• Title/Summary/Keyword: mass production phase

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Purification and Characterization of the Bacteriocin Thuricin Bn1 Produced by Bacillus thuringiensis subsp. kurstaki Bn1 Isolated from a Hazelnut Pest

  • Ugras, Serpil;Sezen, Kazim;Kati, Hatice;Demirbag, Zihni
    • Journal of Microbiology and Biotechnology
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    • v.23 no.2
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    • pp.167-176
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    • 2013
  • A novel bioactive molecule produced by Bacillus thuringiensis subsp. kurstaki Bn1 (Bt-Bn1), isolated from a common pest of hazelnut, Balaninus nucum L. (Coleoptera: Curculionidae), was determined, purified, and characterized in this study. The Bt-Bn1 strain was investigated for antibacterial activity with an agar spot assay and well diffusion assay against B. cereus, B. weinhenstephenensis, L. monocytogenes, P. savastanoi, P. syringae, P. lemoignei, and many other B. thuringiensis strains. The production of bioactive molecule was determined at the early logarithmic phase in the growth cycle of strain Bt-Bn1 and its production continued until the beginning of the stationary phase. The mode of action of this molecule displayed bacteriocidal or bacteriolytic effect depending on the concentration. The bioactive molecule was purified 78-fold from the bacteria supernatant with ammonium sulfate precipitation, dialysis, ultrafiltration, gel filtration chromatography, and HPLC, respectively. The molecular mass of this molecule was estimated via SDS-PAGE and confirmed by the ESI-TOFMS as 3,139 Da. The bioactive molecule was also determined to be a heat-stable, pH-stable (range 6-8), and proteinase K sensitive antibacterial peptide, similar to bacteriocins. Based on all characteristics determined in this study, the purified bacteriocin was named as thuricin Bn1 because of the similarities to the previously identified thuricin-like bacteriocin produced by the various B. thuringiensis strains. Plasmid elution studies showed that gene responsible for the production of thuricin Bn1 is located on the chromosome of Bt-Bn1. Therefore, it is a novel bacteriocin and the first recorded one produced by an insect originated bacterium. It has potential usage for the control of many different pathogenic and spoilage bacteria in the food industry, agriculture, and various other areas.

Comparison of Aviary, Barn and Conventional Cage Raising of Chickens on Laying Performance and Egg Quality

  • Ahammed, M.;Chae, B.J.;Lohakare, J.;Keohavong, B.;Lee, M.H.;Lee, S.J.;Kim, D.M.;Lee, J.Y.;Ohh, S.J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.8
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    • pp.1196-1203
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    • 2014
  • This study intended to compare the productive performance of three different layer raising systems; conventional cage (CC), barn (BR) and aviary (AV). The AV is welfare bestowed housing that allows free locomotion for birds within the BR. The BR allows bird's free locomotion inside BR but without multilevel structures. Both pullets and cockerels were housed together in both AV and BR, but only pullets in CC. Seventeen weeks old Lohmann Brown Lite (n = 800) pullets were housed in AV during this study. The same age layer pullets were simultaneously assigned to either at CC or BR to compare egg production performance with AV. The duration of experiment was 40 weeks (from 21st to 60th week). There were no remarkable differences in egg production, hen day egg production (HDEP) and average egg weight among three rearing systems. First 20 weeks (phase-1) average HDEP (%) of AV, CC, and BR were 85.9, 88.8, 87.1 and average egg weights (g) were 57.5, 59.9, and 56.9 respectively. Those of the remaining 20 weeks (phase-2) were 87.1, 87.9, 85.5 and 64.2, 63.0 62.1, respectively. Daily feed intakes (122 g, 110 g, 125 g); feed conversion ratio (2.4, 2.1, 2.5) and daily egg mass (53.9 g, 54.4 g, 52.8 g) data from AV, CC, and BR were not influenced significantly by the respective raising systems. Daily feed intake of layers in both AV (124 g) and BR (127 g) tended to be higher than that in CC (113 g) during phase-2. Overall, exterior egg quality (dirty and cracked eggs) in both phases was superior in BR compared with AV and CC, whereas CC generated intermediate results. This study indicated that the HDEP per se in AV and BR were not significantly different from that in CC. The study implied that the facility depreciation cost for AV and cost for increased feed intake in AV compared to CC are believed to be critical to evaluate the cost effectiveness of egg production in AV.

