• Title/Summary/Keyword: a-amylase activity

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Raw Starch Degrading Amylase Production by Various Fungal Cultures Grown on Cassava Waste

  • Pothiraj, C.;Balaji, P.;Eyini, M.
    • Mycobiology
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    • v.34 no.3
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    • pp.128-130
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    • 2006
  • The solid waste of sago industry using cassava was fermented by Aspergillus niger, Aspergillus terreus and Rhizopus stolonifer in solid state fermentation. Cassava waste contained 52 per cent starch and 2.9 per cent protein by dry weight. The amylase activity was maintained at a high level and the highest amylase activity was observed on the $8^{th}$ day in R. stolonifer mediated fermentation. R. stolonifer was more efficient than Aspergillus niger and Aspergillus terreus in bioconverting cassava waste into fungal protein (90.24 mg/g) by saccharifying 70% starch and releasing 44.5% reducing sugars in eight days of solid state fermentation.

IRRADIATION EFFECT ON SECRETING FUNCTION, AMYLASE ACTIVITY AND NUCLEIC ACID CONTENTS OF RAT PAROTID GLAND (방사선 조사가 이하선 기능에 미치는 영향에 관한 연구)

  • Cho Yong Jin;Park Tae-Won
    • Journal of Korean Academy of Oral and Maxillofacial Radiology
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    • v.20 no.1
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    • pp.53-62
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    • 1990
  • This experiment was performed to clarify the effects of /sup 60/Co gamma irradiation on secretory function, amylase activity and contents of nucleic acids of parotid gland in rat. Experimental animals were divided into 6th hours, 3rd, 7th, 14th and 28th days after irradiation and control. The experimental animals are singly irradiated with 20Gy (2,000rad) through protective lead block. Secretory function of parotid gland was evaluted by uptake and clearance of /sup 99m/TcO₄. /sup 99m/TcO₄. 0.2μ ci/gm, was injected into peritonium in uptake groups. Rats were sacrified with cervical dislocation after 30 minutes and gland was excised. In the clearance group. pilocarpine nitrate (8㎎/㎏) was intraperitoneally injected at 30 minutes after /sup 99m/TcO₄ injection and rats were sacrified 30 minutes after pilocarpine injection. Radioactivity of excised parotid gland was measured by using of gamma counter and stimulation-secretion coefficients, uptake radioactivity divided by clearance radioactivity, was calculated. Amylase activity and contents of DNA and RNA were determined by spectrophotometry. The results obtained were as follows: 1. In the uptake test, the radioactivity of /sup 99m/TcO₄ per unit weight increase in experimental group except 6th hours group, compared with control groups and showed a peak at 3rd days after irradiation. 2. In the clearance test, the radioactivity of /sup 99m/cO₄per unit weight rose to a peak at 3rd days after irradiation and gradually recovered thereafter. 3. Stimulation-secretion coefficient of parotid gland decreased at 6th hours, 3rd and 7th days after irradiation, and gradually increased. 4. Amylase activity of parotid gland decreased in 3rd and 7th days group, and especially lowest in 3rd days after irradiation. 5. The contents of DNA showed no definite difference in all the experimental groups, but RNA was seemed to decrease with time after irradiation.

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Developmental Changes in Digestive Enzymes Activity of Black Rockfish Sebastes inermis (볼락, Sebastes inermis 자.치어의 성장에 따른 소화효소 활성 변화)

  • Gwak Woo-Seok;Park Dae-Won
    • Journal of Aquaculture
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    • v.19 no.2
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    • pp.125-132
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    • 2006
  • Black rockfish larvae and juveniles were reared for 95 days after parturition (DAP) in order to determine four enzyme activities (trypsin, pepsine-like enzyme, lipase, amylase) during ontogeny. Larvae were fed rotifers Brachionus plicatilis from 1 to 25 DAP, Artemia nauplii from 10 to 78 DAP and then gradually changed to pelleted feed from 30 DAP. Temperature was kept between $13.5{\sim}14.9^{\circ}C$. Trypsin and lipase activities were found in 2 DAP larvae ($7.0{\pm}1.5$ unit and $4.5{\pm}1.4$ unit, $mean{\pm}SD$, respectively). The evolution of both enzymes activities showed a profile marked by drastic increases between postflexion and juvenile stage. There is an increment on specific trypsin activity at 10 DAP, corresponding with the beginning of Artemia feeding. Pepsin-like enzyme activity was found at 11 DAP and increased drastically from 56 DAP, cor-responding with the initiation of juvenile stage. Amylase activity was also found at 11 DAP and maintained at a low level up to 38 DAP followed by a drastic increase from 39 DAP to 50 DAP. Considering our results of both trypsin and pepsin-like enzyme activities, it might be concluded that higher somatic growth of Sebastes inermis could be possible with the initiation juvenile of stage and the early juvenile stage is a suit-able period for feeding an artificial diet for fish.

