• 제목/요약/키워드: raw-starch-digesting enzyme

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Isolation of Soil Bacteria Secreting Raw-Starch-Digesting Enzyme and the Enzyme Production

  • Sung, Nack-Moon;Kim, Keun;Choi, Sung-Ho
    • Journal of Microbiology and Biotechnology
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    • 제3권2호
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    • pp.99-107
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    • 1993
  • Two strains (No. 26 and 143) of bacteria which secrete both pectinase and raw-starch-digesting amylase simultaneously, were isolated from various domestic soil samples. The two bacteria were identified as Pasteurella ureae judging by their morphological and physiological characteristics. The optimal culture conditions for the production of raw-starch-digesting enzyme by the Pasteurella ureae 26 were using $NH_4NO_3$ as the nitrogen source at $37^{\circ}C$ with the pH of 7.5, and 15 of C/N ratio. Since the enzyme was produced only when raw or soluble starch was used as a carbon source, but not when glucose or other sugars was used, the enzyme was considered to be an inducible enzyme by starch. Thin layer chromatography of the hydrolyzed product of starch by the raw-starch-digesting enzyme of the strain No. 26 showed that glucose, maltose and other oligosaccharides were present in the hydrolyzates, and therefore the enzyme seemed to be ${\alpha}-amylase$. The enzyme had adsorbability onto raw com starch in the pH range of 3 to 9.

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Bacillus sp.가 생산하는 전분 분해효소의 활성에 미치는 Alum첨가의 영향 (Effect of Alum on the Activity of Raw Starch-Digesting Enzyme Produced by Bacillus sp.)

  • 이신영;이상귀;강태수;이명열
    • 한국식품과학회지
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    • 제27권5호
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    • pp.773-775
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    • 1995
  • The effect of alum$(Al{\cdot}K(SO_4)_2{\cdot}12H_{2}O)$ on the activity of raw starch-digesting enzyme produced by alkali-tolerant Bacillus sp. was investigated. In adding alum of 0.5%(w/w), activities of raw starch-digesting enzyme on the gelatinized and raw rice starches have not been inhibited. In case of adding alum of 5%(w/w), competitive and uncompetitive inhibition were observed for the gelatinized and raw rice starches, respectively. The inhibitory effect on the raw starch was much higher than that on the gelatinized starch.

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

  • 이신영;조택상
    • KSBB Journal
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    • 제13권5호
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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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Improvement of a Fungal Strain by Repeated and Sequential Mutagenesis and Optimization of Solid-State Fermentation for the Hyper-Production of Raw-Starch-Digesting Enzyme

  • Vu, Van Hanh;Pham, Tuan Anh;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • 제20권4호
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    • pp.718-726
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    • 2010
  • A selected fungal strain, for production of the raw-starchdigesting enzyme by solid-state fermentation, was improved by two repeated sequential exposures to ${\gamma}$-irradiation of $Co^{60}$, ultraviolet, and four repeated treatments with Nmethyl-N'-nitrosoguanidine. The mutant strain Aspergillus sp. XN15 was chosen after a rigorous screening process, with its production of the raw-starch-digesting enzyme being twice that of usual wild varieties cultured under preoptimized conditions and in an unsupplemented medium. After 17 successive subculturings, the enzyme production of the mutant was stable. Optimal conditions for the production of the enzyme by solid-state fermentation, using wheat bran as the substrate, were accomplished for the mutant Aspergillus sp. XN15. With the optimal fermentation conditions, and a solid medium supplemented with nitrogen sources of 1% urea and 1% $NH_4NO_3$, 2.5 mM $CoSO_4$, 0.05% (v/w) Tween 80, and 1% glucose, the mutant Aspergillus sp. XN15 produced the raw-starch-digesting enzyme in quantities 19.4 times greater than a typical wild variety. Finally, XN15, through simultaneous saccharification and fermentation of a raw rice corn starch slurry, produced a high level of ethanol with $Y_{p/s}$ of 0.47 g/g.