Importance of Strain Improvement and Control of Fungal cells Morphology for Enhanced Production of Protein-bound Polysaccharides(β-D-glucan) in Suspended Cultures of Phellinus linteus Mycelia (Phellinus linteus의 균사체 액상배양에서 단백다당체(β-D-glucan)의 생산성 향상을 위한 균주 개량과 배양형태 조절의 중요성)

  • Shin, Woo-Shik;Kwon, Yong Jung;Jeong, Yong-Seob;Chun, Gie-Taek
    • Korean Chemical Engineering Research
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    • v.47 no.2
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    • pp.220-229
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    • 2009
  • Strain improvement and morphology investigation in bioreactor cultures were undertaken in suspended cultures of Phellinus linteus mycelia for mass production of protein-bound polysaccharides(soluble ${\beta}$-D-glucan), a powerful immuno-stimulating agent. Phellineus sp. screened for this research was identified as Phellinus linteues through ITS rDNA sequencing method and blast search, demonstrating 99.7% similarity to other Phellinus linteus strains. Intensive strain improvement program was carried out by obtaining large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because large numbers of protoplasts ($1{\times}10^5{\sim}10^6\;protoplasts/ml$) formed using the banding filtration method with the cell wall-disrupting enzymes could be regenerated in relatively high regeneration frequency($10^{-2}{\sim}10^{-3}$) in the newly developed regeneration medium. It was demonstrated that the strains showing high performances in the protoplast regeneration and solid growth medium were able to produce 5.8~6.4%(w/w) of ${\beta}$-D-glucan and 13~15 g/L of biomass in stable manners in suspended shake-flask cultures of P. linteus mycelia. In addition, cell mass increase was observed to be the most important in order to enhance ${\beta}$-D-glucan productivity during the course of strain improvement program, since the amount of ${\beta}$-D-glucan extracted from the cell wall of P. linteus mycelia was almost constant on the unit biomass basis. Therefore we fully investigated the fungal cell morphology, generally known as one of the key factors affecting cell growth extent in the bioreactor cultures of mycelial fungal cells. It was found that, in order to obtain as high cell mass as possible in the final production bioreactor cultures, the producing cells should be proliferated in condensed filamentous forms in the growth cultures, and optimum amounts of these filamentous cells should be transferred as active inoculums to the production bioreactor. In this case, ideal morphologies consisting of compacted pellets less than 0.5mm in diameter were successfully induced in the production cultures, resulting in shorter period of lag phase, 1.5 fold higher specific cell growth rate and 3.3 fold increase in the final biomass production as compared to the parallel bioreactor cultures of different morphological forms. It was concluded that not only the high-yielding but also the good morphological characteristics led to the significantly higher biomass production and ${\beta}$-D-glucan productivity in the final production cultures.

Effects of the High Pressure Sodium Lamp Lighting on the Dynamics of Growth and Dry Mass Partitioning in Sweet Pepper Plant (고압나트륨등 조사가 파프리카의 동적 생장과 건물분배율에 미치는 영향)

