• Title/Summary/Keyword: Optimal pH

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Improvement of the Optimum pH of Aspergillus niger Xylanase towards an Alkaline pH by Site-Directed Mutagenesis

  • Li, Fei;Xie, Jingcong;Zhang, Xuesong;Zhao, Linguo
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.11-17
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    • 2015
  • In an attempt to shift the optimal pH of the xylanase B (XynB) from Aspergillus niger towards alkalinity, target mutation sites were selected by alignment between Aspergillus niger xylanase B and other xylanases that have alkalophilic pH optima that highlight charged residues in the eight-residues-longer loop in the alkalophilic xylanase. Multiple engineered XynB mutants were created by site-directed mutagenesis with substitutions Q164K and Q164K+D117N. The variant XynB-117 had the highest optimum pH (at 5.5), which corresponded to a basic 0.5 pH unit shift when compared with the wild-type enzyme. However, the optimal pH of the XynB-164 mutation was not changed, similar to the wild type. These results suggest that the residues at positions 164 and 117 in the eight-residues-longer loop and the cleft's edge are important in determining the pH optima of XynB from Aspergillus niger.

Optimization of Semi-Batch Process for Ethanol Production (에타놀 생산을 위한 Semi-batch 발효 공정의 최적화)

  • Lee, Jae-Heung
    • Microbiology and Biotechnology Letters
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    • v.11 no.1
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    • pp.33-38
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    • 1983
  • As flocculent strains are likely to have considerable potential for internal cell recycle, kinetic studies on glucose medium with flocculent Saccharomyces uvarum were carried out in batch and continuous culture. Using a mathematical model, the kinetic parameters at each temperature and pH were estimated in order to establish optimal conditions. It was found that an overall optimum temperature for growth and ethanol production in the range 33-35$^{\circ}C$ was desirable. With regard to the effect of pH, ethanol production by S. uvarum was found to be relatively insensitive to pH value between 4 and 6, with an optimum pH of around 5. At these optimal conditions a maximum ethanol productivity of 12 g/$\ell$/h was determined using semi-batch process together with 5. uvarum.

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The Optimal Conditions for Fibrinolytic Enzyme Production from Streptomyces sp. JK-20 (Streptomyces sp. JK-20유래 혈전용해효소의 생산조건)

  • 정영기;전홍기;김유정
    • Journal of Life Science
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    • v.12 no.1
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    • pp.43-48
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    • 2002
  • An actinomycetes which produces fibrinolytic enzyme was isolated from soil. Characteristics of the isolated strain and the optimal conditions for the productions of fibrinolytic enzyme were summarized as follows; The fibrinolytic enzyme production strain generates gray airmycelium and had about 0.6~0.8$\times$0.4~0.8${\mu}{\textrm}{m}$ cylindrical spore, smooth surface and formed spore chain of 10~40 spores. We have identified this strain as Streptomyces sp. JK-20. This strain was able to grow up at 20~32$^{\circ}C$ and its optimum growth temperature and pH was 24$^{\circ}C$ and pH 6.0, respectively. The optimal conditions for porducing fibrinolytic enzyme; carbon source, nitrogen source, metal ions and phosphorous sources was 1% xylose, 0.5% yeast extract, 0.5% polypepton, 0.1% MgSO$_4$.7$H_2O$ and 0.1% NaH$_2$PO$_4$.2$H_2O$, respectively. This strain showed the highest productivity of fibrinolytic enzyme after the fourth day under such optimal culture conditions.

THE EFFECT OF QUADRATURE ERRORS IN PRACTICE

  • Kim, Chang-Geun
    • Korean Journal of Mathematics
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    • v.6 no.2
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    • pp.195-203
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    • 1998
  • In [3], we showed that overintegration may be needed to obtain the optimal $H^1$ error rate for the $p$ version. In this paper, we examine the convergence of the $p$ version in practice, and comment on the implementation of the $p$ version in commercial codes. Also, we give an example of a problem with extremely rough coefficients, for which overintegration is necessary to obtain the optimal $H^1$ convergence rate.

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Activities of the Hydrolytic Enzymes Produced by Plant Pathogenic Fungi, Sclerotium rolfsii, Sclerotinia Sclerotinia and Sclerotiorum, and Helminthosporium sigmoideum var. irregulare (수종의 식물병원균(흰비단병균$\cdot$균핵병균 및 좀검은 균핵병균)이 생산하는 가수분해효소의 활성)

