• Title/Summary/Keyword: $\beta$-type

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Cloning and Sequencing of the ${\beta}-Amylase$ Gene from Paenibacillus sp. and Its Expression in Saccharomyces cerevisiae

  • Jeong, Tae-Hee;Kim, Hee-Ok;Park, Jeong-Nam;Lee, Hye-Jin;Shin, Dong-Jun;Lee, Hwang-Hee Blaise;Chun, Soon-Bai;Bai, Suk
    • Journal of Microbiology and Biotechnology
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    • v.11 no.1
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    • pp.65-71
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    • 2001
  • A gene from Paenibacillus sp. KCTC 8848P encoding ${\beta}-amylase$ was cloned and expressed in Escherichia coli. The Paenibacillus ${\beta}-amylase$ gene cosisted of a 2,409-bp open reading frame without a translational stop codon, encoding a protein of 803 amino acids. The presumed ribosime-binding site, GGAGG, was located 10 bp upstream from the TTG initiation codon. The deduced amino acid sequence of the ${\beta}-amylase$ gene had a 95% similarity to the ${\beta}-amylase$ of Bacillus firmus. The ${\beta}-amylase$ gene was introduced into wild-type strains of Saccharomyces cerevisiae using a linearized yeast integrating vector containing a geneticin resistance gene and its product was secreted into the culture medium.

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The Reaction Conditions of $\beta$-Galactosidases from Aspergillus oryzae, Bovine Liver, and Saccharomyces fragilis to Asialofetuin (Asialofetuin에 대한 Aspergillus oryzae, bovine liver Saccharomyces fragilis 유래 $\beta$-galactosidase의 반응 조건)

  • 윤재경;이영재;구본웅;윤상영;유창수;김하영
    • YAKHAK HOEJI
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    • v.44 no.2
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    • pp.197-203
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    • 2000
  • The enzymatic properties of $\beta$-galactosidases from Aspergillus oryzae, bovine liver and Saccharomyces pragilis have been studied using enzyme-linked lectin assay based on the RC $A_{120}$ and BS-II lectins which specifically bind to terminal galactose and GlcNAc residue, respectively. Asialofetuin, a monomeric glycoprotein with approximately 48 kDa in molecular weight, was used as a substrate. This glycoprotein contains three N-linked triantennary complex type carbohydrate chains with each of which terminating in Ga1$\beta$P1 longrightarrow4G1cNAc (74%). Their optimal pHs were 3.5 and 6.5 (A. oryzae), and 3.5~5.5 (bovine liver and S. fragilis) at 37$^{\circ}C$ during 24 hrs, and the effective concentrations were 0.9, 2.9, and 1.7 mg/ml, respectively The enzyme from A oryzae requires 100 mM N $a^{+}$ or $K^{+}$, while the enzyme from bovine liver requires $Ba^{2+}$ for activity. However all of the three $\beta$-galactosidases were inactivated by SDS and C $u^{2+}$. These results indicate that the hydrolysis of glycoprotein such as asialofetuin depends on the reaction conditions of $\beta$-galactosidases and some metal ions. ions.

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Kinetic Study on Michael-type Reactions of 1-Phenyl-2-propyn-1-one with Alicyclic Secondary Amines: Effect of Medium on Reactivity and Mechanism