  • Kim, Eun Jeong;Lee, Sang Hyun;Lee, Jeong Hyun
    • Horticultural Science & Technology
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    • v.31 no.5
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    • pp.565-572
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    • 2013
  • The objective of this study was to describe and analyze the effect of high pressure sodium lamp lighting (HPS) on dynamics of growth and dry matter partitioning, and light use efficiency of sweet pepper crop grown over winter season. Young sweet pepper seedlings were planted at 3.75 plants per $m^2$ on September 29, 2010 and treated with HPS for 16 hours from December 14, 2010 until March 18, 2011. The number of leaves per plant were significantly increased with HPS, whereas the number of internodes and leaf area were less affected. HPS reduced the plant height with higher number of fruits per stem compared to those of without HPS lighting (CON). There were large differences in total dry mass production, stem and fruit dry mass between HPS and CON and those with HPS increased by 67.8%, 28.5%, and 97.1% compared to CON, respectively. Each organs of dry mass partitioning was calculated by leaf, stem or fruit growth rate divided by total plant growth rate. Dynamics of dry mass partitioning to leaf and stem between HPC and CON was measured in range of 45-47% at beginning of growth phase and drastically decreased after starting fruit growth in both treatments. Dry matter partitioning to vegetative organs was 4% higher compared to the plant grown under HPS lighting. Averaged dry matter partitioning to fruit with HPS, however, was largely increased by 14.2% compared to CON. Dynamics of the plant growth were well described by expolinear growth equation with three parameters of maximum relative growth rate, absolute growth rate and lost time to reach linear phase. The maximum growth rate of leaf, stem and fruit with HPS was increased by 18.6%, 74.7%, and 143.5% compared to CON. There was a linear relationship between intercepted light integral and vegetative organs (leaf and stem), fruit or total dry mass production. Light use efficiency (LUE, $g{\cdot}MJ^{-1}$) of total dry mass was $4.90g{\cdot}MJ^{-1}$ for HPS and $3.84g{\cdot}MJ^{-1}$ for CON, LUE of vegetative organs was $1.56g{\cdot}MJ^{-1}$ for HPS and $1.61g{\cdot}MJ^{-1}$ for CON and LUE of fruit dry mass was $3.34g{\cdot}MJ^{-1}$ for HPS and $2.23g{\cdot}MJ^{-1}$ for CON. The difference in LUE of total dry mass between treatments, therefore, occurred mainly from the different in LUE of fruit dry mass.

Co-administration of erythropoietin and iron complex improves late-phase liver regeneration

  • Kim, Ji-Yoon;Choi, Dongho;Kim, Joohwan;Kim, Young-Myeong;Lim, Hyunyoung;Sung, Jeong Min;Lee, Min Kyu;Choung, Yoo Jin;Chang, Ji Hee;Jeong, Mi Ae
    • BMB Reports
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    • v.53 no.3
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    • pp.148-153
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    • 2020
  • Erythropoietin and iron have individually shown beneficial effects on early-phase liver regeneration following partial hepatectomy (PHx); however, there are limited data on the combined effect on late-phase liver regeneration after PHx. Here we examined combined effects of recombinant human erythropoietin (rhEPO, 3,000 IU/kg) and iron isomaltoside (IIM, 40 mg/kg) on late-phase liver regeneration following PHx and investigated the possible underlying mechanism. Rats administrated with rhEPO showed significantly higher liver mass restoration, interleukin-6 (IL-6, a hepatocyte mitogen) levels, and Ki-67-positive hepatocytes on day 7 after PHx than saline-treated controls. These beneficial effects were further enhanced on days 7 and 14 by co-treatment with IIM. This combination also significantly improved liver function indices, such as increased albumin production and decreased bilirubin levels, but did not alter serum levels of toxic parameters, such as aspartate transaminase and alanine transaminase. This study demonstrates that the combination of rhEPO and IIM synergistically improves late-phase liver regeneration and function after PHx, probably by promoting IL-6-mediated hepatocyte proliferation without adverse effects. Thus, this combination treatment can be a potential therapeutic strategy for patients undergoing resection for hepatic malignancies.

Production of Biosurfactant Lipopeptides Iturin A, Fengycin, and Surfactin A from Bacillus subtilis CMB32 for Control of Colletotrichum gloeosporioides

  • Kim, Pyoung-Il;Ryu, Jae-Won;Kim, Young-Hwan;Chi, Youn-Tae
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.138-145
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    • 2010
  • A bacterial strain isolated from soil for its potential to control the anthracnose disease caused by Colletotrichum gloeosporioides was identified as a Bacillus subtilis. Bacillus subtilis CMB32 produced antifungal agents on M9 broth at $30^{\circ}C$. Biosurfactant lipopeptides produced by Bacillus subtilis CMB32 were precipitated by adjusting to pH 2 and extracting using chloroform/methanol, and then were purified using column chromatography and reverse-phase HPLC. The molecular masses of the lipopeptides were estimated by MALDI-TOF mass spectrometry as (a) 1,080, (b) 1,486, and (c) 1,044 Da, respectively. They had cyclic structures and amino acid compositions of (a) Pro, Asx, Ser, Tyr, Glx, (b) Glx, Tyr, Thr, Ala, Pro, lie, and (c) Glx, Leu, Val, Asx, respectively. Further analysis revealed that Bacillus subtilis CMB32 produced three antifungal lipopeptides: (a) iturin A, (b) fengycin, and (c) surfactin A.

