• Mycobiology
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• v.48 no.2
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• pp.122-132
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• 2020

Citric acid is a commercially valuable organic acid widely used in food, pharmaceutical, and beverage industries. In this study, 260 yeast strains were isolated from soil, bread, juices, and fruits wastes and preliminarily screened using bromocresol green agar plates for their ability to produce organic acids. Overall, 251 yeast isolates showed positive results, with yellow halos surrounding the colonies. Citric acid production by 20 promising isolates was evaluated using both free and immobilized cell techniques. Results showed that citric acid production by immobilized cells (30-40 g/L) was greater than that of freely suspended cells (8-19 g/L). Of the 20 isolates, two (KKU-L42 and KKU-L53) were selected for further analysis based on their citric acid production levels. Immobilized KKU-L42 cells had a higher citric acid production rate (62.5%), while immobilized KKU-L53 cells showed an ~52.2% increase in citric acid production compared with free cells. The two isolates were accurately identified by amplification and sequence analysis of the 26S rRNA gene D1/D2 domain, with GenBank-based sequence comparison confirming that isolates KKU-L42 and KKU-L53 were Candida tropicalis and Pichia kluyveri, respectively. Several factors, including fermentation period, pH, temperature, and carbon and nitrogen source, were optimized for enhanced production of citric acid by both isolates. Maximum production was achieved at fermentation period of 5 days at pH 5.0 with glucose as a carbon source by both isolates. The optimum incubation temperature for citric acid production by C. tropicalis was 32 ℃, with NH₄Cl the best nitrogen source, while maximum citric acid by P. kluyveri was observed at 27 ℃ with (NH₄)₂ SO₄ as the nitrogen source. Citric acid production was maintained for about four repeated batches over a period of 20 days. Our results suggest that apple and banana wastes are potential sources of novel yeast strains; C. tropicalis and P. kluyveri which could be used for commercial citric acid production.

• KSBB Journal
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• v.8 no.2
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• pp.156-163
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• 1993

Strains degrading ethylene glycol(EG) and terephthalic acid(TPA) were isolated from water systems, and identified as Pseudomonas sp. They were named as Pseudomonas sp. EAW for EG and as Pseudomonas sp. TS2 for TPA. The optimal culture conditions of temperature, pH and nitrogen source were found to be $35^{\circ}C$, 7.5 and ammonium sulfate, respectively. The growth of strains and removal efficiency was slightly promoted by trace elements such as niacin and biotin in case of EG, and by trace elements such as $Na_2MoO_4{\cdot}2H_2O$ and thiamin i case of TPA. With increasing inoculation sloe for batch culture, the removal efficiency of EG by the strain EAW was conspicuously increased, while the removal efficiency of TPA by the strain TS2 was not changed as much as that of EG. The growth rate of the strain EAW was much more decreased than that of the strain TS2 in the enrichment medium, as the frequency of repeated-batch culture in the rich-medium increased. in case of real wastewater, growth rate and removal efficiencies of EG and TPA were lower than those in the enrichment medium. $COD_{Mn}\;and\;COD_{Cr}$ removal efficiencies after 48 hrs batch culture in real wastewater were 89% and 93%, respectively. The specific growth rate was inhibited when the initial concentration of EG or TPA was more than 25g/L.

• Membrane and Water Treatment
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• v.11 no.1
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• pp.79-85
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• 2020