  • Hwang, So-Jeong;Park, Youn-Min;Um, Ik-Hwan
    • Bulletin of the Korean Chemical Society
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    • v.29 no.10
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    • pp.1911-1914
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    • 2008
  • Second-order rate constants (kN) have been measured for Michael-type addition reactions of a series of alicyclic secondary amines to 1-phenyl-2-propyn-1-one (2) in MeCN at 25.0 ${\pm}$ 0.1 ${^{\circ}C}$. All the amines studied are less reactive in MeCN than in $H_2O$ although they are more basic in the aprotic solvent by 7-9 p$K_a$ units. The Bronsted-type plot is linear with $\beta_{nuc}$ = 0.40, which is slightly larger than that reported previously for the corresponding reactions in $H_2O$ ($\beta_{nuc}$ = 0.27). Product analysis has shown that only E-isomer is produced. Kinetic isotope effect is absent for the reactions of 2 with morpholine and deuterated morpholine (i.e., $k^H/k^D$ = 1.0). Thus, the reaction has been concluded to proceed through a stepwise mechanism, in which proton transfer occurs after the rate-determining step. The reaction has been suggested to proceed through a tighter transition state in MeCN than in H2O on the basis of the larger $\beta_{nuc}$ in the aprotic solvent. The nature of the transition state has been proposed to be responsible for the decreased reactivity in the aprotic solvent.

Comparision of Some Structural Characteristics of Japonica and Tongil Rice Starches and their Fractions (일반계와 통일계 쌀 전분과 분획의 몇가지 구조적 성질 비교)

  • Kim, Kwan;Kang, Kil-Jin;Kim, Sung-Kon
    • Korean Journal of Food Science and Technology
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    • v.24 no.2
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    • pp.187-192
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    • 1992
  • Some structural characteristics of Japonica and Tongil type rice starches and their fractions were investigated to elucidate the differences between the two rice types. Japonica rice starches showed broader gelatinization temperature ranges and had higher ${\lambda}_{max}$ than those of Tongil ones. Intrinsic viscosity of starch, amylose and amylopectin for Japonica type were lower compared with those of Tongil type. ${\beta}-amylolysis$ limit of rice starch were not different between two rice types, but those of amy_lose and amylopectin of Japonica ty_p_e were lower than Tongil type. The average unit chain length(${\overline}CL$) and average outer chain length(${\overlne}OCL$) of Japonica type amylopectin were shorter than those of Tongil one. Sepharose CL-2B chromatography of hot water soluble starch revealed that each starch had unique elution profile.

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On Possible Application of Microorganism for Chemical Pulping (화학(化學)펄프 제조(製造)에 미생물(微生物)의 응용(應用) 가능성(可能性))

  • Lee, Seon-Ho;Yoon, Byong-Ho;Lee, Won-Yong
    • Journal of Forest and Environmental Science
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    • v.13 no.1
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    • pp.143-152
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    • 1997
  • The decayed wood by Fomes pini (Thore) Lloyd required a smaller H factor than the sound wood for pulping to permanganate number 20. The H factors for the decayed wood pulping by the kraft and soda processes were reduced by 15% and 17%, respectively, in the presence of 1% anthraquinone. The wood components degraded by fungi are normally more readily solubilized in alkali than the corresponding components in sound wood. The nonphenolic ${\beta}$-O-4 type lignin model compound, veratrylglycerol-${\beta}$-guaiacyl ether(I), and phenolic model compound, syringylglycerol-${\beta}$-syringyl ether(III), were degraded by the white-rot fungi to yield ${\alpha}$-guaiacoxy-${\beta}$-hydroxypropioveratrone(II) from the former and ${\alpha}$-syringyloxy-${\beta}$-hydroxypropiosyringone(IV) from the latter. Structures of the degradation products indicated that C ${\alpha}$-oxidation could occur with white-rot fungi. It has been shown that the alkaline cleavage of ${\beta}$-aryl ether bonds in the lignin units is accelerated by the presence of ${\alpha}$-carbonyl groups.