Recombinant production of human glucagon-like peptide-1 mutant (인간 Glucagon-like Peptide-1 변이체의 재조합 생산)

  • Kim, Sung-Gun;Park, Jong-Tae
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.237-243
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    • 2014
  • Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

Optimization of HPLC-tandem mass spectrometry for chlortetracycline using response surface analysis

  • Bae, Hyokwan;Jung, Hee-Suk;Jung, Jin-Young
    • Environmental Engineering Research
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    • v.23 no.3
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    • pp.309-315
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    • 2018
  • Chlortetracycline (CTC) is one of the most important compounds in antibiotic production, and its distribution has been widely investigated due to health and ecological concerns. This study presents systematic approach to optimize the high-performance liquid chromatography-tandem mass spectrometry for analyzing CTC in a multiple reaction monitoring mode ($479{\rightarrow}462m/z$). One-factor-at-a-time (OFAT) test with response surface analysis (RSA) was used as optimization strategy. In OFAT tests, the fragmentor voltage, collision energy, and ratio of acetonitrile in the mobile phase were selected as major factors for RSA. The experimental conditions were determined using a composite in cube design (CCD) to maximize the peak area. As a result, the partial cubic model precisely predicted the peak area response with high statistical significance. In the model, the (solvent composition) and (collision $energy^2$) terms were statistically significant at the 0.1 ${\alpha}$-level, while the two-way interactions of the independent variables were negligible. By analyzing the model equation, the optimum conditions were derived as 114.9 V, 15.7 eV, and 70.9% for the fragmentor voltage, collision energy, and solvent composition, respectively. The RSA, coupled with the CCD, offered a comprehensive understanding of the peak area that responds to changes in experimental conditions.

Membrane Technologies in Biotechnology (생물공학에서의 막분리기술)

  • 구윤모
    • Membrane Journal
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    • v.8 no.2
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    • pp.69-76
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    • 1998
  • Membrane technologies have been used frequently in industries, taking advantage of that it is energy-saving and employable in relatively large scale. The fact that a non-mass separating agent is used in mild conditions without phase change in membrane separation makes it a method of choice in the recovery of biological materials. Recently, the development of noble separating modules has been solving the inherent problems in membrane separation, the fouling and the concentration polarization. In addition, membrane separation has broadened its applications from the conventional crude separation to the purificational use by the advent of the new and functional membrane materials. The role of membrane technologies is expected to be enormous in the production and recovery of biological products, considering the excellent applicability of membrane in the fields of integrated separation and in-situ separation, the two trends in modem bioseparation.

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Lab-scale experimental setup to evaluate the performance of band driers (통기밴드식 건조기의 성능 평가 실험 장치)

  • Seongmin, Park;Sang Hyun, Oh;Sung Il, Kim;Wonjung, Kim
    • Journal of the Korean Society of Visualization
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    • v.20 no.3
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    • pp.36-41
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    • 2022
  • Drying process is involved in the production of various products including food, textiles, paper, pharmaceuticals, and batteries. Phase change of liquid to vapor generally requires enormous thermal energy, so in order to save energy, it is advantageous to develop an appropriate drier and use it under appropriate operating conditions, depending on the characteristics of materials. However, due to the complex, multiscale heat and mass transfer occurring during drying processes, predictions of appropriate drying conditions before actual operation are not easily achieved, leading to challenges in designing driers. Here, we developed a lab-scale experimental setup to evaluate the performance of band dries. The experimental setup was used to measure the moisture content and temperature change in the materials being dried in a belt dryer. Experimental results obtained using our lab-scale setup allow us to predict the performance of a full-scale band drier, thus suggesting a practical framework for predicting the drying process of various materials and developing band driers.