• Title/Summary/Keyword: H-Beta

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Extracellular Overproduction of $\beta$-Cyclodextrin Glucanotransferase in a Recombinant E. coli Using Secretive Expression System

  • Lee, Kwang-Woo;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.12 no.5
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    • pp.753-759
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    • 2002
  • $\beta$-Cyclodextrin glucanotransferase ($\beta$-CGTase) was overproduced extracellularly using recombinant E. coli by transforming the plasmid pECGT harboring a secretive signal peptide. The $\beta$-CGTase gene of alkalophilic Bacillus firmus var alkalophilus was inserted into the high expression vector pET20b(+) containing a secretive pelB signal peptide, and then transformed into E. coli BL2l(DE3)pLysS. The optimum culture conditions fer the overproduction of $\beta$-CGTase were determined to be TB medium containing 0.5% (w/v) soluble starch at post-induction temperature of $25^{\circ}C$. A significant amount of $\beta$-CGTase, up to 5.83 U/ml, which was nine times higher than that in the parent strain B. firmus var. alkalophilus, was overproduced in the extracellular compartment. A pH-stat fed-batch cultivation of the recombinant E. coli was also performed to achieve the secretive overproduction of $\beta$-CGTase at a high cell density, resulting in production of up to 21.6 U/ml of $\beta$-CGTase.

The Action Mode of $\beta$-glucosidase Purified from Trichoderma koningii (Trichoderma koningii에서 분리한 $\beta$-glucosidase의 작용양상)

  • 정춘수;최지영;이헌주;맹필재;한인섭;강사욱;하영칠
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.8-13
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    • 2000
  • We have examined the mode of transglycosylation, catalyzed by an extracellular $\beta$-glucosidase purified from Trichoderma koningii ATCC 26113, using cellobiose, sophorose, laminaribiose and gentiobiose as substrates. The dimers separated from the reaction mixture by HPLC were analyzed by $^(1)H$-NMR spectroscopy. When cellobiose was subjected to the action of the $\beta$-glucosidase, the products included laminaribiose, sophorose and gentiobiose. When laminaribiose, sophorose or gentiobiose was used as a substrate, the $\beta$-glucosidase accumulated transglycosylation products possessing different types of $\beta$-glycosidic linkages from the original one. The amount of dimers accumulated as reaction proceeded seemed to be dependent on the velocity of hydrolysis but not on that of formation.

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Synthesis of Nucleophilic Adducts of Thiols (Ⅰ). Addition of Cysteine to $\beta$-Nitrostyrene Derivatives

  • Kim, Tae-Rin;Choi, Sung-Yong
    • Bulletin of the Korean Chemical Society
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    • v.2 no.4
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    • pp.125-129
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    • 1981
  • The addition reactions of cysteine without blocking amino and carboxyl groups to substituted and unsubstituted ${\beta}$-nitro-styrene derivatives were investigated. ${\beta}$-Nitrostyrene(1a), p-methyl-${\beta}$-nitrostyrene(1b), 3,4,5-trimethoxy-$[\beta}$ -nitrostyrene(1c), $[\varpi}$-3,4-methylenedioxy-${\beta}$ -nitrostyrene(1d), o-, m- and p-chloro-${\beta}$ -nitrostyrene (1e, 1f, 1g) and o-, m- and p-methoxy-${\beta}$-nitrostyrene (1h, 1i, 1j) easily undergo addition reactions with cysteine to form S-(2-nitro-1-phenylethyl)-L-cysteine(3a), S-[2-nitro-1-(p-methyl)phenyl-ethyl]-L-cysteine(3b), S-[2-nitro-1-(3',4',5'-trimethoxy) phenylethyl]-L-cysteine(3c), S-[2-nitro-1-($[\vatpi}$ -3',4'-methylenedioxy)phenylethyl]-L-cysteine(3d), S-[2-nitro-1-(o-chloro)phenylethyl]-L-cysteine(3e), S-[2-nitro-1-(m-chloro)-phenylethyl]-L-cysteine(3f), S-[2-nitro-1-(p-chloro)phenylethyl]-L-cysteine(3g), S-[2-nitro-1-(o-methoxy)phenylethyl]-L-cysteine(3h), S-[2-nitro-1-(m-methoxy)phenylethyl]-L-cysteine(3i) and S-[2-nitro-1-(p-methoxy)phenylethyl]-L-cysteine(3j), respectively. The structure of adducts were confirmed by means of UV-spectrum, IR-spectrum, molecular weight measurement and elemental analysis. The various factors effecting the yield of cysteine adducts to ${\beta}$-nitrostyrene derivatives were also studied.

