• Journal of Plant Biotechnology
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• v.4 no.3
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• pp.109-115
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• 2002

Submerged cultures of Ganoderma lucidum, a valuable mushroom in traditional Chinese medicine, were used for production of bioactive Banoderic acids and Ganoderma polysaccharides. The significant effects of oxygen supply were demonstrated in both shake flasks and bioreactors. By changing the medium loading volume in a shake flask, a different value of initial volumetric oxygen transfer coefficient ($K_L$a) was obtained, and a higher $K_L$a value led to a higher biomass density and a higher productivity of both intracellular polysaccharide and ganoderic acid. In a stirred bioreactor, at an initial $K_L$a of 78.2 $h^{-1}$, a maximal cell concentration of 15.6 g/L by dry weight was obtained, as well as a maximal intracellular polysarcharide (IPS) production of 2.2 g/L and its maximal productivity of 220 mg/(L.d). An increase of initial $K_L$a led to a higher production and productivity of GA, and the GA production and productivity at an initial $K_L$a of 96.0 $h^{-1}$ was 1.8-fold those at an initial $K_L$a of 16.4 $h^{-1}$. The fundamental information obtained in this study may be useful for efficient large-scale production of these valuable bioactive products by the submerged cultures.

• 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.

• KSBB Journal
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• v.21 no.4
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• pp.298-301
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• 2006

A gene(GeneBank AF 135796) coding for a trehalose synthase from Thermus caldophilus GK24 was cloned into Escherichia coli K12 using five vector systems. The constitutive expression system(pHCETS) which shows the highest trehalose synthase activity from flask culture of recombinant E. coli was selected for the production of trehalose from maltose. For the shake flask culture, the final dry cell weight was 0.9 g/L and the trehalose synthase activity was 25 U/mL. Fed-batch culture of recombinant E. coli harboring plasmid pHCETS which uses the glycerolas a carbon source was performed in jar fermentor: the dry cell weight of 20 g/L and the trehalose synthase activity of 13.7 U/mL were attained in 48 h.

• KSBB Journal
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• v.15 no.6
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• pp.566-572
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• 2000

Experimental studies were carried out to develop an osmotolerant mutant of Candida magnoliae JH and to determine the optimum concentrations of carbon and nitrogen sources to improve erythritol yield and productivity. Mutants of C. magnoliae JH were prepared by treatment with ethylmethane sulfonate, and osmotolerant mutants were isolated from the agar plate colonies containing 2.5 M KCI. Among the mutants isolated, one mutant M26 was finally selected based on erythritol yield and productivity. In shake flask culture, glucose proved to be the best carbon source and the optimum yeast extract concentration was 5 g/L based on 100 g/L glucose. The erythritol yield and productivity of mutant M26 were greater than wild type in 100 g/L glucose medium. in the fermentation experiments, erythritol production increased with increased glucose concentration, up to a limit of 250 g/L. The maximum concentration of erythritol achieved 127.5 g/L, and the yield and productivity of erythritol production were 51.0% and 0.63 g/L-h, respectively.

• Journal of Microbiology
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• v.39 no.1
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• pp.79-82
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• 2001

The influence of levulinic acid (LA) on the production of copolyester consisting of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) by Ralstonia eutropha was investigated. Addition of LA into the culture medium greatly increased the molar fraction of 3HV in the copolyester, indicating that LA can be utilized as a precursor of 3HV. In shake flask culture, the 3HV content in the copolyester increased from 7 to 75 mol% by adding 0.5 to 4.0 g/L LA to the medium containing fructose syrup as a main carbon source. A maximal copolyester concentration of 3.6 g/L (69% of dry cell weight) was achieved with a 3HV content of 40 mo1% in a jar fermentor culture containing 4.0 g/L of LA. When LA (total concentration, 4 g/L) was added repeatedly into a fermentor culture to maintain its concentration at a low level, the copolyester content and the 3HV yield from LA reached up to 85% of dry cell weight and 5.0 g/g, respectively, which were significantly higher than those when the same concentration of the LA was supplied al1 at once. The present results indicated that LA is more effective than propionate or valerate as a cosubstrate fur the production of copolyesters with varying molar fractions of 3HV by R. eutropha.

• Journal of Microbiology
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• v.41 no.3
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• pp.252-258
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• 2003

The carbon utilizations of Bacillus species and Pythium species were investigated by using a Biolog$^{(R)}$ microplate assay to determine if there are differences in the carbon utilizations of selected strains of these species. It may be possible to afford a competitive advantage to bacterial biological control agents by providing them with a substrate that they can readily use as a carbon source, for example, in a seed coating formulation. Microplates, identified as SFP, SFN and YT were used to identify spore-forming bacteria, nonspore-forming bacteria, and yeast, respectively. Bacterial and mycelial suspensions were adjusted to turbidities of 0.10 to 0.11 at 600 nm. One hundred microliters of each of the bacterial and mycelial suspension were inoculated into each well of each of the three types of microplates. L-arabinose, D-galactose, D-melezitose and D-melibiose of the 147 carbohydrates tested were found to be utilized only by bacteria, and not by Pythium species, by Biolog$^{(R)}$ microplate assay, and this was confirmed by traditional shake flask culture. Thus, it indicated that the Biolog$^{(R)}$ microplate assay could be readily used to search for specific carbon sources that could be utilized to increase the abilities of bacterial biological control agents to adapt to contrived environments.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.390-393
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• 2001

The optimal temperature, pH and aeration rate for spore production by Bacillus polyfermenticus SCD in 500 ml shake flask and 5-1 jar fermenter were found to be $32^{\circ}C$, 7.0 and 1.0 vvm. respectively. When batch culture processes was performed under optimized culture conditions. viable cells were $3.3{\times}10^{10}$ CFU/ml and spore cells were $3.3{\times}10^{10}$ CFU/ml. Fed-batch culture processes were also examined with regard to higer maximum viable cell and spore production. The highe viable cells and spores were obtained in 5-1 jar fermenter at 72 h cultivation time by strategy in an intermediate feeding mode with 60% glucose solution 150 ml and 5% soybean flour solution 150 ml fed to the fermenter twice, and the productivity of spore cells was significantly increased. Finally. volumetric productivity of spore cells on fed-batch culture indicated $9.9{\times}10^8$ CFU/ml/h, which was approximately 2 times higher than batch culture. Thus, fed-batch culture show a promise as an industrial production method.

• 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.

• 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.

• Fashion & Textile Research Journal
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• v.9 no.1
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• pp.119-123
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• 2007

Dyeing properties of soybean fabrics on gromwell colorants were studied for the effect of dyeing conditions, such as colorants concentration, temperature, time and pH on the dye uptake and effect of mordants on color change, dye uptake and various colorfastness. Antimicrobial activity of soybean fabrics dyed and sim-mordanted with gromwell colorants was examined by shake flask method. Gromwell colorants showed considerably affinity to soybean fabric and its isotherm adsorption curve was Freundlich type. Therefore, it is considered that hydrogen bonding and Van der Waals force were involved in the adsorption of gromwell colorants to soybean fabric. Soybean fabrics showed R color on Al, Cu and Sn mordant, RP color on Cr and Fe mordant, but soybean fabrics showed low dye uptake depending on mordanting treatment. Light colorfastness was increased for Cr and Fe mordants. Staphylococcus aureus reduction rates were above 90% for Cr and Cu mordanted soybean fabrics, and the others were poor. Klebsiella pneumoniae reduction rates soybean fabrics did not show reduction rate hardly.