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${\beta}-Glucan$ Enrichment from Pearled Barley and Milled Barley Fractions (보리의 도정 및 제분분획을 이용한 ${\beta}-Glucan$의 강화)

  • Lee, Young-Tack;Seog, Ho-Moon;Cho, Mi-Kyung
    • Korean Journal of Food Science and Technology
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    • v.29 no.5
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    • pp.888-894
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    • 1997
  • Two hulled and two hull-less barley varieties were investigated for ${\beta}-glucan$ enrichment. Hull-less barleys contained higher levels of total ${\beta}-glucan$ than hulled barleys, and were thus suitable as starting materials for preparing ${\beta}-glucan-rich$ fractions. Particularly, a waxy type (Suweon-291) of hull-less barley was found to have high soluble dietary fiber content containing primarily ${\beta}-glucan$, compared to the other non-waxy barley varieties. ${\beta}-Glucan$ content of barley during pearling process was measured, and the highest value was observed at the pearling yield of approximately $70{\sim}75%$. The pearled barley grains were ground and sieved to yield ${\beta}-Glucan$ enriched fractions containing up to 22% ${\beta}-glucan$. In the meanwhile, whole barley samples were directly milled by $B{\ddot{u}}hler$ mill to produce bran, shorts, break flour and reduction flour. ${\beta}-Glucan$ contents in the bran and shorts from the milled stream were relatively high, and further concentration of ${\beta}-glucan$ could be accomplished by successive sieving of the bran and shorts fractions. Pearled barley and milled stream could be used to prepare barley fractions with ${\beta}-glucan$ concentrations $2.4{\sim}3.1$ times those of the original barley grain. Water solubility of barley ${\beta}-glucan$ from pearled barley and the milled stream was in the range of $40{\sim}81%$.

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Screening of Hemicellulose Oligosaccharides and Preparation of the Recipe for Modified MRS Medium by the Replacement of Carbon Source (Hemicellulose계열 올리고당 탐색 및 탄소원 대체에 의한 장내세균 생육활성용 신규 MRS배지의 조제)

  • Lee, Hee-Jung;Park, Gwi-Gun
    • Journal of Applied Biological Chemistry
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    • v.51 no.6
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    • pp.272-276
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    • 2008
  • Purification and some properties of Xylogone sphaerospora ${\beta}$-mannanase were reprevious previous paper. Locust bean gum galactomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 4 and 6 galactosyl mannooligosaccharides. For elucidate the structure of D.P 4 and 6 galactosyl mannooligosaccharides, sequential enzymatic action was performed. D.P 4 and 6 were identified as ${Gal^2}{Man_3}\;(6^2-mono-O-{\alpha}-D-galactopyranosyl-4-O-{\beta}-D-mannotriose)$ and ${Gal^2}{Man_5}\;(6^2-mono-O-{\alpha}-D-galacto- pyranosyl-4-O-{\beta}-D-mannopentaose)$. To investigate the effects of locust bean gum galactosyl mannooligosaccharides on in vitro growth of Bifidobacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 4 and D.P. 6 galactosyl mannooligosaccharides, respectively. B. longum and B. bifidum grew up to-fold and 6.6-fold more effectively by the treatment of D.P. 6 galactosyl mannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 6 was more effective than D.P. 4 galactosyl mannooligosaccharide on the growth of Bifidobacterium spp.

Purification of Xylogone sphaerospora ${\beta}$-mannanase and Growth Activity of Bifidobacterium spp. by Konjac Glucomannan Hydrolysates (Xylogone sphaerospora 유래 ${\beta}$-mannanase 정제 및 Konjac Glucomannan 가수분해 올리고당의 중합도별 Bifidobacterium spp.에 대한 증식활성)

  • Lee, Hee-Jung;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.159-163
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    • 2008
  • Xylogone sphaerospora ${\beta}$-mannanase was purified by Sephadex G-100 column chromatography. The specific activity of the purified enzyme was 8.44 units/ml protein, representing an 56.27-folds purification of the original crude extract. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 42kDa. Konjac glucomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 3 and 4 glucomannooligosaccharides. For elucidate the structure of D.P 3 and 4 glucomannooligosaccharides, sequential enzymatic action was performed. D.P 3 and 4 were identified as M-G-M and M-M-G-M (G- and M- represent glucosidic and mannosidic link-ages). To investigate the effects of konjac glucomannooligosaccharides on in vitro growth of Bifido-bacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 3 and D.P. 4 glucomannooligosaccharides, respectively. B. longum and B. bifidum grew up 3.9-fold and 2.8-fold more effectively by the treatment of D.P. 4 glucomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 4 was more effective than D.P. 3 glucomannooligosaccharide on the growth of Bifidobacterium spp.

