• Title/Summary/Keyword: biological stability

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Temperature Effect on the Productivity of Recombinant Protein in a Lysis and DNA packaging-deficient and Temperature-sensitive Bacteriophage $\lambda$System (용균과 DNA 패키징 유전자가 결핍된 온도 민감성 박테리오 파아지 람다 시스템에서 재조합 단백질 생산성에 미치는 온도의 영향)

  • Oh, Jeong-Seok;Park, Tai-Hyun
    • KSBB Journal
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    • v.20 no.2 s.91
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    • pp.112-115
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    • 2005
  • E. coli in combination with bacteriophage $\lambda$ was used to overcome the intrinsic plasmid instability that is frequently found in recombinant fermentation especially in long-term operation. In order to enhance the stability and productivity, the bacteriophage ${\lambda}NM1070$ was used in this study. It is a $\lambda$ mutant, which is deficient in the synthesis of protein related to DNA packaging and cell lysis. The ${\lambda}NM1070$ is also a temperature-sensitive mutant. To optimize the production of recombinant protein in this temperature-sensitive system, the temperature effects on growth and cloned gene expression were investigated for stable and efficient recombinant gene expression. The induction to the lytic state was not complete at $36^{\circ}C$ while the temperature above $40^{\circ}C$ induced the lytic state completely. However, the productivity was decreased at $42^{\circ}C$ by temperature inhibition. The L-free cell concentration increased with the increase of temperature until $40^{\circ}C$. In conclusion, ${\lambda}NM1070$ has the optimal temperature at $38^{\circ}C$ for stability and at $40^{\circ}C$ for expression.

Batch Kinetics of Exo-polysaccharide Production by Submerged Cultivation of Ganoderma lucidum (영지의 액체배양에 의한 세포외 다당 생산의 동력학적 특성)

  • Lee, Shin-Young;Lee, Hak-Su;Park, Heung-Cho
    • The Korean Journal of Mycology
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    • v.27 no.4 s.91
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    • pp.304-311
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    • 1999
  • Batch kinetics during the exo-polysaccharide (EPS) fermentation of Ganoderma lucidum was investigated as a function of different substrates (glucose and starch), substrate concentration $(1{\sim}7%,\;w/v)$ and subculture (3 times). Logistic model for mycelial growth fitted the experimental data better than Monod and two thirds power model. The Luedeking-Pirt equation was adequate to fit the kinetic data of product formation and substrate consumption. The EPS production was strongly non-growth associated, although it was mixed type. The product formation and sustrate consumption by growth associated mechanism decreased as the concentration of glucose increased, while those of the non-growth associated mechanism increased. However, starch medium increased the growth associated and non-growth associated substrate consumption indicating higher availability of substrate. Also, batch culture in starch medium showed the higher specific growth rate and stability during subculture than those in glucose medium. In conclusion, the enhanced EPS production and stability in the subculture was found to be remarkably improved by use of starch as sole carbon source in medium. The maximum mycelium dry weight and EPS production of 9.463 and 10.410 g/l, respectively, were obtained after shake culture of 7 days at $30^{\circ}C$ from the media containing 7% starch.

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Studies on the Production of Fermented Feeds from Agricultural Waste Products (Part IV) -On the Production of Cellulase by Aspergillus niger and Trichoderma viride- (농산폐기물(農産廢棄物)에서 발효사료(醱酵飼料)의 생산(生産)에 관(關)한 연구(硏究)[제4보(第四報)] -Aspergillus niger와 Trichoderma viride에 의(依)한 Cellulase의 생산성(生産性)에 관(關)하여-)

