• Title/Summary/Keyword: batch culture

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Tylosin Production by Mutant Resistant to Oleic Acid (Oleic Acid 내성균주로부터 Tylosin 생산)

  • Choi, Du-Bok;Choi, On-You;Moon, Ok-Ran;Yoon, Mi-Ran;Ji, Sung-Nam;Shin, Dae-Yewn
    • Journal of Environmental Health Sciences
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    • v.31 no.5 s.86
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    • pp.360-364
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    • 2005
  • When rapeseed oil as the carbon source was used for tylosin production from Streptomyces fradiae TP-1239 was very sensitive to oleic acid. Cell growth was restrained by adding 0.8 g/l of oleic acid to the culture broth. Mutant strain TM-224-1 resistant to 1.2 g/l of oleic acid was obtained by screening in solid and liquid media containing oleic acid. The uptake rate of oleic acid by TM-224-1 was approximately 3.8 fold higher than the parent strain. For comparing the TM-224-1 and the parent strain, batch cultures were carried out in a jar fermentor. Cell growth of TM-224-1 strain was higher than the parent strain after two days of culturing. However, after four days of culturing, it was similar to that of the parent strain. The amount of rapeseed oil consumed by TM-224-1 and the parent strain were 60.5 and 78.2 g/l, respectively. The production and yield of tylosin was aproximately 2.0 and 3.2 fold higher than the parent strain, respectively. From these results, it was concluded that this mutant, which was resistant to oleic acid, has improved tylosin production.

Effect of Cell Density on Xylitol Fermentation by Candida parapsilosis (Candida parapsilosis에 의한 Xylitol 생산시 균체농도가 미치는 영향)

  • Kim, Sang-Yong;Yoon, Sang-Hyun;Kim, Jung-Min;Oh, Deok-Kun
    • Korean Journal of Food Science and Technology
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    • v.28 no.5
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    • pp.970-973
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    • 1996
  • Effect of cell density on the xylitol production from xylose by Candida parapsilosis KFCC 10875 was investigated. The concentrated cells were obtained by centrifugation of culture broth. The xylitol production rate was maximum at the cell concentration of 20 g/l and the specific xylitol production rate decreased when the cell concentration was increased due to oxygen limitation. Effect of the initial concentration of xylose on the xylitol production was also examined using the concentrated cells of 20 g/l. The xylitol production rate, specific xylitol production rate, and xylitol yield from xylose were maximum at 170 g/l xylose. Above 170 g/l xylose, the xylitol production rate was remarkably decreased. The concentrated cells could also be obtained by adjusting the dissolved oxygen (DO) during fermentation. The rapid accumulation of cells up to 20 g/l was achieved by maintaining an increased level of DO during the exponential growth phase and then, for the efficient xylitol production, the DO was changed to a low level in the range of 0.7-1.5%. A fed-batch fermentation of xylose by adjusting the DO level was carried out in a fermentor and the final xylitol concentration of 140 g/l from xylose of 200 g/l could be obtained for 56 h fermentation.

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A Study on the Growth Characteristics of Commercially Developed Nitrifying Bacteria and its Application to Activated Sludge Process (상업용 질산화 박테리아의 성장특성과 활성슬러지 공정에서의 적용 방법에 따른 연구)

