• Title/Summary/Keyword: a-glucoamylase

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Effect of Rice Pre-treatment on Enzymatic Saccharification in the Brewing Process (주류 제조를 위한 효소 당화에 쌀의 전처리가 미치는 영향)

  • An, Jin-Ok;Chung, Chang-Ho;Lee, Seung-Joo
    • Microbiology and Biotechnology Letters
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    • v.45 no.4
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    • pp.277-283
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    • 2017
  • To produce sweet liquor without artificial sweeteners, 8 traditional rice pre-treatment methods (juk, beombeok, seolgitteok, gumeongtteok, mulsongpyeon, injeolmi, gaetteok, and godubap) were analyzed in this study. The formation of sugars with the help of ${\alpha}$-amylase, ${\beta}$-amylase, and glucoamylase using nuruk as a substrate has been previously confirmed. During the early stages of the pre-treatment processes, the amount of maltose produced (in descending order of its concentration) by ${\alpha}$-amylase was observed to be as follows: gaetteok > seolgitteok > beombeok > mulsongpyeon > juk > injeolmi > gumeongtteok > godubap. However, changes in maltose concentrations with respect to the pre-treatment processes after 48 hours were observed to be as follows: injeolmi > beombeok = godubap > gumeongtteok > gaetteok = mulsongpyeon > seolgitteok > juk. Maltose produced using either ${\alpha}$-amylase or ${\beta}$-amylase showed similar results. Glucoamylase produced 10 mg/ml of glucose during the godubap process, which was the highest amount of glucose among all the methods. Moreover, when ${\alpha}$-amylase, ${\beta}$-amylase, and glucoamylase were used concurrently, glucoamylase increased glucose production in the later stages. Therefore, the possibility of producing sweet liquor without employing artificial sweeteners was confirmed, even if the amount of sugar in the liquor varied with the pre-treatment process.

Characterization of the Starch Degradation Activity of recombinant glucoamylase from Extremophile Deinococcus geothermalis (극한성 미생물Deinococcus geothermalis 유래 재조합 글루코아밀레이즈의 전분 분해 활성 특징)

  • Jang, Seung-Won;Kwon, Deok-Ho;Park, Jae-Bum;Jung, Jong-Hyun;Ha, Suk-Jin
    • Journal of Industrial Technology
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    • v.39 no.1
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    • pp.15-19
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    • 2019
  • This work focused on characterization of the starch degradation activity from extremophile strain Deinococcus geothermalis. Glucoamylase gene from D. geothermalis was cloned and overexpressed by pET-21a vector using E. coli BL21 (DE3). In order to characterize starch degrading activity of recombinant glucoamylase, enzyme was purified using HisPur Ni-NTA column. The recombinant glucoamylase from D. geothermalis exhibited the optimum temperature as $45^{\circ}C$ for starch degradation activity. And highly acido-stable starch degrading activity was shown at pH 2. For further optimization of starch degrading activity with metal ion, various metal ions ($AgCl_2$, $HgCl_2$, $MnSO_4{\cdot}4H_2O$, $CoCl_2{\cdot}6H_2O$, $MgSO_4$, $ZnSO_4{\cdot}7H_2O$, $K_2SO_4$, $FeCl_2{\cdot}4H_2O$, NaCl, or $CuSO_4$) were added for enzyme reaction. As results, it was found that $FeCl_2{\cdot}4H_2O$ or $MnSO_4{\cdot}4H_2O$ addition resulted in 17% and 9% improved starch degrading activity, respectively. The recombinant glucoamylase from D. geothermalis might be used for simultaneous saccharification and fermentation (SSF) process at high acidic conditions.

Effect of Aeration and Agitation Conditions on the Production of Glucoamylase with Aspergillus niger No. PFST-38

  • Oh, Sung-Hoon;O, Pyong-Su;Lee, Cherl-Ho
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.292-297
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    • 1993
  • Aspergillus niger No. PFST-38 was grown on complex media in 30L agitated fermentors at various aeration rates and stirrer speeds. We could correlate the mixing time as a function of the Reynolds number and the apparent viscosity, as follows. ${\theta}_M=2.95\;\NRe^{-0.52},\;{\theta}_M=1.88\;{\eta_a}^{0.57}$ Also, the effects of the apparent viscosity (${\theta}_a$), the impeller rotational speed (N), the air flow rate ($V_s$), and the mixing time (${\theta}_M$) on the oxygen transfer coefficient, $K_L a$ were determined experimentally, and equated as follows. $K_La=12.04N^{0.88}Vs^{0.71}{n_a}^{-0.83},\;K_La=30.2N^{0.88}Vs^{0.71}{\theta_M}^{-1.45}$ $K_La$ increased as the agitation speed and the air flow rate increased. The rate of $K_La$ increase was dependent more on the rotational speed of impeller than on the air flow rate. The glucoamylase production increased with the increase of the agitation speed upto at 500 rpm and increased with the increase of air flow rate upto at 1.0 vvm. The values calculated from the above equation confirmed that the experimental maximum production of glucoamylase was achieved when the $K_La$ and the apparent viscosity of the broth were $260\;hr^{-1}$ and 1800 cps, respectively.