  • Lee, Ke-Ho;Koh, Jeong-Sam;Lee, Kang-Hup
    • Applied Biological Chemistry
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    • v.19 no.3
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    • pp.139-144
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    • 1976
  • In order to investigate the properties of enzymes from two strains of mold, reported in the previous paper, (1) studies have been made concerning the characteristics of cellulase of Aspergillus niger-SM6 and Trichoderma viride-SM10, and summarized as follows. 1. In the semi-purification the recovery of ${\beta}-glucosidase$ was the highest when 80-90% ethanol was used and 0.8 saturation of $(NH_4)_2SO_4$. 2. The characteristics of the semi-purified enzyme were as follows. Aspergillus niger-SM6 Trichoderma viride-SM10 Optimum pH 3.5 4.0 pH stability 3.0-6.0 3.0-6.0 Optimum temperature $60^{\circ}C$ $60^{\circ}C$ Heat stability below $60^{\circ}C$ below $50^{\circ}C$ Optimum reaction time 30 min. 60 min. Optimum CMC concentration 3% 3% 3. The Km values of CMCase were 0.8% and 1.01 for Aspergillus niger-SM6 and Trichoderma viride-SM10, respectively. 4. In the strain of Aspergillus niger-SM6, there were high activity of xylanase and pectinase.

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Physicochemical Characteristics Based on Fining and Filtering of Muscat Bailey A Grape Wine (Muscat Bailey A 품종 포도주의 청징과 여과에 따른 이화학적 특성)

  • Bang, Byung-Ho;Paik, Jean Kyung;Lee, Seung-Wook;Jeong, Eun-Ja;Rhee, Moon-Soo;Yi, Dong-Heui
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.11
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    • pp.1687-1692
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    • 2015
  • Since the domestic grape wine industry has been stagnant, the quality of Korean wine must be improved to compete with imported wines. For improving the quality of domestic wine up to 'unfined' or 'unfiltered' wine, this study investigated the possibility of producing 'unfined' or 'unfiltered' wine and their physicochemical characteristics. Muscat Bailey A as a domestic grape was selected to make wine. Prior to refining process after alcohol fermentation, the wine samples were divided into three specimens depending on the treatment of bentonite for clarification and filter (sample A treated with both bentonite and filter, sample B treated with bentonite only, and sample C treated without bentonite and filter). The physicochemical characteristics of each specimen after one week matureness were then investigated. Sample C showed highest values in total phenol and organic acid content among others. Samples B and C showed more stability in intensity, hue, and color. Although it is hard to compare domestic wine with imported wine, it may be possible to improve the quality of domestic wine and contribute to the development of the domestic wine industry when further studies are conducted on in-depth fermentation of wine and improvement of microbiological stability.

Synthesis of L-threo-3,4-Dihydroxyphenylserine(L-threo-DOPS) with Thermostabilized Low-Specific L-Threonine Aldolase from Streptomyces coelicolor A3(2)

  • Baik, Sang-Ho;Yoshioka, Hideki;Yukawa, Hideaki;Harayama, Shigeaki
    • Journal of Microbiology and Biotechnology
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    • v.17 no.5
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    • pp.721-727
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    • 2007
  • Stability-enhanced mutants, H44, 11-94, 5A2-84, and F8, of L-threonine aldolase(L-TA) from Streptomyces coelicolor A3(2)(SCO1085) were isolated by an error-prone PCR followed by a high-throughput screening. Each of these mutant, had a single amino acid substitution: H177Y in the H44 mutant, A169T in the 11-94 mutant, D104N in the 5A2-84 mutant and F18I in the F8 mutant. The residual L-TA activity of the wild-type L-TA after a heat treatment for 20 min at $60^{\circ}C$ was only 10.6%. However, those in the stability-enhanced mutants were 85.7% for the H44 mutant, 58.6% for the F8 mutant, 62.1% for the 5A2-84 mutant, and 67.6% for the 11-94 mutant. Although the half-life of the wild-type L-TA at $63^{\circ}C$ was 1.3 min, those of the mutant L-TAs were longer: 14.6 min for the H44 mutant, 3.7 min for the 11-94 mutant, 5.8 min for the 5A2-84 mutant, and 5.0 min for the F8 mutant. The specific activity did not change in most of the mutants, but it was decreased by 45% in the case of mutant F8. When the aldol condensation of glycine and 3,4-dihydroxybenzaldehyde was studied by using whole cells of Escherichia coli containing the wild-type L-TA gene, L-threo-3,4-dihydroxyphenylserine(L-threo-DOPS) was successfully synthesized with a yield of 2.0 mg/ml after 20 repeated batch reactions for 100 h. However, the L-threo-DOPS synthesizing activity of the enzyme decreased with increased cycles of the batch reactions. Compared with the wild-type L-TA, H44 L-TA kept its L-threo-DOPS synthesizing activity almost constant during the 20 repeated batch reactions for 100 h, yielding 4.0 mg/ml of L-threo-DOPS. This result showed that H44 L-TA is more effective than the wild-type L-TA for the mass production of L-threo-DOPS.

