• Title/Summary/Keyword: amylolytic enzyme

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Activity and stability of purified amylase produced by streptomyces aureofaciens 77

  • Ibrahim, A.N.;Ahmed, F.H.;Ibrahim, M.M.K.;Arafa, M.A.I.
    • Archives of Pharmacal Research
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    • v.13 no.1
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    • pp.33-37
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    • 1990
  • The effects of pH values, temperature and some elements on the amylolytic activity and stability of the purified S. aureofacienc 77 amylase were studied in this investigation. The purified enzyme showed its maximum activity at pH 6 within 8 min incubation at $40^{\circ}C$. None of the tested 6 metals showed on stimulatory effect on the enzymatic activity, $Fe^{+++}$, $Cu^{++}$ and $Hg^{++}$ at high dose inhibited the enzyme activity to great extent as compared with $Zn^{++}$, $Mn^{++}$ and $Fe^{++}$ whih gave less effect in this respect. The enzyme liquor was found to be thermolabile, since it lost completely its activity after 4 days incubation under room temperature and showed maximum activity during this period as a result of additions of $Ca^{++}$and NaCl, Gradual reduction was however recorded until activity reached 30% after 60 days of incubation.

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Amylolytic Activity of Brown Rice and Black Rice during Germination (현미와 흑미의 발아과정 중 amylolytic activity)

  • Lee, Hyang-Mi;Im, Ji-Soon;Park, Jong-Dae;Kum, Jun-Seok;Lee, Hyun-Yu;Lee, Young-Tack
    • Korean Journal of Food Science and Technology
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    • v.45 no.3
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    • pp.333-338
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    • 2013
  • Enzymatic activity in germinated cereal grains is important for the saccharification of starch materials. This study was conducted to investigate the amylolytic activities of germinating brown rice and black rice that have different amylose contents. Brown rice and black rice were steeped at room temperature for 24 h and germinated at 20 and $30^{\circ}C$ for 1, 2, and 3 days. ${\alpha}$- and ${\beta}$-Amylase activities in normal brown rice increased very slightly during the 3-day germination period, but the enzymatic activities were slightly higher in low-amylose (waxy type) brown rice. Diastatic power (DP), a measure of starch-saccharifying enzyme, was higher in the germinating brown rice with low amylose than in those with normal amylose content. ${\alpha}$- and ${\beta}$-Amylase activities in black rice increased gradually during germination, and DP of low-amylose black rice appeared to be higher than that of normal brown rice. Amylase activities in brown rice and black rice germinated at $30^{\circ}C$ were higher than those germinated at $20^{\circ}C$. Compared to brown rice, the overall amylolytic activity of germinated black rice was observed to be higher than that of brown rice.

Isolation and Characteristics of an Amylase-producing Fungus for Saccharifying Food Wastes (음식물쓰레기 당화를 위한 Amylase 생산균의 분리 및 특성조사)

  • Li, Hong-Xian;Kim, Seong-Jun
    • KSBB Journal
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    • v.22 no.2
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    • pp.114-118
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    • 2007
  • In this study, an amylase-producing fungus, strain 15 was isolated from soil in order to saccharify food wastes with cellulolytic and amylolytic enzymes. The amylase production cultures were performed in Mandel's medium with 1% rice straw and 1% paper wastes as carbon sources. The strain produced various cellulolytic (FPase 0.25, xylanase 20.09, CMCase 3.15 U/mL-supernatant) and amylolytic ($\alpha$-amylase 1.20, gluco-amylase 0.70, $\beta$-amylase 2.40 U/mL-supernatant) enzymes in Mandel's medium. In 10 L jar fermenter, maximum amylase and FPase activities, 3.25 and 0.23 U/mL, were obtained when the culture was grown at 30$^{\circ}C$, 200 rpm and 0.6 vvm for 3 days. In 100 mL flask level and 10 L jar fermenter, amylase produced by the strain 15 showed similar cellulolytic and amylolytic enzyme activities with Trichoderma inhamatum KSJ1 isolated from rotten woods by previous researcher. The ability of saccharification to food wastes also showed similar degree. However, the isolate 15 appeared to be yellowish in YMEA plate comparing to Trichoderma inhamatum KSJ1 in greenish.