Construction of a Shuttle Vector for Heterologous Expression of a Novel Fungal α-Amylase Gene in Aspergillus oryzae

  • Yin, Yanchen;Mao, Youzhi;Yin, Xiaolie;Gao, Bei;Wei, Dongzhi
    • Journal of Microbiology and Biotechnology
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    • v.25 no.7
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    • pp.988-998
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    • 2015
  • The filamentous fungus Aspergillus oryzae is a well-known expression host used to express homologous and heterologous proteins in a number of industrial applications. To facilitate higher yields of proteins of interest, we constructed the pAsOP vector to express heterologous proteins in A. oryzae. pAsOP carries a selectable marker, pyrG, derived from Aspergillus nidulans, and a strong promoter and a terminator of the amyB gene derived from A. oryzae. pAsOP transformed A. oryzae efficiently via the PEG-CaCl2-mediated transformation method. As proof of concept, green fluorescent protein (GFP) was successfully expressed in A. oryzae transformed by pAsOP-GFP. Additionally, we identified a novel fungal α-amylase (PcAmy) gene from Penicillium sp. and cloned the gene into the vector. After transformation by pAsOPPcAmy, the α-amylase PcAmy from Penicillium sp. was successfully expressed in a heterologous host system for the first time. The α-amylase activity in the A. oryzae transformant was increased by 62.3% compared with the untransformed A. oryzae control. The PcAmy protein produced in the system had an optimum pH of 5.0 and optimum temperature of 30oC. As a cold-adapted enzyme, PcAmy shows potential value in industrial applications because of its high catalytic activity at low temperature. Furthermore, the expression vector reported in this study provides promising utility for further scientific research and biotechnological applications.

Inactivation of Microorganisms and Enzymes in Foxtail Millet Takju by High Hydrostatic Pressure Treatment (초고압 처리에 의한 좁쌀탁주의 미생물 살균 및 효소 불활성화)

  • Jwa, Mi-Kyung;Lim, Sang-Bin;Mok, Chul-Kyoon;Park, Young-Seo
    • Korean Journal of Food Science and Technology
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    • v.33 no.2
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    • pp.226-230
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    • 2001
  • High hydrostatic pressure was applied to Foxtail Millet Takju to investigate the effects of high pressure on inactivation of microorganisms and enzymes. Total bacteria, lactic acid bacteria and yeast in untreated Takju were $6.8{\times}10^7,\;1.3{\times}10^8\;and\;8.4{\times}10^7\;CFU/mL$, respectively. Total bacterial count in Takju reduced to $2.2{\times}10^5\;CFU/mL$, while lactic acid bacteria and yeast were sterilized completely when heated at $65^{\circ}C$ for 30 min. Lactic acid bacteria and yeast decreased with the increase of treatment pressure, and pressurization of 400 MPa for 10 min at room temperature sterilized completely the lactic acid bacteria and yeast in Takju. Total bacteria were not sterilized with pressurization of even 600 MPa at room temperature. Total bacteria were completely sterilized at $66^{\circ}C/400\;MPa/60\;min\;and\;66^{\circ}C/600\;MPa/10\;min$. Pressurization of Takju caused a partial inactivation of ${\alpha}-amylase$, and after pressurization at 600 MPa for 10 min at room temperature, 73.2% of the original activity remained. The activity of glucoamylase increased with the increase of treatment pressure. Treatment at $66^{\circ}C/400\;MPa/10\;min$ reduced the activity of ${\alpha}-amylase$ by 59.7% and glucoamylase by 20.5%. ${\alpha}-Amylase$ was inactivated to less than 1.2% of the original activity at $66^{\circ}C/600\;MPa/30\;min$.

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Epigallocatechin 3-gallate Binds to Human Salivary α-Amylase with Complex Hydrogen Bonding Interactions

  • Lee, Jee-Young;Jeong, Ki-Woong;Kim, Yang-Mee
    • Bulletin of the Korean Chemical Society
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    • v.32 no.7
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    • pp.2222-2226
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    • 2011
  • Amylase is a digestive enzyme that catalyses the starch into sugar. It has been reported that the green tea flavonoid (or polyphenols) (-)-epigallocatechin 3-gallate (EGCG) inhibits human salivary ${\alpha}$-amylase (HSA) and induced anti-nutritional effects. In this study, we performed docking study for seven EGCG-like flavonoids and HSA to understand the interaction mechanism of HSA and EGCG and suggest new possible flavonoid inhibitors of HSA. As a result, EGCG and (-)-epicatechin gallate (ECG) bind to HSA with complex hydrogen bonding interactions. These hydrogen bonding interactions are important for inhibitory activity of EGCG against HSA. We suggested that ECG can be a potent inhibitor of HSA. This study will be helpful to understand the mechanism of inhibition of HSA by EGCG and give insights to develop therapeutic strategies against diabetes.

