• Title/Summary/Keyword: bioreactor system

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Secretory Expression and Purification of the Recombinant Duck Interleukin-2 in Pichia pastoris

  • Du, Cuihong;Han, Long;Xiao, Anfeng;Cao, Minjie
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1264-1269
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    • 2011
  • Interleukin-2 (IL-2) is a vital cytokine secreted by activated T lymphocytes, and plays an important role in the regulation of cellular functions and immunity of animals. In this study, the recombinant duck IL-2 (rduIL-2) was secretory expressed in Pichia pastoris (P. pastoris). The recombinant P. pastoris strain was cultured in shake flasks and then scaled up in a 5.0-l bioreactor. The result showed that the maximal fresh-cell-weight of 594.1 g/l and the maximal $OD_{600}$ of 408 were achieved in the bioreactor. The rduIL-2 was purified by two steps of purification procedures, and approximately 311 mg of rduIL-2/L fermentation supernatant was obtained. SDS-PAGE showed that the purified rduIL-2 constituted a homogeneous band of ~16 kDa or ~14 kDa corresponding to the glycosylated or non-glycosylated duIL-2 protein in size, respectively. The bioactivity of rduIL-2 was determined by lymphocyte proliferation assay. The result indicated that the rduIL-2 greatly promoted the proliferation of ConA-stimulated lymphocytes in vitro. The P. pastoris expression system described here could provide promising, inexpensive, and large-scale production of the rduIL-2, which lays the foundation for development of novel immunoadjuvants to enhance both the immunity of ducks against various infectious pathogens and vaccine efficacy.

Immobilization on Chitosan of a Thermophilic Trehalose Synthase from Thermus thermophilus HJ6

  • Kim, Hyun-Jung;Kim, Ae-Ran;Jeon, Sung-Jong
    • Journal of Microbiology and Biotechnology
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    • v.20 no.3
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    • pp.513-517
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    • 2010
  • A thermostable trehalose synthase (TtTSase) from Thermus thermophilus HJ6 was immobilized on chitosan activated with glutaraldehyde. The yield of immobilization was evaluated as 39.68%. The optimum pH of the immobilized enzyme was similar to that of the free enzyme. However, the optimal temperature ranges were shifted by about $4^{\circ}C$ owing to better thermal stability after immobilization. The half-life of heat inactivation for free and immobilized enzymes was 5.7 and 6.3 days at $70^{\circ}C$, respectively, thus showing a lager thermostability of the immobilized enzyme. When tested in batch reaction, the immobilized enzyme retained its relative activity of 53% after 30 reuses of reaction within 12 days, and still retained 82% of its initial activity even after 150 days at $4^{\circ}C$. A packed-bed bioreactor with immobilized enzyme showed a maximum yield of 56% trehalose from 100 mM maltose in a continuous recycling system (bed volume: 10 ml) under conditions of pH 7.0 and $70^{\circ}C$.

Multi-Dimension Scaling as an exploratory tool in the analysis of an immersed membrane bioreactor

  • Bick, A.;Yang, F.;Shandalov, S.;Raveh, A.;Oron, G.
    • Membrane and Water Treatment
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    • v.2 no.2
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    • pp.105-119
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    • 2011
  • This study presents the tests of an Immersed Membrane BioReactor (IMBR) equipped with a draft tube and focuses on the influence of hydrodynamic conditions on membrane fouling in a pilot-scale using a hollow fiber membrane module of ZW-10 under ambient conditions. In this system, the cross-flow velocities across the membrane surface were induced by a cylindrical draft-tube. The relationship between cross-flow velocity and aeration strength and the influence of the cross-flow on fouling rate (under various hydrodynamic conditions) were investigated using Multi-Dimension Scaling (MDS) analysis. MDS technique is especially suitable for samples with many variables and has relatively few observations, as the data about Membrane Bio-Reactor (MBR) often is. Observations and variables are analyzed simultaneously. According to the results, a specialized form of MDS, CoPlot enables presentation of the results in a two dimensional space and when plotting variables ratio (output/input) rather than original data the efficient units can be visualized clearly. The results indicate that: (i) aeration plays an important role in IMBR performance; (ii) implementing the MDS approach with reference to the variables ratio is consequently useful to characterize performance changes for data classification.

MBR technology for textile wastewater treatment: First experience in Bangladesh

  • Saha, Pradip;Hossain, Md. Zakir;Mozumder, Md. Salatul I.;Uddin, Md. Tamez;Islam, Md. Akhtarul;Hoinkis, Jan;Deowan, Shamim A.;Drioli, Enrico;Figoli, Alberto
    • Membrane and Water Treatment
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    • v.5 no.3
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    • pp.197-205
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    • 2014
  • For the first time in Bangladesh, a bench scale membrane bioreactor (MBR) unit was tested in treating a textile wastewater in the industry premises of EOS Textile Mills LTD, Dhaka for three months. The performance of the unit was compared with that of the conventional activated sludge treatment plant, which is in operation in the same premises. The COD and BOD removal efficiency of the MBR unit reached to around 90% and 80% respectively in 20 days whereas the removal efficiency of the conventional treatment plant was as low as 40-50% and 38-40% respectively. The outlet COD and the BOD level for the MBR unit remained stable in spite of the fluctuation in the feed value, while the conventional effluent treatment plant (ETP) failed to keep any stabilized level. The performance of the MBR unit was much superior to that of the functional ETP and the water treated by the MBR system can meet disposal standard.

