• Title/Summary/Keyword: Amylase activity

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Studies on Improved Amylases Developed by Protoplast Fusion of Aspergillus species

  • Adeleye, Tolulope Modupe;Kareem, Sharafadeen Olateju;Olufunmilayo, Bankole Mobolaji;Atanda, Olusegun;Osho, Michael Bamitale;Dairo, Olawale
    • Microbiology and Biotechnology Letters
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    • v.49 no.1
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    • pp.45-56
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    • 2021
  • Improved amylases were developed from protoplast fusants of two amylase-producing Aspergillus species. Twenty regenerated fusants were screened for amylase production using Remazol Brilliant Blue agar. Crude enzyme extracts produced by solid state fermentation of rice bran were assayed for activity. Three variable factors (temperature, pH and enzyme type) were optimized to increase the amylase activity of the parents and selected fusants using rice bran medium and solid state fermentation. Analysis of this optimization was completed using the Central Composite Design (CCD) of the Response Surface Methodology (RSM). Amylase activity assays conducted at room temperature and 80℃ demonstrated that Aspergillus designates, T5 (920.21 U/ml, 966.67 U/ml), T13 (430 U/ml, 1011.11 U/ml) and T14 (500.63 U/ml, 1012.00 U/ml) all exhibited improved function making them the preferred fusants. Amylases produced from these fusants were observed to be active over the entire pH range evaluated in this study. Fusants T5 and T14 demonstrated optimal activity under acidic and alkaline conditions, respectively. Fusants T13 and T14 produced the most amylase at 72 h while parents TA, TC and fusant T5 produced the most amylase after 96 h of incubation. Response surface methodology examinations revealed that the enzyme from fusant T5 was the optimal enzyme demonstrating the highest activity (1055.17 U/ml) at pH 4 and a temperature of 40℃. This enzyme lost activity with further increases in temperature. Starch hydrolysis using fusant T5 gave the highest yield of glucose (1.6158 g/100 ml). The significant activities of the selected fusants at 28 ± 2℃ and 80℃ and the higher sugar yields from cassava starch hydrolysis over their parental strains indicate that it is possible to improve amylase activity using the protoplast fusion technique.

Activities of Hydrolytic Enzymes in Barley Malts Prepared under Different Germination Conditions (발아조건별 당화용 엿기름의 분해효소 활성도)

  • Mok, Chul-Kyoon;Lee, Young-Tack
    • Applied Biological Chemistry
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    • v.42 no.4
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    • pp.324-329
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    • 1999
  • Barley malts were prepared at 15, 18 and $21^{\circ}C$ for $3{\sim}6$ days, and assayed for ${\beta}-glucanase$, ${\alpha}-amylase$ and ${\beta}-amylase$ activities. ${\beta}-Glucanase$ activity increased markedly during earley germination and reached maximum at the 6th day of germination. ${\beta}-Glucanase$ activity in six-rowed barley malt was much higher than that in two-rowed malt. ${\beta}-Glucanase$ activity was associated with reduction in ${\beta}-glucan$ content during germination. ${\beta}-amylase$ activity was also considerably higher in two-rowed barley, and increased continuously during 6-day germination. ${\beta}-Amylase$ activity was the lowest at $15^{\circ}C$, the highest at $18^{\circ}C$, and intermediate at $21^{\circ}C$ of germination temperature. Considerable amount of ${\beta}-amylase$ was detected in ungerminated raw barley, and this enzymatic activity tended to increase during 6-day germination. Diastatic power, measure of starch-saccharifying enzyme, in six-rowed malt was $1.4{\sim}1.6$ fold higher than in two-rowed malt. Germination at $18^{\circ}C$ for $5{\sim}6$ days was suggested to be the optimum condition for manufacturing good quality malts, in terms of enhanced starch-degrading enzymatic activity.

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Effect of the Dietary Composition (quantitative ratio between carbohydrate and protein) and the Rearing Temperature during the 4th Moulting Period on Amylase activity of Digestive Juice in the Silkworm, Bombyx mori L. (가잠유충의 소화액 Amylase 활성에 미치는 사료조성(당과 단백질과의 량비) 및 4면기보호온도의 영향)

  • 문재유
    • Journal of Sericultural and Entomological Science
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    • v.15 no.1
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    • pp.1-7
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    • 1973
  • his study has been carried out to investigate amylase activity of digestive juice in the 5th day of the 5th instar influenced by the dietary composition (quantitative ratio between carbohydrate and protein) and the rearing temperature during the 4th moulting period. The larvae grew on three kinds of semi-synthetic diets. The A-diet has more carbohydrate than the others, the B-diet has carbohydrate in 1 : 2 with protein, and the C-diet has more protein than the others. All the diets were kept at 16$^{\circ}C$, 25$^{\circ}C$ and 32$^{\circ}C$ during the 4th moulting period. Amylase activity of digestive juice at the 5th day of the 5th instar was analyzed by Fuwa's method. The results were as follows. 1. Both A and C-diets were worse than B-diet in the larval growth and development. 2. The dietary composition influencing amylase activity of digestive juice was not related to the rearing temperature during the 4th moulting period. Amylase activity was stronger in C-diet, B-diet and A-diet order. 3. It was found that amylase activity at 32$^{\circ}C$ was stronger than that at 16$^{\circ}C$ in all kinds of diets. 4. There was an inter-action in amylase activity of male larval digestive juice between the dietary composition and the rearing temperature during 4th the moulting period.

