• Journal of Marine Bioscience and Biotechnology
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• v.10 no.2
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• pp.73-82
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• 2018

Chlorella are source of valuable compounds as lipids, proteins, carbohydrates, bioactive compounds. To continuous obtain the high-value materials of Chlorella protothecoides, we performed continuous cultivation after applying milking techniques to C. protothecoides grown with culture for 7 days in optical panel bioreactor (OPBR) system. Fatty acid and lutein in extracts from pre- and post-milking of C. protothecoides were analyzed using gas chromatography and high performance liquid chromatography, respectively. C. protothecoides were rich in unsaturated fatty acids with a high content of oleic acid(C18:1), which is suitable as a biofuel feedstock. The fatty acid content in pre- and post-milking of C. protothecoides was decreased from 126.424mg/g d.w. to 119.341mg/g d.w, and the lutein content decreased from 0.258mg/g d.w. to 0.178mg/g d.w. The results of this study demonstrate the feasibility of milking C. protothecoides for production of lipids for biofuels production. It was confirmed that microalgae can continuously obtain lutein present in a trace amount through a continuous culture from milking.

• KSBB Journal
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• v.8 no.5
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• pp.495-503
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• 1993

To investigate the characteristics of growth and oil production of peppermint cells during a batch culture, cells derived from peppermint callus was cultivated in an air bubble reactor. During the batch culture, effects of inoculum size, abiotic stress, yeast elicitor, and two stage culture on the cell growth, the productivity of oleolesin, and the formation of flavor components were determined and also the sugar concentrations and kinetics of cell growth were analyzed. Among the various sizes of inoculum, the culture with 2.0% packed cell volume inoculum showed the optimum condition for cell growth in the proposed bioreactor, and the cell yield and essential oil production reached to 5.7g/1 and 0.109g/1, respectively. When the abiotic stress of daily 8hr dark and $10^{\circ}C$ cold treatments were given to the culture cell growth decreased but essential oil production increased to 0.546g/l. In a modified Lin-Staba medium in which 100mg/l yeast extract as an elicitor was added to the culture, the cell growth and oil production increased, and menthol content was 22.5% of oil. In the two stage culture, in which the basic culture conditions of 27$^{\circ}C$, light, and without elicitor were employed during the first six days followed by the second stage with daily 8hr treatment of cold and dark condition, and also with yeast extract as an elicitor, cell growth decreased after eight days, essential oil production was not increased, and menthol was not detected. Dry cell yield was 0.38g dry cell/g sugar and specific growth rate was 0.25 day-1. The major terpenoid in the oil was not the menthol but pulegone and piperitone, precursors of menthol were accumulated. However, when yeast elicitor was added, menthol was produced to the level of 22.5% which was the highest value in the peppermint cell culture reported so far.

• Korean Journal of Medicinal Crop Science
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• v.14 no.6
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• pp.371-375
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• 2006

To investigate effects on the growth and ginsenosides accumulation in ginseng hairy root, potassium phosphate was supplemented with 1.25, 2.5, and 5.0 mM concentration in 1/2 MS medium, respectively. Potassium phosphate supplement was increased the biomass and ginsenosides accumulation when it was dose at the concentration of 1.25 mM. And the growth rate of hairy root in the light condition was higher than in dark condition. The highest contents and productivity of ginsenosides were observed at the supplement of 1.25 mM potassium phosphate at the 7th day after the culture onset. Ginseng hairy root cultured in 20 L bioreactor supplemented with 1.25 mM KH$_2$PO$_4$ was increased the growth with 1,609 g (F${\cdot}$W) and ginsenosides content with 11.09 mg than those in control.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.6
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• pp.806-813
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• 2010

Animal bioreactors have been regarded as alternative tools for the production of limited human therapeutic proteins. The mammary glands of cattle are optimal tissues to produce therapeutic proteins that cannot be produced in large amounts in traditional systems based on microorganisms and eukaryotic cells. In this study, two knock-in vectors, pBCTPOKI-6 and pBCTPOKI-10, which target the hTPO gene on the bovine beta-casein locus, were designed to develop cloned transgenic cattle. The pBCTPOKI-6 and pBCTPOKI-10 vectors expressed hTPO protein in culture medium at a concentration of 774 pg/ml and 1,867 pg/ml, respectively. Successfully, two targeted cell clones were obtained from the bovine fibroblasts transfected with the pBCTPOKI-6 vector. Cloned embryos reconstructed with the targeted nuclei showed a lower in vitro developmental competence than those with the wild-type nuclei. After transfer of the cloned embryos into recipients, 7 pregnancies were detected at 40 to 60 days of gestation, but failed to develop to term. The results are the first trial for targeting of a human gene on the bovine milk protein gene locus, providing the potential for a large-scale production of therapeutic proteins in the animal bioreactor system.

