• Title/Summary/Keyword: Substrate culture

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Continuous Ethanol Fermentation by Immobilized Kluyveromyces marxianus F043 Using Jerusalem Arichoke Powder (돼지감자 분말을 이용한 고정화 Kluyveromyces marxianus sp.의 에탄올 연속발효)

  • 신지현;최언호
    • Microbiology and Biotechnology Letters
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    • v.23 no.3
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    • pp.346-351
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    • 1995
  • To produce ethanol from Jerusalem artichoke powder efficiently, Kluyveromyces marxianus F043 cells were encapsulated in 2% sodium alginate and were cultured in a countinuous reactor to investigate the fermentation properties. Immobilized K. marxianus F043 cells were activated for 48 hours in a fermentor for continuous ethanol production. The culture in a CSTR using a Jerusalem artichoke substrate treated with 2% cellulase showed a decrease in ethanol concentration and an increase in residual saccharide concentration with a increasing dilution rate. Optimum conditions for high ethanol productivity and low residual saccharide output were clarified to be given at a dilution rate of 0.2 h$^{-1}$ and a Jerusalem artichoke medium concentration of 75 g/l. Ethanol productivity of 3.1 g/l-h and saccharide utilization of 62.6% were obtained under the optimum condition. When the fermentation was performed for 3 weeks under these conditions, the effluent medium showed stable ethanol concentrations of 16.3 - 17.9 g/l and viable cells of 6.60-7.16 log cells/ml without contamination. Trace amounts of methyl, n-propyl, iso-butyl, isoamyl alcohols besides ethanol were detected.

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Ethanol Fermentation Characteristics in Chemostat Culture of Kluyveromyces fragilis (효모 Kluyveromyces fragilis의 연속 에탄올 발효 특성에 관한 연구)

  • 허병기;김진한류장수목영일
    • KSBB Journal
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    • v.4 no.3
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    • pp.253-258
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    • 1989
  • It was investigated that the influent sugar concentration and the dilution rate have an influence on the ethanol fermentation characteristics at steady state in a chemostat culture of K. fragilis using Jerusalem Artichoke juice as substrate. And also the optimum condition of high ethanol productivity and low residual sugar output in the ethanol production by K. fragilis, was clarified to be given at a dilution rate of 0.22/hr and at an influent sugar concentration of 85g/l.

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The Biodegradation of Mixtures of Benzene,Phenol,and Toluene by Mixed and Monoculture of Bacteria (단일배양 및 혼합배양에 의한 Benzene, Phenol 및 Toluene 혼합물의 생분해)

  • Lee, Chang-Ho;Oh, Hee-Mock;Kwon, Tae-Jong;Kwon, Gi-Seok;Kim, Seong-Bin;Kho, Yung-Hee;Yoon, Byung-Dae
    • Microbiology and Biotechnology Letters
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    • v.22 no.6
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    • pp.685-691
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    • 1994
  • The biodegradation of aromatic compounds by mixed and monoculture was investigated in an artificial wastewater containing 500 mg/l of benzene(B), phenol(P), and toluene(T) in various combinations. None of three strains utilized P-xylene(X) as a carbon source, but they grew well on p-xylene in mixtures with benzene and toluene. In the mixed culture on mixed substrate, the length of lag phase was different depending on the nature of mixture. Cell growths of Flavobac- terium sp. BEN2 and Acinetobacter sp. GEM63 were inhibited in the presence of a 500 mg/l of phenol. When the mixed culture of three strains was cultured in a bench-scale reactor containing artificial wastewater, each of benzene, phenol, and toluene was not detected at 30 hrs, 50 hrs, and 12 hrs after incubation in the treatment. The removal rates of COD$_{t}$(total COD) and COD$_{s}$,(soluble COD) of upper phase after centrifugation during early 50 hrs were ca. 80% and ca. 93.8%, respectively.

