• 한국생물공학회:학술대회논문집
• /
• 2003.04a
• /
• pp.253-256
• /
• 2003

In batch culture of A. pullulans HP-2001, maximal production of pullulan was 57.90 g/l at 96 hr. The maximal production of pullulan with 76.55 g/l occurred when substituted solution was 10% (w/v) sucrose, 0.25% (w/v) yeast extract and mineral salts. The maximal production of pullulan was 75.00 g/l with a dilution rate of $0.015\;h^{-1}$. Productivity of pullulan in the continuous culture fed with the fresh medium was higher than that fed with the fresh medium without nitrogen source.

• Journal of Soil and Groundwater Environment
• /
• v.17 no.5
• /
• pp.49-55
• /
• 2012

A 1.28 L-batch reactor and continuous-flow stirred tank reactor (CFSTR) fed with formate and trichloroethene (TCE) were operated for 120 days and 56 days, respectively, to study the effect of formate as electron donor on anaerobic reductive dechlorination (ARD) of TCE to cis-1,2-dichloroethylene (c-DCE), vinyl chloride (VC), and ethylene (ETH). In batch reactor, injected 60 ${\mu}mol$ TCE was completely degraded in the presence of 20% hydrogen gas ($H_2$) in less than 8 days by anaerobic dechlorination mixed-culture (300 mg-soluble protein), Evanite Culture with ability to completely degrade tetrachloroethene (PCE) and -TCE to ETH under anaerobic conditions. Once the formate was used as electron donor instead of hydrogen gas in batch or chemostat system, the TCE-dechlorination rate decreased and acetate production rate increased. It indicates that the concentration of hydrogen produced in both systems is possibly more close to threshold for homoacetogenesis process. Soluble protein concentration of Evanite culture during the batch test increased from 300 mg to 688 mg for 120 days. Through the protein monitoring, we confirmed an increase of microbial population during the reactor operation. In CFSTR test, TCE was fed continuously at 9.9 ppm (75.38 ${\mu}mol/L$) and the influent formate feed concentration increased stepwise from 1.3 mmol/L to 14.3 mmol/L. Injected TCE was accumulated at 18 days of HRT, but TCE was completely degraded at 36 days of HRT without accumulation of the injected-TCE during the left of experiment period, getting $H_2$ from fermentative hydrogen production of injected formate. Although c-DCE was also accumulated for 23 days after beginning of CFSTR operation, it reached steady-state in the presence of excessive formate. We also evaluated microbial dynamic of the culture at different chemical state in the reactor by DGGE (denaturing gradient gel electrophoresis).

• KSBB Journal
• /
• v.25 no.2
• /
• pp.179-186
• /
• 2010

We investigated the optimal strategy for ethanol production using flocculent Sacchromyces cerevisiae ATCC 96581. Considering the characteristic of flocculent yeast, a repeated fed-batch ethanol fermentation was designed, in which non-sterile glucose powder was fed every 12 hours and, after cell flocculation, new feeding medium was exchanged every 24 or 36 hours. We particularly compared this fermentation process with those when cell flocculation was not carried out. Finally, the maximal total ethanol production was 825 g-ethanol during 120 hours, in which the time interval of withdrawal-fill of feeding medium was 24 hours and cell flocculation was carried out.

• KSBB Journal
• /
• v.16 no.5
• /
• pp.516-522
• /
• 2001

The large-scale production of antibiotics by filamentous mycelial organism requires and adequate supply of dissolved oxygen. In terms of productivity, it means that oxygen transfer is the rate-limiting step. Therefore, the oxygen transfer coefficients(K$\_$L/A) were determined in a broth involving a filamentous mycelial organism such as Streptomyces avermitilis for use in fermentations. To determine (K$\_$L/A) inn a stirred vessel, a great deal of effort is required to provide all the cells with a sufficient oxygen supply. To overcome the oxygen limitation in a batch culture, a fed-batch culture was applied to control the growth rate by an intermittent supply of nutrients. Thus, it was possible to maintain a suitable dissolved oxygen concentration at a low agitation rate. The optimal agitation speed was 350 rpm at low cell concentrations (below 7 g/L) by considering the efficiency of agitation and shear stress. The (K$\_$L/A) was found to decrease from 64.26 to 29.21h.$\^$-1/ when the biomass concentration was increased from 9.82 to 12.06 g/L. In addition, and increase in viscosity was also observed during the growth phase. By comparing the (K$\_$L/A) values for the various agitation and aeration rates, it was found that the effect of an increase in (K$\_$L/A) by aeration was reduced dramatically at high biomass concentrations. However, this effect was not observed when altering the agitation rate. This suggests that controlling the dissolved oxygen concentration by altering the agitation rate was more efficient than increase the aeration rate.

