• Title/Summary/Keyword: bioreactor process

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Integrated Whole-Cell Biocatalysis for Trehalose Production from Maltose Using Permeabilized Pseudomonas monteilii Cells and Bioremoval of Byproduct

  • Trakarnpaiboon, Srisakul;Champreda, Verawat
    • Journal of Microbiology and Biotechnology
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    • v.32 no.8
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    • pp.1054-1063
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    • 2022
  • Trehalose is a non-conventional sugar with potent applications in the food, healthcare and biopharma industries. In this study, trehalose was synthesized from maltose using whole-cell Pseudomonas monteilii TBRC 1196 producing trehalose synthase (TreS) as the biocatalyst. The reaction condition was optimized using 1% Triton X-100 permeabilized cells. According to our central composite design (CCD) experiment, the optimal process was achieved at 35℃ and pH 8.0 for 24 h, resulting in the maximum trehalose yield of 51.60 g/g after 12 h using an initial cell loading of 94 g/l. Scale-up production in a lab-scale bioreactor led to the final trehalose concentration of 51.91 g/l with a yield of 51.60 g/g and productivity of 4.37 g/l/h together with 8.24 g/l glucose as a byproduct. A one-pot process integrating trehalose production and byproduct bioremoval showed 53.35% trehalose yield from 107.4 g/l after 15 h by permeabilized P. moteilii cells. The residual maltose and glucose were subsequently removed by Saccharomyces cerevisiae TBRC 12153, resulting in trehalose recovery of 99.23% with 24.85 g/l ethanol obtained as a co-product. The present work provides an integrated alternative process for trehalose production from maltose syrup in bio-industry.

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.

Can a Fermentation Gas Mainly Produced by Rumen Isotrichidae Ciliates be a Potential Source of Biohydrogen and a Fuel for a Chemical Fuel Cell?

  • Piela, Piotr;Michalowski, Tadeusz;Miltko, Renata;Szewczyk, Krzysztof W.;Sikora, Radoslaw;Grzesiuk, Elzbieta;Sikora, Anna
    • Journal of Microbiology and Biotechnology
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    • v.20 no.7
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    • pp.1092-1100
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    • 2010
  • Bacteria, fungi, and protozoa inhabiting the rumen, the largest chamber of the ruminants' stomach, release large quantities of hydrogen during the fermentation of carbohydrates. The hydrogen is used by coexisting methanogens to produce methane in energy-yielding processes. This work shows, for the first time, a fundamental possibility of using a hydrogen-rich fermentation gas produced by selected rumen ciliates to feed a low-temperature hydrogen fuel cell. A biohydrogen fuel cell (BHFC) was constructed consisting of (i) a bioreactor, in which a hydrogen-rich gas was produced from glucose by rumen ciliates, mainly of the Isotrichidae family, deprived of intra- and extracellular bacteria, methanogens, and fungi; and (ii) a chemical fuel cell of the polymer-electrolyte type (PEFC). The fuel cell was used as a tester of the technical applicability of the fermentation gas produced by the rumen ciliates for power generation. The average estimated hydrogen yield was ca. 1.15 mol $H_2$ per mole of fermented glucose. The BHFC performance was equal to the performance of the PEFC running on pure hydrogen. No fuel cell poisoning effects were detected. A maximum power density of $1.66\;kW/m^2$ (PEFC geometric area) was obtained at room temperature. The maximum volumetric power density was $128\;W/m^3$ but the coulombic efficiency was only ca. 3.8%. The configuration of the bioreactor limited the continuous operation time of this BHFC to ca. 14 h.

Optimization of Environmental Parameters for Extracellular Chitinase Production by Trichoderma harzianum SJG-99721 in Bioreactor (Trichoderma harzianum SJG-99721의 체외 분비 chitinase 생산에 미치는 생물 반응기에서의 반응 최적화 연구)

  • 이호용
    • Korean Journal of Environmental Biology
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    • v.22 no.1
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    • pp.167-170
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    • 2004
  • A self-directing optimization procedure was applied to determine the best environmental factors in operating the bioreactor. The self-directing optimization process was employed to determine the best conditional combination of multi parameters, pH, temperature, aeration rate and mixing rate toy maximal production of chitinase by Trichoderma harzianum SJG-99721 in batch mode fermentation. Among these factors, the parameters of pH and aeration rate were found to be particularly important on mycellial growth and chitinase activity. pH 4.89, an aeration rate of 3.22 ι per minute and an agitation rate of 225 rpm was found to be the best combination. By the optimization, chitinase activity was dramatically increased from an initial value of 4.221 U under basic conditions to n final value of 16.825 U.

