• Title/Summary/Keyword: scale-up fermentation

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Microbial Metabolism of the Environmental Estrogen Bisphenol A

  • Yim, Soon-Ho;Kim, Hyun-Jung;Lee, Ik-Soo
    • Archives of Pharmacal Research
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    • v.26 no.10
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    • pp.805-808
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    • 2003
  • Preliminary microbial metabolism studies of bisphenol A (BPA) (1) on twenty six microorganisms have shown that Aspergillus fumigatus is capable of metabolizing BPA. Scale-up fermentation of 1 with A. fumigatus gave a metabolite (2) and its structure was established as bisphenol $A-O-{\beta}-D-glucopyranoside$ (BPAG) based on spectroscopic analyses.

Linear Correlation between Online Capacitance and Offline Biomass Measurement up to High Cell Densities in Escherichia coli Fermentations in a Pilot-Scale Pressurized Bioreactor

  • Knabben, Ingo;Regestein, Lars;Schauf, Julia;Steinbusch, Sven;Buchs, Jochen
    • Journal of Microbiology and Biotechnology
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    • v.21 no.2
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    • pp.204-211
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    • 2011
  • To yield high concentrations of protein expressed by genetically modified Escherichia coli, it is important that the bacterial strains are cultivated to high cell density in industrial bioprocesses. Since the expressed target protein is mostly accumulated inside the E. coli cells, the cellular product formation can be directly correlated to the bacterial biomass concentration. The typical way to determine this concentration is to sample offline. Such manual sampling, however, wastes time and is not efficient for acquiring direct feedback to control a fedbatch fermentation. An E. coli K12-derived strain was cultivated to high cell density in a pressurized stirred bioreactor on a pilot scale, by detecting biomass concentration online using a capacitance probe. This E. coli strain was grown in pure minimal medium using two carbon sources (glucose and glycerol). By applying exponential feeding profiles corresponding to a constant specific growth rate, the E. coli culture grew under carbon-limited conditions to minimize overflow metabolites. A high linearity was found between capacitance and biomass concentration, whereby up to 85 g/L dry cell weight was measured. To validate the viability of the culture, the oxygen transfer rate (OTR) was determined online, yielding maximum values of 0.69 mol/l/h and 0.98mol/l/h by using glucose and glycerol as carbon sources, respectively. Consequently, online monitoring of biomass using a capacitance probe provides direct and fast information about the viable E. coli biomass generated under aerobic fermentation conditions at elevated headspace pressures.

Development of an Enrichment Culture Growing at Low Temperature used for Ensiling Rice Straw

  • Yang, Hong Yan;Wang, Xiao Fen;Gao, Li Juan;Haruta, Shin;Ishii, Masaharu;Igarashi, Yasuo;Cui, Zong Jun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.711-717
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    • 2008
  • To speed up the conversion of rice straw into feeds in a low-temperature region, a start culture used for ensiling rice straw at low temperature was selected by continuous enrichment cultivation. During the selection, the microbial source for enrichment was rice straw and soil from two places in Northeast China. Lab-scale rice straw fermentation at $10^{\circ}C$ verified, compared with the commercial inoculant, that the selected start culture lowered the pH of the fermented rice straw more rapidly and produced more lactic acid. The results from denatured gradient gel eletrophoresis showed that the selected start culture could colonize into the rice straw fermentation system. To analyze the composition of the culture, a 16S rRNA gene clone library was constructed. Sequencing results showed that the culture mainly consisted of two bacterial species. One (A) belonged to Lactobacillus and another (B) belonged to Leuconostoc. To make clear the roles of composition microbes in the fermented system, quantitative PCR was used. For species A, the DNA mass increased continuously until sixteen days of the fermentation, which occupied 65%. For species B, the DNA mass amounted to 5.5% at six days of the fermentation, which was the maximum relative value during the fermentation. To the authors' best knowledge, this is the first report on ensiling rice straw with a selected starter at low temperature and investigation of the fermented characteristics.