Protoplast Formation of the Amylolytic Yeast and Saccharomyces cerevisiae by Snail Lytic Enzyme from Helix pomatia (Snail Lytic Enzyme에 의한 전분리용성 효모 및 Saccharomyces cerevisiae의 원형질체 형성)

  • 구영조;박완수;신동화;유태종
    • Microbiology and Biotechnology Letters
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    • v.13 no.2
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    • pp.137-144
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    • 1985
  • Studies were conducted on the conditions for preparation of yeast protoplasts utilizing Hansenula anomala var. anomala FRI YO-32 as well as Saccharomyces cerevisiae KFCC 32356 and a lytic enzyme from the snail Helix pomatia. The cell wails of the strain FRI YO-32 and S cerevisiae were found to be resistant to activity of the snail lytic enzyme if they were not treated with thiol compounds. Dithiothreitol was found to be more effective than 2-mercaptoethanol, but the latter was considered to be practical. As factors influencing the formation of yeast protoplast, it was considered to be concentration and incubation time of 2-mercaptoethanol or the lytic enzyme, growth stages in yeast cultivation, initial number of yeast cells, and concentration of osmotic stabilizer (KCI). Optimum conditions for the preparation of yeast protoplasts were determined.

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Biochemical Properties of Starch Granule Non-Digestive Enzyme(SGNA) of Bacillus polymyxa No.26

  • Sohn, Cheon-Bae;Kim, Myung-Hee;Bae, Jung-Surl
    • Journal of Microbiology and Biotechnology
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    • v.2 no.3
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    • pp.189-196
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    • 1992
  • A $\alpha$-l, 4-D-glucan maltohydrolase $(\beta$-amylase), secreted by the mesophilic aerobic bacterium Bacillus polymyxa No.26, was purified and characterized. The enzyme production was increased after a logarithmic phase of bacterial growth and paralleled with the onset of bacterial sporulation. By applying anion exchange chromatography and gel filtration the enzyme was purified 16.7-fold and had a specific activity of 285.7 units/mg. Two enzyme activities were eluted on a column of DEAE-Sephadex chromatography, and they were designated as E-I for a major enzyme peak and E-II for a minor peak. Of them, E-I enzyme peak was further purified by using gel chromatography. The molecular mass of this enzyme was determined to be 64, 000 daltons and consisted of a single subunit, showing an isoelectric point of 8.9. The enzyme was able to attack specifically the $\alpha$-l, 4-glycosidic linkages in soluble starch and caused its complete hydrolysis to maltose and $\beta$-limited dextrin. This amylolytic enzyme displayed a temperature optimum at $45^\circ{C}$ and a pH optimum at 7.0. The amino acid composition of the purified enzyme was quite similar to the other bacterial $\beta$-amylases reported. Surprisingly, the purified enzyme from this aerobe only exhibited hydrolytic activity on soluble starch, not on starch granules. The degradation of from starch by $\beta$-amylase was greatly stimulated by pullulanase addition. These results differentiated from other $\beta$-amylases reported. Based on a previous result that showed the enzyme system involves in effective degradation of raw starch granules, this result strongly suggested that the purified enzyme (E-I) can be a synergistic part of starch granule-digestion and E-II plays a crucial role in digestion of starch granules.