Nitrogen Removal Performance at Various DO Concentrations in the Bioreactor Packed with Submerged Cilia Media and Granular Sulfur (DO농도 조절에 따른 황 충전 섬모상 반응조의 질소제거 성능 변화)

  • Moon, Jin-Young;Hwang, Yong-Woo
    • Journal of Korean Society of Water and Wastewater
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    • v.20 no.4
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    • pp.519-526
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    • 2006
  • In this study, the major operating factors in SND(simultaneous nitrification and denitrification) using bioreactor packed with submerged cilia media and granular sulfur such as variation of nitrification rate, organic matter removal efficiency and denitrification efficiency in different DO concentration were mainly evaluated. Synthetic wastewater and actual sewage were used as influent wastewater. Experiment with synthetic wastewater as influent wastewater was divided into three phases with the adjustment of DO concentration. As the results, nitrification efficiency and T-N removal efficiency in the Phase 3(DO 1.0~2.0 mg/L) were 99% and 52.3%, which is significantly greater than those in other two phases. Also, loading rate and denitrification efficiency of SCPGS(Submerged Cilia media Packed with Granular Sulfur) were calculated as $0.44kg\;NO_3^--N/m^3-day$ and 50%, respectively. On the other hand, nitrification rate was decreased from 99% to 64% according to the DO concentration with the variation from 3.0~3.5 mg/L(phase1) to 0.4~0.6mg/L(phase2). Although the nitrification rate was decreased in 64% according to the variation of the DO concentration, T-N removal rate was rapidly increased to 49% by increasing of the denitrification efficiency. Experiment with actual sewage as influent wastewater was carried out to evaluate efficiency of SCPGS in real operation condition of full-scale sewage water treatment plant. At the time, T-N removal rate in this experiment and full-scale wastewater treatment plants were given by 43% and 20%, respectively. The above results indicate that SCPGS can be used as an advanced treatment process for economical efficiency considered.

Optimization of Tannase Production by Aspergillus niger in Solid-State Packed-Bed Bioreactor

  • Rodriguez-Duran, Luis V.;Contreras-Esquivel, Juan C.;Rodriguez, Raul;Prado-Barragan, L. Arely;Aguilar, Cristobal N.
    • Journal of Microbiology and Biotechnology
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    • v.21 no.9
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    • pp.960-967
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    • 2011
  • Tannin acyl hydrolase, also known as tannase, is an enzyme with important applications in the food, feed, pharmaceutical, and chemical industries. However, despite a growing interest in the catalytic properties of tannase, its practical use is very limited owing to high production costs. Several studies have already demonstrated the advantages of solid-state fermentation (SSF) for the production of fungal tannase, yet the optimal conditions for enzyme production strongly depend on the microbial strain utilized. Therefore, the aim of this study was to improve the tannase production by a locally isolated A. niger strain in an SSF system. The SSF was carried out in packed-bed bioreactors using polyurethane foam as an inert support impregnated with defined culture media. The process parameters influencing the enzyme production were identified using a Plackett-Burman design, where the substrate concentration, initial pH, and incubation temperature were determined as the most significant. These parameters were then further optimized using a Box-Behnken design. The maximum tannase production was obtained with a high tannic acid concentration (50 g/l), relatively low incubation temperature ($30^{\circ}C$), and unique low initial pH (4.0). The statistical strategy aided in increasing the enzyme activity nearly 1.97-fold, from 4,030 to 7,955 U/l. Consequently, these findings can lead to the development of a fermentation system that is able to produce large amounts of tannase in economical, compact, and scalable reactors.

Effect of Electrolysis on Bacterial Activity in Electrokinetic Bioremediation (동전기 생물학적 복원에서 전기분해반응이 미생물 활성에 미치는 영향)

  • Kim, Sang-Joon;Park, Ji-Yeon;Lee, You-Jin;Yang, Ji-Won
    • Journal of Korean Society of Environmental Engineers
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    • v.28 no.7
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    • pp.764-769
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    • 2006
  • In the electrokinetic(EK) process, oxygen production by electrolysis was proportional to current density. The dissolved oxygen (DO) concentration in anode tank and bioreactor increased with the circulation rate of electrolyte. The bacterial population in bioreactor rapidly increased by the supplement of current, but the DO concentration deceased by the increased bacterial oxygen consumption. From the results of EK bioremediation for pentadecane-contaminated soil, the bacterial population and removal efficiency at 1.88 $mA/cm^2$ were lower than those at 0.63 $mA/cm^2$. This is because the high oxygen production rate largely increased the production rate of organic acids, which reduced the electrolyte pH and bacterial activity. At 0.63 $mA/cm^2$, the highest bacterial population and removal efficiency could be obtained due to the appropriate oxygen production and small decrease in pH.