  • Cho B. H.;Kim K.
    • Korean journal of applied entomology
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    • v.16 no.4 s.33
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    • pp.199-208
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    • 1977
  • Activities of various hydrolytic enzymes produced by three plant pathogenic fungi, Sclerotium rolfsii Sacc., Sclerotinia sclerotiorum (Lieb.) deBary and Helminthosporium sigmoideum var. irregulare Crallery et Tullius, were measured. Activties and amounts of the enzymes in mycelia, cultural filtrates, and sclerotia(except of sclerotia of H. sigmoideum var. irregulare) were estimated at various pH levels in order to find out optimal pH for their enzymatic activities. Enzymes such as cellulase (ex), invertase, xylanase, $\beta-amylase$, polymethylgalacturonase, polygalacturonase, phosphatase and protease were estimated. Culture solution for production of enzymes was prepared by adding of 10g, D-glucose, 1.3g $NH_4NO_3,\; 0.5g\; MgSO_4,\;7H_2O,\; and\; 1.0g\; KH_2PO_4$ into 1 liter of potato decoction plus 2ml of micro element solution consisting of 0.2mg. Fe, 0.2mg Zn, and 0.1mg Mn as the sulphates into 1 liter of distilled water. All tested mycelia and cultural filtrates were obtained from the cultures incubarted in previous solution for ten days at $25^{\circ}C$, and sclerotia were harvested from PDA plates of 3. days old, The crude enzyme solutions were prepared according to the method of Miyazaki etal. Ten days after incubation, activities of Cx produced by Scl. sclerotiorum were higher than those of the other fung and each of Cx from three fungi showed different pH optima, such as S. rolfsii and Scl. schlerotiorum in acid side (around pH 3.0), H. sigmoideum var. irregulare in neutral side (around pH 6.3). Invertase activities of S. rolfsii were 20 times higher than those of the other fungi in all samples. All tested fungi, however, showed no significant difference between the enzymatic activities of their cultural filtrate and mycelia and the activities in sclerotia of S. rolfsii and Scl. sclerotiorum were hardly recognized. There were multiple peaks on the xylanase activity curves of three fungi in terms of pH values. High activities of the xylanase were revealed in sclerotia of S. rolfsii and Scl. sclerotiorum, and in mycelia of H. sigmoideum var. irregulare. The highest activities of $\beta-amylase$ were shown both in mycelia and cultural filtrate of H. sigmoideum var. irregulae among the tested fungi, and their optimal pH was 6.2 in both mycelia and cultural filtrate. In the S. rofsii and Sel. sclerotiorum, however, the activities of cultural filtrates were higher than those of the other fungi, and optimal pH was 3.0 and 6.2 for cultural filtrate and both mycelia and sclerotia, respectively. Activities of PMG were high in cultural filtrates of all tested fungi, especially in Scl. sclerotiorum and H. sigmoideum var. irregulare. Mycelia of themalso showed the considerable activities. Optimal pH for enzymatic activities were variable with thekind of fungi or with the samples measured. The highest activities of PG were presented by mycelia of S. rolfsii and Scl. sclerotiorum. $9.l\mu /min.\; and\; 9.5\mu g/min.$, respectively. Optimal pH for activity of PG in mycelia was around 4.5 in S. rolfsii and around 3.0 in Scl. sclerotiorum. Phosphatase of S. rolfsii and Scl. sclerotiorum was more active in acid side (optimal PH3. 5) and that of H. sigmoideum var. irregulare showed one peak each in acid, neutral and alkaline side. But the highest peak was at pH 9.5. Protease of all tested fungi was more active at pH 10.0, especially that of the cultural filtrate of H. sigmoideum var. irregualre.

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Studies on Naringinase Produced from Aspergillus nidulans (Part 4) Immobilization of Naringinase on DEAE-Sephadex A-25 (Aspergillus nidulans가 생산하는 Naringinase에 관한 연구 (제4보) DEAE-Sephadex A-25에 의한 Naringinase의 고정화)

  • 송충석;변유량;유주현
    • Microbiology and Biotechnology Letters
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    • v.7 no.3
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    • pp.141-147
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    • 1979
  • Naringinase from Atpergillus nidulans was immobillized on DEAE-Sephadex A-25 and its characteristics were studied. The optimal conditions for the preparation of the immobilized enzyme were as follow; optimal pH, incubation time and the suitable amount of enzyme were 6.0, 30 min. and 110 units per gram of the dried ion exchage resin, respectively. The optimal pH of the immobilized enzyme was higher than that of the native enzyme. The optimal temperature increased from 4$0^{\circ}C$ to 5$0^{\circ}C$. The heat and pH stability of the immobillized enzyme were better than those of the native enzyme. No significant difference in the Michaelis constant was detected. Activation energy of the immobilized enzyme was 7.96 Kcal/mole, and the apparent Michaelis rate equation was used to describe the action of this material. The degree of hydrolysis was dependant on the flow rate at low rate of perfusion through the column. As the flow rate increased, the value of the apparent Km decreased.

