• Title/Summary/Keyword: bioreactor optimization

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Fermentation and Metabolic Pathway Optimization to De Novo Synthesize (2S)-Naringenin in Escherichia coli

  • Zhou, Shenghu;Hao, Tingting;Zhou, Jingwen
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1574-1582
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    • 2020
  • Flavonoids have diverse biological functions in human health. All flavonoids contain a common 2-phenyl chromone structure (C6-C3-C6) as a scaffold. Hence, in using such a scaffold, plenty of high-value-added flavonoids can be synthesized by chemical or biological catalyzation approaches. (2S)-Naringenin is one of the most commonly used flavonoid scaffolds. However, biosynthesizing (2S)-naringenin has been restricted not only by low production but also by the expensive precursors and inducers that are used. Herein, we established an induction-free system to de novo biosynthesize (2S)-naringenin in Escherichia coli. The tyrosine synthesis pathway was enhanced by overexpressing feedback inhibition-resistant genes (aroGfbr and tyrAfbr) and knocking out a repressor gene (tyrR). After optimizing the fermentation medium and conditions, we found that glycerol, glucose, fatty acids, potassium acetate, temperature, and initial pH are important for producing (2S)-naringenin. Using the optimum fermentation medium and conditions, our best strain, Nar-17LM1, could produce 588 mg/l (2S)-naringenin from glucose in a 5-L bioreactor, the highest titer reported to date in E. coli.

Characterization of HEK293 and Namalwa Cell Cultures by Using Design of Experiment (실험계획법을 이용한 HEK293 및 Namalwa 세포배양 특성 규명)

  • Kang, Kyung-Ho;Seo, Joon-Serk;Kim, Dong-Il
    • KSBB Journal
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    • v.27 no.3
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    • pp.186-194
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    • 2012
  • Various human host cell lines, which are more effective than the other original human cell lines, have been developed and used. Highly efficient human cell line can be obtained from the fusion between human embryonic kidney 293 (HEK293) and human Burkitt's lymphoma cells (Namalwa). Fused cell line has the advantages of both cell lines such as the high transfection efficacy of HEK293 cells and the constitutive expression of Epstein-Barr virus (EBV) genome which is related with high expression of target protein and anti-apoptotic growth of Namalwa cells. In this study, characterization of two original cell lines was performed by using design of experiment (DOE) considering cell maintenance, media development, optimization of culture condition, and scale-up. The formation of aggregates was apparent with high glutamine concentration at more than 6 mM. Supplementation of hydrolysates showed positive effects on the growth performances of HEK293 cells. On the contrary, Namalwa cells showed negative results. It was confirmed that Namalwa cells were more sensitive to lower temperature at $35^{\circ}C$ and hyperosmotic condition over 260 mOsm/kg. In addition, both cell lines showed limited growth in 3-L bioreactor due to shear stress.

ENGINEERING A BIOARTIFICIAL LIVER DEVICE

  • Park, Jae-Sung;Yarmush, Martin L.;Tilles, Arno W.
    • Proceedings of the KSME Conference
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    • 2008.11a
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    • pp.1419-1426
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    • 2008
  • Fulminant hepatic failure is a clinical syndrome associated with a high mortality rate. Orthotopic liver transplantation is the only clinically proven effective treatment for patients with end-stage liver disease who do not respond to medical management. A major limitation of this treatment modality is the scarcity of donor organs available, resulting in patients dying while waiting for a donor liver. An extracorporeal bioartificial liver (BAL) device containing viable hepatocytes has the potential to provide temporary hepatic support to liver failure patients, serving as a bridge to transplantation while awaiting a suitable donor. In some patients, providing temporary hepatic support may be sufficient to allow adequate regeneration of the host liver, thereby eliminating the need for a liver transplant. Although the BAL device is a promising technology for the treatment of liver failure, there are several technical challenges that must be overcome in order to develop systems with sufficient processing capacity and of manageable size. In this overview, the authors describe the critical issues involved in developing a BAL device. They also discuss their experiences in hepatocyte culture optimization within the context of a microchannel flat-plate BAL device.

