• Title/Summary/Keyword: Metabolic Engineering

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Effect of Methionine on Cephalosporin C Production in a Fluidized- bed Bioreactor (유동층 생물반응기에서 세파로스포린 C 생산에 대한 메치오닌의 영향)

  • Kim, Eui-Yong;Yoo, Young-Je;Park, Young-Hoon
    • Microbiology and Biotechnology Letters
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    • v.17 no.6
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    • pp.611-618
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    • 1989
  • Effects of methionine on cephalosporin C(CPC) production in a fluidized-bed bioreactor were investigated using bioparticles of Cephalosporium acremonium. Since methionine was found to be an important metabolic regulator on the synthesis of cephalosporin C, the effects of its concentration in the cuture broth and feeding mode to the bioreactor were studied. It was observed that the presence of initial methionine was essential for higher cephalosporin C production and there existed an optimal content of methionine. Carbon consumption rate also increased significantly under the presence of methionine. Production of cephalosporin C was most active when methionine was exhausted in the broth; however its additional feeding did not enhance the antibiotic production in the fluidized-bed bioreactor as much as expected. It was therfore considered important to feed an optimal content of methionine at the early operating stage for a higher cephalosporin C production in a fluidized-bed bioreactor. An interesting thing to note was that titre of the antibiotic with reused bioparticles was about 2 times higher in the methionine containing medium than that without methionine. Therefore repeated use of bioparticles, with an optimal content of methionine, was believed to be very useful to enhance to process productivity.

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Production of Fructose 6-Phoschate from Starch Using Thermostable Enzymes (내열성 효소를 이용한 전분으로부터 6-인산과당의 제조)

  • Kwun, Kyu-Hyuk;Cha, Wol-Suk;Kim, Bok-Hee;Shin, Hyun-Jae
    • KSBB Journal
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    • v.22 no.5
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    • pp.345-350
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    • 2007
  • Phosphosugars are found in all living organisms and are commercially valuable compounds with possible applications in the development of a wide range of specialty chemicals and medicines. In carbohydrate metabolism, fructose 6-phosphate (F6P) is an essential intermediate formed by phosphorylation of 6' position of fructose in glycolysis, gluconeogenesis, pentose phosphate pathway and Calvin cycle. In glycolysis, F6P lies within the glycolysis metabolic pathway and is produced by isomerisation of glucose 6-phosphate. For large-scale production, F6P could be produced from starch using many enzymes such as pullulanase, starch phosphorylase, isomerase and mutase. In enzymatic reactions carried out at high temperatures, the solubility of starch is increased and microbial contamination is minimized. Thus, thermophile-derived enzymes are preferred over mesophile-derived enzymes for industrial applications using starch. Recently, we reported the production of glucose 1-phosphate (G1P) from starch by Thermus caldophilus GK24 enzymes. Here we report the production of F6P from starch through three steps; from starch to glucose 1-phosphate (glucan phosphorylase, GP), then glucose 6-phosphate (phosphoglucomutase, GM) and then F6P (phosphoglucoisomerase, GI). Using 200 L of 1.2% soluble starch solution in potassium phosphate buffer, 1,253 g of G1P were produced. Then, 30% yields of F6P were attained at the optimum reaction conditions of GM : G1 (1 : 2.3), 63.5$^{\circ}C$, and pH 6.85. The optimum conditions were found by response surface methodology and the theoretical values were confirmed by the experiments. The optimum starch concentrations were 20 g/L under the given conditions.

