• Journal of Microbiology and Biotechnology
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• v.16 no.8
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• pp.1185-1191
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• 2006

The expression of a pullulanase gene in Pichia pastoris was investigated. The gene encoding pullulanase was cloned by PCR using the chromosomal DNA of Bacillus naganoensis as the template. The expression vector pPIC9K-Pu was constructed by inserting the pullulanase gene into plasmid pPIC9K and then transformed into Pichia pastoris SMD 1168 by electroporation. Activity determination, SDS-PAGE, and PCR amplification indicated that the gene of the pullulanase from B. naganoensis had successfully been expressed in SMD 1168 and the molecular size of the expressed recombinant product was about 119.9 kDa. This is the first report on the successful expression of the pullulanase from B. naganoensis in P. pastoris. The transformant secreted recombinant pullulanase with the activity of 350.8 IU/ml in shake-flask culture. The properties of the recombinant pullulanase were characterized.

• Proceedings of the Korean Environmental Health Society Conference
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• 2003.06a
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• pp.106-110
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• 2003

The study was carried out immobilized Aspergillus niger used of milk-wastewater. The purpose of investigation is to optimize the ermentational conditions of milk-wastewater The optimal pH, temperature and dilution rate were 3.0, 30$^{\circ}C$ and 0.025 h$\^$-1/. The maximum amount and yield of citric acid produced by immobilizes Aspergillus niger ATCC 9142 were 4.5g/1 and 70.3%. Compared to shake-flask culture, yield was increased about 20%.

• 한국생물공학회:학술대회논문집
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• 2005.10a
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• pp.364-367
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• 2005

This study investigated the production of antioxidant from Actinomyces culture supernatant. For the research of the natural marine antioxidant, several bacteria were isolated from the coast of Je-ju in Korea. An actinomycetes strains, S-1 was identified to a genus level 16S ribosomal DNA sequence and fatty acid analysis. From these results and other characteristics described in the Bergey's Manual, this strain was identificated as a Nocardiopsis dassonvillei. Strain S-1 showed high activity of 1,1-diphenyl-2-prcrylhydrazyl radical scavenging. The hydroxyl radical scavenging ability of Nocardiopsis sp. S-1 supernatant was 53%. Nutritional and cultural conditions for the production of antioxidant by this organism under shake-flask conditions have optimized. Similary initial medium pH 7.6, incubation temperature of $25^{\cicr}C$, sodium chloride concentration 2.5 and incubation time of 8 day were found to be optimal.

• Journal of the Korean Society of Food Science and Nutrition
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• v.19 no.6
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• pp.636-640
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• 1990

For a studyon the production of ${\gamma}$-linolenic acid(GLA) by fungi 3 fungal strains were isolated from soil. Their cell growth lipid content and fatty acid composition were compared in shake flask culture. Among these fungi the fungus designated as FA-007 has high lipid content(21.1%) and GLA content(15.6% of total fatty acids) The fungal strain FA-007 was tentatively identified as Mucor sp. on the basis of morphological characteristics, Fungal oil produced by this fungus was composed of 75.2% neutral lipid 5.3% glycolipid and 19.5% phospholipid. Although the GLA content in phospholipid was higher than it in neutral lipid the GLA content in neurtal lipid was high as 15.5%.

• Journal of Plant Biotechnology
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• v.3 no.2
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• pp.107-112
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• 2001

The kinetics of Ti-transformed Salvia miltiorrhiza cell cultures was studied in 250-$m\ell$ shake flasks by using B5 medium with addition of 30 gfL of sucrose. In the cell cultures, the maximum cell mass obtained was 11.5 g DW/L on day 15. The highest amount of phenolic compounds - rosmarinic acid (RA) and lithospermic acid B (LAB) reached 871.3 mg/L (day 15) and 121.3 mg/L (day 13), respectively. The total tanshinone production, i.e., intracellular plus extracellular cryptotanshinone, tanshinone 1, and tanshinone IIA, was 5.3 mg/L on day 13. For the cultivations in 2.4-L stirred bioreactors, the residual sugar level and medium conductivity were a little higher in a small turbine impeller reactor ($T_s$) than those in a large turbine impeller reactor ($T_L$), while a higher cell density was obtained in the $T_L$. For the production of tanshinones and phenolics, better results were obtained in the $T_L$ than in the $T_s$. In the $T_L$, similar or even a little higher production titers of tanshinones and phenolic compounds were achieved compared to those in the flasks. The results suggest that the shake flask results could be successfully scaled up to the $T_L$ reactor. Such a large impeller reactor like $T_L$ may be better than a small impeller one for the large-scale production of the valuable metabolites by the suspension cultures of Ti transformed S.miltiorrhiza cells. This is considered due to the beneficial culture environment in the $T_L$, such as low shear rates as estimated theoretically.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.140-143
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• 2005