Real-time Control of Biological Animal Wastewater Treatment Process and Stability of Control Parameters (생물학적 축산폐수 처리공정의 자동제어 방법 및 제어 인자의 안정성)

  • Kim, W.Y.;Jung, J.H.;Ra, C.S.
    • Journal of Animal Science and Technology
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    • v.46 no.2
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    • pp.251-260
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    • 2004
  • The feasibility and stability of ORP, pH(mV) and DO as a real-time control parameter for SBR process were evaluated in this study. During operation, NBP(nitrogen break point) and NKP(nitrate knee point), which reveal the biological and chemical changes of pollutants, were clearly observed on ORP and pH(mV)-time profiles, and those control points were easily detected by tracking the moving slope changes(MSC). However, when balance of aeration rate to loading rate, or to OUR(oxygen uptake rate), was not optimally maintained, either false NBP was occurred on ORP and DO curves before the appearance of real NBP or specific NBP feature was disappeared on ORP curve. Under that condition, however, very distinct NBP was found on pH(mV)-time profile, and stable detection of that point was feasible by tracking MSC. These results might mean that pH(mV) is superior real-time control parameter for aerobic process than ORP and DO. Meanwhile, as a real-time control parameter for anoxic process, ORP was very stable and more useful parameter than others. Based on these results, a stable real-time control of process can be achieved by using the ORP and pH(mv) parameters in combination rather than using separately. A complete removal of pollutants could be always ensured with this real-time control technology, despite the variations of wastewater and operation condition, as well as an optimization of treatment time and capacity could be feasible.

Immobilization of $\beta-galactosidase$ with High Transgalactosylation Activity from Bacillus sp. A4442 Mutant (변이주 Bacillus sp. A4442가 생산하는 갈락토스 전이활성이 높은 $\beta-galactosidase$의 고정화)

  • Kim, Min-Hong;Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.39 no.5
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    • pp.333-337
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    • 1996
  • For continuous production of galactooligosaccharides(GOS), $\beta-galactosidase$ with h1gh transgalactosylation activity from Bacillus sp. A 4442 was Immobilized onto $Diaion^{TM}$ HPA 75(styrene-divinylbenzene resin). The parameters influencing enzyme immobilization were scrutinized in order to maximize immobilization yield while minimizing enzyme inactivation. The optimum conditions turned out to be: Tris buffer concentration 30 mM, pH 8.0, contact time at room temperature 3 hr, and enzyme loading 25 mg protein/g resin. Both the thermal stability and the operational stability of immobilized enzyme were markedly enchanced by the treatment with 0.5% glutaraldehyde as a cross-linker. Under the experimental conditions established, the yield of ${\beta}-galactosidase$ immobilization was 40% or more and the activity of the immobilized enzyme ca. 200 U/g resin. When a packed-bed reactor was employed to continuously convert lactose to GOS, the specific production, which refers to as the amount of commercially valuable GOS produced by a unit amount of immobilized ${\beta}-galactosidase$, was found to be ca. 300 g GOS/g carrier.

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Thermal Stability of the Major Color Component, Cyanidin 3-glucoside, from a Korean Pigmented Rice Variety in Aqueous Solution (한국산 유색미의 주요성분인 Cyanidin 3-glucoside의 수용액에서의 열안정성)

