• Title/Summary/Keyword: a-amylase activity

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Screening, Gene Cloning, and Characterizations of an Acid-Stable α-Amylase

  • Liu, Xinyu;Jia, Wei;An, Yi;Cheng, Kun;Wang, Mingdao;Yang, Sen;Chen, Hongge
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.828-836
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    • 2015
  • Based on its α-amylase activity at pH 5.0 and optimal pH of the crude enzyme, a strain (named B-5) with acid α-amylase production was screened. The B-5 strain was identified as Bacillus amyloliquefaciens through morphological, physiological, and biochemical characteristics analysis, as well as 16S rDNA phylogenetic analysis. Its α-amylase gene of GenBank Accession No. GU318401 was cloned and expressed in Escherichia coli. The purified recombinant α-amylase AMY-Ba showed the optimal pH of 5.0, and was stable at a pH range of 4.0-6.0. When hydrolyzing soluble starch, amylose, and amylopectin, AMY-Ba released glucose and maltose as major end products. The α-amylase AMY-Ba in this work was different from the well-investigated J01542-type α-amylase which also came from B. amyloliquefaciens. AMY-Ba exhibited notable adsorption and hydrolysis ability towards various raw starches. Structure analysis of AMY-Ba suggested the presence of a new starch-binding domain at its C-terminal region.

Effects of Raw Materials and Various Molds on the Production of Koji

  • Yi, Sang-Duk;Yang, Jae-Seung;Lee, Gyu-Hee;Park, Seong-Hyun;Oh, Man-Jin
    • Preventive Nutrition and Food Science
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    • v.6 no.2
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    • pp.101-106
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    • 2001
  • Alpha-amylase and glucoamylase activities were higher in koji with 40% water than that with 30 and 50% water, and A. oryzae exhibited very high alpha-amylase and glucoamylase activities compared to A. sojae and A. niger. Acidic, neutral and alkaline protease activities also showed higher activities in koji prepared with flour, Korean wheat powder and soybean powder with 40% water based on the weight of the sample. Alpha-amylase, acidic, neutral and alkaline protease activities of all the koji samples according to incubation periods increased until 3~4 days of incubation and maintained nearly the same level or slightly decreased after 5 days of incubation. The protease activities of A. oryzae and A. sojae showed nearly the same trend regardless of differences in substrate conditions and koji materials, but those of A. niger showed a lower activity than those of A. oryzae and A. sojae. These results suggest that the preparation of koji is possible with Korean wheat powder and soybean powder and A. sojae can be utilized as a new strain for fermented foods using soybean as the main materials to increase functional properties and produce products having a new taste and flavor.

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Quality Characteristics of Fermented Rice Beverage Prepared with Green Glutinous Rice (찰녹미 첨가 발효음료 품질 특성)

  • Min Jeong Cho ;Hee Sun Jeong
    • The Korean Journal of Food And Nutrition
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    • v.36 no.5
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    • pp.341-353
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    • 2023
  • In this study, we aimed to develop a fermented green rice beverage with a unique flavor and physiological function activity. With glutinous green rice and rice nuruk as independent variables, we modeled the antioxidant characteristics and α-glucosidase, α-amylase inhibitory activity of glutinous green rice fermented beverage to verify its significance. The total flavonoid content and α-glucosidase inhibitory activity were selected as Quadratic models, and DPPH radical scavenging ability and α-amylase inhibitory activity were selected as linear models. For the sensory characteristics of glutinous green rice fermented beverage, sweetness, sourness, savory taste, bitterness, throat feel, nuruk scent, and overall preference increased in preference as the amount of glutinous green rice and rice nuruk increased, but significantly decreased after the center point (p<0.01). A blending ratio of 180.00 g of glutinous green rice and 400.00 g of rice nuruk had the highest preference among all the sensory items. Based on these results, we developed a green rice fermented beverage with unique flavor and physiological function activity of rice using glutinous green rice and rice nuruk, and the optimal blending ratio was determined to be 164.04 g of white rice, 195.96 g of glutinous green rice, and 414.61 g of rice nuruk.

Cloning and Expression of an Acidophilic $\alpha$-Amylase Gene from Bacillus circulans in Escherichia coli (Bacillus circulans의 호산성 $\alpha$-amylase 유전자의 클로닝 및 발현)

  • 이종석;김지연;김한복;이동석
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.112-118
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    • 2000
  • A new gene encoding an acidophilic TEX>$\alpha$-amylase of Bacillus cil-culans KCTC3004 was cloned into Eschericlzia coli using pUC19 as a vector. The gene localized in the 5.8 kb PstI DNA fragment was expressed independently of its orientation in the cloning vector showing enzyme activity about 40 times greater than that produced by the original B, circulans The optimum pH and temperature of the cloned enzyme were pH 3.6 and 45^{\circ}C.$ respectively. The enzyme hydrolyzed starch to produce maltotriose and maltooligosaccharides. The SDS-PAGE and zymopram of the enzyme produced in E coli(p.4L850) indicated a molecular weight of 55,000.