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Production of Liquiritigenin with Cell-based Biotransformation and Its Anti-Aging Activity (균사체 생물전환기술을 이용한 리퀘리티게닌 생산과 항노화 활성)

  • Hwang, Hye Jin;Jeong, Sang Chul;Park, Jong Pil
    • KSBB Journal
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    • v.30 no.4
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    • pp.166-174
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    • 2015
  • In this study, an efficient whole cell-based biotransformation for the production of liquiritigenin was developed using Laetiporus sulphureus CS0218 as biocatalyst and aqueous extracts of Glycyrrhiza uralensis as co-substrate, respectively. In order to determine the efficacy of this method, the optimal bioconversion conditions including mycelial growth, three important enzyme activities (${\beta}$-glucosidase, ${\alpha}$-rhamnosidase and ${\beta}$-xylosidase), and apparent viscosity of culture broth were monitored. After optimization, aqueous extracts of G. uralensis were added to the culture medium to directly produce algycone liquiritigenin. By applying this strategy, 67.5% of liquiritin was converted to liquiritigenin at pH 3.0 after 9 days of incubation and finally liquiritigenin was purified from the reaction mixture. And then, their biological activities including anti-oxidant and superoxide dismutase were observed. In fact, purified liquiritigenin was capable of bi-directional functions (i.e., either up-regulation or down-regulation of SIRT1 which is associated with aging). The results indicate that this strategy would be beneficial to produce biologically active liquiritigenin and could be used in pharmaceutical, cosmetic and food applications.

Bio-gas Production from Nemopilema nomurai Using Anaerobic Digestion (혐기성 소화를 이용한 노무라입깃 해파리로부터 바이오 가스 생산)

  • Kim, Ji-Youn;Lee, Sung-Mok;Kim, Jong-Hun;Lee, Jae-Hwa
    • KSBB Journal
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    • v.25 no.6
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    • pp.547-552
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    • 2010
  • The recent bloom of a very large jellyfish Nemopilema nomurai has caused a danger to sea fishery and sea bathers. Presently, Nemopilema nomurai is thrown away through a separator system in the sea. The objective of this work was to produce bio-gas from Nemopilema nomurai by using anaerobic digestion. The bio-gas includes the hydrogen or the methane gases. It relates that Nemopilema nomurai is effectually changed into the renewable energy. When the jellyfish biomass was used as an organic carbon source the bio-gases were evolved. The aim of this study was to determine the optimal conditions for hydrogen and methane gases production according to the substrate concentrations of Nemopilema nomurai, optimal culture condition and the sludge-pretreatment without pH control. The optimal culture condition was found to be $35^{\circ}C$ and the heat-treatments of jellyfish was done at $120^{\circ}C$ for 30 min. The production rate of hydrogen and methane gas were found to be 8.8 mL/L/h, 37.2 mL/L/h from 1.5 g of dry Nemopilema nomurai.

Steroid modification with aspergillus phoenices (Aspergillus phoenicis를 이용한 steroid의 변형)

  • 김말남;이영종
    • Korean Journal of Microbiology
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    • v.23 no.4
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    • pp.297-301
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    • 1985
  • The dependence of activities of Aspergillus phoenicis on the culture conditions in the progesterone transformation reaction was investigated. In the beginning of the reaction, $6{\beta},\;11{\alpha}-dihydroxyprogesterone$ was not produced even at high concentration of $11{\alpha}-hydroxyprogesterone$. However, large amount of the product was obtained after the complete exhaustion of progesterone. When spores of A.phoenicis replaced mycelia as enzyme source, $11{\alpha}-hydroxyprogesterone$ was produced after a considerably long indyction period, and its maximum production rate followed the exponential growth phase. The $6{\beta}-hydroxylation\;of\;11{\alpha}-hydroxyprogesterone$ continued, even after the stationary growth phase. A. phoenicis showed high enzyme activity for these reactions when the phosphate buffer solutions were used in place of the ordinary culture medium. The buffer solutions of low pH gave more yield of $11{\alpha}-hydroxyprogesterone$ than those of high pH. However, the addition of flucose to the buffer solutions did not activate the transformation reaction. The presence of progesterone seems to be necessary for the induction of enzymes for the $6{\beta}-hydroxylation\;of\;11{\alpha}-hydroxyprogesterone\;since\;6{\beta},\;11{\alpha}-dihydroxyprogesterone$ is not produced in the reaction medium containing only $11{\alpha}-hydroxyprogesterone$ as a substrate.