Effect of Water-Activity Depressor on the Enzymatic Synthesis of Maltosyl-$\beta$-Cyclodextrin through the Reverse Reaction of Pullulanase (Water-Activity Depressor가 Pullulanase의 역반응에 의한 Maltosyl-$\beta$-Cyclodextrin의 합성에 미치는 영향)

  • 이용현;한일근
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.422-429
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    • 1992
  • The effect of various water-activity depressors, such as pol yo Is, sugars, and polymers, on the conversion yields of the enzymatic synthesis of maltosyl-$\beta$-cyclodextrin from $\beta$-cyc1odextrin and maltose through reverse reaction of pullulanase was investigated. PEG 6000 of concentration of 10% (w/w) was found to be the most acceptable water-activity depressor resulting for increment of conversion yield from 43.0% to 55.9%, corresponding maltosyl-$\beta$-cyc1odextrin concentration of 3.02 g/100 ml H20. Water activity was changed from initial 0.966 to 0.914 upon addition of 20% (w/w) of PEG 6000. The conversion yields were inversely proportional to the water activities, and the increased conversion yield was caused by water activity depression which inhibited the hydrolysis reaction of maltosyl-$\beta$-CD to maltose and $\beta$-cyc1odextrin. The changes of enthalpy ($\Delta$H), entropy ($\Delta$S), and Gibbs free energy ($\Delta$G) were calculated to be 36.788 kJ/mole, 0.067 kJ/mole K. and 14.433 kJ/mole, respectively. The synthesis of maltosyl-$\beta$-CD could be increased substantially by the intermittent feeding of $\beta$-cyclodextrin. PEG 6000 could be separated effectively from reaction mixture using ultrafiltration membrane for reutilization.

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Studies on Rheological Characterization of Barley ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$] (보리 ${\beta}-Glucan$ [mixed-linked $(1-3),(1-4)-{\beta}-D-Glucan$의 리올로지 특성)

  • Kim, Mi-Ok;Cha, Hee-Sook;Koo, Sung-Ja
    • Korean Journal of Food Science and Technology
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    • v.25 no.1
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    • pp.15-21
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    • 1993
  • Crude ${\beta}-glucan$ extracted from Barley was purified by stepwise enzyme treatment (Thermostable ${\alpha}-amylase$, amyloglucosidase, protease). The Intrinsic Viscosity $[{\eta}]$ of the purified ${\beta}-glucan$ was determined by Cannon Fenske Capillary Viscometer (size 50, Cannon Instruments, State, College pa.) at different pH (2, 4, 7, 9, 11) and various salt concentration (0.01 M, 0.03 M, 0.05 M, 0.07 M, 0.1 M and 0.2 M). The $[{\eta}]$ of purified ${\beta}-glucan$ was ranged from $0.997{\sim}2.290\;dl/g$. The $[{\eta}]$ of purified ${\beta}-glucan$ at both alkali, acid condition were lower than those at pH 7. However, the alkali condition of puified ${\beta}-glucan$ solution showed less $[{\eta}]$ than the acid condition of this solution. From 0 M to 0.2 M salt concentration, the $[{\eta}]$ of purified ${\beta}-glucan$ solution was decreased to 0.03 M then increased to 0.05 M NaCl and remained constant to 0.2 M NaCl. The chain stiffness parameter of purified ${\beta}-glucan$ was not affected by temperature from $15^{\circ}C$ to $65^{\circ}C$. The shear rates of various ${\beta}-glucan$ conditions were determined by Bohlin Rheometer (Lund, Sweden). The ${\beta}-glucan$ concentration of 1.0 g/dl and 2.0 g/dl behaved as Newtonian fluid. However, above the concentration of 3.0 g/dl ${\beta}-glucan$ solution, it showed thixotropic and psedoplastic characteristics. Barley ${\beta}-glucan$ appears a damping at 0.5 frequency for the 4.0 g/dl solution. Below 0.5 frequency, it appears a viscous behavior property and above 0.5 frequency, it appears a elastic behavior property.

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Effective Production of $\beta$-Glucan by the Liquid Cultivation of Agaricus blazei (Agaricus blazei 균사체 배양기술을 통한 효율적인 $\beta$-glucan의 생산)

  • 이승현;임환미;김태영;조남석;박준성;유연우;김무성
    • Korean Journal of Microbiology
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    • v.40 no.1
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    • pp.54-59
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    • 2004
  • $\beta$-Glucan has been efficiently produced with higher yield by the optimization of liquid cultivation conditions. The optimal composition of medium for batch culture was 5% (w/v) of glucose as a carbon source, 0.5% (w/v) of yeast and 0.5% (w/v) of malt extract as a nitrogen source, 0.1% (w/v) of $KH_2PO_4$ and 0.05% (w/v) $MgSO_4{\cdot}7H_2O$, which had been the base medium for determination of other conditions. The set-up conditions are pH 5.0, $28^{\circ}C$, 1 vvm for aeration and 300 rpm for agitation. In order to minimize the inhibition effect of glucose on the initial growth of mycelia and to maximize the production of extracellular $\beta$-glucan, we have reduced the initial glucose feed to 4% and added 2nd feed at the point of 70 hr from the initial feed. The 2nd feed was composed of glucose 3%, yeast extract 0.1 % and malt extract 0.1 %. It improved the $\beta$-glucan yield upto 5.2 g/L in comparison with 2.8 g/L resulted from batch cultivation. Moreover, the serial treatment of a cell wall lytic enzyme and bromelain to the mycelia was effective for extraction of the cell wall bound $\beta$-glucan. The yield of $\beta$-glucan extraction by the enzyme treatment was 3.5 g/L, which was almost 4 times higher than that by hot-water extraction.

