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

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

  • Jung, Jin-Woo;Jung, Jong-Hyun;Seo, Dong-Ho;Kim, Byung-Yong;Park, Cheon-Seok
    • Korean Journal of Food Science and Technology
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    • v.43 no.3
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    • pp.369-374
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    • 2011
  • A putative maltogenic amylase gene (DGMA) was cloned from the Deinococcus geothermalis DSM 11300 genome using the polymerase chain reaction. The gene encoded 608 amino acids with a predicted molecular mass of 68,704 Da. The recombinant DGMA was constitutively expressed using the pHCXHD plasmid. As expected, the recombinant DGMA hydrolyzed cyclodextrins and starch to maltose and pullulan to panose by cleaving the ${\alpha}$-(1,4)-glycosidic linkages, as observed for typical maltogenic amylases. Characterization of the recombinant DGMA revealed that the highest maltogenic amylase activity occurred at $40^{\circ}C$ and pH 6.0. The half-life of catalytic activity at $65^{\circ}C$ and $55^{\circ}C$ were 8.2 min and 187.4 min, respectively. DGMA mainly hydrolyzed ${\beta}$-cyclodextrin, soluble starch, and pullulan and its efficient ratio of those substrates was 9:4.5:1.

Effects of Calcium Ion Concentration on Starch Hydrolysis of Barley ${\alpha}$-Amylase Isozymes

  • Yuk, Jeong-Bin;Choi, Seung-Ho;Lee, Tae-Hee;Jang, Myoung-Uoon;Park, Jung-Mi;Yi, Ah-Rum;Svensson, Birte;Kim, Tae-Jip
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.730-734
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    • 2008
  • Barley ${\alpha}$-amylase genes, amy1 and amy2, were separately cloned into the expression vector of $pPICZ{\alpha}A$ and recombinant Pichia strains were established by homologous recombination. Both AMYs from Pichia shared almost identical hydrolysis patterns on short maltooligosaccharides to result in glucose, maltose, or maltotriose. Against insoluble blue starch, AMY1 showed the highest activity at 0.1-5 mM calcium concentration, whereas 15-20 mM was optimal for AMY2. On the hydrolysis of soluble starch, unexpectedly, there was no significant difference between AMYs with increase of calcium. However, the relative activity on various starch substrates was significantly different between AMYs, which supports that the isozymes are clearly distinguished from each other on the basis of their unique preferences for substrates.

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.

Effects of opioid and non-opioid antagonists, pH and enzymes on Corchorus olitorius antinociception in mice

  • Zakaria Zainul Amiruddin;Neelendran M;Pubalan S;Sulaiman MR;Fatimah CA
    • Advances in Traditional Medicine
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    • v.6 no.3
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    • pp.186-195
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    • 2006
  • The present study was carried out to determine the involvement of opioid and non-opioid receptor and the effect of pH and enzymes on the recently reported antinociceptive activity of aqueous extract of Corchorus olitorius (AECO) leaves using the abdominal constriction test. The extract was prepared by soaking the dried powdered leaves of Corchorus (C.) olitorius in distilled water overnight, and the supernatant obtained was considered as a stock solution with 100% concentration/ strength. The extract, administered subcutaneously in the concentrations/ strength of 10, 50 and 100%, was found to show a significant concentration-independent antinociception. The 50% concentration AECO were further used to study on the above mentioned parameters. The extract exhibited: significant (P < 0.05) decreased in activity when pre-treated (s.c.) against 10 mg/kg naloxonazine, bicuculine (10 mg/kg), phenoxybenzamine (10 mg/kg), 10 mg/kg pindolol, and 5 mg/kg mecamylamme, but not 10 mg/kg naltrindole, 10 mg/kg atropine, respectively; significant (P < 0.05) decreased in activity after pre-treatment against 10% a-amylase, but not 1 % protease or 10% lipase and; significant (P < 0.05) decreased in activity after exposure to alkaline condition (pH between 9 and 13) while maintaining the activity at acidic condition, respectively. The C. olitorius leaves antinociception, which involved, at least in part, activation of $\mu-opioid,\;\alpha-and\;\beta-adrenergic$, and nicotinic receptors, was found to decrease under alkaline condition and in the presence of $\alpha-amylase$.