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The Study on Amylolytic Enzyme and Protease Activities of Kimchi (김치에 있어서의 amylolytic enzyme과 protease 활성에 관한 연구)

  • Hahn, Young-Sook;Oh, Ji-Young;Song, Joo-Eun
    • Korean Journal of Food Science and Technology
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    • v.34 no.2
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    • pp.269-273
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    • 2002
  • The amlyolytic enzymes $({\alpha}-amlyase,\;{\beta}-amlyase,\;glucoamlyase)$ and protease activities were studied during Kimchi fermentation. The optimum pH of Kimchi was 4.1 within 2 days at $20^{\circ}C$. The optimum acidity calculated as lactic acid was 0.44% within 2 days at $20^{\circ}C$. On the first day of fermentation, ${\alpha}-amlyase$ activity was reduced from 0.49 unit/mg protein to 0.20 unit/mg protein but increased in the later stage of fermentation. In case of ${\beta}-amlyase,\;glucoamlyase$ and protease showed the highest activity of 505.73, 13.43 and 1.72 unit/mg protein at the 2nd day of fermentation at $20^{\circ}C$. In the sensory evaluation of Kimchi were estimated taste, color, texture and overall acceptability. Overall acceptability of kimchi showed the highest score value on the 2nd day of fermentation, respectively.

Effect of Anaerobic Treatment on Carbohydrate-Hydrolytic Enzyme Activities and Free Amino Acid Contents in Barley Malt

  • Yun, Song-Joong;Choi, Kyeong-Gu;Kim, Jin-Key
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.1
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    • pp.19-22
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    • 1998
  • Effects of acute anoxia on carbohydrate hydrolytic enzyme activities and free amino acid contents in malt were examined. Malts were prepared with barley grains germinated for 7 days which contained the highest levels of amylolytic and(1-3,1-4)-$\beta$-glucanase activities. $\alpha$-Amylase and $\beta$-amylase activities in malts were not significantly affected by anoxia for 5 or 10 h.(1-3,1-4)-$\beta$-Glucanase activity, however, decreased about 7 to 10% by anoxia for 5 or 10 h. Alanine and $\gamma$-aminobutyric acid content changed drastically. Alanine contents in malts increased by 2.2- and 2-fold, and $\gamma$-aminobutyric acid contents by 1.4- and 1.9-fold under anoxia for 5 and 10 h, respectively.

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Purification and Characterization of Branching Specificity of a Novel Extracellular Amylolytic Enzyme from Marine Hyperthermophilic Rhodothermus marinus

  • Yoon, Seong-Ae;Ryu, Soo-In;Lee, Soo-Bok;Moon, Tae-Wha
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.457-464
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    • 2008
  • An extracellular enzyme (RMEBE) possessing ${\alpha}-(1{\rightarrow}4)-(1{\rightarrow}6)$-transferring activity was purified to homogeneity from Rhodothermus marin us by combination of ammonium sulfate precipitation, Q-Sepharose ion-exchange, and Superdex-200 gel filtration chromatographies, and preparative native polyacrylamide gel electrophoresis. The purified enzyme had an optimum pH of 6.0 and was highly thermostable with a maximal activity at $80^{\circ}C$. Its half-life was determined to be 73.7 and 16.7 min at 80 and $85^{\circ}C$, respectively. The enzyme was also halophilic and highly halotolerant up to about 2M NaCl, with a maximal activity at 0.5M. The substrate specificity of RMEBE suggested that it possesses partial characteristics of both glucan branching enzyme and neopullulanase. RMEBE clearly produced branched glucans from amylose, with partial ${\alpha}-(1{\rightarrow}4)$-hydrolysis of amylose and starch. At the same time, it hydrolyzed pullulan partly to panose, and exhibited ${\alpha}-(1{\rightarrow}4)-(1{\rightarrow}6)$-transferase activity for small maltooligosaccharides, producing disproportionated ${\alpha}-(1{\rightarrow}6)$-branched maltooligosaccharides. The enzyme preferred maltopentaose and maltohexaose to smaller maltooligosaccharides for production of longer branched products. Thus, the results suggest that RMEBE might be applied for production of branched oligosaccharides from small maltodextrins at high temperature or even at high salinity.

