• Biotechnology and Bioprocess Engineering:BBE
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• v.9 no.6
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• pp.459-464
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• 2004

The objective of this study is to improve cephalosporin C (CPC) production byoptimization of medium and culture conditions. A statistical method was introduced to optimize the main culture medium. The main medium for CPC production was optimized using a statistical method. Glucose and corn steep liquor (CSL) were found to be the most effective factors for CPC production. Glucose and CSL were optimized to 2.84 and $6.68\%$, respectively. CPC produc­tion was improved $50\%$ by feeding of $5\%$ rice oil at day 3rd and 5th day during the shake flask culture of C acremonium M25. The effect of agitation speeds on CPC production in a 2.5-L bio­reactor was also investigated with fed-batch mode. The maximum cell mass (54.5 g/L) was obtained at 600 rpm. However, the maximum CPC production (0.98 g/L) was obtained at 500 rpm. At this condition, the maximum CPC production was improved about $132\%$ compared to the re­sult with batch flask culture.

• Journal of Microbiology
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• v.43 no.3
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• pp.301-307
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• 2005

The feasibility of laccase production by immobilization of Pleurotus ostreatus 1804 on polyurethane foam (PUF) cubes with respect to media composition was studied in both batch and reactor systems. Enhanced laccase yield was evidenced due to immobilization. A relatively high maximum laccase activity of 312.6 U was observed with immobilized mycelia in shake flasks compared to the maximum laccase activity of free mycelia (272.2 U). It is evident from this study that the culture conditions studied, i.e. biomass level, pH, substrate concentration, yeast extract concentration, $Cu^{2+}$ concentration, and alcohol nature, showed significant influence on the laccase yield. Gel electrophoretic analysis showed the molecular weight of the laccase produced by immobilized P. ostreatus to be 66 kDa. The laccase yield was significantly higher and more rapid in the packed bed reactor than in the shake flask experiments. A maximum laccase yield of 392.9 U was observed within 144 h of the fermentation period with complete glucose depletion.

• Journal of Microbiology and Biotechnology
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• v.27 no.1
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• pp.101-111
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• 2017

The task of improving a fungal strain is highly time-consuming due to the requirement of a large number of flasks in order to obtain a library with enough diversity. In addition, fermentations (particularly those for fungal cells) are typically performed in high-volume (100-250 ml) shake-flasks. In this study, for large and rapid screening of itaconic acid (IA) high-yielding mutants of Aspergillus terreus, a miniaturized culture method was developed using 12-well and 24-well microtiter plates (MTPs, working volume = 1-2 ml). These miniaturized MTP fermentations were successful, only when highly filamentous forms were induced in the growth cultures. Under these conditions, loose-pelleted morphologies of optimum sizes (less than 0.5 mm in diameter) were casually induced in the MTP production cultures, which turned out to be the prerequisite for the active IA biosynthesis by the mutated strains in the miniaturized fermentations. Another crucial factor for successful MTP fermentation was to supply an optimal amount of dissolved oxygen into the fermentation broth through increasing the agitation speed (240 rpm) and reducing the working volume (1 ml) of each 24-well microtiter plate. Notably, almost identical fermentation physiologies resulted in the 250 ml shake-flasks, as well as in the 12-well and 24-well MTP cultures conducted under the respective optimum conditions, as expressed in terms of the distribution of IA productivity of each mutant. These results reveal that MTP cultures could be considered as viable alternatives for the labor-intensive shake-flask fermentations even for filamentous fungal cells, leading to the rapid development of IA high-yield mutant strains.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.43-47
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• 2006

Ginseng (Panax ginseng CA. Meyer) hairy root cultures, which are established via the infection of ginseng root discs with Rhizobium rhizogenes, have been used to construct profiles of both biomass growth and nutrient consumption in flask cultures. In a 250 mL shake flask culture, the maximum biomass was observed on the 59th day of the culture period, at 216.8 g (fresh wt) per liter or 11.4 g (dry wt) per liter. The hairy roots were determined to have a growth rate of 0.355 g-DW/g cells/day during the exponential growth phase and a maximum specific growth rate on day 7. Total ginseng saponin and phenolic compound contents were noted to have increased within the latter portion of the culture period. Linear correlations between increases in biomass weight and nutrient uptake were used to imply the conductivity yield $2.60g-DW/(L{\cdot}mS)$ and carbon yield 0.45 g-DW/(g sugar) in the 250 mL flask cultures. The biomass yield when two different nitrogen sources were used (ammonia and nitrate) was shown to remain approximately constant. at $0.47g-DW/(L{\cdot}mM\;NH_4$) and $0.33g-DW/(L{\cdot}mM\;NO_3$); it remained at these levels for 16 days with the ammonia. and for 24 days with the nitrate. The biomass yield when a phosphate source was used was also shown to remain approximately constant for 9 days, at $3.17g-DW/(L{\cdot}mM\;PO_4$), with an $R^2$ of 0.99.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.317-320
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• 2000

The optimal temperature and pH for both mycelial growth and exe-biopolymer production by Cordyceps millitaris in shake flask culture were found to be $20^{\circ}C$ and 6.0, respectively. Sucrose (4%) and corn steep powder (1%) were the most suitable carbon and nitrogen source for mycelial growth and exo-biopolymer production. The maximum specific growth rate $(0.142h^{-1})$ was achieved when sucrose was used as the sole carbon source. Exo-biopolymer production was increased with the increase in C/N molar ratio concentration, probably due to the facilitated carbon uptake. Under the optimal culture conditions, the maximum mycelial growth exe-biopolymer concentration were reached to around 13.3 g dry cell weigh/l and 3.33 g/l, respectively.