A single-stage deammonification with a sequencing batch reactor (SBR) that simultaneous nitritation, anaerobic ammonia oxidation (anammox), and denitrification (SNAD) occur in one reactor has been widely applied for sidestream of wastewater treatment plant. For the stable and well-balanced SNAD, a feeding strategy of influent wastewater is one of the most important operating factors in the single-stage deammonification SBR. In this study, single-stage deammonification SBR (working volume 30L) was operated to treat a high-strength ammonium wastewater (1200 mg NH₄⁺-N/L) with different feeding strategies (single feeding and nine-step feeding) under the condition without COD. Each cycle of the step feeding involved 6 sub-cycles consisted of aerobic and anoxic periods for partial nitritation (PN) and anammox, respectively. Contrary to unstable performance in the single feeding, the step feeding showed better deammonification performance (0.565 kg-N/m³/day). Under the condition with COD, however, the nitrogen removal rate (NRR) decreased to 0.403 kg-N/m³/day when the Nine-step feeding strategies had an additional denitrification period before sub-cycles for PN and anammox. The NRR was recovered to 0.518 kg-N/m³/day by introducing an enhanced multiple-step feeding strategy. The strategy had 50 cycles consisted of feed, denitrification, PN, and anammox, instead of repeated sub-cycles for PN and anammox. The multiple-step feeding strategy without sub-cycle showed the most stable and excellent deammonification performance: high nitrogen removal efficiency (98.6%), COD removal rate (0.131 kg-COD/m³/day), and COD removal efficiency (78.8%). This seemed to be caused by that the elimination of the sub-cycles might reduce COD oxidation during aerobic condition but increase the COD utilization for denitrification period. In addition, among various sensor values, the ORP pattern appeared to be applicable to monitor and control each reaction step for deammonification in the multiple-step feeding strategy without sub-cycle. Further study to optimize the number of multiple-step feeding is still needed but these results show that the multiple-step feeding strategy can contribute to a well-balanced SNAD for deammonification when treating high-strength ammonium wastewater with COD in the single-stage deammonification SBR.

• Journal of Microbiology and Biotechnology
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• v.18 no.4
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• pp.725-729
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• 2008

An enzyme reactor installed with ultrafiltration membrane was developed to produce ${\alpha}-,\;{\beta}-$, and ${\gamma}$-cyclodextrins (CDs) from soluble starch by Bacillus macerans cyclodextrin glycosyltransferase (CGTase) tagged with 10 lysines at its C-terminus (CGTKIOase). Ultrafiltration membrane YM10 with 10,000 of molecular cutoff was chosen for membrane modification and CD production. A repeated-batch type of the enzyme reaction with free CGTK10ase resulted in a ${\alpha}$-CD yield of 24.0 (${\pm}1.5$)% and a productivity of 4.68 (${\pm}0.88$) g/l-h, which were 7 times higher that those for CGTK10ase immobilized on modified YM10 membrane. Addition of 1-nonanol increased CD yields by 30% relative to the control, which might be due to prevention of the reversible hydrolysis of CDs.

• 한국생물공학회:학술대회논문집
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• 2000.04a
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• pp.121-124
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• 2000

Organic acids were produced from wastes streams in food industries by cell-recycle fermentation using Propionibacterium acidipropionici ATCC 4965. As a results of continuous fermentation, maximum productivity was 3.32g organic acid/L/hr at the dilution rate of 0.2/hr. Compared to batch fermentation, maximum productivity was improved by as much as 13 times and cell mass production was increased by as much as 22 times. The diluted organic acids in the fermenter were selectively separated by liquid-liquid extraction using 30%(w/w) trioctylamine(TOA) dissolved in methylisobutylketone(MIBK). The degree of extraction was reached above 90% for both acetic and propionic acid through repeated extraction of organic acids in fermentation broth.

• KSBB Journal
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• v.23 no.3
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• pp.199-204
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• 2008

Culture conditions for biological hydrogen production were investigated in wastewater of Takju manufacturing factory. Rhodobacter spaeroides KCTC1425, photosynthesis bacteria, and Enterobacter cloacae YJ-1, anaerobic bacteria were used. The hydrogen production were $195.3m{\ell}{\cdot}H_2/{\ell}$ broth for Rhodobacter spaeroides KCTC1425 and $271.8m{\ell}{\cdot}H_2/{\ell}$ broth for Enterobacter cloacae YJ-1 during 36 h. The hydrogen production increased with light intensity, and were highest over 12000Lux. In mixed culture of Rhodobacter spaeroides KCTC1425 and Enterobacter cloacae Y J-1, the optimum mixing ratio of hydrogen production was 20 and 80. Adding volume of yeast extract for maximum hydrogen production was 15 $g/{\ell}$, but there was no effect over that. $Na_2MoO_4$ was most effective among the inorganic salts, and the optimum volume was 0.4 $g/{\ell}$. In semi-continuous culture, total hydrogen production was $13086m{\ell}{\cdot}H_2/{\ell}$ broth for 144 h with operating period of 24 h.