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Kinetic and Thermodynamic Analysis of AChE Inhibition of Solvent Extract Fractions from Inonotus obliquus (차가버섯 용매추출분획의 Acetylcholinesterase 저해활성에 대한 동역학 및 열역학적 해석)

  • Kim, Hak-Kyu;Hur, Won;Hong, Eok Kee;Lee, Shin-Young
    • Food Engineering Progress
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    • v.15 no.4
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    • pp.289-296
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    • 2011
  • Twenty four fractions by solvent extraction and/or acid precipitation from fruit body and culture broth of Inonotus obliquus were prepared, and their inhibitory effect against acetylcholinesterase (AChE) was investigated. Among these fractions, acid (1 M HCl) precipitates from cell-free culture broth and fruit body exhibited the highest inhibitory effect on AChE in vitro. Acid precipitates inhibited AChE activity in a concentration-dependant manner and $IC_{50}$ values of both acid precipitates were 0.53 mg/mL. The inhibition pattern was general non-competitive inhibition. The energetic parameters were also determined by dual substrate/temperature design. Both acid precipitates increased the values of Ea, ${\Delta}H,/;{\Delta}G$ and ${\Delta}H^{\ast}$ decreasing the value of ${\Delta}S$ for AChE. The results implied that the acid precipitates from I. obliquus increased the thermodynamic barrier, leading to the breakdown of ES complex and the formation of products as inhibitory mechanism.

Enhancing anaerobic digestion of vegetable waste and cellulose by bioaugmentation with rumen culture

  • Jo, Yeadam;Hwang, Kwanghyun;Lee, Changsoo
    • Membrane and Water Treatment
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    • v.10 no.3
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    • pp.213-221
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    • 2019
  • Anaerobic digestion (AD) has been widely used to valorize food waste (FW) because of its ability to convert organic carbon into $CH_4$ and $CO_2$. Korean FW has a high content of fruits and vegetables, and efficient hydrolysis of less biodegradable fibers is critical for its complete stabilization by AD. This study examined the digestates from different anaerobic digesters, namely Rs, Rr, and Rm, as the inocula for the AD of vegetable waste (VW) and cellulose (CL): Rs inoculated with anaerobic sludge from an AD plant, Rr inoculated with rumen fluid, and Rm inoculated with anaerobic sludge and augmented with rumen fluid. A total of six conditions ($3\;inocula{\times}2\;substrates$) were tested in serial subcultures. Biogas yield was higher in the runs inoculated with Rm than in the other runs for both VW (up to 1.10 L/g VS added) and CL (up to 1.05 L/g VS added), and so was biogas production rate. The inocula had different microbial community structures, and both substrate type and inoculum source had a significant effect on the formation and development of microbial community structures in the subcultures. The overall results suggest that the bioaugmentation with rumen microbial consortium has good potential to enhance the anaerobic biodegradability of VW, and thereby can help more efficiently digest high fiber-content Korean FW.