This study was concerned with the optimization of liquid culture conditions for mycelial growth and polysaccharide production and its physicochemical properties in Cordyceps militaris. The one factor at a time method was adopted to investigate the effects of medium composition, environmental factors and C/N ratio. Among the these varialbles, glucose 80g/L, yeast extract 10g/L, $MgSO_4{\cdot}7H_{2}O\;0.5g/L$, $KH_{2}PO_4\;0.5g/L$ were proved to be the most suitable carbon, nitrogen, and mineral sources, respectively. The optimal temperature, initial pH, working volume were identified to be $24^{\circ}C$, 7.0 and 100ml, respectively. Under the optimal conditions, the strategies in shake flask culture and 5L jar fermentor led to mycelial growth of 29.43 g/L, 28.88g/L and polysaccharide production of 2.53g/L, 6.38 g/L, respectively. Among the phisicochemical properties, relative concentrations(w/v) of total sugar, uronic acid, protein and hexoseamine were identified to be 74.07%, 1.13%, 0.91%, and 0.46%, respectively. The fraction of neutral and acidic polysaccharide were identified to be 81.9% and 18.1%, respectively.

• Journal of Microbiology and Biotechnology
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• v.4 no.1
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• pp.49-55
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• 1994

A fungal strain capable of producing extracellular cellulase was isolated from farmland. It was identified as Aspergillus niger, and named Aspergillus niger KKS. Production of cellulase and xylanase by the A. niger KKS was studied through a shake-flask culture. The effects of culture conditions such as inoculum size, temperature, pH, and medium composition on the cellulase and xylanase production were examined. The optimum temperature and pH for the enzyme production were $30^{\circ}C$ and pH 7.0, respectively. The optimized medium was composed of 2.0% (w/v) rice straw, 0.5% (w/v) proteose peptone, 0.5% (w/v) $KH_2 PO_4$, 0.05% (w/v) yeast extract, 0.01% (w/v) $CoSO_4 \cdot 7H_2O$, and 0.05% (w/v) $CuSO$_4$\cdot 5H_2O$. When the strain was incubated with the optimized medium, it gave the activities of endoglucanase, $\beta$-glucosidase, $\beta$-xylosidase, xylanase were 3.80, 4.20, 4.00, 80.0 (IU/mL), respectively. Filter paper and cotton activities were 0.68 and 0.045 (IU/mL), respectively. The results of this study show that A. niger KKS is a potential organism with a wide spectrum of enzyme activities, such as those of $\beta$-glucosidase, $\beta$-xylosidase, and xylanase.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.380-383
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• 2000

The effect of on the cell growth, the elaboration of pullulan, the morphology and were the effect of on the molecular weight of pullulan were investigated. A. pullulans showed maximum pullulan production when initial pH 6.5 was 11.98 g/l in shake-flask culture. In batch culture, the maximum pullulan production of 15.16 g/l was obtained at an aeration rate of 0.5 vvm. The mixture of yeast-like form and mycelial form of cells was found at the constant pH 4.5, at which condition, the elaboration of pullulan was high, about 13.31 g/l. However, pullulan with its higher molecular weight (>1,000,000) was produced at the constant pH 6.5.

• Mycobiology
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• v.37 no.2
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• pp.114-120
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• 2009

Streptomyces albidoflavus C247 was isolated from the soil of the Gyeongsan golf course in Korea. Physiological, biochemical and 16S rDNA gene sequence analysis strongly suggested that the isolate belonged to Streptomyces albidoflavus. Preliminary screening revealed that the isolate was active against fungi and bacteria. Self-directing optimization was employed to determine the best combination of parameters such as carbon and nitrogen source, pH and temperature. Nutritional and culture conditions for the production of antibiotics by this organism under shake-flask conditions were also optimized. Maltose (5%) and soytone (5%) were found to be the best carbon and nitrogen sources for the production of antibiotics by S. albidoflavus C247. Additionally, 62.89% mycelial growth inhibition was achieved when the organism was cultured at $30^{\circ}C$ and pH 6.5. Ethyl acetate (EtOAc) was the best extraction solvent for the isolation of the antibiotics, and 100 ${mu}$/ml of EtOAc extract was found to inhibit 60.27% of the mycelial growth of Rhizoctonia solani AG2-2(IV) when the poison plate diffusion method was conducted.

• Applied Chemistry for Engineering
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• v.8 no.4
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• pp.660-666
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• 1997

The production of doxorubicin by a mutant of Streptomyces peucetius subsp. caesius was studied. The optimal culture conditions, such as inoculum size and medium composition were established to improve the productivity of doxorubicin. The optimal medium composition was found to be 4% maltose, 0.5% HEPES, 0.02% $K_2HPO_4$, 0.01% $MgSO_4$. As an antiform agent, 0.001% KG(10% Adekanol+10% Silicone) was suitable one among various agents. Culture was carried out in 2.5 L jar-fermenter with different aeration rates of 1.5, 1.0, and 1.5 v/v min. The maximum production of doxorubicin(29 mg/l) was obtained at 1.5 v/v min of aeration rate, and even at 1.0 v/v min, the production of doxorubicin was increased up to 15% compared with that of shake-flask culture.