Heterologous Expression of ${\alpha}$-Amylase Gene of Bifidobacterium adolescentis Int57 in Bacillus polyfermenticus SCD

  • Paik, Hyun-Dong;Kim, Il-Gi;Lee, Jin-Hyoung;Lee, Jang-Hyun;Park, Kyu-Yong;Ji, Geun-Eog;Jin, Tae-Eun;Rhim, Seong-Lyul
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.655-658
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    • 2007
  • Bacillus polyfermenticus SCD was transformed by the recombinant shuttle vector for Bacillus and Escherichia coli containing 3 antibiotic resistant genes and an ${\alpha}$-amylase gene from Bifidobacterium adolescentis Int57. The ${\alpha}$-amylase gene fused to a secretion sequences was expressed under the control of the promoter of amylase gene from B. subtilis var. natto. The recombinant plasmid was maintained stably in the transformants producing the ${\alpha}$-amylase. The enzyme was secreted to outside of the cell and showed the similar enzyme activity as that of Bacillus subtilis BD170 under the same conditions of pH and growth temperature. Because of the relatively easy transformation and the secretion of the enzyme, the transformants of B. polyfermenticus SCD may give a new strategy in the production of foreign genes.

Production of Maltopentaose and Biochemical Characterization of Maltopentaose-Forming Amylase

  • Kim, Young-Min;Ryu, Hwa-Ja;Lee, Sun-Ok;Seo, Eun-Seong;Lee, So-Young;Yoo, Sun-Kyun;Cho, Dong-Lyun;Kim, Do-Man;Kimura, Atsuo;Chiba, Seiya;Lee, Jin-Ha
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.636-643
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    • 2001
  • Bacillus sp. AIR-5, a strain from soil, produced an extracellular maltopentaose-forming amylase from amylose and soluble starch. This bacterium produced 8.9 g/l of maltopentaose from 40 g/l of soluble starch in a batch fermentation and the maltopentaose made up 90 % of the maltooligosaccharides produced (from maltose to maltoheptaose). The culture supernatant was concentrated using a 30 K molecular weight cut-off membrane and purified by DEAE-Cellulose and Sephadex G-150 column chromatographies. The purified protein showed one band on a native-PAGE and its molecular mass was estimated as 250 kDa. The 250-kDa protein was composed of tetramers of a 63-kDa protein. the isoelectric point of the purified protein was pH 6.9, and the optimum temperature for the enzyme activity was $45^{\circ}C$. The enzyme was quickly inactivated above $55^{\circ}C$, and showed a maximum activity at pH 8.5 and over 90% stability between a pH of 6 to 10. The putative N-terminal amino acid sequence of AIR-5 amylase, ATINNGTLMQYFEWYVPNDG, showed a 96% sequence similarity with that of BLA, a general liquefying amylase.

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Optimization of Rice (Oryza Sativa) Malting Process by Second-Order Experimental Design

  • Nguyen, Thach Minh;Nguyen, Xich Lien;Hoang, Kim Anh;Lee, Soo
    • Journal of the Korean Applied Science and Technology
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    • v.25 no.3
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    • pp.282-290
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    • 2008
  • The malting process of rice (OM4080 variety from Mekong Delta Rice Research Institute) was studied under pilot condition plan by means of the second-order experimental design. Processing parameters, such as the steeping time (0-60 hrs), steeping temperature ($5-45^{\circ}C$), germination time (0-8 days), germination temperature ($5-45^{\circ}C$) and gibberellin concentration (0-2 mg/kg) were investigated. As a result, all germination conditions, especially germination time, germination temperature, and gibberellin concentration had a significant effect on the malting loss, amylase activity and starch content. The protein content was not clearly affected by any conditions. The optimum conditions for malting process (with highest amylase activity) were as follows: 30 hrs of steeping time, $30-35^{\circ}C$ of steeping temperature, 5-5.5 days of germination time, $25^{\circ}C$ of germination temperature, and 1.5 mg/kg of giberrellin concentration.

Physiological Characteristics and Anti-Obesity Effect of Lactobacillus plantarum K6 isolated from Kimchi (김치에서 분리한 Lactobacillus plantarum K6의 생리적 특성 및 비만억제효과)

  • Kim, Seulki;Lim, Sang-Dong
    • Journal of Dairy Science and Biotechnology
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    • v.35 no.4
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    • pp.221-231
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    • 2017
  • This study aimed to investigate the physiological characteristics and anti-obesity effects of a newly isolated bacterium, Lactobacillus plantarum K6. L. plantarum K6 showed good ${\alpha}-amylase$ inhibitory activity ($96.78{\pm}3.29%$), ${\alpha}-glucosidase$ inhibitory activity ($92.55{\pm}9.62%$), and lipase inhibitory activity ($85.17{\pm}0.79%$), and the strain inhibited the adipocyte differentiation of 3T3-L1 cells ($27.4{\pm}1.4%$) when present at a concentration of $100{\mu}g/mL$. L. plantarum K6 was isolated from kimchi and its physiological characteristics were investigated. A comparison of the sensitivity of the isolate to 15 different antibiotics showed that L. plantarum K6 is highly sensitive to erythromycin and highly resistant to vancomycin, ampicillin, and polymyxin B. This strain also showed high arylamidase and ${\beta}-galactosidase$ activities. Moreover, it was relatively tolerant to bile acid and low pH, and displayed resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus, with rates of 51.8%, 42.4%, 61.6%, and 54.9%, respectively. No bio genic amines were produced. L. plantarum K6 also showed high adhesion activity to HT-29 cells compared to L. rhamnosus GG. These results demonstrate that Lactobacillus plantarum K6 has potential as a probiotic with anti-obesity effects.