• Title/Summary/Keyword: Alpha-Amylase

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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.

Changes of α-Amylase Activity and Free Proline Content Under Low Temperature During Germination of Rice (볍씨 저온발아중 α-amylase 활성과 유리 Proline 함량 변화)

  • Kim, Sang-Kuk;Kim, Young-Jong;Won, Jong-Gun;Lee, Sang-Chul
    • Korean Journal of Soil Science and Fertilizer
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    • v.31 no.2
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    • pp.158-161
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    • 1998
  • The study was carried out to examine changes of ${\alpha}$-amylase activity and free proline contents during rice germination under low temperature($13^{\circ}C$). The plant height, root length, and germination rate were investigated during seed germination under the low temperature. Those growth chracteristics were the highest in Dongjin cultiver compared with other rice cultivars. The ${\alpha}$-amylase activity of the 9 days after germination was higher in Dongjin than those of Sangju and Koshihikari cultivars. The content of free proline was increased in all rice cultivars, when the germination of rice seeds was prolonged. As a result, it was concluded that ${\alpha}$-amylase activity and free proline content of Donjin cultivar were relatively increased higher under low temperature stress than those of other rice cultivars.

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Properties of Crude Amylase Isolated from Pine Nut (잣에 존재하는 아밀라제의 특성)

  • Kim, Jong-Sang;Seog, Ho-Moon
    • Korean Journal of Food Science and Technology
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    • v.26 no.4
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    • pp.398-402
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    • 1994
  • The participation of thermostable amylase in the decrease of viscosity of pine nut's porridge was investigated using the crude enzyme obtained from ammonium sulfate fractionation of pine nut extracts. The fraction precipitated at $35{\sim}55%$ saturation of ammonium sulfate had the highest specific activity of the enzyme. ${\alpha}-amylase$ activity was maximal at $75^{\circ}C$, pH 5.4. Amylograph data showed that addition of the enzyme to rice flour resulted in the significant decrease of its viscosity, suggesting the existence of thermostable ${\alpha}-amylase$ in pine nut.

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Studies on $\alpha$-Amylase of Bacillus circulans F-2 (Part I) Purification of $\alpha$-amylase (Bacillus circulans F-2가 생산하는 $\alpha$-Amylase에 관한 연구 (제 1보) $\alpha$-Amylase의 정제)

  • ;Hajime Taniguchi;Yoshiharu Maruyama
    • Microbiology and Biotechnology Letters
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    • v.9 no.4
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    • pp.185-190
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    • 1981
  • 1. $\alpha$-amylase from B. circulans F-2 was purified with specific activity 55.0 u/mg. protein (about 23 times of the original specific activity) and the yield of 25.5%, by means of corn starch absorption, salting out with ammonium sulfate (80% saturation), gel filtration on Bio-Gel P-100 and DE-32 column chromatography. 2 The purified enzyme showed two closely migrated protin bands on polyacrylamide disc gel electrophoresis, both of which have amylase activity judging from the activity staining of the gel. On SDS-polyacrylamide disc gel electrophoresis, however, the purified enzyme showed a single band suggesting that those two bands are the charge isomers of an amlyase having the slightly different charge. 3. Plot of log mobility of two bands versus polyacrylamide gel concentration according to Hedrick and Smith gave the parallel lines indicating them to be charge isomers. 4. To confirm the action pattern of two enzyme protein bands, each band was separated and was eluted from the gel and eluates were incubated with soluble starch. Oligosaccharide pattern produced by each eluate was examined by paper chromatography. The eluates of two bands showed the same action pattern. 5. The maltohexaose was the only hydrolysis product of soluble starch in the early stage of hydrolysis.

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$\alpha$- and $\beta$-Amylase Isozyme Expresser Native Proteins in Tropical Silkworm Bombyx mori L.

