• Title, Summary, Keyword: ${\alpha}$-amylase

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Studies on $\alpha$-amylase of Bacillus circulans F-2 (Part 3) Hydrolysis of Various Substrates by Purified $\alpha$-amylase (Bacillus circulans F-2가 생산하는 $\alpha$-amylase에 관한 연구 (제3보) 정제 $\alpha$-amylase에 의한 각종 기질의 분해)

  • ;Hajime Taniguchi;Yoshiharu Maruyama
    • Microbiology and Biotechnology Letters
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    • v.10 no.4
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    • pp.259-265
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    • 1982
  • These experiments were conducted to investigate the hydrolysis products on the various oligosaccharides of Bacillus cirulans F-2 $\alpha$-amylase, and the hydrolysis rate on the various raw starches of Bacillus circulans F-2 $\alpha$-amylase, Bacillus amylotiquefaciens $\alpha$-amylase and Rhizopus niveus glucoamylase. The results obtained were as follows : 1. Maltotetraose, maltopentaose, maltohexaose, maltoheptaose and maltooctaose were hydrolyzed, but maltose and maltotriose were not hydrolyzed by Bacillus circulans F-2 $\alpha$-amylase. Among maltotetraose, maltopentaose, maltohexaose, maltoheptaose and maltooctaose, especially maltotetraose was hydrolyzed weakly by Bacillus circulans F-2 $\alpha$-amylase. 2. The Hydrolysis rate of oyster glycogen was slightly lower than soluble starch, amylose and amylopectin. 3. The hydrolysis rate of com starch was higher in shaking incubation than in stationary incubation, but the hydrolysis rate of potato starch was not definite according to kinds of enzyme. 4. On com, rice, arrowroot, high amylose corn, banana, sago, yam and potato starch, Bacillus circulans F-2 $\alpha$-amylase exhibited a remarkably higher hydrolysis rate than Bacillus amyloquefaciems $\alpha$-amylase and Rhizopus niveus glucoamylase.

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The Activity and Characteristics of $\alpha$-Amylase Present in Soy Milk and Jeungpyun Batters (증편 제조시 콩물과 반죽 내의 $\alpha$-amylase활성 및 특성에 관한 연구)

  • Na, Han-Na;Yoon, Sun;Kim, Jung-Soo;Kim, Bo-Young
    • Korean journal of food and cookery science
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    • v.14 no.3
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    • pp.261-265
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    • 1998
  • The activity and characteristics of ${\alpha}$-amylase in soy milk as well as in Jeungpyun batters were determined to investigate the enzyme system related to Jeungpyun preparation. ${\alpha}$-Amylase activity was detected in soy milk as well as in Jeungpyun batters. Soy milk had ${\alpha}$-amylase activity of 0.79 units/mg protein for gelatinized starch and 0.036 units/mg protein for raw starch. ${\alpha}$-Amylase in soy milk showed maximum activities at pH 5.92∼6.87 and at 60$^{\circ}C$ for both gelatinized starch and raw starch. ${\alpha}$-Amylase activities of Jeungpyun batters containing soy milk were 25.59 units/mg protein for gelatinized starch and 1.37 units/mg protein for raw starch. Jeungpyun batters without soy milk demonstrated ${\alpha}$-amylase activities of 3.37 units/mg protein for gelatinized starch and 0.49 units/mg protein for raw starch. ${\alpha}$-Amylase of Jeungpyun batters showed an optimal activity at pH 5.25 and at 60$^{\circ}C$ for both gelatinized and raw starch. The results demonstrated that Jeungpyun batters with soy milk showed significantly higher ${\alpha}$-amylase activity than the ones without soy milk.

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Thermostable ${\alpha}$-Amyalse of Bacillus licheniformis YB-1234 Isolated from the Fermented Soybean of a Korean Buddhist Temple (사찰의 된장에서 분리된 Bacillus licheniformis YB-1234의 내열성 ${\alpha}$-Amyalse)

  • Lee, Eun Ji;Yoon, Ki-Hong
    • Microbiology and Biotechnology Letters
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    • v.40 no.4
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    • pp.296-302
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    • 2012
  • A bacterial strain was isolated from soybean paste fermented in a Korean Buddhist temple as a producer of the extracellular thermostable ${\alpha}$-amylase. The isolate YB-1234 has been identified as Bacillus licheniformis on the basis of its 16S rDNA sequence, morphology and biochemical properties. A gene encoding the thermostable ${\alpha}$-amylase of B. licheniformis YB-1234 was cloned into Escherichia coli and its nucleotide sequence was determined. The deduced amino acid sequence of ${\alpha}$-amylase was very highly homologous to those of the thermostable ${\alpha}$-amylases of B. licheniformis belonging to the glycosyl hydrolase family 13. The ${\alpha}$-amylase produced by recombinant E. coli carrying the ${\alpha}$-amylase gene exhibited maximal activity at pH 6.0, identical to ${\alpha}$-amylase in the culture filtrate of B. licheniformis, while the temperature profile was somewhat different between the two. Particularly, ${\alpha}$-amylase produced from B. lcheniformis is much more thermostable than that from recombinant E. coli. The predominant products resulting from the ${\alpha}$-amylase hydrolysis were glucose, maltose and maltotriose for maltotetraose and maltohexaose.