Hydrogen Sulfide Removal by Immobilized Thiobacillus novellas on $SiO_2$ in a Fluidized Bed Reactor

  • Cha, Jin-Myung;Shin, Hyun-Jae;Roh, Sung-Hee;Kim, Sun-Il
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.320-324
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    • 2007
  • The removal of hydrogen sulfide ($H_2S$) from aqueous media was investigated using Thiobacillus novellas cells immobilized on a $SiO_2$ carrier (biosand). The optimal growth conditions for the bacterial strain were $30^{\circ}C$ and initial pH of 7.0. The main product of hydrogen sulfide oxidation by T. novellus was identified as the sulfate ion. A removal efficiency of 98% was maintained in the three-phase fluidized-bed reactor, whereas the efficiency was reduced to 90% for the two-phase fluidized-bed reactor and 68% for the two-phase reactor without cells. The maximum gas removal capacity for the system was 254 g $H_2S/m^3/h$ when the inlet $H_2S$ loading was $300g/m^3/h(1,500ppm)$. Stable operation of the immobilized reactor was possible for 20 days with the inlet $H_2S$ concentration held to 1,100 ppm. The fluidized bed bioreactor appeared to be an effective means for controlling hydrogen sulfide emissions.

Strain Improvement and Bioprocess Optimization for Enhanced Production of Haluronic Acid(HA) in Bioreactor Cultures of Streptococcus zooepidemicus (히알루론산 생산성 향상을 위한 Streptococcus zooepidemicus 균주 개량 및 발효조 배양공정 최적화)

  • Kim, Soo Yeon;Chun, Gie-Taek
    • Microbiology and Biotechnology Letters
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    • v.48 no.3
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    • pp.344-357
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    • 2020
  • Strain improvement and bioprocess development were undertaken to enhance hyaluronic acid(HA) production by Streptococcus zooepidemicus cells. Using a high-yielding mutant strain, statistical medium optimization was carried out in shake flask cultures, resulting in 52% increase in HA production (5.38 g/l) at the optimal medium composition relative to the parallel control cultures. For sufficient supply of dissolved oxygen (DO), which turned out to be crucial for enhanced production of HA, agitation system and speed were intensively investigated in 5 L bioreactor cultures. Increase in oxygen mass transfer coefficient (kLa) through increment of agitation speed (rpm) and 35% expansion of diameter of the newly-designed impellers showed significantly positive effects on HA production. By installing an expanded Rushton-turbine impeller for efficient break-down of sparged air, and an extended marine impeller above the Rushton-turbine impeller for efficient mixing of the air-born viscous fermentation broth, maximum amount of HA (9.79 g/l) was obtained at 450 rpm, 1.8 times higher level than that of the corresponding flask culture. Subsequently, the possibility of bioprocess scale-up to a 50 L bioreactor was investigated. Despite almost identical maximum HA production (9.11 vs 9.25 g/l), the average HA volumetric productivity (rp) of the 50 L culture turned out only 74% compared to the corresponding 5 L culture during the exponential phase, possibly caused by shear damages imposed on the producing cells at the high stirring in the 50 L culture. The scale-up process could be successfully achieved if a scale-up criterion of constant oxygen mass transfer coefficient (kLa) is applied to the 50 L pilot-scale bioreactor system.

Continuous high cell density culture of Anaerobiospirillum succiniciproducens with membrane filtration for the production of succinic acid

  • Lee, Pyeong-Cheon;Lee, U-Gi;Lee, Sang-Yeop;Jang, Ho-Nam
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.338-341
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    • 2000
  • An internal membrane bioreactor system was employed for continuous succinic ac id production from glucose in order to prove its performance and practicality. Succinic acid-producing Anaerobiospirillum succiniciproducens required more $CO_2$ for the proper growth and succinic acid production in cell recycled continuous culture than in batch culture. The maximum productivity obtained in cell recycled continuous culture was about 3.3 g/L-h which was ca. 3.3 times higher than that obtained in batch culture.

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Optimization of recombinant E. coli fermentation through biological manipulation and engineering control

  • Kim, Jeong-Yoon
    • The Microorganisms and Industry
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    • v.19 no.4
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    • pp.14-26
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    • 1993
  • Optimizing protein production in recombinant E. coli strains involves manipulation of genetic and environmental factors. In designing a production system, attention must be paid to gene expression efficiency, culture conditions and bioreactor configuration. Although not much emphasis was given to the physiology of host strains in this review, an understanding of the relationship between the physiology of host cell growth and the overproduction of a cloned gene protein is of primary importance to the improvement of the recombinant fermentation processes. Sometimes it is desirable to make use of gene fusion systems, e.g. protein A, polypeptide, gutathione-S-transferase, or pneumococcal murein hydrolase fusion, to facilitate protein purification.

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