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Enzymatic Reactions in Citric Acid Fermentation of Mandarin Orange Peel by Aspfrgillus niger (만다린 오렌지 과피를 기질로 한 Aspergillus niger의 구연산 발효에 관련된 효소적 반응)

  • 강신권;노종수;성낙계
    • Microbiology and Biotechnology Letters
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    • v.21 no.1
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    • pp.13-17
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    • 1993
  • When mandarin orange peel was used for a substrate of citric aCid fermentation by Aspergillus niger, principal enzyme activities were investigated. Not only the activity of polygalacturonase and pectin esterase being capable of digesting pectin and crude fiber of mandarin orange peel. but also that of carboxymethyl cellulase, xylanase and amylase was high. In carbohydrate metabolism, the activity of enzymes related in HMP pathway was higher than that in EMP pathway at the orange peel medium designed hereby rather than synthetic medium. Productivity of citric acid was significantly increased when the activity of citrate synthetase was high and 5imultaneously those of aconitase and NADP-dependent dehydrogenase were low.

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Antioxidative activity, including Inhibitory activities of ACE, APN and $\alpha$-amylase, in Theaceae Plants Native to Jeju Island (제주도 자생 차나무과 식물의 ACE, APN, $\alpha$-amylase 저해 활성 및 항산화활성에 대한 연구)

  • Oh, Soon-Ja;Lee, Jin-Ho;Ko, Kwang-Sup;Shin, Dong-Bum;Koh, Seok-Chan
    • Korean Journal of Plant Resources
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    • v.23 no.5
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    • pp.406-414
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    • 2010
  • Antioxidative activity, including inhibitory activities of angiotensin I converting enzyme(ACE), aminopeptidase N(APN) and $\alpha$-amylase, was investigated in the methanol extracts from Theaceae plants native to Jeju island, in order to select the plant species containing bioactive materials for functional food or medicines. ACE inhibitory activity was above 50% in Ternstroemia japonica(stem bark) and Cleyera japonica(leaf), and APN inhibitory activity was low to be positive only in C. japonica(leaf, stem bark) and T. japonica(stem bark). $\alpha$-Amylase inhibitory activity was above 30% in Camellia japonica(fruit), Eurya emarginata(stem), T. japonica(stem bark) and Thea sinensis(stem). The antioxidative activity, estimated by the DPPH radical scavenging capacity, was above 30% in C. japonica(stem bark), T. japonica(stem bark) and T. sinensis(leaf). Particularly, the antioxidative activity analyzed by dot-blot test was very high in C. japonica(stem bark) relatively to those of other plants, and remained high in the low concentration($1.25\;{\mu}g/m{\ell}$). From the TLC analysis of antioxidative compounds, EGC(Rf 0.26) was found to have high activity in stem bark of C. japonica and EGCG(Rf 0.09) was found to have high activity in stem bark of C. japonica, E. emarginata, and T. japonica. Five bands (Rf 0.54, 0.46,0.44, 0.16, 0.03) which were not identified as compared with catechins were detected as polyphenolic compounds on the TLC plates sprayed with the Folin-Ciocalteu solution or the Ferric chloride-alcohol solution. These results suggests that Theaceae plants except E. japonica could be potentially used as a resource of bioactive materials for functional foods or medicines and further research is reguired to identify the bioactive substances and determine the functions of them.

Molecular Cloning and Characterization of Maltogenic Amylase from Deinococcus geothermalis (Deinococcus geothermalis 유래 maltogenic amylase의 유전자 발현 및 특성확인)

  • Jung, Jin-Woo;Jung, Jong-Hyun;Seo, Dong-Ho;Kim, Byung-Yong;Park, Cheon-Seok
    • Korean Journal of Food Science and Technology
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    • v.43 no.3
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    • pp.369-374
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    • 2011
  • A putative maltogenic amylase gene (DGMA) was cloned from the Deinococcus geothermalis DSM 11300 genome using the polymerase chain reaction. The gene encoded 608 amino acids with a predicted molecular mass of 68,704 Da. The recombinant DGMA was constitutively expressed using the pHCXHD plasmid. As expected, the recombinant DGMA hydrolyzed cyclodextrins and starch to maltose and pullulan to panose by cleaving the ${\alpha}$-(1,4)-glycosidic linkages, as observed for typical maltogenic amylases. Characterization of the recombinant DGMA revealed that the highest maltogenic amylase activity occurred at $40^{\circ}C$ and pH 6.0. The half-life of catalytic activity at $65^{\circ}C$ and $55^{\circ}C$ were 8.2 min and 187.4 min, respectively. DGMA mainly hydrolyzed ${\beta}$-cyclodextrin, soluble starch, and pullulan and its efficient ratio of those substrates was 9:4.5:1.