Streptomyces sp. 4M-2가 생산하는 생전분 분해효소의 특성 (Characteristics of Raw Starch-Digesting Enzyme from Streptomyces sp. 4M-2)

  • 최성현;김찬조;오만진;이종수
    • 한국미생물·생명공학회지
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    • 제17권2호
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    • pp.136-141
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    • 1989
  • Streptomyces sp. 4M-2의 생전분 분해효소를 황산 암모니움 염석과 이온교환 크로마토그라피 및 Gel 여과 등으로 비활성이 51.22 RSU/mg, 수율 4.5%로 정제할 수 있었다. 정제효소의 분자량은 102,000이었으며 생옥수수 전분에 대한 Km값은 44.44mg/m1 이었다 정제효소의 작용 최적온도는42$^{\circ}C$, pH는 5.5였고 $Ca^{2+}$$Ba^{2+}$의 첨가에 의해 효소활성이 증가되었다. 정제효소는 옥수수 amylose를 가장 잘 분해하고 감자전분도 비교적 잘 분해하였다. 이 효소에 의한 옥수수 생전분의 분해산물은 주로 maltose와 maltotriose였고 소량의 glucose와 oligosaccharide도 검출되었으므로 $\alpha$-amylase 계통의 효소로 추정된다.

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Ethanol Production from Rice Winery Waste - Rice Wine Cake by Simultaneous Saccharification and Fermentation Without Cooking

  • Vu, Van Hanh;Kim, Keun
    • Journal of Microbiology and Biotechnology
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    • 제19권10호
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    • pp.1161-1168
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    • 2009
  • Ethanol production by the simultaneous saccharification and fermentation (SSF) of low-value rice wine cake (RWC) without cooking was investigated. RWC is the filtered solid waste of fermented rice wine mash and contains 53% raw starch. For the SSF, the RWC slurry was mixed with the raw-starch-digesting enzyme of Rhizopus sp. and yeast, where the yeast strain was selected from 300 strains and identified as Saccharomyces cerevisiae KV25. The highest efficiency (94%) of ethanol production was achieved when the uncooked RWC slurry contained 23.03% starch. The optimal SSF conditions were determined as 1.125 units of the raw-starch-digesting enzyme per gram of RWC, a fermentation temperature of $30^{\circ}C$, slurry pH of 4.5, 36-h-old seeding culture, initial yeast cell number of $2{\times}10^7$ per ml of slurry, 17 mM of urea as the nitrogen additive, 0.25 mM of $Cu^{2+}$ as the metal ion additive, and a fermentation time of 90 h. Under these optimal conditions, the ethanol production resulting from the SSF of the uncooked RWC slurry was improved to 16.8% (v/v) from 15.1% (v/v) of pre-optimization.

Rhizopus oryzae가 생성하는 생전분 분해효소의 정제 및 특성 (Purification and Characterization of Raw Starch-Digesting Enzyme from Rhizopus oryzae)

  • 김찬조;오만진;이종수
    • 한국식품과학회지
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    • 제18권4호
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    • pp.288-293
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    • 1986
  • 유안염석과 column chromatography 및 gel여과 등으로 비활성이 45.2U/mg가 되는 정제된 Rhizopus oryzae의 생전분 분해효소를 16.2%의 수율로 얻었다. 정제효소는 전기영동상에서 그 순도가 인정되었고 분자량은 67000, Km값은 4.082mg/ml이었다. 정제효소는 $50^{\circ}C$, pH $4.0{\sim}5.0$에서 잘 작용하였으며 옥수수amylose가 가장 적합한 전분이었고 옥수수 생전분에 작용한 분해산물은 거의 glucose였다.