Immobilization of a Mediator onto Carbon Cloth Electrode and Employment of the Modified Electrode to an Electroenzymatic Bioreactor

  • Jeong, Eun-Seon;Sathishkumar, Muthuswamy;Jayabalan, Rasu;Jeong, Su-Hyeon;Park, Song-Yie;Mun, Sung-Phil;Yun, Sei-Eok
    • Journal of Microbiology and Biotechnology
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    • v.22 no.10
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    • pp.1406-1411
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    • 2012
  • 5,5'-Dithiobis(2-nitrobenzoic acid) (DTNB) was selected as an electron transfer mediator and was covalently immobilized onto high porosity carbon cloth to employ as a working electrode in an electrochemical $NAD^+$-regeneration process, which was coupled to an enzymatic reaction. The voltammetric behavior of DTNB attached to carbon cloth resembled that of DTNB in buffered aqueous solution, and the electrocatalytic anodic current grew continuously upon addition of NADH at different concentrations, indicating that DTNB is immobilized to carbon cloth effectively and the immobilized DTNB is active as a soluble one. The bioelectrocatalytic $NAD^+$ regeneration was coupled to the conversion of L-glutamate into ${\alpha}$-ketoglutarate by L-glutamate dehydrogenase within the same microreactor. The conversion at 3 mM monosodium glutamate was very rapid, up to 12 h, to result in 90%, and then slow up to 24 h, showing 94%, followed by slight decrease. Low conversion was shown when substrate concentration exceeding 4 mM was tested, suggesting that L-glutamate dehydrogenase is inhibited by ${\alpha}$-ketoglutarate. However, our electrochemical $NAD^+$ regeneration procedure looks advantageous over the enzymatic procedure using NADH oxidase, from the viewpoint of reaction time to completion.

Microbial Desulfurization of a Bituminous Coal by Iron-Oxidizing Bacteria Thiobacillus ferooxidans (철산화 박테리아 Thiobacillus ferrooxidans에 의한 역청탄의 생물학적 탈황)

  • 류희욱
    • KSBB Journal
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    • v.11 no.2
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    • pp.238-245
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    • 1996
  • Microbial desulfurization characterlstics of a bituminous coal have been determined by using Thiobacillus ferrooxidans. The effects of process variables (such as coal pulp density, particle size and addition of surfactants) on pyrite removal have been investigated in both shake and airlift-bioreactor culture experiments. In shake experiments, pyrite could be removed over 78% for pulp densifies below 20% (w/v) and removed below 40% for pulp densities over 30% (w/v) in 8 days. Pyrite removal decreased with increasing pulp densities, and it also decreased sharply with increasing particle sizes. In airlift bioreactor experiments, pyrite at 50% (w/v) pulp density could be removed about 50%. Its value is much higher than 15% at the same pulp density in a shake experiment. With addition of surfactants, pyrite removal was enhanced in shake experiments significantly, whereas it was slightly decreased in an airlift bioreactor experiment.

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Effects of Alkalinity on the Nitrification Capability of Nonwoven Fabric Filter Bioreactor (부직포 여과막 생물반응조에서 알칼리도가 질산화 성능에 미치는 영향)

  • Bae, Min-Su;Ahn, Yoon-Chan;Jang, Myung-Bae;Cho, Yun-Kyung;Cho, Kwang-Myeung
    • Journal of Korean Society of Environmental Engineers
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    • v.29 no.7
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    • pp.783-792
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    • 2007
  • To investigate the effects of alkalinity on the nitrification capability of the nonwoven fabric filter bioreactor(NFBR), an experiment was performed for 641 days at a hydraulic retention time of approximately 11 hours by changing the influent concentration of $NH_3-N$ from 54 mg/L to 1,400 mg/L and alkalinity from 43 mg/L to 10,480 mg/L. The MLSS concentration reduced from an initial value of 2,650 mg/L down to 830 mg/L, then increased up to 8,340 mg/L. Though the volumetric loading rate varied in a range of $0.120\sim3.130$ kg $NH_3-N/m^3-day$, the F/M ratio showed a narrow range of $0.067\sim0.414$ kg $NH_3-N/kg$ MLSS-day. The average nitrification efficiency at each experimental stage resulted in the range of $35.2\sim100%$, and the maximum nitrification rate was 2.970 kg $N/m^3-day$ or 0.489 g N/g MLVSS-day. The nitrifiers' fraction of the MLVSS increased up to 100% from an initial value of 7.1% and the biofilm formed on the nonwoven fabric filter showed a very low nitrifiers' fraction of mere 2.2%. The growth yield of the MLSS and the alkalinity consumption rate were computed to be 0.117 g VSS/g N removed and 7.08 g alkalinity/g $NO_x^--N$ produced, respectively. Results of the research suggest that NFBR could be an adequate process for nitrification of wastewaters with high ammonia concentrations.