  • Chattopadhyay, G.K.;Verma, A.K.;Sengupta, A.K.;Das, S.K.;Urs, S.Raje
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.2
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    • pp.189-194
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    • 2004
  • Amylase isozyme based three multivoltine viz., N+p, Np, N+ $p^{cho}$ and two bivoltine-D6+p, D6p syngenic lines (Syn. L) were developed from germplasm (GP) stocks Nistari (N) and D6 respectively. haemolymph isozyme pattern at pH 7.0 and 8.5 depicted a total 11 number (Am $y_{1 to 6}$ at pH 7.0 and Am $y^{l to 5}$ at pH 8.5) of native proteins (NP) of various sizes are amylase isozyme expressers. Among eleven NPs, two NPs of 770 kDa (Am $y^{6}$ at pH 7.0) and 376 kDa (Am $y^3$ at pH 8.5) are $\alpha$-amylase expressers and remaining NPs of 370, 364, 350, 329 and 274 kDa at pH 7.0 and 206, 292, 416, 725 kDa at pH 8.5 are $\beta$-amylase expressers. Accordingly, digestive juice amylase isozyme pattern at aforesaid pH also depicted a total number of 10 NPs (Am $y^{1 to 5}$) at each pH 7.0 and 8.5 are amylase expressers of which NP of 387 kDa (Am $y^4$ at pH 7.0) and 780 kDa (Am $y^{5}$ at pH 8.5) are a-amylase expresser. Remaining NPs of 338,297 & 216 kDa at pH 7.0 and 370, 341, 329 &302 kDa at pH 8.5 are $\beta$-amylase expresser. Recurrent backcross lines (RBL) viz., N+pRBL and NpRBL were developed through introgression of high shell weight character (a multigenic trait) to be used further for congenic line (Con. L) development and to understand any association with introgressed character. Isozyme pattern in haemolymph of RBLs depicted only one $\alpha$-amylase of 770 kDa at pH 7.0 and 376 kDa at pH 8.0 with three and four respective $\beta$-amylase bands but in bivoltine lines numbers of $\beta$-amylase bands vary between 1 to 2 at aforesaid pH. Variability was also observed in digestive juice of multivolitine and its RBLs but bivoltine lines express null activity at both pH except appearance of one very week $\alpha$-amylase band D6+p at pH 8.5. Overall study suggests that not a single NP at both pH is common for expression of any band of amylase isozyme i.e., a totally different set of proteins are the amylase isozyme expresser at specific pH and no molecular factor of amylase is associated in developed RBLs which showed improvement on survival, single cocoon shell weight (SCSW) and single filament length over receptor parents.s.s.s.

Screening of Biological Activities in Fern Plants Native to Jeju Island (제주도에 자생하는 양치식물의 생리활성 검색)

  • Oh, Soon-Ja;Hong, Sung-Soo;Kim, Yeon-Hee;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.21 no.1
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    • pp.12-18
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    • 2008
  • Antioxidative activity and inhibitory activity of angiotensin I converting enzyme(ACE), aminopeptidase N(APN) and $\alpha$-amylase were investigated in the methanol extracts from 25 fern plants native to Jeju Island, in order to screen the plant species containing bioactive materials for functional foods or medicines. The antioxidative activity was higher in Cytomium fortunei(41.9%) and Rumohra standishii(34.1%) than in leaves of Thea sinensis(30.9%), a small tree for antioxidative beverage. Inhibitory activities of ACE and APN were relatively high in Cytomium fortunei as 26.7% and 28.2% respectively. $\alpha$-Amylase inhibitory activity was higher than 50% in 10 species. Particularly, Cytomium fortunei(87.4%) and Dryopteris erythrosora(71.6%) showed the inhibitory activities higher than those of other form plants. Of 25 fern plants investigated here, Cytomium fortunei showed not only the highest antioxidative activity but also the highest inhibitory activity of ACE, APN and $\alpha$-amylase. It suggests that Cytomium fortunei could be potentially used as a resource of bioactive materials for fuctional foods or medicines.

Effects of Salting Process on Ascorbic Acid Contents, ${\alpha}-Amylase$ Activity, Seasoning Penetration and Microbial Counts of Radish Cubes for Kakdugi (간절임이 무우 Cube의 Ascorbic Acid 함량, ${\alpha}-Amylase$ 활성, 양념류 침투성, 생균수에 미치는 영향)

  • Kim, Joong-Man;Shin, Mi-Kyung;Hwang, Ho-Sun;Kim, Hyung-Tae
    • Korean Journal of Food Science and Technology
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    • v.22 no.4
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    • pp.492-495
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    • 1990
  • In succession to the previous paper(Korean J. Food Sci. and Techn., 21(2), 300, 1989), to investigate effect of salting process on physico-chemical properties of Korean radish cubes processed for kakdugi Ascorbic acid content, ${\alpha}-amylase$ activity, the penetration rate of glucose and red-pepper juice into the radish cubes, and aerobic microbial counts (bacteria, yeast and mold) were investigated during salting (packout ratio : 1:1g/ml) of radish-cube$(2{\times}2{\times}2cm)$ for kakdugi in the 15%(w/v) sodium chloride solution. For 6hr salting, ascorbic acid was reduced from 38.5mg% to 15.7 mg%, ${\alpha}-amylase$ activity was gradually increased until 5hr salting, and then deceased, Amount and rate of penetration of glucose and red-pepper juice(on base of redness) into the salted radish cube were both higher than raw cubes, and microbial counts were decreased, and mostly within one hour of salting.