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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.

Effect of Ginseng Saponin on The Growth of Heat-Stressed Yeast Saccharomyces rouxii - Protective Effect of Saponin on The Heat Denaturation of The Amylase Produced by Saccharomyces rouxii- (열처리된 효모의 생육에 미치는 인삼 Saponin의 영향 -Saccharomyces rouxii가 생산한 Amylase의 열변성에 대한 보호작용-)

  • 오영숙;이갑랑
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.19 no.6
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    • pp.591-595
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    • 1990
  • A promoting effect of ginseng saponin on the growth recovery rate of Saccharomyces rouxii which was treated by heat was confirmed in previous report(22). In order to deduce the promoting effect on the growth recovery of the heat stressed yeast, the effect of ginseng saponin on the activity and the heat stability of the amylase produced by Sacchirromyces rouxii were observed. The amylase showed the highst activity at 0.01% of saponin. At this concentration, the activity increased about 23% compared to the control. Furthermore, the ginseng saponin showed a protective effect against thermal inactivation of the amylase produced by Saccharomyces rouxii.

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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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Studies on the Amylase Rhizopus(II) (Rhizopus의 아밀라아제에 관한 연구 2)

  • 이영녹;윤경하
    • Korean Journal of Microbiology
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    • v.11 no.2
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    • pp.89-100
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    • 1973
  • Dextrinogenic and saccharogenic amylase activities of Rhizopus isolated from various substrates collected throughout South Korea are measured, and their amylase activities are surveyed from taxnomical, ecological, and physiological viewpoints. Among the 151 strains of Rhizopus, strain number 49 exhibiting most prominent saccharogenic amylase activity is selected as a best strain, and strain number 17 is selected as a best strain for dextrinogenic amylase activity. Amylase activities of Rhizopus are so variable in different strains even in the same species. In general, however, R. japonicus, R. liquefaciens, and R. shanghaiensis show high saccharogenic amylase activities, and R.chiuniang, R.achlamydosporus, and R.bahrnensis show high dextrinogenic amylase activities, while R.nigricans exhibits lower activities for both amylase. The strains isolated from kneaded cereals, such as "meju" and "kockja", exhibited very strong dextrinogenic and saccharogenic amylase activities as compared with the strains isolated from other sunstrates. The mean value of dextrinogenic amylase activities of the strains from southern inland region and saccharogenic amylase activities of the strains from middle coast region are, more or less, higher than those of the strains from the other regions. In the culture at 30.deg.C, those strains which exhibit better mycelial and air mycelial growth than sporangial formation show very high dextrinogenic and saccharogenic amylase activities. amylase activities.

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Identification and Characterization of Useful Fungi with ${\alpha}$-Amylase Activity from the Korean Traditional Nuruk

  • Kim, Hye-Ryun;Kim, Jae-Ho;Bai, Dong-Hoon;Ahn, Byung-Hak
    • Mycobiology
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    • v.39 no.4
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    • pp.278-282
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    • 2011
  • The objective of this study was to find useful fungi with ${\alpha}$-amylase activity from the Korean traditional nuruk for the quality of traditional Korean alcoholic beverage. In this study, 165 samples of traditional nuruk were collected from 170 regions throughout Korea and the fungi were isolated to a total of 384 strains. In order to investigate the effect of microflora on nuruk, ${\alpha}$-amylase activity, saccharogenic power (SP), starch hydrolysis activity and acid producing activity were evaluated. Ten strains were selected by ${\alpha}$-amylase activity, which ranged from 458.47 to 1,202.75 U/g. The size of the discolored zone for the starch hydrolysis activity of each fungus ranged from 0.3 to 2 cm. The SP of the 10 strains ranged from 228.8 to 433.4 SP. Of the 10 stains, three were identified as Aspergillus oryzae, two as Aspergillus flavus, two as Lichtheimia sp., one as Rhizopus oryzae and two as other strains. The total aflatoxins present in the nuruks were examined using enzyme-linked immunosorbent assay. The 10 nuruks had less than 1.11 ppb of aflatoxins.

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.

Hydrolysis Characteristics of Amylase from Alkaline-Tolerant Bacillus sp. on the Raw Starch (알칼리 내성 Bacillus sp.가 생산하는 Amylase의 생전분 분해 특성)

  • 이신영;조택상
    • KSBB Journal
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    • v.13 no.5
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    • pp.621-625
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    • 1998
  • The raw starch hydrolysis by amylase prepared from alkaline-tolerant Bacillus sp. were investigated. Degree of hydrolysis(%) of 5%(w/v) raw rice, corn and potato starch by this enzyme were about 40, 25 and 20%, respectively. The hydrolysis action on raw starch by change of blue value was similar to the action pattern of exo ${\beta}$-amylase. The hydrolysis products of rice starch were mainly glucose and maltose. Oligosaccarides were also detected. From the above results, this enzyme was considered as exo type ${\alpha}$-amylase. This enzyme activity on the raw starch and the gelatinized starch were 28.40 and 86.60 IU/mg protein, respectively, and the ratio of raw starch-digesting activity to gelatinized starch-digesting activity (raw starch digestivity) was about 32%. The Km values for the raw and the gelatinized starch were 4.22 and 3.0mg/mL, respectively, and the VmaX values were 0.20 and 0.31mg/mL/min, respectively.

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