• KSBB Journal
• /
• v.19 no.6 s.89
• /
• pp.457-461
• /
• 2004

DO-stat fed-batch cultures of Agaricus blazei were carried out using various feeding solutions, for the production of mycelial biomass and exopolysaccharides (EPS). It was observed to be more effective to use a feeding solution containing both carbon and nitrogen sources than that containing only carbon source. The best result was obtained when a feeding solution containing 450 g/l glucose, 60 g/l yeast extract, 30 g/l soytone peptone was used. The maxium mycelial biomass and EPS concentrations were 36.5 g/l and 10.9 g/l, respectively, at 100 hours of cultivation. The mycelial and EPS productivities were 0.37 g/l-h and 0.11 g/l-h, respectively. As compared with the batch culture, the mycelial biomass concentration and its productivity were 6.0- and 2.2-folds increased, respectively. The EPS concentration and its productivity were increased by 4.7 times and 1.8 times, respectively.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.25 no.4
• /
• pp.708-714
• /
• 1996

Ethanol production from raw starch was performed using the co-immobilized culture system of Rhizopu japonicus and zymomonas mobilis(R-Z). Glucose Production in immobilized R. japonicus culture was 2-fold higher than that in free cell culture. Ethanol production was 1.67g/L(Yp/s, 0.094) and 6.549/L(Yp/s, 0.38) in R-Z and R-Z 24 culture system, respectively. R-Z system was modified and designated as R-Z 24 system by replacing cotton plug with silicon check valve after 24h fermentation with R-Z system. Optimal substrate concentration for ethanol production in batch culture was 5%(w/v) and ethanol concentration produced was 15.02g/L(Yp/s, 0.36). Ethanol yield(Yp/s, 0.38) in fed-batch culture of 5 times with 2%(w/v) substrate was equal to that in batch culture of 2%(w/v) substrate.

• KSBB Journal
• /
• v.5 no.3
• /
• pp.307-313
• /
• 1990

A microcomputer-aided fermentation system was constructed for high density fed-batch culture using dissolved oxygen(DO) as a substrate feeding indicator. DO signal was processed prior to aquisition to computer. Agitation speed and oxygen flow rate was changed stepwisely to maintain DO value at a constant level. Agitation speed was controlled by the output signal of D/A converter. Oxygen flow rate was controlled by a flow rate control valve connected to a stepping motor. Substrate was fed with a feeding pump operated by the abrupt increase of DO signal. Methylobacillus sp. SK1 was cultivated to test the system and 16.53g/l of cell density was obtained after 10 hr.

• KSBB Journal
• /
• v.17 no.1
• /
• pp.93-99
• /
• 2002

Two-stage fed-batch culture of Candide tropicalis that was designated primarily to cultivate the cell in the glucose medium (1st stage) and then produced the xylitol from xylose medium (2nd stage) was developed to improve a xylitol yield and productivity. In the growth stage, glucose was automatically supplied to the fermentor by pH-stat mode when the pH was up 5.7, When a feeding medium was added in order to reach the glucose and yeast extract concentrations up to 100 and 40 g/L, respectively, a high cell concentration and a relatively low ethanol concentration were obtained in 18.5 h culture. In the production stage, initial xylose concentration of 150 g/L was the most favorable for obtaining the final xylitol concentration and productivity. The addition of mineral salts was also enhanced a xylitol production. But the aeration rate was not significantly affected a xylitol production. When the addition of 16 g yeast extract and 232.5 g xylose powder at the production stage was used, xylitol yield and productivity were significantly increased. With these conditions, xylitol concentration, yield and productivity of 108.9 g/L, 74%) and 3.3 g/L·h, respectively, were obtained in a final volume of 1.58 L. The further addition of 16 g yeast extract and 232.5 g xylose powder increased the working volume partly (1.67 L) and resulted in a relatively high xylitol concentration, yield and productivity of 193 g/L, 70% and 3.6 g/L·h, respectively.

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

• Journal of Microbiology and Biotechnology
• /
• v.32 no.9
• /
• pp.1178-1185
• /
• 2022

Steroids are a class of compounds with cyclopentane polyhydrophenanthrene as the parent nucleus, and they usually have unique biological and pharmacological activities. Most of the biosynthesis of steroids is completed by a series of enzymatic reactions starting from cholesterol. Synthetic biology can be used to synthesize cholesterol in engineered microorganisms, but the production of cholesterol is too low to further produce other high-value steroids from cholesterol as the raw material and precursor. In this work, combinational strategies were established to increase the production of cholesterol in engineered Saccharomyces cerevisiae RH6829. The basic medium for high cholesterol production was selected by screening 8 kinds of culture media. Single-factor optimization of the carbon and nitrogen sources of the culture medium, and the addition of calcium ions, zinc ions and citric acid, further increased the cholesterol production to 192.53 mg/l. In the 5-L bioreactor, through the establishment of strategies for glucose and citric acid feeding and dissolved oxygen regulation, the cholesterol production was further increased to 339.87 mg/l, which was 734% higher than that in the original medium. This is the highest titer of cholesterol produced by microorganisms currently reported. The fermentation program has also been conducted in a 50-L bioreactor to prove its stability and feasibility.