Isolation and Characterization of An Alcohol Fermentation Strain from Anaerobic Acid Fermentor to Treat Food Wastes (음식폐기물 처리용 혐기성 산 발효조로부터 알코올발효 균주의 분리 및 특성)

  • Kim, Jung-Kon;Han, Gui-Hwan;Yoo, Jin-Cheol;Seong, Chi-Nam;Kim, Seong-Jun;Kim, Si-Wouk
    • KSBB Journal
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    • v.21 no.6 s.101
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    • pp.451-455
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    • 2006
  • An efficient pilot scale (10 ton) three-stage methane fermentation system to digest food waste has been developed in this laboratory. This system consisted of three stages: semianaerobic hydrolysis, anaerobic acidogenesis and strictly anaerobic methanogenesis. From the secondary acidogenesis reactor, a novel strain KA4 responsible for alcohol fermentation was isolated and characterized. The cell was oval and its dimension was $5.5-6.5{\times}3.5-4.5\;{\mu}m$. This strain was identified as Saccharomyces cerevisiae KA4 by 26S rDNA D1/D2 rDNA sequence. Optimal culture temperature was $30-35^{\circ}C$. Cells were tolerant to 5% (v/v) ethanol concentration, however, were inhibited significantly by higher ethanol concentration up to 7%. The strain could grow well up to 50% (w/v) initial glucose concentration in the YM liquid medium, however, optimal concentration for ethanol fermentation was 10%. It could produce ethanol in a broad initial pH range from 4 to 10, and optimal pH was 6. In this condition, the strain converted 10% glucose to 7.4% ethanol during 24 hr, and ethanol yield was estimated to be 2.87 moi EtOH/mol glucose.

Fermentation characteristics of mulberry (Cudrania tricuspidata) fruit vinegar produced by acetic acid bacteria isolated from traditional fermented foods (전통발효식품으로부터 분리한 초산균을 이용한 꾸지뽕 열매 발효식초 제조 및 발효특성)

  • Yim, Eun-Jung;Jo, Seung-Wha;Lee, Eun-Sil;Park, Hae-Suk;Ryu, Myeong-Seon;Uhm, Tai-Boong;Kim, Hyoun-Young;Cho, Sung-Ho
    • Food Science and Preservation
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    • v.22 no.1
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    • pp.108-118
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    • 2015
  • To raise the added value of the fruits of Cudrania tricuspidata, Cudrania tricuspidata vinegar was produced and examined for its fermentation conditions. Forty nine acetic acid bacteria with resistance against acetic acid, ethanol, and sulfide as high acetic acid producers were isolated from fermented foods and identified as Acetobacter indonesiensis, A. cerevisiae, A. orientalis, A. tropicalis, A. fabarum, A. pasteurianus, and A. syzygii based on the results of the analysis of the 16S rRNA gene sequences. Among them, two GRAS strains, A. pasteurianus SCMA5 and SCMA6, were finally selected for the production of acetic acid. Optimal vinegar productions were obtained from the medium containing 40% (v/v) fruit juice of Cudrania tricuspidata and 5% (v/v) ethanol at $25^{\circ}C$ for 72 hr. The sensory panel preferred the vinegar fermented with the SCMA06 to that with the SCM05 strain. The radical scavenger capacity of DPPH was 53% higher than that of the control in the vinegar fermented with the SCMA06 strain. The ${\alpha}$-glucosidase inhibitor activity as an index of the antidiabetic drug showed 91% inhibition, which is higher than that of acabose. This study will be helpful for the scale-up production of vinegar with the fruit of Cudrania tricuspidata.

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.

Enhancement of L-Threonine Production by Controlling Sequential Carbon-Nitrogen Ratios during Fermentation

  • Lee, Hyeok-Won;Lee, Hee-Suk;Kim, Chun-Suk;Lee, Jin-Gyeom;Kim, Won-Kyo;Lee, Eun-Gyo;Lee, Hong-Weon
    • Journal of Microbiology and Biotechnology
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    • v.28 no.2
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    • pp.293-297
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    • 2018
  • Controlling the residual glucose concentration is important for improving productivity in $\text\tiny{L}$-threonine fermentation. In this study, we developed a procedure to automatically control the feeding quantity of glucose solution as a function of ammonia-water consumption rate. The feeding ratio ($R_{C/N}$) of glucose and ammonia water was predetermined via a stoichiometric approach, on the basis of glucose-ammonia water consumption rates. In a 5-L fermenter, 102 g/l $\text\tiny{L}$-threonine was obtained using our glucose-ammonia water combined feeding strategy, which was then successfully applied in a 500-L fermenter (89 g/l). Therefore, we conclude that an automatic combination feeding strategy is suitable for improving $\text\tiny{L}$-threonine production.