  • Whang, Gyu-Dae;Lee, Bong-Hee
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.4
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    • pp.595-604
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    • 2006
  • The growth characteristics of Commercially Developed Nitrifying Bacteria (CDNB) were studied in laboratoryscale. CDNB, a pure, artificially isolated bacterium, was cultivated to produce Cultivated Nitrifying Bacterium Group (CNBG). The average ammonia removal rate of CDNB was 0.0234g $NH_4^+-N/g$ MLSS/hr. CNBG was produced in the batch reactor and Specific Nitrification Rate (SNR) was determined at 0.0107g $NH_4^+-N/g$ MLSS/hr. The SNR of CNBG was lower than the SNR of CDNB because the diverse and multi-cultured microbial growth took place during cultivation. The effect of the temperatures and the mixing ratios of sewage and culture solution on the SNR of CNBG was studied. The SNR of CNBG, 0.0107g $NH_4^+-N/g$ MLSS/hr at $27^{\circ}C$, decreased to 0.0048g $NH_4^+-N/g$ MLSS/hr at $15^{\circ}C$, and temperature coefficient (${\Theta}$) was calculated to be 1.07. With the varied sewage mixing ratios, the SNR of CNBG remained unchanged. Activated sludge reactors maintaining an MLSS of 2,000mg/L at HRT of 4 h were operated under conditions in which dosage of Concentrated CNBG Solution (CCNBGS, 10,000mg MLSS/L) and application method of CNBG were varied. The reactor with 20mL of CCNBGS took shorter time to oxidize $NH_4^+-N$ reaching 1mg/L than the reactor with 5mL of CCNBGS showing that higher dosages were associated with greater mass removal of $NH_4^+-N$. However, the total removal was not great. In terms of different methods of CNBG application, reactor seeded with 20mL of CCNBGS took 3days to reach 1mg/L of effluent ammonia concentration while reactor dosed with 20% (v/v) CNBG implanted media took 2days. Both the control reactor and the reactor dosed with 20% (v/v) media only did not reach 1mg $NH_4^+-N/L$ after operating 18days. The reactor with CNBG implanted media had the highest $NH_4^+-N$ removal rate because of maintaining high concentration of Nitrifying Oxidizing Bacteria (NOM), and is regarded as an appropriate method for the activated sludge process.

Optimization of Hydrogen Production using Clostridium beijerinckii KCTC 1785 (Clostridium beijerinckii KCTC 1785를 이용한 수소생산 최적화 조건 탐색)

  • Kim, Jung-Kon;Nhat, Le;Kim, Seong-Jun;Kim, Si-Wouk
    • KSBB Journal
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    • v.20 no.6
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    • pp.401-407
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    • 2005
  • Optimum culture conditions and medium composition for hydrogen production by Clostridium beijerinckii KCTC 1785 were investigated. Initial pH and temperature for growth were 7.0 and $35^{\circ}C$, respectively. Agitation accelerated the hydrogen production. Although C. beijerinckii KCTC 1785 could grow up to 6%(w/v) glucose in the medium, the optimum glucose concentration for hydrogen production was 4% and hydrogen content in the biogas was 37%(v/v). However, the economical glucose concentration for hydrogen production was 1% regarding to the residual glucose which was not used in the medium. During hydrogen fermentation, acetic and butyric acid were produced simultaneously. High concentrations of acetic(>5,000 mg/L) or butyric(>3,000 mg/L) acid inhibited hydrogen production. When pH was maintained at 5.5 in the batch fermentation, 1,728 mL of hydrogen was produced from 0.5% glucose within 15 hr. $H_2$ yield was estimated to be 1.23 mol $H_2/mol$ glucose. It was found that yeast extract or tryptose in the medium was essential for hydrogen production.

Development of Alginate-Celite Immobilization Technique for the Improvement of Ethanol Productivity (에탄올 생산성 향상을 위한 Alginate-Celite 고정화 방법의 개발)

  • 김승욱;김은영홍영기
    • KSBB Journal
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    • v.11 no.1
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    • pp.77-85
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    • 1996
  • The optimal initial pH for the ethanol production by Saccharomyces K35 was found to be 5.0, and about 80% of yield was obtained when 200g/$\ell$ of glucose was used as a substrate, which showed sugar tolerant. As the additives and cross-linking agent, the addition of 1.67%(w/v) Celite R-634 together with 0.33%(v/v) of glutaraldehyde(ACG bead) resulted in better stability, ethanol productivity and cell viability than Ca-alginate bead. Also, ACG bead seemed to be more resistant to phosphate ion than Ca-alginate bead, considering outgrowing cell concentration in the media. Scanning electron microscopic observation depicted that the surface of ACG bead was almost similar to the original state but not for Ca-alginate bead. When repealpd-batch culture was performed with Ca-alginate bead for 60 days in a 500m1 Erlenmeyer flask, ethanol and cell concentration were maintained about 138g/$\ell$-gel and 29~30g/$\ell$-gel, respectively, up to 40 days(7th run number), and then both were rapidly decreased. In the case of ACG bead, ethanol and cell concentration were maintained about 130~150g/$\ell$-gel and 32~35g/$\ell$-gel, respectively, up to 60days(10th run number). Cell viability was maintained about 70%, and outgrowing cell concentration was below 5.8% of total cell concentration.