Thermophilic Biohydrogen Production from Glucose with a Long-term Operation of CSTR (CSTR의 장기운전을 통한 포도당으로부터의 고온 수소생산)

  • Ahn, Yeong-Hee;Oh, You-Kwan;Park, Sung-Hoon
    • KSBB Journal
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    • v.20 no.6
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    • pp.425-430
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    • 2005
  • Thermophilic $H_2$ was produced for 1 year using a bench-scale continuous stirred tank reactor(CSTR). The CSTR was inoculated with anaerobically digested sludge after heat treatment and fed with a glucose-based medium. The reactor showed relatively short start-up period(30 days) and high maximal $H_2$ yield(2.4 mol $H_2/mol$ glucose). Keeping pH 5.0 or less suppressed methanogenic activity. Bacteria affiliated with Thermoanaerobacterium thermosaccharolyticum kept being dominant from approximately 40 days as determined by DGGE. Environmental perturbation(pH or temperature) caused the decrease of biomass concentration in the reactor and the instability of reactor performance, $H_2$ production rate and $H_2$ yield. The unstable performance was accompanied with high concentration of lactate in the effluent. Taken together, the poor recovery of CSTR after perturbations could be partly explained by low biomass concentration and/or metabolic shift of the major population in the CSTR.

A New Shock Index for Predicting Survival of Rats with Hemorrhagic Shock Using Perfusion and Lactate Concentration Ratio (흰쥐의 출혈성 쇼크에서 관류와 젖산 농도 비를 이용한 새로운 생존 예측 지표 개발)

  • Choi, Jae-Lim;Nam, Ki-Chang;Kwon, Min-Kyung;Jang, Kyung-Hwan;Kim, Deok-Won
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.48 no.4
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    • pp.1-9
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    • 2011
  • Hemorrhagic shock is a clinically widespread syndrome characterized by inadequate oxygenation and supply. It is important to diagnose hemorrhagic shock in its early stage for improving treatment effects and survival rate. However, an accurate diagnosis and treatment could be delayed in the early stage of hemorrhagic shock by evaluating only vital signs such as heart rate and blood pressure. There have been many studies for the early diagnosis of hemorrhagic shock, reporting that lactate concentration and perfusion were useful variables for tissue hypoxia and metabolic acidosis. In this study, we measured both perfusion using a laser Doppler flowmeter and lactate concentration from the volume controlled hemorrhagic shock using rats. We also proposed a new shock index which was calculated by dividing lactate concentration by perfusion for early diagnosis. As a result of the survival prediction by the proposed index with the receiver operating characteristic curve method, the sensitivity, specificity, and accuracy of survival were 90.0, 96.7 and 94.0%, respectively. The proposed index showed the fastest significant difference among the other parameters such as blood pressure and heart rate. It could offer early diagnosis and effective treatment for human hemorrhagic shock if it is applicable to humans.

Characterization of Microbial Community in the Leachate Associated with the Decomposition of Entombed Pigs

  • Yang, Seung-Hak;Hong, Sun Hwa;Cho, Sung Back;Lim, Joung Soo;Bae, Sung Eun;Ahn, Heekwon;Lee, Eun Young
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1330-1335
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    • 2012
  • Foot and mouth disease (FMD) is one of the acute infectious diseases in hoofed and even-toed mammals, including pigs, and it occurs via acute infection by Aphthovirus. When FMD is suspected, animals around the location of origin are typically slaughtered and buried. Other methods such as rendering, composting, and incineration have not been verified in practice in Korea. After the FMD incident, the regular monitoring of the microbial community is required, as microorganisms greatly modify the characteristics of the ecosystem in which they live. This is the result of their metabolic activities causing chemical changes to take place in the surrounding environment. In this study, we investigated changes in the microbial community during a 24 week period with DNA extracts from leachate, formed by the decomposition of buried pigs at a laboratory test site, using denaturing gradient gel electrophoresis (DGGE) with a genomic DNA. Our results revealed that Bacteroides coprosuis, which is common in pig excreta, and Sporanaerobacter acetigenes, which is a sulfur-reduced microbe, were continuously observed. During the early stages (0~2 weeks) of tissue decomposition, Clostridium cochlearium, Fusobacterium ulcerans, and Fusobacterium sp., which are involved in skin decomposition, were also observed. In addition, various microbes such as Turicibacter sanguinis, Clostridium haemolyticum, Bacteroides propionicifaciens, and Comamonas sp. were seen during the later stages (16~24 weeks). In particular, the number of existing microbial species gradually increased during the early stages, including the exponential phase, decreased during the middle stages, and then increased again during the later stages. Therefore, these results indicate that the decomposition of pigs continues for a long period of time and leachate is created continuously during this process. It is known that leachate can easily flow into the neighboring environment, so a long-term management plan is needed in burial locations for FMD-infected animals.