  • Jo, Man-Ho;Yoon, Hye-Hyun;Hahn, Tae-Ryong
    • Applied Biological Chemistry
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    • v.39 no.3
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    • pp.245-248
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    • 1996
  • Thermal stability of the major color component, cyanidin 3-glucoside, isolated from Korean pigmented rice (Oryza sativa var. Suwon 415) were investigated to explore possible application of value-added natural colors as food additives. The anthocyanin showed red and blue color with maximum absorption peaks at 511 nm and 572 nm in acidic (pH 2.0) and alkaline (pH 9.0) buffer solutions, respectively, and the thermal degradation reactions were carried out with different temperature ranges at $50{\sim}95^{\circ}C$. Degree of degradation was determined with UV/Vis spectra which indicate characteristic absorption patterns with sharp isosbestic points at 350 nm (pH 2.0), and 275, 310, and 405 nm (pH 9.0). Thus the reaction follows simple first-order kinetics. The anthocyanin was very stable against heat at acidic pH and relatively stable at alkaline pH with half-life values of 50.3 hr and 0.6 hr at $70^{\circ}C$, respectively. The activation energies and Arrhenius frequency factors of the pigment were 26.9 kcal $mol^{-1}\;and\;6.0{\times}10^{11}\;s^{-1}$, at pH 2.0, and 15.2 kcal $mol^{-1}\;and\;1.4{\times}10^{6}\;s^{-1}$, pH 9.0, and respectively.

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Comparison of the Gel Formation Ability and Stability of Encapsulated Microbial Inoculant Using Extractable Alginate from Sea Tangle (다시마 추출 Alginate를 이용한 미생물 캡슐화제의 겔 형성능 및 생균력 비교)

  • Choi, So-Young;Yoon, Min-Ho;Whang, Kyung-Sook
    • Applied Biological Chemistry
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    • v.49 no.3
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    • pp.170-174
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    • 2006
  • For the purpose of developing a high quality agricultural microbial inoculant, methods and materials for improving encapsulation were investigated. Preparation of capsule was conducted by improving extrusion system with micro-nozzle and peristaltic pump. The sodium alginate was selected because of its cheapness, stability of cells, and gel formation ability. The yields, physical properties and gel formation abilities of extractable alginate from sea tangle were investigated by hot water extractable and alkali soluble methods. The extraction yields of hot water extractable alginate (HWEA) and alkali soluble alginate (ASA) from sea tangle were 8 and 20%, respectively. The HWEA was almost not viscous even in 1.5% of the sample solution, whereas the ASA was very highly viscous in above 3% sample solution. The gel formation ability of each samples varied from 1.5% to 5% and the ASA showed a good gel formation ability at 3% solution as commercial alginate (CA). The soil microbial inoculant, Bacillus thuringiensis, Bacillus subtilis, Lactobacillus plantarum and Geotrichum candidum encapsulated sodium alginate with starch and zeolite for stabilizer. The survivability of encapsulated soil microbial inoculant using alginate without stabilizer appeared to be 66, 52, 70 and 50%, respectively. Inclusion of starch and zeolite with alginate bead increased viabilities in Bacillus sp. and Geotrichum candidum by 81-83% and 89%, respectively.

Application of L-Arginine as a Substitute for Inorganic Polyphosphate in Pork Sausage Production (소시지 제조시 인산염 대체 소재로 L-Arginine의 활용)

  • Kim, Yong-Duk;Lee, Jun-Soo;Park, Joon-Hyun;Park, Dong-Cheol;Jeon, Young-Seung;In, Man-Jin;Oh, Nam-Soon
    • Journal of Applied Biological Chemistry
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    • v.57 no.2
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    • pp.171-174
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    • 2014
  • $\small{L}$-Arginine was applied to processed pork sausages and evaluated as a substitute for commonly used inorganic polyphosphate. Processed pork sausage was prepared with an addition of 0.25, 0.5, and 1.0% (w/w) $\small{L}$-arginine. Quality characteristics of the processed pork sausages were than evaluated in terms of pH, moisture content, emulsion stability and textural properties. The pH of sausage increased with increasing $\small{L}$-arginine concentration. Moisture content of 0.25% $\small{L}$-arginine sausage, emulsion stability of 0.5% $\small{L}$-arginine sausage, and textural properties of 1.0% $\small{L}$-arginine sausage were similar to those of 0.5% polyphosphate sausage, respectively. In terms of sensory evaluation, taste and overall acceptability of 0.5% $\small{L}$-arginine sausage were slightly higher than those of 0.5% polyphosphate sausage and textural properties were reversed. According to sensory score and quality characteristics, $\small{L}$-arginine had a potential as a substitute for inorganic polyphosphate and the optimum concentration of $\small{L}$-arginine was around 0.5%.