• Journal of Microbiology and Biotechnology
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• v.21 no.9
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• pp.960-967
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• 2011

Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett-Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature ($30^{\circ}C$), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

• Journal of Microbiology and Biotechnology
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• v.22 no.2
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• pp.199-206
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• 2012

Naturally immobilized tannase (tannin acyl hydrolase, E.C. 3.1.1.20) has many advantages, as it avoids the expensive and laborious operation of isolation, purification, and immobilization, plus it is highly stable in adverse pH and temperature. However, in the case of cell-associated enzymes, since the enzyme is associated with the biomass, separation of the pure biomass is necessary. However, tannic acid, a known inducer of tannase, forms insoluble complexes with media proteins, making it difficult to separate pure biomass. Therefore, this study optimizes the production of cell-associated tannase using a "protein-tannin complex" free media. An exploratory study was first conducted in shake-flasks to select the inducer, carbon source, and nitrogen sources. As a result it was found that gallic acid induces tannase synthesis, a tryptose broth gives higher biomass, and lactose supplementation is beneficial. The medium was then optimized using response surface methodology based on the full factorial central composite design in a 3 l bioreactor. A $2^3$ factorial design augmented by 7 axial points (${\alpha}$ = 1.682) and 2 replicates at the center point was implemented in 17 experiments. A mathematical model was also developed to show the effect of each medium component and their interactions on the production of cell-associated tannase. The validity of the proposed model was verified, and the optimized medium was shown to produce maximum cell-associated tannase activity of 9.65 U/l, which is 93.8% higher than the activity in the basal medium, after 12 h at pH 5.0, $30^{\circ}C$. The optimum medium consists of 38 g/l lactose, 50 g/l tryptose, and 2.8 g/l gallic acid.

• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.717-725
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• 2021

This study aimed to optimize medium composition and culture conditions for enhancing the biomass of Lactobacillus plantarum 200655 using statistical methods. The one-factor-at-a-time (OFAT) method was used to screen the six carbon sources (glucose, sucrose, maltose, fructose, lactose, and galactose) and six nitrogen sources (peptone, tryptone, soytone, yeast extract, beef extract, and malt extract). Based on the OFAT results, six factors were selected for the Plackett-Burman design (PBD) to evaluate whether the variables had significant effects on the biomass. Maltose, yeast extract, and soytone were assessed as critical factors and therefore applied to response surface methodology (RSM). The optimal medium composition by RSM was composed of 31.29 g/l maltose, 30.27 g/l yeast extract, 39.43 g/l soytone, 5 g/l sodium acetate, 2 g/l K₂HPO₄, 1 g/l Tween 80, 0.1 g/l MgSO₄·7H₂O, and 0.05 g/l MnSO₄·H₂O, and the maximum biomass was predicted to be 3.951 g/l. Under the optimized medium, the biomass of L. plantarum 200655 was 3.845 g/l, which was similar to the predicted value and 1.58-fold higher than that of the unoptimized medium (2.429 g/l). Furthermore, the biomass increased to 4.505 g/l under optimized cultivation conditions. For lab-scale bioreactor validation, batch fermentation was conducted with a 5-L bioreactor containing 3.5 L of optimized medium. As a result, the highest yield of biomass (5.866 g/l) was obtained after 18 h of incubation at 30℃, pH 6.5, and 200 rpm. In conclusion, mass production by L. plantarum 200655 could be enhanced to obtain higher yields than that in MRS medium

• Journal of Mushroom
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• v.21 no.4
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• pp.209-214
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• 2023

In this study, Pleurotus ostreatus No.42 was cultured in glucose-peptone-yeast-wheat bran medium using a previously reported novel rotary draft tube bioreactor. Versatile peroxidase (VP), a lignin-degrading enzyme, was isolated from a pellet-type mycelium culture grown in the medium for seven days. The VP was purified by sequentially applying ultra-filtration, DEAE-Sepharose CL-6B column, and Mono Q column. SDS-PAGE analysis revealed the molecular weight of VP to be 36.4 KDa with an isoelectric point of 3.65. The amino acid sequence was confirmed as VTCATGQTT. The purified VP was observed to possess the property of not only oxidizing Mn ions but also decomposing veratryl alcohol, a non-phenolic compound. The catalytic ability of VP is a subject for future research.

• Applied Biological Chemistry
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• v.36 no.5
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• pp.332-338
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• 1993

For the production of riboflavin, strain development of Ashbya gossypii NRRL Y-1056 was attempted by NTG(N-methyl-N'-nitro-N-nitrosoguanidne) treatment. The optimum composition of culture medium and other culture conditions for the production of riboflavin by selected mutant Ashbya gossypii JAG-13 were determined. The optimum composition of medium was 9% of corn oil, 3% of gellatone, 4% of CSL, 0.3% of glycine, 0.2% of S770. The optimum culture temperature and initial pH of medium was $28^{\circ}C$ and 6.5, respectively. oxygen was essential for the production of riboflavin, but excess oxygen inhibit the production of riboflavin. When Ashbya gossypii JAG-13 was cultured under above conditions for 12 days with a bioreactor, 6.9 mg/ml of riboflavin was produced.

• KSBB Journal
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• v.22 no.1
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• pp.1-6
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• 2007

The optimum liquid culture conditions were investigated for cell growth and polysaccharide production from liquid culture of Lentinus edodes. In flask culture, the optimal medium compositions for the polysaccharide production contained glucose 60 g/L, yeast extract 10 g/L, $KH_2PO_4$ 2.0 g/L, and $MgSO_4{\cdot}7H_2O$ 1.0 g/L. The maximum mycelial growth and polysaccharide production were 11.01 g/L and 1.64 g/L, respectively. In bioreactor, through the variation of aeration in order to increase mycelial growth and polysaccharide production, the maximum mycelial growth and polysaccharide production were 55.9 g/L at 8th day and 7.34 g/L at 7th day of cultivation with 1.5 vvm, respectively.