• The Korean Journal of Mycology
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• v.32 no.2
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• pp.95-100
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• 2004

The main objectives of the study were to investigate cultural characteristics of Phellinus linteus. The optimum culture media for mycelial growth of P. linteus were MYA (malt yeast agar) and SMS (soybean powder malt Sucrose). Similarly, optimum temperature and pH were $30^{\circ}C$ and 6.0, respectively. Malt extract (2%, v/v) and yeast extract (0.2%, v/v) were optimum carbon and nitrogen sources. Similarly, 0.1% $KH_2PO_4$ was optimum mineral salt. Highest mycelial growth was observed when C/N ratio was 10 : 1. Optimum inoculum amount for flask culture was $5{\sim}6$ mycelial discs (6 mm diameter) per 100 ml of liquid medium, Highest mycelial dry weight was obtained when cultured in 100 ml liquid medium in 300 ml shaking flask after 20 days of shaking culture, For mass liquid culture (8 l), flask culture was homogenized and used as an inoculum. Optimum culture period and aeration rate for 8l fermentation culture were 12 days and 2.0 vvm, respectively.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.3
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• pp.198-204
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• 2005

Seeds of neem were collected from different parts of India and analyzed for their azadirachtin content by High Performance Liquid Chromatography (HPLC). In order to assess the effects of genotypic and geographical variation on azadirachtin content in cell cultures, callus development was attempted in the seeds containing high and low concentration of azadirachtin. The concentration of azadirachtin in callus cultures was significantly affected by the explant source. Seed kernels with higher azadirachtin content produced higher azadirachtin content in callus cultures and lower azadirachtin content was seen in callus cultures produced from seed kernels with low azadirachtin content. The protocol for development of elite stock culture of Azadirachta indica was established with the objective of selecting a high azadirachtin-producing cell line. The highest azadirachtin-producing cell line was selected and the effects of different media and illumination conditions on growth and azadirachtin production were studied in shake flask suspension culture. Detailed batch growth kinetics was also established. These studies provided elite starter culture and associated protocols for cultivation of A. indica plant cell culture in the bioreactor.

• KSBB Journal
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• v.14 no.3
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• pp.365-370
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• 1999

To optimize the culture conditions for Beauveria bassiana 726, the effects of culture medium, pH, and temperature on mycelium and spore production were investigated. The optimum temperature and pH for the cultivation of B. bassiana 726 were 28 $^{\circ}C$ and 5.0, respectively. The optimized medium was composed of 1.0~2.0% total sugar from molasses, 0.5% corn steep liquor and 0.05% KH$_2$PO$_4$. In the cultivation of B. bassiana 726 with the optimum medium, the specific growth rate and substrate utilization were well-fitted with the proposed kinetic model in the shake flask and stirred tank reactor. When the fed-batch cultivation using carbon suorce, nitrogen source, and mineral salt as a feeding medium was compared with batch cultivation in stirred tank reactor, mycelium (12.7 g/L) and spore production (5.4$\times$$10^8/mL$) were enhanced up to 110% and 85%, respectively.

• Microbiology and Biotechnology Letters
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• v.21 no.5
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• pp.486-493
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• 1993

A gram(+) bacteria that produced microbial flocculant was isolated from soil and classified as a Bacillus species and named as Bacillus sp. A56. The culture conditions of the strain for fluocculant production were studied in a shake flask. Optimum temperature and initial pH for flcculant production were 30C and 6.5, respectively. The optimized medium has gollowing composition: glucose 20g/l, NH4NO3 0.5g/l, K2HPO4 1.0g/l, KH2PO4 0.8g/l, MgSO4.7H2O 0.2g/l, MnSO4.4-6H2O 0.3g/l, CaCO3 0.5g/l, yeast extract 0.3g/l, tryptone 0.3g/l in tap water.

• Mycobiology
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• v.37 no.2
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• pp.89-93
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• 2009

The effect of fermentation parameters and medium composition on the simultaneous mycelial growth and exo-polymer production from submerged cultures of Ganoderma applanatum was investigated in shake-flask cultures. The optimum initial pH for mycelial growth and exo-polymer production was 5.0 and 6.0, respectively. The optimum temperature was $25^{\circ}C$ and the optimum inoculum content was 3.0% (v/v). The optimal carbon and nitrogen sources were glucose and corn steep powder, respectively. After 12 days fermentation under these conditions, the highest mycelial growth was 18.0 g/l and the highest exo-polymer production was 3.9 g/l.