Culture Conditions of E. coli Harboring Human O-Linked N-Acetyl-${\beta}$-Glucosaminidase Gene and Enzymatic Properties (사람의 O-linked-N-acetyl-${\beta}$-D-glucosaminidase 유전자를 함유한 대장균의 배양조건과 효소학적 특성)

  • 강대욱;조용권;서현효
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.147-153
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    • 2004
  • Protein modification by N-acetyl-${\beta}$-D-glucosamine (O-G1cNAc) on the hydroxyl groups of Ser or Thr ubiq-uitously occurs in eukaryotic cells and is involved in many cellular phenomena. The level of O-G1cNAc-mod-ified protein is regulated by OGT and O-GlcNAcase enzymes. We have tried to produce recombinant O-GlcNAcase in E. coli as an effort to establish in vitro screening system for modulators of O-GlcNAcase. The culture conditions for improvement of O-GlcNAcase productivity, were as follows: induction temperature, $30^{\circ}C$; the concentration of L-arabinose, 0.02% and induction time, 5 hr. Under these culture conditions, E. coli cells containing O-GlcNAcase gene had no enzyme activity until up to 3 hr culture. However, O-GlcNAcase activity dramatically increased from 3 to 5 hr culture. It almost maintained the same level after 5 hr culture. Western blot analysis verified the amount of expressed O-GlcNAcase increased with culture time, being con-sistent with activity data. The optimal reaction condition determined in this study was as follows: protein quan-tity, $5{\mu}g$; reaction time, 30 min; reaction temperature, $45^{\circ}C$; substrate concentration, 2 mM; reaction pH, 6.5. Methanol had little effect on O-GlcNAcase activity and 90% of activity were retained at 10%. Only 15% resid-ual activity were detected at 5% of chloroform.

Field Studios of In-situ Aerobic Cometabolism of Chlorinated Aliphatic Hydrocarbons