Improved Production, and Purification of Aclacinomycin A from Streptomyces lavendofoliae DKRS

  • Kim, Wan-Seop;Youn, Deok-Joong;Cho, Won-Tae;Kim, Myung-Kuk;Kim, Hak-Ryul;Rhee, Sang-Ki;Choi, Eui-Sung
    • Journal of Microbiology and Biotechnology
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    • v.5 no.5
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    • pp.297-301
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    • 1995
  • An anthracycline antibiotic, aclacinomycin A (aclarubicin), was produced from a mutant strain of Streptomyces lavendofoliae. The mutant strain which showed a 4-fold higher productivity of aclacinomycin A compared with the parent strain was also found to produce a significantly higher amount of aclacinomycin A than the reported production strain, Streptomyces galilaeus. The aclacinomycin A was produced up to 125 mg/l using potato starch and soybean meal as carbon and nitrogen sources, respectively, on a 3 liter scale fermentation in a 5 liter jar fermentor. The mutant strain also produced significant amount of aclacinomycins Band Y. Aclacinomycin A was isolated from the culture broth by solvent extractions and further purified by silica gel column chromatography. The yield of aclacinomycin A with over 99$%$ purity was found to be over 60$%$ starting from 3 liters of culture broth.

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Production of Organic Acids from Food By-Products - Mass Production of Organic Acids by Continuous Flow Ceil Recycling Fermentation - (식품부산물로부터 유기산의 대량생산공정에 관한 연구 - 세포재순환식 연속발효를 이용한 유기산의 대량 생산 -)

  • Ju Yun-Sang;Jin Sun-Ja;Hwang Pil-Gi;Choi Chul-Ho;Lee Eui-Sang
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.484-488
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    • 2004
  • Fermentation studies were conducted in batch and continuous flow cell-recycle reactors with food by-products as substrates. The genus Propionibacterium acidipropionici ATCC 4965 was utilized in the production of organic acids. Good performance was achieved in the batch fermentation using hydrol as a carbon source and corn steep liquor (CSL) as nitrogen and vitamin sources. Product yields and productivity based on maximum values were 0.80 g total acids/g glucose and 0.26 g total acids/L/h, respectively, when $3\%$, (w/v) of hydrol and $2.5\%$, (w/v) of CSL were utilized. Continuous fermentation with cell-recycling system using the optimum amounts of substrates resulted in dramatic increase in cell concentration (X) and maximum productivity (P). Compared to the batch fermentation, X and P were increased by as much as 21 and 13 times, respectively, at the dilution ratio of $0.2\;hr^{-1}$, indicating that cell recycling fermentation of food by-products provides valuable means for the mass production of organic acids as well as utilizing cell mass as good nutrient resources.

Study of the Rheological Properties of a Fermentation Broth of the Fungus Beauveria bassiana in a Bioreactor Under Different Hydrodynamic Conditions

  • Nunez-Ramirez, Diola Marina;Medina-Torres, Luis;Valencia-Lopez, Jose Javier;Calderas, Fausto;Lopez-Miranda, Javier;Medrano-Roldan, Hiram;Solis-Soto, Aquiles
    • Journal of Microbiology and Biotechnology
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    • v.22 no.11
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    • pp.1494-1500
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    • 2012
  • Fermentation with filamentous fungi in a bioreactor is a complex dynamic process that is affected by flow conditions and the evolution of the rheological properties of the medium. These properties are mainly affected by the biomass concentration and the morphology of the fungus. In this work, the rheological properties of a fermentation with the fungus Beauveria bassiana under different hydrodynamic conditions were studied and the rheological behavior of this broth was simulated through a mixture of carboxymethyl cellulose sodium and cellulose fibers (CMCNa-SF). The bioreactor was a 10 L CSTR tank operated at different stir velocities. Rheological results were similar at 100 and 300 rpm for both systems. However, there was a significant increase in the viscosity accompanied by a change in the consistence index, calculated according to the power law model, for both systems at 800 rpm. The systems exhibited shear-thinning behavior at all stir velocities, which was determined with the power law model. The mixing time was observed to increase as the cellulose content in the system increased and, consequently, the efficiency of mixing diminished. These results are thought to be due to the rheological and morphological similarities of the two fungal systems. These results will help in the optimization of scale-up production of these fungi.