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Formulation of a novel bacterial consortium for the effective biodegradation of phenol

  • Dhanya, V.
    • Advances in environmental research
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    • v.9 no.2
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    • pp.109-121
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    • 2020
  • Phenol is frequently present as the hazardous pollutant in petrochemical and pesticide industry wastewater. Because of its high toxicity and carcinogenic potential, a proper treatment is needed to reduce the hazards of phenol carrying effluent before being discharged into the environment. Phenol biodegradation with microbial consortium offers a very promising approach now a day's. This study focused on the formulation of phenol degrading bacterial consortium with three bacterial isolates. The bacterial strains Bacillus cereus strain VCRC B540, Bacillus cereus strain BRL02-43 and Oxalobacteraceae strain CC11D were isolated from detergent contaminated soil by soil enrichment technique and was identified by 16s rDNA sequence analysis. Individual cultures were degrade 100 μl phenol in 72 hrs. The formulated bacterial consortium was very effective in degrading 250 μl of phenol at a pH 7 with in 48 hrs. The study further focused on the analysis of the products of biodegradation with Fourier Transform Infrared Spectroscopy (FT/IR) and Gas Chromatography-Mass Spectroscopy (GC-MS). The analysis showed the complete degradation of phenol and the production of Benzene di-carboxylic acid mono (2-ethylhexyl) ester and Ethane 1,2- Diethoxy- as metabolic intermediates. Biodegradation with the aid of microorganisms is a potential approach in terms of cost-effectiveness and elimination of secondary pollutions. The present study established the efficiency of bacterial consortium to degrade phenol. Optimization of biodegradation conditions and construction of a bioreactor can be further exploited for large scale industrial applications.

The Mutant Lactobacillus plantarum GNS300 Showed Improved Exopolysaccharide Production and Antioxidant Activity

  • Jae-Youn Jung;Deok-Ho Kwon;Yoo Jin Lee;Young Keun Song;Moon Sik Chang;Suk-Jin Ha
    • Microbiology and Biotechnology Letters
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    • v.51 no.1
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    • pp.18-25
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    • 2023
  • After random mutagenesis, the mutant Lactobacillus plantarum GNS300 showed improved exopolysaccharide production as determined by the quantification of total sugar. The mutant L. plantarum GNS300 produced 2.82 g/l of exopolysaccharide which showed 79.62% improved exopolysaccharide production compared with the parental strain. When exopolysaccharide of L. plantarum GNS300 was analyzed, the exopolysaccharide is composed of galactose (93.35%) and glucose (6.65%). Through the optimization of fermentation conditions using a bioreactor, 2.93 g/l of exopolysaccharide was produced from 20 g/l of glucose at 35℃, 500 rpm, and 0.1 vvm for 12 h. The mutant L. plantarum GNS300 exhibited 69.18% higher antioxidant activity than that from the parental strain, which might be caused by higher exopolysaccharide production. The concentrated supernatant of the mutant L. plantarum GNS300 inhibited the growth of gram-positive bacteria (Bacillus cereus and Staphylococcus aureus) and gram-negative bacteria (Escherichia coli, Vibrio parahaemolyticus, and Salmonella typhimurium).

Optimization of Interesterification Reaction for the Continuous Production of trans-Free Fat in a Packed Bed Enzyme Bioreactor with Immobilized Lipase (고정화 리파제를 이용한 충진형 효소생물반응기 내에서의 무-트랜스 유지 연속 생산을 위한 에스테르 교환 반응의 최적화)

  • Kim, Sang-Woo;Park, Kyung-Min;Ha, Jae-Uk;Lee, Jae-Hwan;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
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    • v.41 no.2
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    • pp.173-178
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    • 2009
  • Epidemiological studies showed that high trans-fat consumption is closely associated with getting the risks of cardiovascular disease. The objective of this study was to produce trans-free fat through lipase-catalyzed interesterification, as a substitute for the cream margarine commonly used in industry. Optimum conditions for interesterification in a packed bed enzyme bioreactor (PBEB) were determined using response surface methodology (RSM) based on central composite design. Three kinds of reaction variables were chosen, such as substrate flow rate (0.4-1.2 mL/min), reaction temperature (60-70$^{\circ}C$), and ratio of fully hydrogenated canola oil (FHCO, 35-45%) to evaluate their effects on the degree of interesterification. Optimum conditions from the standpoint of solid fat content (SFC) were found to be as follows: 0.4 mL/min flow rate, 64.7$^{\circ}C$ reaction temperate, and 42.8% (w/w) ratio of FHCO, respectively. The half-life of immobilized lipase in PBEB with two stages at 60$^{\circ}C$ ($1^{st}$ stage) and 55$^{\circ}C$ ($2^{nd}$ stage) was about more than 30 days as estimated by extrapolating the incubation time course of tristearoyl glycerol (TS) conversion, whereas the half-life of the enzyme in PBEB with single stage at 65$^{\circ}C$ was only about 15 days. Finally, the results from SFC analysis suggest that trans-free fat produced in this study seems to be a suitable substitute for the cream margarine commonly used in industry.