Molecular Cloning of ${\alpha}$-Amylase Gene from Schwanniomyces CBS 2863 (Schwanniomyces castellii CBS 2863으로부터 ${\alpha}$-Amylase 유전자 Cloning)

  • Park, Jong-Chun;Bai, Suk;Chun, Bai-CHun
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.34-39
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    • 1994
  • The gene encoding ${\alpha}$-amylase of Schwanniomyces castellii was cloned in Saccharomyces cerevisiae. The 5.0-kilobase insert was shown to direct the synthesis of ${\alpha}$-amylase. Southern blot analysis confirmed that this ${\alpha}$-amylase gene was derived from the genomic DNA of Sch. castellii. Immunoblot analysis showed that ${\alpha}$-amylase production from S. cerevisiae transformant was less than that of donor strain. The ${\alpha}$-amylase secreted from S. cerevisiae transformant was shown to be indistinguishable from that of Sch. castellii on the basis of molecular weight and enzyme properties.

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Molecular Cloning of a Thermostable $\alpha$-Amylase Gene from Bacillus stearothermophilus and Its Expressions in E. coli (Bacillus stearothermophilus의 열안정성 $\alpha$-amylase 유전자의 E. coli내에서의 cloning과 발현)

  • Huh, Tae-Lin;Koh, Suk-Hoon;Lee, Se-Yong
    • Microbiology and Biotechnology Letters
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    • v.13 no.4
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    • pp.349-354
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    • 1985
  • A 4.7 kb Hind III fragment containing $\alpha$-amylase gene of Bacillus stearothermophilus IAM 11062 was cloned in Escherichia coil HB101, using plasmid pBR322 and runaway plasmid pSY343 as a vector. The cloned gene was stably maintained and expressed In E.coli. The constructed strain of E. coli have at least 3 times higher amylase activity than the donor strain, of B. stearothermophilus. About 75% of the $\alpha$-amylase produced by the constructed strain of E. coli was localized in the periplasm and it was found that the enzymes can be released by an osmotic shock using EDTA. The enzymatic properties of L-amylase produced in E. coli were very similar to those produced by B. stearothermophilus in terms of optimum temperature, heat stability and molecular weight.

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Molecular Cloning and Expression of $\alpha$-Amylase Gene from Bacillus stearothermophilus in Zymomonas mobilis ZM4

  • Song, Ki-Bang
    • Journal of Microbiology and Biotechnology
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    • v.2 no.2
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    • pp.115-121
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    • 1992
  • In order to broaden the spectrum of substrate utilization of a Gram negative bacterium Zymomonas mobilis which has a great potential as an industrial ethanol producing microorganism, cloning of $\alpha$-amylase gene into Z. mobilis ZM4 was tried. The $\alpha$-amylase gene was isolated from Bacillus stearothermophilus. By Southern blot analysis, it was proven that the $\alpha$-amylase gene fragment was originated from a naturally occuring plasmid of B. stearothermophilus ATCC 31195. To place $\alpha$-amylase gene under the control of Z. mobilis promoter, two different Z. mobilis expression vectors, pZA26 and pLOI204, were used. The truncated $\alpha$-amylase gene was then introduced into these vectors. Both qualitative and quantitative activities of $\alpha$-amylase were observed in Z. mobilis cells harboring these plasmids with the $\alpha$-amylase gene inserted. Gas chromatographic analysis of ethanol showed that one of the Z. mobilis transconjugants was capable of producing 67 mM ethanol from rich medium(RM) containing 5% soluble starch as a sole carbon source.