  • Semprini, Lewts
    • Proceedings of the Korean Society of Soil and Groundwater Environment Conference
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    • 2004.04a
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    • pp.3-4
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    • 2004
  • Results will be presented from two field studies that evaluated the in-situ treatment of chlorinated aliphatic hydrocarbons (CAHs) using aerobic cometabolism. In the first study, a cometabolic air sparging (CAS) demonstration was conducted at McClellan Air Force Base (AFB), California, to treat chlorinated aliphatic hydrocarbons (CAHs) in groundwater using propane as the cometabolic substrate. A propane-biostimulated zone was sparged with a propane/air mixture and a control zone was sparged with air alone. Propane-utilizers were effectively stimulated in the saturated zone with repeated intermediate sparging of propane and air. Propane delivery, however, was not uniform, with propane mainly observed in down-gradient observation wells. Trichloroethene (TCE), cis-1, 2-dichloroethene (c-DCE), and dissolved oxygen (DO) concentration levels decreased in proportion with propane usage, with c-DCE decreasing more rapidly than TCE. The more rapid removal of c-DCE indicated biotransformation and not just physical removal by stripping. Propane utilization rates and rates of CAH removal slowed after three to four months of repeated propane additions, which coincided with tile depletion of nitrogen (as nitrate). Ammonia was then added to the propane/air mixture as a nitrogen source. After a six-month period between propane additions, rapid propane-utilization was observed. Nitrate was present due to groundwater flow into the treatment zone and/or by the oxidation of tile previously injected ammonia. In the propane-stimulated zone, c-DCE concentrations decreased below tile detection limit (1 $\mu$g/L), and TCE concentrations ranged from less than 5 $\mu$g/L to 30 $\mu$g/L, representing removals of 90 to 97%. In the air sparged control zone, TCE was removed at only two monitoring locations nearest the sparge-well, to concentrations of 15 $\mu$g/L and 60 $\mu$g/L. The responses indicate that stripping as well as biological treatment were responsible for the removal of contaminants in the biostimulated zone, with biostimulation enhancing removals to lower contaminant levels. As part of that study bacterial population shifts that occurred in the groundwater during CAS and air sparging control were evaluated by length heterogeneity polymerase chain reaction (LH-PCR) fragment analysis. The results showed that an organism(5) that had a fragment size of 385 base pairs (385 bp) was positively correlated with propane removal rates. The 385 bp fragment consisted of up to 83% of the total fragments in the analysis when propane removal rates peaked. A 16S rRNA clone library made from the bacteria sampled in propane sparged groundwater included clones of a TM7 division bacterium that had a 385bp LH-PCR fragment; no other bacterial species with this fragment size were detected. Both propane removal rates and the 385bp LH-PCR fragment decreased as nitrate levels in the groundwater decreased. In the second study the potential for bioaugmentation of a butane culture was evaluated in a series of field tests conducted at the Moffett Field Air Station in California. A butane-utilizing mixed culture that was effective in transforming 1, 1-dichloroethene (1, 1-DCE), 1, 1, 1-trichloroethane (1, 1, 1-TCA), and 1, 1-dichloroethane (1, 1-DCA) was added to the saturated zone at the test site. This mixture of contaminants was evaluated since they are often present as together as the result of 1, 1, 1-TCA contamination and the abiotic and biotic transformation of 1, 1, 1-TCA to 1, 1-DCE and 1, 1-DCA. Model simulations were performed prior to the initiation of the field study. The simulations were performed with a transport code that included processes for in-situ cometabolism, including microbial growth and decay, substrate and oxygen utilization, and the cometabolism of dual contaminants (1, 1-DCE and 1, 1, 1-TCA). Based on the results of detailed kinetic studies with the culture, cometabolic transformation kinetics were incorporated that butane mixed-inhibition on 1, 1-DCE and 1, 1, 1-TCA transformation, and competitive inhibition of 1, 1-DCE and 1, 1, 1-TCA on butane utilization. A transformation capacity term was also included in the model formation that results in cell loss due to contaminant transformation. Parameters for the model simulations were determined independently in kinetic studies with the butane-utilizing culture and through batch microcosm tests with groundwater and aquifer solids from the field test zone with the butane-utilizing culture added. In microcosm tests, the model simulated well the repetitive utilization of butane and cometabolism of 1.1, 1-TCA and 1, 1-DCE, as well as the transformation of 1, 1-DCE as it was repeatedly transformed at increased aqueous concentrations. Model simulations were then performed under the transport conditions of the field test to explore the effects of the bioaugmentation dose and the response of the system to tile biostimulation with alternating pulses of dissolved butane and oxygen in the presence of 1, 1-DCE (50 $\mu$g/L) and 1, 1, 1-TCA (250 $\mu$g/L). A uniform aquifer bioaugmentation dose of 0.5 mg/L of cells resulted in complete utilization of the butane 2-meters downgradient of the injection well within 200-hrs of bioaugmentation and butane addition. 1, 1-DCE was much more rapidly transformed than 1, 1, 1-TCA, and efficient 1, 1, 1-TCA removal occurred only after 1, 1-DCE and butane were decreased in concentration. The simulations demonstrated the strong inhibition of both 1, 1-DCE and butane on 1, 1, 1-TCA transformation, and the more rapid 1, 1-DCE transformation kinetics. Results of tile field demonstration indicated that bioaugmentation was successfully implemented; however it was difficult to maintain effective treatment for long periods of time (50 days or more). The demonstration showed that the bioaugmented experimental leg effectively transformed 1, 1-DCE and 1, 1-DCA, and was somewhat effective in transforming 1, 1, 1-TCA. The indigenous experimental leg treated in the same way as the bioaugmented leg was much less effective in treating the contaminant mixture. The best operating performance was achieved in the bioaugmented leg with about over 90%, 80%, 60 % removal for 1, 1-DCE, 1, 1-DCA, and 1, 1, 1-TCA, respectively. Molecular methods were used to track and enumerate the bioaugmented culture in the test zone. Real Time PCR analysis was used to on enumerate the bioaugmented culture. The results show higher numbers of the bioaugmented microorganisms were present in the treatment zone groundwater when the contaminants were being effective transformed. A decrease in these numbers was associated with a reduction in treatment performance. The results of the field tests indicated that although bioaugmentation can be successfully implemented, competition for the growth substrate (butane) by the indigenous microorganisms likely lead to the decrease in long-term performance.

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