The Synthesis and Characterization of Beta Alumina Powder by Microwave Extraction and Calcination Method (마이크로파 추출 및 하소법에 의한 베타 알루미나 분말 합성 및 특성)

  • 박희찬;박성수;정기석;김병우;이상은
    • Journal of the Korean Ceramic Society
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    • v.38 no.8
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    • pp.749-754
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    • 2001
  • 마이크로파 추출 및 하소 공정을 통하여 카올린으로부터 베타 알루미나 전구체 및 베타 알루미나 분말을 합성하였다. 황산 알루미늄 및 가성소다의 혼합 용액을 교반되는 에탄올 용액 중에 적하시켜서 $Al_2$(SO$_4$)$_3$.17$H_2O$$Na_2$SO$_4$.10$H_2O$의 균일한 침전물을 얻었다. $\beta$- 및 $\beta$\"-Al$_2$O$_3$상으로 구성된 베타 알루미나를 얻기 위하여 재래식 및 마이크로파 하소법으로 주어진 온도에서 약 2h 동안 침전물 시료를 하소하였다. 재래식 하소된 시료에 비하여 마이크로파 하소된 시료는 $\beta$\"-Al$_2$O$_3$상이 상대적으로 많이 존재하였다. 또한, 재래식 하소된 시료와 마이크로파 하소된 시료의 미세구조는 매우 상이하였다. 매우 상이하였다.

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Pharmaco-Constituents of Korean Cultivated Rhubarb Leaves -The Flavonoids from Leaves- (한국산 재배대황엽의 약효성분 -엽의 후라보노이드-)

  • Ham, In-Hye;Oh, In-Se;Whang, Wan-Kyunn;Kim, Il-Hyuk
    • YAKHAK HOEJI
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    • v.38 no.4
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    • pp.469-475
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    • 1994
  • As the continued studies for Korean cultivated rhubarb, MeOH extract of the leaves was fractionated with ether, ethylacetate, and n-butanol. From the ethyl acetate fraction of MeOH extract, one flavone glycoside, apigenin-8-${\beta}$-D-glucopyranoside(vitexin, $C_{21}H_{20}O_{10}$) and from the n-BuOH fraction of MeOH extract, two flavonol glycosids, kaempferol-3-O-(2,6-di-O-rhamnopyranosyl)-${\beta}$-D-galactopyranoside$(C_{33}H_{40}O_{19})$and quercetin-3-O-rutinoside(rutin, $C_{27}H_{30}O_{16}$) were isolated and identified through the physico-chemical properties and spectroscopic evidences(UV, IR, NMR, Mass) respectively.

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Enhanced Production of L-Aspartate ${\beta}-Decarboxylase$ by Nitrogen Source in Pseudomonas dacunhae

  • Kim, Dong-Chung;Lee, Sung-Dong;In, Man-Jin
    • Journal of Applied Biological Chemistry
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    • v.49 no.3
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    • pp.106-109
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    • 2006
  • Improvement of L-asparate ${\beta}-decarboxylase$ production from Pseudomonas dacunhae ATCC 21192 was attempted by optimizing fermentation conditions. Optimum carbon and nitrogen sources for cell growth and enzyme production were determined. L-Glutamate (2%) was the most suitable carbon source, and D-glucose, D-glycerol and fumarate repressed enzyme production. Yeast extract (2%) was the most effective as nitrogen source. A slight change of pH to 6.5 from medium pH resulted in a meaningful increase in the production of enzyme. The production of the enzyme was highly improved by using 2% yeast extract and 2% L-glutamate in culture media. Maximum L-asparate ${\beta}-decarboxylase$ activity reached up to over 24 U/mL-broth by 15 h flask fermentation.

Effect of Anaerobic Treatment on Carbohydrate-Hydrolytic Enzyme Activities and Free Amino Acid Contents in Barley Malt

  • Yun, Song-Joong;Choi, Kyeong-Gu;Kim, Jin-Key
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.1
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    • pp.19-22
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    • 1998
  • Effects of acute anoxia on carbohydrate hydrolytic enzyme activities and free amino acid contents in malt were examined. Malts were prepared with barley grains germinated for 7 days which contained the highest levels of amylolytic and(1-3,1-4)-$\beta$-glucanase activities. $\alpha$-Amylase and $\beta$-amylase activities in malts were not significantly affected by anoxia for 5 or 10 h.(1-3,1-4)-$\beta$-Glucanase activity, however, decreased about 7 to 10% by anoxia for 5 or 10 h. Alanine and $\gamma$-aminobutyric acid content changed drastically. Alanine contents in malts increased by 2.2- and 2-fold, and $\gamma$-aminobutyric acid contents by 1.4- and 1.9-fold under anoxia for 5 and 10 h, respectively.

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