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Ethanol Fermentation of Corn Starch by a Recombinant Saccharomyces cerevisiae Having Glucoamylase and $\alpha$-Amylase Activities

  • Lee, Dae-Hee;Park, Jong-Soo;Ha, Jung-Uk;Lee, Seung-Cheol;Hwang, Yong-Il
    • Preventive Nutrition and Food Science
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    • v.6 no.4
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    • pp.206-210
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    • 2001
  • Starch is an abundant resource in plant biomass, and it should be hydrolyzed enzymatically into fermentable sugars for ethanol fermentation. A genetic recombinant yeast, Saccharomyces cerevisiae GA-7458, was constructed by integrating the structural gene of both $\alpha$-amylase from Bacillus stearothermophilus and the gene (STA1) encoding glucoamylase from S. diastaticus into the chromosome of S. cerevisiae SH7458. The recombinant yeast showed active enzymatic activities of $\alpha$-amylase and glucoamylase. The productivity of ethanol fermentation from the pH-controlled batch culture (pH 5.5) was 2.6 times greater than that of the pH-uncontrolled batch culture. Moreover, in a fed-batch culture, more ethanol was produced (13.2 g/L), and the production yield was 0.38 with 2% of corn starch. Importantly, the integrated plasmids were fully maintained during ethanol fermentation.

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Studies on the Production of Acid Digestive Enzyme -Isolation and Characterization of a Fungal Strain Which Produces Acid Enzymes- (내산성(耐酸性) 소화효소제(消化酵素劑)의 생산(生産)에 관(關)한 연구(硏究) -내산성(耐酸性) 효소생산균(酵素生産菌)의 분리(分離)와 효소(酵素) 생산조건(生産條件)에 관(關)하여-)

  • Sohn, Cheon-Bae;Park, Yoon-Joong
    • Korean Journal of Food Science and Technology
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    • v.13 no.3
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    • pp.241-246
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    • 1981
  • A fungal strain which produced high levels of acid protease and amylase was isolated from the atmosphere for application to the manufacture of digestive enzme preparation. This study was carried out to elucidate its microbiological characteristics, environmental conditions for production of the enzymes, and relationships between the enzyme activity and acidity. 1. The isolate was identified as a fungal strain which belonged to Aspergillus niger by the manual of Rafer and Fennel, and was found to be a strain producing high levels of acid protease and amylase. 2. The optimal pH of tile enzymes produced by the strain were: protease, 2.0;, ${\alpha}-amylase$, 4 to 5; and glucoamylase, 3 to 5. 3. The optimal culture conditions for production of the enzymes were: protease (at pH 2.5), 2 to 3 days incubation on wheat bran at $30^{\circ}C$; ${\alpha}-amylase$ and glucoamylase(at pH 3.0), 3 days incubation at $30^{\circ}C$. 4. The production of acid protease and glucoamylase was increased approximately by 20 percent when 2 percent of corn starch was added to the wheat bran medium. 5. The addition of 0.3 percent ammonium sulfate to the wheat bran medium resulted in enhancing the enzyme production, especially of acid prctease.

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Purification and Characteristics of Glucoamylase in Aspergillus oryzae NR 3-6 Isolated from Traditional Korean Nuruk

  • Yu, Tae-Shick;Kim, Tae-Hyoung;Joo, Chong-Yoon
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.80-85
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    • 1999
  • The purification system of glucoamylase (glucan 1,4-${\alpha}$-glucosidase, EC 3. 2. 1. 3), some characteristics of the purified enzyme and hydrolysis rate of various raw starch were investigated through several experiments. The enzyme was produced on a solid, uncooked wheat bran medium of Aspergillus oryzae NR 3-6 isolated from traditional Korean Nuruk. The enzyme was homogeneously purified 6.8-fold with an overall yield of 28.3% by the criteria of disc- and SDS-polyacrylamide gel electrophoresis. The molecular weight was estimated to be 48 kDa by SDS-PAGE. The optimum temperature and pH were 55$^{\circ}C$ and 4.0, respectively. The enzyme was stable at a pH range of 3.0∼10.0 and below 45$^{\circ}C$. Enzyme activity was inhibited about 27% by 1mM Hg2+. The hydrolysis rate of raw wheat starch was shown to be 17.5-fold faster than the hydrolysis rate of soluble starch. The purified enzyme was identified as glucoamylase because the product of soluble starch by the purified enzyme was mainly glucose by thin layer chromatography.