Inherent Strength Anisotropy of the Shale in Daegu Region (대구지역 셰일 압축강도의 고유이방성에 관한 연구)

  • Lee, Younghuy;Kim, Heedong
    • Journal of the Korean GEO-environmental Society
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    • v.9 no.7
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    • pp.45-51
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    • 2008
  • Triaxial compression tests on anisotropic rock specimens are carried out to investigate the failure strength characteristic of anisotropic rocks. The test core specimens were obtained in Daegu region. Test specimens are rock cores with the 7 different angles of bedding plane. The applied confining pressures were 5, 10, 20, 30, 40 MPa, and the rate of displacement was adopted 0.1%/min to fail the specimen within 5-15 min. The results were analyzed by using the failure criteria for anisotropic rocks proposed by Hoek & Brown (1980) and Jaeger (1960). The results of this study are summerised as follows: The results of inherent anisotropy show the shoulder type of anisotropy, and the effect of anisotropy is reduced as the confining pressure increases. The compressive strength of anisotropic rock shows the highest value at the ${\beta}$ (the angle of bedding plane) = $0^{\circ}$ and $90^{\circ}$ and the lowest value at $30^{\circ}$. The Hoek & Brown failure criterion for anisotropic rocks gives a relatively good agreement with the measured strength in all the range of ${\beta}$ angles, but the theory of Jaeger shows a reasonable agreement only in the range of ${\beta}=15^{\circ}$ and $45^{\circ}$.

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Identification of Dammarane-type Triterpenoid Saponins from the Root of Panax ginseng

  • Lee, Dong Gu;Lee, Jaemin;Yang, Sanghoon;Kim, Kyung-Tack;Lee, Sanghyun
    • Natural Product Sciences
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    • v.21 no.2
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    • pp.111-121
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    • 2015
  • The root of Panax ginseng, is a Korea traditional medicine, which is used in both raw and processed forms due to their different pharmacological activities. As part of a continued chemical investigation of ginseng, the focus of this research is on the isolation and identification of compounds from Panax ginseng root by open column chromatography, medium pressure liquid chromatography, semi-preparative-high performance liquid chromatography, Fast atom bombardment mass spectrometric, and nuclear magnetic resonance. Dammarane-type triterpenoid saponins were isolated from Panax ginseng root by open column chromatography, medium pressure liquid chromatography, and semi-preparative-high performance liquid chromatography. Their structures were identified as protopanaxadiol ginsenosides [gypenoside-V (1), ginsenosides-Rb1 (2), -Rb2 (3), -Rb3 (4), -Rc (5), and -Rd (6)], protopanaxatriol ginsenosides [20(S)-notoginsenoside-R2 (7), notoginsenoside-Rt (8), 20(S)-O-glucoginsenoside-Rf (9), 6-O-[$\alpha$-L-rhamnopyranosyl(1$\rightarrow$2-$\beta$-D-glucopyranosyl]-20-O-$\beta$-D-glucopyranosyl-$3\beta$,$12\beta$, 20(S)-dihydroxy-dammar-25-en-24-one (10), majoroside-F6 (11), pseudoginsenoside-Rt3 (12), ginsenosides-Re (13), -Re5 (14), -Rf (15), -Rg1 (16), -Rg2 (17), and -Rh1 (18), and vinaginsenoside-R15 (19)], and oleanene ginsenosides [calenduloside-B (20) and ginsenoside-Ro (21)] through the interpretation of spectroscopic analysis. The configuration of the sugar linkages in each saponin was established on the basic of chemical and spectroscopic data. Among them, compounds 1, 8, 10, 11, 12, 19, and 20 were isolated for the first time from P. ginseng root.