Alleviating Effects of Baechu Kimchi Added Ecklonia cava on Postprandial Hyperglycemia in Diabetic Mice

  • Lee, Hyun-Ah;Song, Yeong-Ok;Jang, Mi-Soon;Han, Ji-Sook
    • Preventive Nutrition and Food Science
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    • v.18 no.3
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    • pp.163-168
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    • 2013
  • In this study, we investigated the inhibitory effects of Baechu kimchi added Ecklonia cava on the activities of ${\alpha}$-glucosidase and ${\alpha}$-amylase and its alleviating effect on the postprandial hyperglycemia in STZ-induced diabetic mice. Baechu kimchi added Ecklonia cava (BKE, 15%) was fermented at $5^{\circ}C$ for 28 days. Optimum ripened BKE was used in this study as it showedthe strongest inhibitory activities on ${\alpha}$-glucosidase and ${\alpha}$-amylaseby fermentation time among the BKEs in our previous study. The BKE was extracted with 80% methanol and the extract solution was concentrated, and then used in this study. The BKE extract showed higher inhibitory activities than Baechu kimchi extract against ${\alpha}$-glucosidase and ${\alpha}$-amylase. The $IC_{50}$ values of the BKE extract against ${\alpha}$-glucosidase and ${\alpha}$-amylase were 0.58 and 0.35 mg/mL, respectively; BKE exhibited a lower ${\alpha}$-glucosidase inhibitory activity but a higher ${\alpha}$-amylase inhibitory activity than those of acarbose. The BKE extract alleviated postprandial hyperglycemia caused by starch loading in normal and streptozotocin- induced diabetic mice. Furthermore, the BKE extract significantly lowered the incremental area under the curve in both normal and diabetic mice (P<0.05). These results indicated that the BKE extract may delay carbohydrate digestion and thus glucose absorption.

Practice of industrial strain improvement (제 1차 한.중 생명공학 심포지움)

  • Lei, Zhao-zu
    • The Microorganisms and Industry
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    • v.19 no.2
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    • pp.34-41
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    • 1993
  • Industrial strain improvement is concerned with developing or modifying microorganisms used in production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific characteristics such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empirical approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids, organic acids and enzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is a homoserine auxotroph with AEC, TA double metabolic analogue resistant markers. The yield reaches 100 g/l. Besides, the citric acid-producing organism Aspergillus niger, Co827, its productivity reaches the advanced level in the world, is also the result of a series mutations especially with $^60Co{\gamma}$-radiation. The thermostable .alpha.-amylase producing strain A 4041 is the third example. By combining physical and chemical mutations, the strain A 4041 becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The .alpha.-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus niger SP56, its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV-11. Recently, recombinant DNA approach provides a worthwhile alternative strategy to industrial strain improvement. This technique had been used by us to increase the thermostable .alpha.-amylase production and on some genetic researches.

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Effects of Seed Soaking of Kinetin with Zinc Treatment on ${\alpha}-amylase$ Activity and Free Proline Content during Germination of Rice(Oryza sative L.) (아연처리 볍씨의 발아중 ${\alpha}-Amylase$ 활성과 유리 Proline 함량 변화에 미치는 Kinetin 침종의 영향)

  • Kim, Sang-Kuk;Lee, Sang-Chul
    • Korean Journal of Environmental Agriculture
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    • v.16 no.3
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    • pp.245-248
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    • 1997
  • The study was carried out to elucidate the changes of ${\alpha}-amylase$ activity and free proline content for zinc toxicity in two rice cultivars(Ilpumbyeo and Namchunbyeo) during germination and early growth stages. Plant height in all kinetin treatments was promoted but zinc 120ppm treatment was decreased. Soaking treatment of kinetin $10^{-3}M$ increased germination rate of both cultivars, Ilpumbyeo and Namchunbyeo by 95% and 96% as compared with zinc 120ppm. Chlorophyll content of Ilpumbyeo was higher than that of Namchunbyeo. Activity of ${\alpha}-amylase$ in kinetin $10^{-3}M$ of both rice cultivars was most highest in the 3days after treatment of zinc 120ppm. Free proline content in all rice cultivars of zinc 120ppm treatment was sharply increased at the 3days after treatment of zinc 120ppm. As a result, the effects of kinetin treatment were recognized to promote the plant height and germination rate under zinc toxicity(120ppm) during rice seed germination and early growth stages.

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