Effect of PVA-Encapsulation on Hydrogen Production and Bacterial Community Structure (수소 생산과 세균 군집구조에 미치는 PVA-포괄고정화의 영향)

  • Yun, Jeonghee;Kim, Tae Gwan;Cho, Kyung-Suk
    • Microbiology and Biotechnology Letters
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    • v.42 no.1
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    • pp.41-50
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    • 2014
  • In this study, the performances of PVA-encapsulation and non-encapsulation in a fed-batch bioreactor system were compared for biohydrogen production. Hydrogen production in the PVA-encapsulation bioreactor was not significantly different in comparison to the non-encapsulation bioreactor. However, the hydrogen gas in the encapsulation bioreactor could be stably produced when it was exposed to environmental difficulties such as pH impact by the accumulation of organic acids as fermentative metabolic products. Bacterial communities by DGGE analysis were differently shifted between the PVA-encapsulation and non-encapsulation bioreactors from the initial sludge. The community of hydrogen producing bacteria was stable during the experimental period in the PVA-encapsulation bioreactor compared to the non-encapsulation method. The absolute quantitation of the DNA copy number by a high-throughput droplet digital PCR system for six genera contributed to hydrogen production showing that the numbers of dominant bacteria existed at similar levels in the two bioreactors regardless of encapsulation. In both of two bioreactors, not only Clostridium and Enterobacter, which are known as anaerobic hydrogen producing bacteria, but also Firmicutes, Ruminococcus and Escherichia existed with $1{\times}10^5-1{\times}10^6$ copy numbers of ml-samples exhibiting rapid growth during the initial operation period.

Natural Origin Polymers: Applications as Wound Care Materials (자연 고분자 : 상처 치료 재료로 활용)

  • Karadeniz, Fatih;Sung, Hye Kyeong;Kim, Han Seong
    • Journal of Life Science
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    • v.29 no.3
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    • pp.382-393
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    • 2019
  • Wound care is a health industry concern affecting millions worldwide. Recent increase in metabolic disorders such as diabetes comes with elevated risk of wound-based complications. Treatment and management of wounds are difficult practices due to complexity of the wound healing process. Conventional wound dressings and treatment applications only provide limited benefits which are mainly aimed to keep wound protected from external factors. To improve wound care, recent developments make biopolymers to be of high interest and importance to researchers and medical practitioners. Biopolymers are polymers or natural origin produced by living organisms. They are credited to be highly biocompatible and biodegradable. Currently, studies reported biopolymers to exhibit various health beneficial properties such as antimicrobial, anti-inflammatory, hemostatic, cell proliferative and angiogenic activities which are crucial for effective wound management. Several biopolymers, namely chitosan, cellulose, collagen, hyaluronic acid and alginic acid have been already investigated and applied as wound dressing agents. Different derivatives of biopolymers have also been developed by cross-linking with other molecules, grafting with other polymers, and loading with bioactive agents or drugs which showed promising results towards wound healing without any undesired outcome such as scarring and physiological abnormalities. In this review, current applications of common biopolymers in wound treatment industry are highlighted to be a guide for further applications and studies.

Physico-chemical Properties and Antibacterial Activities of Lactonic Sophorolipid (락톤형 소포로리피드의 물리화학적 특성 및 항균효과)

