. A continuous culture was able to carry out by adding per every 24 hr the onionic medium supplemented with 10% (v/v) sucrose after 72 hr from the fermenting start. Although S. cerevisiae used showed a little decreased growth, alcohol production maintained roughly the constant level at the maximum yield. To enhance the quality of this alcoholic drink, $2-O-{\alpha}-D-glucopyranosyl$ L-ascorbic acid (AA-2G) was supplemented into the onion extract of the substrate for fermentation. As resulted at this study, this alcoholic drink containing AA-2G should be used as a functional fermented alcohol drink strengthened with vitamin C.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.53-56
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• 2000

Two-stage fed-batch culture was peformed to improve the volumetric productivity of erythritol. In the growth phase dissolved oxygen was maintained to 20% and the feed medium was automatically supplied to the fermenter by pH-stat mode. The cell yield was 0.76 g-cell/g-glucose. In two-stage fed-batch culture, 41% of total erythritol conversion yield with 187 g/L of erythritol concentration and 2.79 g/L-h of maximum erythritol Productivity were obtained when 400 g/L of glucose was directly added in the form of non-sterile powder at production phase. The erythritol productivity increased in parallel with cell mass. The metabolic shift in the biosynthetic pathway of erythritol was caused by dissolved oxygen concentration. The production of gluconic acid was observed when the dissolved oxygen in the medium was maintained over 40% during the production phase, whereas the dissolved oxygen concentration lower than 40% caused the production of citric acid. But the butyric acid was produced independently with dissolved oxygen concentration in the medium. The production of organic acids such as gluconic acid, citric acid, and butyric acid was decreased by addition of mineral salts.

• KSBB Journal
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• v.16 no.5
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• pp.474-479
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• 2001

Fed-batch culture, single stage and two stage continuous cultures of Corynebacterium glutamicum SH 35 for the production of $_{L}$-Iysine were performed and compared. In the case of fed batch culture, $_{L}$-Iysine concentration, $_{L}$-Iysine yield and $_{L}$-Iysine productivity was 129.2 g/L, 47.0% and 3.08 g/L/h, respectively. In a single-stage continuous culture, optimum dilution rate and pH was 0.1 h$^{-1}$ and 6.9, respectively, and optimum concentration of sugar and ammonium sulfate in a medium reservoir was 108 g/L and 25 g/L, respectively. Under the optimized conditions, 67 of cell concentration($OD_{610}$), 44.2 g/L of lysine concentration, 41% of $_{L}$-Iysine yield and 4.39 g$L^{-1}$ of $_{L}$-Iysine productivity were obtained. In a two-stage continuous culture, optimum dilution rate was 0.075 $h^{-1}$. Under the conditions, 103 of cell concentration($OD_{610}$) 84.0g/L of $_{L}$-Iysine concentration and 46% of $_{L}$-Iysine yield were obtained.

• Korean Journal of Microbiology
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• v.46 no.1
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• pp.46-51
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• 2010

Coenzyme Q10 (CoQ10) is an essential lipid-soluble component of membrane-bound electron transport chains. CoQ10 is involved in several aspects of cellular metabolism and is increasingly being used in therapeutic applications for several diseases. Despite the recent accomplishments in metabolic engineering of Escherichia coli for CoQ10 production, the production levels are not yet competitive with those by fermentation or isolation. So we tested several microorganisms obtained from the KCTC of Biological Resource Center to find novel sources of strain-development for CoQ10-production. Then we selected two strains, Paracoccus denitrificans (KCTC 2530) and Asaia siamensis (KCTC 12914), and tested to optimize the CoQ10 production conditions. Among the carbon sources tested, CoQ10 production was the highest when fructose was supplied about 4% concentration. Yeast extract produced the highest CoQ10 production about 2% concentration. The highest CoQ10 production was obtained at pH 6.0 for P. denitrificans and pH 8.0 for A. siamensis. And two strains showed the highest CoQ10 production at $30^{\circ}C$, but the highest DCW was obtained at $37^{\circ}C$. In the fed-batch culture, P. denitrificans yielded $14.34{\pm}0.473$ mg and A. siamensis yielded $12.53{\pm}0.231$ mg of final CoQ10 production.

• Journal of Microbiology and Biotechnology
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• v.16 no.3
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• pp.374-380
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• 2006

In this study, glucose, sucrose, and dextrin were found to be better carbon sources for the production of pullulan by Aureobasidium pullulans HP-2001. Maximal production of pullulan with 200 g/l sucrose as a carbon source was 54.2 g/l. The highest yield of pullulan from sucrose was 0.40, when the sugar concentration was 100 g/1. Optimal conditions for the continuous production of pullulan by A. pullulans HP-2001 in a 7-1 bioreactor were determined by studying the effects of composition of feed solution, dilution rate, and concentration of sucrose in the feed solution. Pullulan concentration and productivity with 100 g/l glucose and 2.5 g/l yeast extract were 38.1 g/l and 0.53 g/l h for 72 h, respectively, in a batch culture of A. pullulans HP-2001. When the substituted medium contained 100 g/l sucrose, 2.5 g/l yeast extract, and mineral salts, which is the same composition as the medium for the production of pullulan, the pullulan concentration and productivity were 74.9 g/l and 0.55 g/l h for 120 h, respectively. The production of pullulan at the steady state increased with a dilution rate up to 0.015/h, and its concentration was 78.4 g/l with a weight average molecular weight ($M_w$) of $4.0{\times}10^5$. Unlike a batch culture, however, the decline of the $M_w$ and the number average molecular weight ($M_n$) of pullulan was not found in the continuous culture of A. pullulans HP-2001. When the concentration of sucrose in the feed solution was 200 g/l, 113.5 g/l of pullulan was obtained at the steady state. The steady state was maintained longer in the continuous culture fed with the feed solution containing 200 g/l sucrose than when fed with the feed solutions containing either 100 or 150 g/l sucrose.