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Production and Preservation of $\alpha$-Amylase from Bacillus sp. Y-124 (Bacillus sp. Y-124로부터 $\alpha$-Amylase의 생산 및 그 보존성에 관한 연구)

  • Jun, Hong-Ki;Yeehn Yeeh;Lee, Jong-Kune
    • Microbiology and Biotechnology Letters
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    • v.9 no.3
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    • pp.145-152
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    • 1981
  • Microorganisms from the waste water of starch industry, were isolated and a strain, Y-124, possessing a powerful enzymic activity was selected and identified as a member of the genus Bacillus. The ideal cultural condition for the formation of $\alpha$-amylase form Bacillus Y-124 and its preservation was investigated in connection with the biotechnological and industrial approach to the bulk enzyme production. High yield of $\alpha$-amylase was observed in medium containing casein as well as calcium pantothenate in this work. Calcium ions were found to have an effect in forming this particular enzyme. Ammonium phosphate dibasic was an important inorganic nitrogen source for the formation of $\alpha$-amylase. And preservation of this enzyme was greatly affected by calcium or sodium salts. The addition of calcium carbonate or sodium sulfate presented the most effective result for the prevention of its denaturation to various factors. The above data was obtained with crude enzyme preparation.

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Screening of Antioxidative Activity and ${\alpha}$-Amylase Inhibitory Activity in Angiosperm Plants Native to Jeju Island (제주도에 자생하는 피자식물의 항산화활성과 ${\alpha}$-amylase 저해활성 검색)

  • Oh, Soon-Ja;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.22 no.1
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    • pp.71-77
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    • 2009
  • Antioxidative activity and ${\alpha}$-amylase inhibitory activity were investigated in the methanol extracts of 289 species of angiosperm plants native to Jeju Island in order to select the plant species containing bioactive materials for functional foods or medicines. The antioxidative activity, estimated by the DPPH radical scavenging capacity, was high in the galla of Rhus chinensis, the fruit of Platycarya strobilacea, and the stem of Cedrela sinensis. Particularly, the antioxidative activity of Rhus chinensis galla (94.2${\pm}$0.8%) was very high even though compared to those of BHA (61.7${\pm}$0.6%) and ascorbic acid (53.6${\pm}$0.7%). The ${\alpha}$-amylase inhibitory activity was 80% or above in 9 species including Cornus macrophylla (stem, leaf), Distylium racemosum (leaf) and Vaccinium oldhami (leaf). These results suggest that these plants could be potentially utilizable to develop bioactive materials for functional foods or medicines.

The Effect on the Culture Condition on the Activity of Amylase used for Alcohol Fermentation (주정발효용(酒精醱酵用) 효소제(酵素劑) 배양제조(培養製造) 조건(條件)이 Amylase 활성(活性)에 미치는 영향(影響))

  • Lee, Sung-Dong;Ryu, Young-Hong
    • Korean Journal of Food Science and Technology
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    • v.5 no.4
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    • pp.224-230
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    • 1973
  • The culture used wheat bran as media for four kind of mold strains such as Aspergillus oryzae, Aspergillus kawachii, Aspergillus usamii and Rhizopus javanicus to examine which strain could higher the activity of amylase most which is used for alcohol fermentation. It also provided three different kind of wheat bran media containing starch of 47%, 51% and 55% respectively for each strain. For each media it also added three different kind of nitrogen sources; ammonium sulfate, casein, and ammonium sulfate and casein equally mixed. Each nitrogen source added was subordinately differentiated into three different percentages, 2%, 4% and 6% respectively, except the 2% for the ammonium sulfate. The results obtained were summarized as follows (1) The activity of ${\alpha}-amylase$ was highest in the media of starch value 47% of wheat bran with 6% of casein added. (2) The activity of ${\beta}-amylase$ was highest in the media of starch value 51% of wheat bran with 2% of the equal mixture of ammonium sulfate and casein added. (3) The activities of both ${\alpha}-amylase$ and ${\beta}-amylase$ of Aspergillus usamii were highest in the media of starch value 47% wheat bran with no addition of nitrogen source. (4) Of the four strains examined, the activities of ${\alpha}-amylase$ and ${\beta}-amylase$ cultured in Rhizopus javanicus were both relatively higher. (5) The activities of ${\alpha}-amylase$ and ${\beta}-amylase$ of the strains examined became lower as the percentage of starch contents increased except in Rhizopus javanicus.

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