Liquid culture condition of Tremella fuciformis mycelia (흰목이 균사 액체배양 조건)

  • Chang, Hyun-You;Lee, Chan;Choi, Sung-Woo;Yun, Jong Won
    • Journal of Mushroom
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    • v.6 no.1
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    • pp.27-31
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    • 2008
  • The optimization of submerged culture conditions for mycelial growth and exopolysaccharide (EPS) production in an edible mushroom Tremella fuciformis were studied in shake flasks and bioreactors. The temperature of $28^{\circ}C$ and pH 8 in the beginning of fermentation in agitated flasks was the most efficient condition to obtain maximum mycelial biomass and EPS. The optimal medium constituents were as follows (g l-1): glucose 20, tryptone 2, $KH_2PO_4$ 0.46, $K_2HPO_4$ 1 and $MgSO_4H_2O$ 0.5. The fungus was cultivated under various agitation and aeration conditions in a 5L stirred-tank bioreactor. The maximum cell mass and EPS production were obtained at a relatively high agitation speed of 200 rpm and at an aeration rate of 2 vvm. The flow behavior of the fermentation broth was Newtonian and the maximum apparent viscosity (35 cP) was observed at a highly aerated condition (2 vvm). The EPS productivity in an airlift reactor was higher than that in the stirred-tank reactor. The EPS was protein-bound polysaccharides consisted of mainly mannose, xylose, and fructose. The molecular weights of EPS were determined to be $1.3{\sim}1.5{\times}10^6$.

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석탄 합성가스로부터 효율적인 생물학적 수소 생산에 관한 연구

  • 강환구;전희진
    • KSBB Journal
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    • v.15 no.3
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    • pp.268-273
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    • 2000
  • A microbiological hydrogen production process was optimized. Anaerobic photosynthetic bacteria like Rhodospirillum rubrum which is known to produce hydrogen from carbon monoxide efficiently and remove sulfur was used. To evaluate the potenital of this microorganism the optimization of media fermentation condition light intensity and light requirement for CO conversionwas tried in batch cultures and the continuous fermenter was also applied for this process. The gas residence time on CO conversion was sought out to get high conversion of carbon monoxide to hydrogen. Through this study the possibility of microbial synthtics gas concersion process was proposed.

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Optimization of Bioreactor Operation by Mass Transfer Coefficient (물질전달계수를 이용한 생물 반응기 운전 최적화)

  • Kim, Hyung-Soon
    • Journal of the Korean Society of Industry Convergence
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    • v.4 no.3
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    • pp.243-251
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    • 2001
  • The effects of various operating parameters(agitation speed, impeller type, antiform agents, impeller spacing etc.) on air-liquid mass transfer was characterized by volumetric mass transfer coefficient($k_La$). Also, the dual-impeller agitated systems are compared with single-impeller agitated systems with a special focus on its applications for bioreactors, $k_La$ was take over a range of 200~450 rpm of agitation speed, and 0.5~2.5 vvm of air flow rates, for four single impeller and impeller combinations consisting of four impeller types, namely rushton, pitched blade, scaba, intermig were tested. The rushton impeller showed the best $k_La$ as compared with other single impellers. The dual impeller system are found to be superior as compared to single impeller in all aspects, The best combination of the dual impeller was a intermig of axial flow type as an upper impeller and a rushton of radial flow type as a lower part. Also, the control of the DO level with the variation of agitation speed was more efficient than that with an increase in air flow rate. The addition of antiform dropped the $k_La$ very large up to 1g/L regardless the type. PPG was less effect on $k_La$ than other antiforms. The impeller spacing and presence of solute are found very effective on $k_La$. When the $NaNO_3$is presented as solute, the $k_La$ increased approximately 50% then control.

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High-Level Production of Human Papillomavirus (HPV) Type 16 L1 in Escherichia coli

  • Bang, Hyun Bae;Lee, Yoon Hyeok;Lee, Yong Jae;Jeong, Ki Jun
    • Journal of Microbiology and Biotechnology
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    • v.26 no.2
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    • pp.356-363
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    • 2016
  • Human papillomavirus (HPV), a non-enveloped, double-stranded DNA tumor virus, is a primary etiological agent of cervical cancer development. As a potential tool for prophylactic vaccination, the development of virus-like particles (VLPs) containing the HPV16 L1 capsid protein is highly desired. In this study, we developed a high-level expression system of the HPV16 L1 in Escherichia coli for the purpose of VLP development. The native gene of HPV16 L1 has many rare codons that cause the early termination of translation and result in the production of truncated forms. First, we optimized the codon of the HPV16 L1 gene to the preferable codons of E. coli, and we succeeded in producing the full-size HPV16 L1 protein without early termination. Next, to find the best host for the production of HPV16 L1, we examined a total of eight E. coli strains, and E. coli BL21(DE3) with the highest yield among the strains was selected. With the selected host-vector system, we did a fed-batch cultivation in a lab-scale bioreactor. Two different feeding solutions (complex and defined feeding solutions) were examined and, when the complex feeding solution was used, a 6-fold higher production yield (4.6 g/l) was obtained compared with that with the defined feeding solution.