• Microbiology and Biotechnology Letters
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• v.21 no.4
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• pp.373-380
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• 1993

When the cells of yeast K35 were immobilized in Ca-alginate gel, cell concentration and viability decreased as alginate concentration increased. Considering the results, 2% (w/v) Ca-alginate concentration would be suitable. Among various concentrations of additives and cross-lin-king agent, the addition of 1.67% (w/v) of bentonite together with 0.33% (v/v) of glutaraldehyde (ABG bead) resulted in the highest ethanol production of 1.8%(w/v), using YPD medium containing 2% glucose. ABG bead seemed to be more resistant to phosphate ion than Ca-alginate bead. 0.33%(w/v) of phosphate was a proper concentration for the ethanol production by ABG bead. Scanning electron microscopic observation depicted that the immobilized cells on the bead surface were coated by alginate gel and that the cells in the internal bead were cross-linked with alginate matrix. When repeated-batch culture was performed with ABG bead for 40 days in a packed-bed reactor, ethanol concentration of about 90~110 g/l-gel was maintained. Cell viability was maintained around 70%, and outgrowing cell concentration was below 6.3% of total cell concentration. Consequently, the results showed that ABG head was a potential carrier for continuous production of ethanol compared to conventional Ca-alginate bead.

• Journal of Soil and Groundwater Environment
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• v.20 no.5
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• pp.26-33
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• 2015

Batch experiments were performed to investigate the feasibility of a surfactant-enhanced soil washing process for soils heavily contaminated with crude oil in Kuwait. TPH concentration of the contaminated soil was 223,754 mg/kg, sampled from the bottom of a vaporized oil extraction pond in the Burgan reservoir field. Commercialized eight nonionic surfactants (Tween and Tergitol series) were used to measure the aqueous solubility for the crude oil. Among them, two Tergitol surfactants were used to evaluate the TPH removal efficiency of the surfactant-enhanced soil washing for heavily contaminated Kuwait soil. The solubility of the crude oil in surfactant solution was in the order Tergitol 15-S-7 > Tergitol 15-S-9 > Tergitol 15-S-12 > Tween-80 > Tween-20 > Tween-60, which showed that the crude oil solubilities of the Tergitol series were higher than those of the Tween series. The TPH removal efficiencies of 2% and 5% Tergitol 15-S-9 solution were 59% and 65%, respectively. Because the residual TPH concentration in the washed soil was still higher than the clean-up level (10,000 mg/kg), the soil washing process was repeated five times. After the fifth washing, the residual TPH concentration in the soil went down to 7,680 mg/kg and its removal efficiency was 97%.

• Microbiology and Biotechnology Letters
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• v.44 no.4
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• pp.488-492
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• 2016

Saccharomycopsis fibuligera is an amylolytic yeast that exhibits raw starch-degrading activity. In this study, adaptive laboratory evolution was performed to improve the tolerance of S. fibuligera to lactic acid by prolonged repeated batch fermentation in which the lactic acid concentration was gradually increased. The evolved S. fibuligera strain exhibited a significantly enhanced tolerance to lactic acid at concentrations up to 2.5% (w/v), as assessed by determining its specific growth rate using a plate assay. Scanning electron microscopy revealed an elongated and perforated morphology of the parent strain under lactic acid stress, indicating that its membrane might be more prone to damage caused by lactic acid than that of the evolved strain.

• KSBB Journal
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• v.21 no.4
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• pp.236-240
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• 2006

IPTG induction caused a sudden drop of alkali consumption rate during cultivation of a recombinant E. coli with ${\beta}$-galactosidase structural gene under T7 promoter on a plasmid. A series of batch cultivations showed the positive correlation of the decrease of alkali consumption and the level of expression. However, repeated IPTG induction did not cause any variation of alkali consumption rate. Supplementation of medium even at stationary phase enhanced the level of ${\beta}$-galactosidase expression. These results suggests that the drop of alkali consumption rate by IPTG induction represents the rate of expression.