  • Cho, Soo A;Eom, Gyeong Tae;Jin, Byung Suk
    • Applied Chemistry for Engineering
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    • v.30 no.3
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    • pp.303-307
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    • 2019
  • Sophorolipid is a biological surfactant of the glycolipid structure produced by Candida bombicola, which generally exists as a mixture of acidic and lactonic forms. In this study, we investigated physico-chemical properties, antibacterial activities, and cytotoxicity of the sophorolipid containing more than 96% of the lactonic form, produced by the gene regulation of production strains and application of a metabolic engineering technique. The lactonic sophorolipid showed a weak acidity in the range of pH 3.2~4.6 when diluted in water at the concentrations from 1 to 0.001 wt%. The $pK_a$ value of the lactonic sophorolipid was estimated to be around 4.3 from the acid-base titration curve. The critical micelle concentration (CMC) of the lactonic sophorolipid was $10^{-2}wt%$, at which the surface tension of aqueous solution was reduced to 36 mN/m. The lactonic sophorolipid showed the minimum inhibitory concentrations (MIC) of $1{\times}10^{-3}$ and $5{\times}10^{-3}g/mL$ against Propionibacterium acnes and Corynebacterium xerosis, respectively. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide] assay showed that cytotoxicity of the lactonic sophorolipid was ten times lower than that of triclosan.

Cardiopulmonary Response during Mechanical Horseback Riding Exercise in Healthy Adult (건강한 성인에서 기계승마 운동 동안의 심폐 반응)

  • Boo, Seung-Hyun;Noh, Se-Eung;Lee, Eun-Sun;Joo, Min-Cheol
    • Journal of rehabilitation welfare engineering & assistive technology
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    • v.10 no.4
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    • pp.243-249
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    • 2016
  • In this paper, we evaluate the effectiveness of mechanical horseback riding exercise training for cardiopulmonary responses in 26 healthy adults (male 14, female 12) and suggest its clinical applicability. Exercise was performed using a mechanical horseback riding machine, JOBA(R) (Panasonic Electric Works, Osaka, Japan) and riding movement speed was increased from level 2 (0.73Hz), to 4 (0.9Hz), to 6 (1.03Hz), to 8 (1.2Hz), to 9 (1.3Hz) and maintained 3minutes in each level. Heart rate, blood pressure, maximum oxygen consumption (VO2max) were measured by respiratory gas analysis system (TruOne 2400, Parvo Medics, USA) and exercise tolerance test system(Q Stress, Cardiac Science, USA). To measure exercise intensity, metabolic equivalent (MET) were measured and percent of Heart rate reserve (%HRR), percent of peak heart rate (%peak HR) and calorie consumption were calculated. The results were analysed by SPSS 18.0. We found that the exercise intensity of mechanical horseback riding exercise using JOBA(R) for 15 minutes is moderate in healthy adult who received pre-training.

Production of Vanillin from Ferulic Acid Using Recombinant Strains of Escherichia coli

  • Yoon Sang-Hwal;Li Cui;Lee Young-Mi;Lee Sook-Hee;Kim Sung-Hee;Choi Myung-Suk;Seo Weon-Taek;Yang Jae-Kyung;Kim Jae-Yeon;Kim Seon-Won
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.378-384
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    • 2005
  • Vanillin is one of the world's principal flavoring compounds, and is used extensively in the food industry. The potential vanillin production of the bacteria was compared to select and clone genes which were appropriate for highly productive vanillin production by E. coli. The fcs (feruloyl-CoA synthetase) and ech (enoyl-CoA hydratase/aldolase) genes cloned from Amycolatopsis sp. strain HR104 and Delftia acidovorans were introduced to pBAD24 vector with $P_{BAD}$ promoter and were named pDAHEF and pDDAEF, respectively. We observed 160 mg/L vanillin production with E. coli harboring pDAHEF, whereas 10 mg/L of vanillin was observed with pDAHEF. Vanillin production was optimized with E. coli harboring pDAHEF. Induction of the fcs and ech genes from pDAHEF was optimized with the addition of 13.3 mM arabinose at 18 h of culture, from which 450 mg/L of vanillin was produced. The feeding time and concentration of ferulic acid were also optimized by the supplementation of $0.2\%$ ferulic acid at 18 h of culture, from which 500 mg/L of vanillin was obtained. Under the above optimized condition of arabinose induction and ferulic acid supplementation, vanillin production was carried out with four different types of media, M9, LB, 2YT, and TB. The highest vanillin production, 580 mg/L, was obtained with LB medium, a 3.6 fold increase in comparison to the 160 mg/L obtained before the optimization of vanillin production.