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Purification of ${\alpha}-Amylase$ Inhibitor from Black Bean in Korea (한국산 검정콩 ${\alpha}-Amylase$ 저해물질의 분리 및 정제)

  • Moon, Ju-Seok;Bae, Young-Il;Shim, Ki-Hwan
    • Korean Journal of Food Science and Technology
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    • v.27 no.5
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    • pp.762-767
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    • 1995
  • The ${\alpha}-Amylase$ inhibitor from black bean(Phaseolus vulgaris) was purified to homogeneity using 70% saturated ammonium sulfate, DEAF-cellulose, Concanavalin-A sepharose chromatography and gel filtration with Superose 6. The purified α-amylase inhibitor showed a single band of 25 KD in molecular weight on the SDS-PAGE. The specific activity of the inhibitor was 544.0 units/mg and the purity was enhanced about 18-fold. The amino acids of ${\alpha}-Amylase$ inhibitor from black bean was mainly glutamic acid, aspartic acid and lysine. The inhibitor was glycoproteins and its carbohydrate contents was 3.2%.

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Identification of Streptomyces DMCJ-49 Producing the alpha-Amylase Inhibitors and the Isolation of the Inhibitor (알파-아밀라제 저해제 생성 Streptomyces DMCJ-49의 동정과 저해제의 분리)

  • Chung, Dong-Jik;Kwak, Jin-Hwan;Choi, Eung-Chil;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.33 no.3
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    • pp.175-182
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    • 1989
  • To find ${\alpha}-amylase$ inhibitors produced by microorganisms from soil, a strain which had a strong inhibitory activity against bacterial ${\alpha}-amylase$ was isolated from the soil sample collected in Korea. The morphological and physiological characteristics of this strain on several media and its utilization of carbon sources showed that it was one of Streptomyces species according to the International Streptomyces Project method. The amylase inhibitor of this strain was purified by active carbon adsorption, silicagel column chromatography, SP-Sephadex C-25 column chromatography, adsorption on Amberlite XAD-2. The inhibitor was oligosaccharide which was composed of glucose. The inhibitor had inhibitory activity against other amylase such as salivary ${\alpha}-amylase$, pancreatic ${\alpha}-amylase$, fungal ${\alpha}-amylase$ and gluco-amylase.

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Cloning of Bacillus amyloliquefaciens amylase gene using YRp7 as a vector I. Expression of cloned amylase gene in Escherichia coli (YRp 7 vector를 이용한 Bacillus amyloliquefaciens amylase gene의 cloning I. Escherichia coli에서의 발현)

  • 서정훈;김영호;전도연;홍순덕;조윤래
    • Microbiology and Biotechnology Letters
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    • v.14 no.2
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    • pp.161-168
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    • 1986
  • A 1.95Kb Sau3Al fragment coding for $\alpha$-amylase from Bacillus amyloliquefaciens was isolated by the shotgun method using Escherichia coli as a host. The genome of Bacillus amyloliquefaciens was partially digested with the restriction endonuclease Sau3Al and joined to plasmid YRp7 cleaved with the restriction endonuclease BamHI. The $\alpha$-amylase gene present in a 1.95Kb insert was stably maintained and expressed in Escherichia coli. The amount of $\alpha$-amylase activity produced by Escherichia coli containing the hybrid plasmid pEA24 was about 65% of the activity produced by the donor Bacillus amyloliquefaciens strain. The properties of $\alpha$-amylase produced by Escherichia coli were very similar to those produced by Bacillus amyloliquefaciens as based on optimum temperature, pH, and effect of CaCl$_2$ concentration. About 70% of the $\alpha$-amylase produced by Escherichia coli was localized in the periplasmic space, whereas the remaining enzyme was localized in the inner part of the cell.

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Effect of Amylase and Emulsifier on the Characteristics of the Bread Dough (Amylase와 유화제의 첨가가 빵 반죽특성에 미치는 영향)

  • Park, Bum-Joon;Hwang, Seong-Yun;Park, Cheon-Seok
    • Korean Journal of Food Science and Technology
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    • v.37 no.5
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    • pp.763-767
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    • 2005
  • Effect of ${\alpha}-amylase$ and various emulsifiers on characteristics of bread dough were examined. Fungal or bacterial ${\alpha}-amylase$ and various emulsifiers including monoglyceride (MG), sodium stearoyl-2-lactylate (SSL), and diacetyltartaric acid ester of mono- and diglycerides (DATEM) were added to bread dough both individually and as mixtures. Rheological characteristics of various bread doughs were examined through falling number, farinograph, alveograph, and rapid visco analysis. Results obtained showed falling number decreased via degradation of starch by ${\alpha}-amylase$. In farinogram, addition of ${\alpha}-amylase$ and emulsifiers in dough decreased consistency, water absorption, mechanical tolerance index, and dough development time. Farinogram characteristic was improved by adding SSL+MG to dough formula. Similar to farinogram addition of ${\alpha}-amylase$ and emulsifiers in alveogram of dough decreased overpressure, extensibility, swelling index, and deformation energy. Whereas addition of ${\alpha}-amylase$ did not affect pasting temperature, viscosity of dough tended to decrease.