Mycelial Production and Amylase Activity of Fungi for Brewing in Different Submerged Culture Conditions (액체배양에서의 양조용 곰팡이의 균체 생산 및 전분분해효소 활성)

  • Noh, Jong-Min;Choi, Ji-Ho;Jung, Seok-Tae;Yeo, Soo-Hwan;Park, Jang-Woo;Lee, Jin-Won;Choi, Han-Seok
    • Journal of the East Asian Society of Dietary Life
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    • v.23 no.6
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    • pp.833-838
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    • 2013
  • In this study, twelve strains of brewing fungi were individually cultivated on wheat extract broth (WEB), potato dextrose broth (PDB) and malt extract broth (MEB) in order to determine the microorganism with good culture characteristics as well as with amylolytic activity. The strain cultured in PDB exhibited mycelia production from 12.6 g/L (Rhizopus oryzae KACC 45714) to a maximum of 48.0 g/L (Aspergillus oryzae KACC 46959), which was 2.3~9.2 times lower than that of the strain cultured in WEB and 1.7~14.6 times lower than that of the strain cultured in MEB. Accorfing the results, We found that the commercial strains of A. oryzae Suwon, CF1001 and CF1003 had a higher dry cell mass than the wild-type strains KACC 46421, 46423, 46424 and 46959. For Rhizopus sp., the acidity levels in WEB, PDB and MEB were 0.12~0.47%, 0.22~1.0% and 0.16~0.68% (equivalent lactic acid concentration) respectively. For A. oryzae, the acidity levels were 0.06~0.11%, 0.03~0.04% and 0.06~0.08% (equivalent lactic acid concentration), respectively. Amylase enzyme from Rhizopus delemar KACC 46419 exhibited an enzyme activity of 0.013 U and 0.019 U in WEB and MEB cultures, respectively. The enzyme activity of the amylase enzyme from A. oryzae was 0.019~0.037, 0.017~0.033 and 0.028~0.046 U in WEB, PDB and MEB cultures, respectively.

Rice Fermentation by Korean Amylolytic Bifidobacterium spp. (한국인유래의 Amylolytic Bifidobacterium 에 의한 쌀발효)

  • Park, Jong-Hyun;Song, Hey-Kyung;Ahn, Jun-Bae;Ji, Geun-Eok;Mok, Chul-Kyoon
    • Korean Journal of Food Science and Technology
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    • v.29 no.3
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    • pp.581-587
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    • 1997
  • For bifidus fermentation food, gelatinized rice solution was fermented without liquefaction/saccharification by amylolytic Bifidobacterium spp. isolated from Korean. Eighteen amylolytic Bifidobcterium on the starch agar were isolated from 38 Korean and four strains were finally selected as good amylase producers. The most enzyme-producing strain of Bif. sp. FBD-12 secreted extracellular amylase of 0.17 U/mg and intracelluar amylase of 1.8 U/mg. Three strains of Bif. sp. FBD-12, Bif. sp. FBD-16 and Bif. sp. FBD-17 also showed good growth on pH controlled media by HCI/acetic acid to pH 5.0 while Bif. sp. FBD-6 was not so tolerant that viable cell counts reduced to $10^2\;CFU/mL$ times on the media. Initial cell number of $10^6\;CFU/mL$ for those strains reached to $10^9\;CFU/mL$ on the rice medium supplemented with yeast extract (0.2%) and cysteine (0.05%). Ascorbic acid instead of cysteine was added to the medium for improving off-flavour and the best growth was shown at 0.1% addition. Isolated soybean proteins (ISP) of 3% accelerated the growth of the strains. Maximum count of $10^9\;CFU/mL$ reached within 12 hour fermentation on the rice medium with ascorbic acid and isolated soybean protein instead of 32 hours on the cysteine medium, and total acidity increased from 0.5% to 1% on each media. Reducing sugar in the ascorbic acid/ISP cultures generally increased especially 2 mg/mL to 15.5 mg/mL for Bif. sp. FBD-6. From sensory evaluation, the products showed good acceptability so that it suggested possibility of development of bifidus-fermented rice food.

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