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Quality Characteristics of High Acidity Apple Vinegar Manufactured Using Two Stage Fermentation (2단계 발효에 의한 고산도 사과식초의 품질특성)

  • Sung, Na-Hye;Woo, Seung-Mi;Kwon, Joong-Ho;Yeo, Soo-Hwan;Jeong, Yong-Jin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.6
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    • pp.877-883
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    • 2014
  • This study investigated the manufacturing conditions of apple vinegar with high acetic acid content following fermentation according to alcohol concentration without any nutrients. We compared and analyzed the quality characteristics of high acetic acid fermentation by varying the initial alcohol content (6%, 7%, 8%, and 9%). In the results, it was possible to manufacture high acetic acid vinegar with 12% titratable acidity and an alcohol content of 6% and 7%. Lower initial alcohol content was associated with higher yield due to a shorter lag phase. For quality characteristics of the high acetic acid apple vinegar, pH was 2.91~3.20, titratable acidity was 12.0%, and organic acid consisted of acetic acid, malic acid, critic acid and oxalic acids. Based on the results, high acetic acid apple vinegar was produced using a two stage fermentation process after alcohol fermentation but the further research is needed to reduce the time of fermentation in fed-batch culture for industrial use.

The Study of TCE Dechlorination using Geobacter lovleyi with Slow Release Substrate Applied (Slow Release Substrate를 이용한 Geobacter lovleyi의 TCE 탈염소화 연구)

  • Cha, Jae Hun;An, Sang Woo;Park, Jae Woo;Chang, Soon Woong
    • Journal of the Korean GEO-environmental Society
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    • v.13 no.9
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    • pp.53-59
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    • 2012
  • This study investigated characteristics of decomposition of tetrabutoxysilane (TBOS) as a slow release substrate (SRS) and on effect of TBOS decompostion compounds (acetate and butylate) for anaerobic dechlorination of trichloroethylene (TCE). In the batch experiment, TCE, cis-dichloroethene (cis-DCE), 1-butanol and TBOS were analysed by GC/FID and acetate and butylate were measured by HPLC. 1M of TBOS transferred and accumulated 4M of 1-butanol by abiotically hydrolysis reaction. The hydrolysis rate was in a range of 0.186 ${\mu}M/day$. On other hand, 1-butanol fermented to butyrate and acetate with indigenous culture from natural sediments. This results showed that TBOS could be used a slow release substrate in the natural sites. The dechlorinated potential of TCE with acetate and butyrate was increased with a decreasing initial TCE concentrations. In addition, first order coefficients of dechlorination with acetate as electron donor was higher then that with butyrate. It is because that dechlorination of Geobacter lovleyi was affected by substrate affinity, biodegradability and microbial acclimation on various substrates. However, dechlorinated potential of Geobacter lovleyi was decreased with accumulation cis-DCE in the anaerobic decholoronation process. The overall results indicated that SRS with Geobacter lovleyi might be a promising material for enhancing dechlorination of TCE on natural site and cis-DCE should be treated by ZVI as reductive material or by coexisting other dechlorinated bacteria.