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Molecular Cloning and Determination of the Nucleotide Sequence of Raw Starch Digesting α-Amylase from Aspergillus awamori KT-11

  • Matsubara, Takayoshi;Ammar, Youssef Ben;Anindyawati, Trisanti;Yamamoto, Satoru;Ito, Kazuo;Iizuka, Masaru;Minamiura, Noshi
    • BMB Reports
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    • 제37권4호
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    • pp.429-438
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    • 2004
  • Complementary DNAs encoding $\alpha$-amylases (Amyl I, Amyl III) and glucoamylase (GA I) were cloned from Aspergillus awamori KT-11 and their nucleotide sequences were determined. The sequence of Amyl III that was a raw starch digesting $\alpha$-amylase was found to consist of a 1,902 bp open reading frame encoding 634 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. On the other hand, the sequence of Amyl I, which cannot act on raw starch, consisted of a 1,500 bp ORF encoding 499 amino acids. The signal peptide of the enzyme was composed of 21 amino acids. The sequence of GA I consisted of a 1,920 bp ORF that encoded 639 amino acids. The signal peptide was composed of 24 amino acids. The amino acid sequence of Amyl III from the N-terminus to the amino acid number 499 showed 63.3% homology with Amyl I. However, the amino acid sequence from the amino acid number 501 to C-terminus, including the raw-starch-affinity site and the TS region rich in threonine and serine, showed 66.9% homology with GA I.

Stabilization of a Raw-Starch-Digesting Amylase by Multipoint Covalent Attachment on Glutaraldehyde-Activated Amberlite Beads

  • Nwagu, Tochukwu N.;Okolo, Bartho N.;Aoyagi, Hideki
    • Journal of Microbiology and Biotechnology
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    • 제22권5호
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    • pp.628-636
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    • 2012
  • Raw-starch-digesting enzyme (RSDA) was immobilized on Amberlite beads by conjugation of glutaraldehyde/polyglutaraldehyde (PG)-activated beads or by crosslinking. The effect of immobilization on enzyme stability and catalytic efficiency was evaluated. Immobilization conditions greatly influenced the immobilization efficiency. Optimum pH values shifted from pH 5 to 6 for spontaneous crosslinking and sequential crosslinking, to pH 6-8 for RSDA covalently attached on polyglutaraldehyde-activated Amberlite beads, and to pH 7 for RSDA on glutaraldehyde-activated Amberlite. RSDA on glutaraldehyde-activated Amberlite beads had no loss of activity after 2 h storage at pH 9; enzyme on PG-activated beads lost 9%, whereas soluble enzyme lost 65% of its initial activity. Soluble enzyme lost 50% initial activity after 3 h incubation at $60^{\circ}C$, whereas glutaraldehyde-activated derivative lost only 7.7% initial activity. RSDA derivatives retained over 90% activity after 10 batch reuse at $40^{\circ}C$. The apparent $K_m$ of the enzyme reduced from 0.35 mg/ml to 0.32 mg/ml for RSDA on glutaraldehyde-activated RSDA but increased to 0.42 mg/ml for the PG-activated RSDA derivative. Covalent immobilization on glutaraldehyde Amberlite beads was most stable and promises to address the instability and contamination issues that impede the industrial use of RSDAs. Moreover, the cheap, porous, and non-toxic nature of Amberlite, ease of immobilization, and high yield make it more interesting for the immobilization of this enzyme.

Streptomyces sp. 4M-2에 의한 생전분 분해효소의 생산 (Production of Raw Starch Digesting Enzyme by Streptomyces sp. 4M-2)

  • 최성현;김찬조;오만진;이종수
    • 한국미생물·생명공학회지
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    • 제16권6호
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    • pp.457-462
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    • 1988
  • 방사균이 생산하는 생전분 분해효소를 식품공업에 이용하기 위한 자료를 얻고자 메주로부터 생전분 분해력이 강한 방사균을 분리하여 Streptomyces sp.로 동정하였다. 밀기울침출액 기본배지에 4%의 호화옥수수전분과 0.16%의 KNO$_3$를 첨가하고 pH를 6.2 로 조정한 다음 분리균주를 접종한 후 3$0^{\circ}C$에서 5~6일간 진탕배양하였을 때 33RSU/$m\ell$의 생전분 분해효소를 생산하였다. 또한 효소생산에 효과적인 금속 이온은 인정되지 않았으며 Hg$^{2+}$ 및 Co$^{2+}$ 등은 저해가 현저하였다.

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