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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Effects of the ratio of raw materials on the quality and taste of soy sauce - (1) Studies on the changes of Enzymatic activities and chemical components during Koji Preparation with various ratio of raw materials - (원료배합(原料配合)이 간장의 품질(品質)과 풍미(風味)에 미치는 영향(影響) - 제1보(第一報) 원료배합(原料配合)을 달리한 고일(一)지제조중(製造中)의 효소역가(酵素力價)및 성분변화(成分變化)에 관(關)한 실험(實驗) -)

  • Kim, Yong-Hwi;Kim, Jae-Uk
    • Applied Biological Chemistry
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    • v.4
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    • pp.17-22
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    • 1963
  • Changes of Enzymatic activities and chemical components during Koji preparation of soy sauce with various ratio of soy bean and wheat were studied as first step for checking the current ratio of raw materials for improved soy sauce and determination of proper ratio of them in the respect to its quality and taste, and following results could be obtained. 1. The Protease in the dryed Koji were mainly conmposed of a part which active at the neutral (about pH 6.0) range, while parts which active at acid and alkaline side were inferior. The more amount of wheat increases as raw materials of Koji, the stronger Protease activities of acid and neural side were, while the weaker alkaline side were. 2. Activity of Enzymes were increased rapidly in earlier stage then gradually in later stage or Protease and ${\beta}-Amylase$ rapidly throughout except drying of ${\beta}-Amylase$ during the course of Koji preparation. The more amount of wheat as raw material increases, the stronger Protease and ${\beta}-Amylase$ activity except. ${\beta}-Amylase$ were. 3. Reducing sugar, amino nitrogen, total nitrogen were increased, while total sugar were decreased during the course of Koji preparation. 4. The more amount of wheat increases as raw materials, the more increase reducing sugar, total sugar were, while the total nitrogen were decreased, no noticeable differences were observed in the amino nitrogen among the dryed Kojies.

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Enzyme Inhibition Activities of Ethanol Extracts from Germinating Rough Rice (Oryza sativar L.) (발아기간에 따른 벼(Oryza sativa L.)의 부위별 효소저해활성)

  • Kim, Min Young;Lee, Sang Hoon;Jang, Gwi Young;Park, Hye Jin;Meishan, Li;Kim, Shinje;Lee, Youn Ri;Lee, Junsoo;Jeong, Heon Sang
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.42 no.6
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    • pp.917-923
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    • 2013
  • This study investigated the enzyme inhibitory effects of ethanol extracts from the different parts of rough rice (Oryza sativar L.) from its germination period. Rough rice was germinated at $37^{\circ}C$ for 6 days, then separated into hull+sprout and brown rice. ${\alpha}$-Glucosidase inhibitory activity had the highest value (39.38%) in hull+sprout extracts after 5 days of germination. ${\alpha}$-Amylase and DPP-4 inhibitory activity had the highest values (75.32% and 47.77%, respectively) in hull+sprout extracts after germination for 5 days. ACE inhibitory activity of hull+ sprout extracts increased from 43.16% at the beginning to 58.60% at 5 days, while brown rice extracts increased this activity from 0.88% at the beginning to 14.50% at 4 days. The xanthine oxidase inhibitory activity of hull+ sprout extracts increased from 62.02% at the beginning to 64.49% at one day, and then decreased. Lipase inhibitory activity had its highest value (55.17%) in hull+sprout extracts after germination for 5 days. These results indicate that the optimal germination period for increasing enzyme inhibitory activities may be 5 days, and that hull+sprout extracts have a higher enzyme inhibition activity than brown rice.