화학독립영양미생물 Aeromonas sp. strain JS-1의 RubisCO 정제 및 특성

  • Na, Suk-Hyeon;Bae, Sang-Ok;Kim, Min-Jeong;Kim, Seong-Jun;Jeong, Seon-Yong
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.461-464
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    • 2002
  • A Chemoautotroph identified as an Aeromonas sp. strain JS-1 was isolated from fresh water. Aeromonas sp. strain JS-1 used the $H_2$ and $CO_2$ as energy and carbon sources, respectively. Growth characteristics for improving the $CO_2$ fixation rate were examined in batch cultivation. Its results shown that the optimal growth appeared at culture conditions of $35^{\circ}C$, pH 7 and NaCl 0.1%(w/v). Some hydrogen-oxidizing bacteria were reported that the enzyme activity of ribulose 1,5-bisphosphate carboxylase- oxygenase (RubisCO-EC 4.1.1.39), in the key enzyme of the Calvin-Benson cycle. A RubisCO was purified from a chemoautotrophic bacterium, Aeromonas sp. strain JS-1. the enzyme was purified by ammonium sulfate precipitation, DEAE-sepharose CL-6B and gel filtration chromatography. The RubisCO showed that molecular mass was about 560KDa from gel filtration chromatography and nondenaturing PAGE, and the RubisCO was confirmed to consist of $L_8S_8$ enzyme structure by sodium dodecyl sulfate polyacrylamide gel electrophoresis. A large subunit was about 56KDa and small one was about 15kDa. The Km values of the enzyme for ribulose 1,5-bisphosphate(RUBP), $NaH^{14}CO_3$, and $Mg^{++}$ were estimated to be 0.25mM, 5.2mM, and 0.91mM, respectively. The optimum temperature for RubisCO enzymatic activity were $50^{\circ}C$, and the enzymatic activity was stable up to $45^{\circ}C$.

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Effects of Seed Sources and Concentration of Ammonia on Anaerobic Digestion (혐기성 소화에 대한 식종원 및 암모니아 농도의 영향)

  • Kim, Yang-Ji;Kim, Sung-Il;Shin, Bum-Shic;Ahn, Ki-Sup;Kim, Jong-Soo
    • Korean Journal of Environmental Agriculture
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    • v.23 no.1
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    • pp.1-6
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    • 2004
  • The seeding sources and concentration of ammonia on anaerobic digestion were investigated by batch culture bioreactors. The sources of seeding on anaerobic digestion were from swine wastewater collection pit of a hog raising farm and from anaerobic digestion sludge of a municipal sewage treatment plant. The inhibition of ammonia on anaerobic microorganisms was initiated at ammonia concentration of $1,500\;mgNH_4-N/L$ and it's effect was increased by increased by increasing ammonia concentration up to $3,500\;mgNH_4-N/L$ regardless the sources of seeding as evidenced by decreases in COD removal efficiencies and biogas yields. The inhibition occurred to not only methanogens but also acidogens since the concentration of volatile fatty acids was maintained at 50 mg/L The COD removal efficiency and biogas yield were Maintained constantly while increasing ammonia concentration up to $3,500\;mgNH_4-N/L$ when swine wastewater collection pit was used as a seeding; however, those were decreased while increasing ammonia concentration when anaerobic digestion sludge was used as a seeding. The results indicate that the seeding acclimated to high concentrations of ammonia for long time was easy in adaptation to high ammonia concentration and less subjective to ammonia inhibitory effects.

Maintenance and Differentiation of Pluripotential Embryonic Cell Lines from Mouse Blastocysts (BCF1 생쥐 배반포기 유래 배아간세포 작성에 관한 연구)

  • 이재원;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.18 no.4
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    • pp.235-244
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    • 1995
  • The present study was designed to demonstrate that ES cell lines efficiently could be isolated from explanted blastocysts of hybrid BCF1 mouse when grown on STO feeder layer derived from mouse fibroblasts in culture medium supplemented with leukemia inhibitory factor (LIF). The expanded blastocysts were attached to mitomycin C-inactivated STO feeder layer and were cultured for 4 days. Four days later the ICM was disaggregated by a short term trypsin treatment (0.25% trypsin / 0.04% EDT A for 2-3 min). The resulting cell suspension was seeded on a new STO feeder layer and covered with DMEM supplemented with 10% FCS, 0.1 mM nonessential amino acid, 0.1 mM sodium pyruvate, 0.1 mM mercaptoethanol and 1,000 U/ml LIF. Colonies of ES-like cells were observed after the second passage. These colonies were repeatedly passaged at approximately 5 day intervals. In this study, five ES-like celllines were isolated by directly explanting blastocysts, but three lines were lost after the 5th passage, possibly due to toxic effects of a new FCS batch. The characterization of developmental potential of isolated cell lines was performed with respect to in vitro differentiation and specific activity of alkaline phosphatase (AP). When cells were cultured in suspension, the aggregates of cell lines were capable of forming simple embryoid bodies (EB), and showed the capacity for forming cystic multilayer EBs. In addition, the cell lines were positive for AP staining, a biochemical marker characteristic of mouse ES cells.

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