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Chemical Components, Antioxidant Activity, and α-Glucoamylase Inhibitory Activity of a New Mushroom Variety 'Dahyang' (신품종 갈색양송이 '다향'의 화학성분과 항산화활성 및 α-Glucoamylase 저해활성)

  • Kim, Hong-Kyu;Yang, Euy-Seog;Park, Gi-Moon;Kim, Gwan-Hou;Kim, Hyun-Ho;Lee, Ka-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.8
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    • pp.1179-1183
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    • 2011
  • This study was conducted to investigate the quality characteristics, antioxidant activity, and ${\alpha}$-glucoamylase inhibitory activity of Dahyang, a Chungnam Agricultural Research & Extension Service's newly bred cultivar of brown button mushroom. Total phenolic compound contents of Dahyang and the no. 705 mushroom were 189${\pm}$12 mg% and 168${\pm}$8 mg%, respectively. The major free sugars in Dahyang were mannitol (3.11%), xylose (0.12%), and trehalose (0.08%). ${\beta}$-Glucan content was 28.34% in Dahyang and 26.55% in the no. 705 mushroom, respectively. Electron donating ability by DPPH in Dahyang and the no. 705 mushroom was 52.14% and 45.27% for the water extract, and 57.81% and 46.93% for the 80% ethanol extract, respectively. ${\alpha}$-Glucoamylase inhibitory activity in a 10 mg/mL concentration of water extract were was 33.25% in Dahyang and 29.22% in the no. 705 mushroom, respectively.

Construction of a Transformed Yeast Strain Secreting Both $\alpha$-Amylase and Glucoamylase for Direct Starch-Fermentation

  • Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • v.4 no.1
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    • pp.7-12
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    • 1994
  • A yeast strain secreting glucoamylase was transformed with an expression vector (pMS12) containing the promoter of yeast alcohol dehydrogenase I gene ADC1, mouse salivary $\alpha$-amylase cDNA, and a segment of yeast $21\mu m$ plasmid. The transformed strain could produce ethanol from starch (4%, w/v) through a direct one-step process with the conversion efficiency of 93.2%, during 5 days of fermentation, while the original, untransformed strain exhibited a conversion efficiency of 38.1% under the same condition. When the regulatory site of the ADC1 promoter region was removed, the production of ethanol increased to 29~37% in the presence of exogenous 3%(v/v) ethanol in the fermentation medium.

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Yeast Cloning Vectors and their Application to the Development of Starch-fermenting Yeast (효모 Cloning Vector와 전분발효 효모의 개발)

  • Kim, Keun
    • Applied Biological Chemistry
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    • v.31 no.3
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    • pp.267-273
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    • 1988
  • Transformed, hybrid strains of the yeast Saccharomyces capable of simultaneous secretion of both glucoamylase and ${\alpha}-amylase$ have been produced. These strains can carry out direct, one-step assimilation of starch with conversion efficiency greater than 93% during a 5 day growth period. One of the transformants converts 92.8% of available starch into reducing sugars in only 2 days. Glucoamylase secretion by these strains results from expression of one or more chromosomal STA genes derived from Saccharomyces diastaticus. The strains were transformed by a plasmid(pMS12) containing mouse salivary ${\alpha}-amylase$ cDNA in an expression vector containing yeast alcohol dehydrogenase promoter and a segment of yeast $2{\mu}$ plasmid. The major starch hydrolysis product produced by crude amylases found in culture broths is glucose, indicating that ${\alpha}-amylase$ and glucoamylase act cooperatively.

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Rheological Properties of Mycelial Broth in Submerged Culture of Aspergillus niger No. PFST-38

  • Oh, Sung-Hoon;O, Pyong-Su
    • Journal of Microbiology and Biotechnology
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    • v.3 no.3
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    • pp.209-213
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    • 1993
  • The flow behavior of the mycelial broth of glucoamylase hyperproducer Asp. niger No. PFST-38 for the production of glucoamylase were studied. The mycelial broth followed Bingham-pseudoplastic flow model described by Herschel-Bulkley equation. The yield stress increased with the increase in mycelial concentration. The dependency of the consistency index and the flow behavior index on the mycelial concentration could be expressed by a linear relationship. The consistency index increased proportionally with the mycelial concentration while the flow behavior index decreased with the increase in mycelial concentration. The flow property of the broth was related to the morphological data obtain in the previous study. The changes in apparent viscosity of the broth could be expressed as a function of the hyphal thickness as shown below.

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