• 제목/요약/키워드: 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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    • 제25권6호
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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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    • 제6권2호
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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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찰녹미 첨가 발효음료 품질 특성 (Quality Characteristics of Fermented Rice Beverage Prepared with Green Glutinous Rice)

  • 조민정;정희선
    • 한국식품영양학회지
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    • 제36권5호
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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.

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

  • 이종석;김지연;김한복;이동석
    • 미생물학회지
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    • 제36권2호
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    • pp.112-118
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    • 2000
  • Bacillus circulans KCT3004 유래의 호산성 $\alpha$-amylase 유전자가 pUC19을 vector로 하여 대장균 내에서 클로닝 되었다. 클로닝된 5.8kb Pst I DNA절편은 pUC19내에서의 삽입방향과는 무관하게 $\alpha$-amylans보다 약40배 정도의 높은 효소활성을 나타내었다. 본 연구에서 클로닝 및 발현된 효소의 최적 pH와 온도는 각각 pH 3.6 $45^{\circ}C$였으며, $40^{\circ}C$에서 1시간 동안의 사전 열처리에도 활성의 감소를 일으키지 않았다. 이 효소는 SDS-PAG와 zymogram을 통해 분석해 본 결과 분자량은 약 55,000으로 추정되었으며 기질로 starch만을 가수분해하여 maltoriose 이상의 올리고당 분자들을 주로 생산할수 있었다.

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

  • 강신권;노종수;성낙계
    • 한국미생물·생명공학회지
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    • 제21권1호
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    • pp.13-17
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    • 1993
  • 만다린 오렌지 과피를 기질로 하여 Asp. niger의 구연산 발효를 행하여 관련된 일련의 효소적 활성을 합성배지와 비교한 결과 만다린 오렌지 과피배지에서는 괴피에 함유된 Pectin이나 조섬유 등의 자화로 인하여 Polygalacturonase와 Pectin의 활성 뿐만 아니라 CMCase, xylannase 및 amylase의 활성이 높게 나타났다.

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

  • 오순자;이진호;고광섭;신동범;고석찬
    • 한국자원식물학회지
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    • 제23권5호
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    • pp.406-414
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    • 2010
  • 본 연구는 제주도에 자생하는 차나무과 식물을 대상으로 식품소재 또는 생약으로의 활용 방안을 모색하고자 angiotensin I converting enzyme(ACE) 저해활성, aminopeptidase N(APN) 저해활성 및 $\alpha$-amylase 저해활성을 조사하고, 항산화활성을 검색하고 TLC를 이용하여 분석하였다. ACE 저해활성은 후피향나무(수피)와 비쭈기나무(잎)에서 50% 이상의 저해활성을 보였으며, APN 저해활성은 비쭈기나무(잎과 수피)와 후피향나무(수피)에서만 양의 활성을 보였다. $\alpha$-amylase 저해활성은 동백나무(열매), 우묵사스레피나무(수피), 후피향나무(수피)와 차나무(줄기)에서 30% 이상의 저해활성을 보였다. 항산화활성은 비쭈기나무(수피), 후피향나무(수피), 차나무(잎)에서 30% 이상의 다소 높은 전자공여능을 나타내었다. 특히, 비쭈기나무(수피)는 dot-blot test에 의해 다른 종에 비해 활성이 높아 $1.25\;{\mu}g/ml$의 낮은 농도에서도 높은 항산화활성을 보였다. TLC 분석에 의해 비쭈기나무(수피)에서 EGC(Rf 0.26) 활성이 높았으며, 비쭈기나무, 우묵사스레피나무, 후피향나무의 수피에서 EGCG(Rf 0.09) 활성이 높게 검출되었다. 그리고, 표준 catechin류와는 다른 것으로 보이는 5개의 밴드(Rf 0.54, 0.46, 0.44, 0.16, 0.03)는 Folin-Ciocalteu Reagent 방법과 Ferric chloride-alcohol 방법을 이용하여 polyphenol류인 것으로 추정되었다. 이상의 결과를 토대로 사스레피나무를 제외한 차나무과 식물들은 생리활성이 높아 식품 소재 또는 생약으로의 활용이 가능할 것으로 보이며, 활성성분의 분리 및 동정 그리고 이들 물질을 이용한 임상실험 등 보다 심도있는 연구가 필요할 것으로 보인다.

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

  • 정진우;정종현;서동호;김병용;박천석
    • 한국식품과학회지
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    • 제43권3호
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    • pp.369-374
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    • 2011
  • D. geothermalis의 dgeo_0475 유전자로부터 만들어지는 효소를 정제하여 그 특성을 확인하였다. DGMA는 분자량이 약 68 kDa 크기의 효소로서 ${\beta}$-CD, soluble starch 및 pullulan을 가수분해하는 CD 분해 효소임을 확인하였다. 효소의 최적 온도는 $40^{\circ}C$ 최적 pH 는 6.0이며 대부분의 기질들을 glucose와 maltose로 가수분해 하였고 pullulan 및 soluble starch로부터 미량의 panose를 생성하였다. ${\beta}$-CD를 가장 잘 가수분해하나 기질간 상대적 활성차이는 다른 CD 분해효소에 비하여 크지 않았다.

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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    • 제18권3호
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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.

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

  • Lei, Zhao-zu
    • 미생물과산업
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    • 제19권2호
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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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아연처리 볍씨의 발아중 ${\alpha}-Amylase$ 활성과 유리 Proline 함량 변화에 미치는 Kinetin 침종의 영향 (Effects of Seed Soaking of Kinetin with Zinc Treatment on ${\alpha}-amylase$ Activity and Free Proline Content during Germination of Rice(Oryza sative L.))

  • 김상국;이상철
    • 한국환경농학회지
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    • 제16권3호
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    • pp.245-248
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    • 1997
  • 본 실험은 식물생장조절제인 kinetin으로 침종 처리된 벼종자에 아연 120ppm을 처리하여 발아중 아연 독성에 대한 초기생육, 엽록소 함량, ${\alpha}-amylase$ 및 유리 proline 함량 등의 변화를 일품벼와 남천벼를 실험 재료로하여 실험한 결과를 요약하면 다음과 같다. 1. 일품벼에서 초장은 무처리보다 모든 kinetin처리구에서 양호한 생육을 보인 반면 아연 120ppm처리구에서는 가장 저조한 생육현상을 보였고 발아율은 두품종 모두 무처리보다 kinetin $10^{-3}M$에서 가장 높게 나타났으나 아연 120ppm처리구에서는 일품벼가 남천벼보다 다소 높았다. 2. 엽록소 함량은 일품벼의 경우 무처리가 1.37mg인 것보다 아연 120ppm에서 매우 낮았고 특히, kinetin $10^{-3}M$에서는 무처리보다 훨씬 높은 함량의 증가를 보였다. 3. ${\alpha}-amylase$함량은 아연 120ppm 처리 후 1일째는 kinetin처리구와 무처리구에서 모두 낮은 활성을 보였으나 아연 120ppm 처리 후 3일째 kinetin $10^{-3}M$에서는 가장 높은 활성을 나타내었다. 4. 유리 proline 함량은 일품벼의 경우 아연 120ppm 처리후 1일째는 무처리를 제외한 모든 처리구에서 함량증가가 완만히 나타났으나 처리 후 3일째에는 무처리보다 아연 120ppm 처리에서 가장 높은 함량 증가를 보였다.

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