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Optimal Production of Protease from Entomopathogenic Fungus Beauveria bassiana (곤충 병원성 곰팡이 Beauveria bassiana로부터 Protease의 최적 생산)

  • Ko, Hwi-Jin;Kim, Hyun-Kyu;Kang, Sun-Chul;Kwon, Suk-Tae
    • Applied Biological Chemistry
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    • v.39 no.6
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    • pp.449-454
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    • 1996
  • We investigated the optimal condition for the production of extracellular pretense(a cuticle-degrading pretense) from entomopathogenic fungus Beauveria bassiana(ATCC7159) in liquid medium by adding of gelatin, bovine serum albumin(BSA), casein and polypeptone. The optimal induction medium for production of extracellular pretenses is composed of 0.5% polypeptone, trace elements and 50 mM potassium phosphate(pH 6.0). In this condition, the production of extracellular pretenses increased rapidly after the 24hrs, peaking at the third day and there was little inductive effect in culture broth more than pH 7.0. The pretenses were inhibited by phenyl methyl sulfonyl fluoride(PMSF). High activity of pretense was showed both range of pH 8.5 and 11.5 and also detected by three different portions of slice gel derived from non-denaturing isoelectricfocusing gel. At least three different extracellular pretenses are produced in optimal production medium when polypeptone is used as the sole carbon and nitrogen source.

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Culture Characteristics and Optimal Conditions for Mycelial Growth of Artomyces microsporus (Artomyces microsporus의 배양적 특성과 균사 적정 배양 조건 설정)

  • Min, Gyeong-Jin;Lee, Een-ji;Park, Hea-sung;Lee, Chan-Jung
    • The Korean Journal of Mycology
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    • v.49 no.1
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    • pp.45-55
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    • 2021
  • Artomyces microspora is a genus of coral fungi from the family Auriscalpiaceae that have sporophores which are clavarioid, profusely and pyxidately branched, and devoid of a conspicuous stipe. These fungi can be found in summer and fall. This study aimed to decipher fundamental information regarding optimal growth conditions of Artomyces microsporus mycelia, including pH, temperature, carbon sources, and nitrogen sources. Based on the assessment of colony diameter and mycelial density, the optimal culture medium, temperature, and pH for mycelial growth were found to be PDA, 25 ℃, and pH 5.0, respectively. Furthermore, the study revealed that the optimal carbon and nitrogen sources for mycelial growth were 1% soluble starch and 2% malt extract, respectively. The other suitable inorganic nitrogen sources were deemed to be 0.1% NH4H4PO4 and 0.1% aspartic acid.

The pH as a Control Parameter for Oxidation-Reduction Potential on the Denitrification by Ochrobactrum anthropi SY 509

  • Kim, Sung-Hong;Song, Seung-Hoon;Yoo, Young-Je
    • Journal of Microbiology and Biotechnology
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    • v.14 no.3
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    • pp.639-642
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    • 2004
  • The pH as a control parameter for oxidation-reduction potential (ORP) was investigated through the denitrification by Ochrobactrum anthropi SY509 under non-growing condition. The optimal pH of nitrate reductase was 7.0, and the minimal ORP level was -250 mV for the denitrification under aerobic condition. In the case of anaerobic condition, the optimal pHs of nitrate and nitrite reductase were shifted to 10.0 and 9.0, respectively, and the minimal ORP levels of nitrate and nitrite reductase were decreased to -370 mV and -340mV, respectively. In the case of alkaline pH and anaerobic condition, the denitrification efficiency of nitrate was increased up to about 2-fold over that of neutral pH and anaerobic condition. Therefore, the combined control of pH and ORP in the anaerobic condition is shown to be an important parameter in the biological denitrification process.

The Production and Properties of Pretense by Serratia sp. 2000-1 Isolated from Clinical Specimes (임상검체에서 분리된 Serratia sp. 2000-1에 의한 Pretense의 생산 및 효소학적 성질)

  • 김태전;김승곤;김상택
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.209-221
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    • 2000
  • The purpose of this study was to investigate the practical availability of pretense production that can be used at home after isolating Serratia sp.2000-1 which produced extracellular pretense from clinical specimen. Basic production conditions and partial enzymatic characteristics of pretense produced by Serratia sp. 2000-1 was as follows: The kind and concentration of carbohydrate, nitrogen and metal salts for optimal enzyme production condition were each identified as the concentration of 1.5% glucose, 2.0% CSP, and 0.1% CaCl$_2$, and the optimal temperature, time and initial pH for culture were each 3$0^{\circ}C$, 72 hours, and pH 8.0. The final enzymatic yeild that was purified by 3 steps with ammonium sulfate precipitation (45~80%), DEAE-cellulose column chromatography, and Sephadex G-200 gel chromatography was 14.4%, and enzyme inactivity rate increased approximately 291314s. The optimal temperature and pH for purified pretense activity were 35$^{\circ}C$ and pH 7.0~8.0, and purified pretense activity was relatively stable by 4$0^{\circ}C$ at pH 6~10 for 30 min, however heating at 6$0^{\circ}C$ for 30 min, it liminated detectable pretense activity. The pretense activity was activated by $Mg^{2+}$, $Ba^{2+}$, $Ca^{2+}$, Mn$^{2+}$, but inactiviaed by Hg$^{2+}$, Ag$^{2+}$, Cu$^{2+}$, and the pretense activity was inhibited strongly by SDS among enzyme activity inhibitors. Further study is required to evaluate the practical availability of pretense production that can be used at home by isolating Serratia sp. from more clinical specimen and examining pretense more in details.

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