Manufacture of Pear Marc Nuruk by Aspergillus oryzae and Characteristics of Makgeolli Fermentation (Aspergillus oryzae를 이용한 배 착즙박 누룩의 제조 및 막걸리 발효특성)

  • Shin, Ja-Won;Kim, Young-Hun;Kim, Jae-Ho;Na, Kwang-Chul;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.41 no.4
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    • pp.255-260
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    • 2013
  • In order to develop new valuable pear marc nuruk and further, reduce environmental pollution by pear marc from pear juice processing, we prepared pear marc nuruk by incubation of Aspergillus oryzae into pear marc containing 50% of moisture at $30^{\circ}C$ for 7 days. ${\alpha}$-Amylase and glucoamylase activities of the pear marc nuruk were 320.2 IU and 442.8 IU, respectively and its acidic protease activity was showed 142.6 IU. After brewed makgeolli by using the pear marc nuruk, cooked rice and Saccharmyces cerevisiae, its physicochemical characteristics was investigated. Ethanol content of pear marc nuruk-makgeolli was 6.8% after fermentation at $25^{\circ}C$ for 10 days and also pear marc makgeolli showed 45.6% of antihypertensive angiotensin I-converting enzyme (ACE) inhibitory activity. In conclusion, pear marc nuruk had high amylase activity and pear marc-makgeolli had also good fermentation characteristics and antihypertensive ACE inhibitory activity. Therefore, it has the potential to become a new nuruk for brewing makgeolli.

Quality characteristics of popped rice Doenjang prepared with Bacillus subtilis strains (Bacillus subtilis 균주를 이용하여 제조한 팽화미 된장의 품질 특성)

  • Lee, Kyung Ha;Kim, Eun Ju;Choi, Hye Sun;Park, Shin Young;Kim, Jae Hyun;Song, Jin
    • Food Science and Preservation
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    • v.22 no.4
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    • pp.545-552
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    • 2015
  • This study investigated the quality characteristics of popped rice Doenjang prepared with different Bacillus strains (Bacillus subtilis KACC 15935, and Bacillus subtilis HJ18-9). The changes in the enzyme activity (protease, cellulase, and ${\alpha}$-amylase), amino-type nitrogen and ammonia-type nitrogen contents, and the reducing sugar were investigated during the fermentation period. Enzymes such as protease, cellulase, and a-amylase plays an important role in the changes in composition of nutrients, and in flavor and taste of popped rice Doenjang. Protease activities of the popped rice deonjang fermented with different Bacillus strains (control, B. subtilis KACC 15935, and B. subtilis HJ18-9) was in the range of 171.77-185.97 unit/g at the beginning of fermentation, and there were no significant differences among the samples. On the other hand, the protease activity in popped rice Doenjang fermented with B. subtilis HJ18-9 increased significantly up to $248.77{\pm}4.53unit/g$ at the end of fermentation (p<0.05). Cellulase activity and a-amylase activity of popped rice Doenjang in HJ18-9 was higher than these of other samples. After 56 days of fermentation, amino-type nitrogen in popped rice deonjang fermented with control, B. subtilis KACC 15935, and B. subtilis HJ18-9 increased significantly up to $174.99{\pm}3.70$, $166.59{\pm}1.40$, $225.39{\pm}3.70mg%$, respectively (p<0.05). These results suggested that B. subtilis HJ18-9 was a suitable starter for the preparation of soybean paste.

Amylolytic activity and Properties of Starch Granules from the Giant Embryonic Rices (발아 거대배아미의 당화력 및 전분입자의 이화학적 특성)

  • Kang, Mi-Young;Lee, Yun-Ri;Nam, Seok-Hyun
    • Applied Biological Chemistry
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    • v.46 no.3
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    • pp.189-194
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    • 2003
  • Rice seeds of 4 cultivars including Whachung-giant embryonic rice and Nampung-giant embryonic rice, as a group of the non-waxy rice cultivars, and Shinsunchal-giant embryonic rice and Whachungchal-giant embryonic rice, as that of the waxy rice cultivars, were germinated at $27^{\circ}C$ for 3 days to compare the changes in some physicochemical properties of the starch granules and the starch-hydrolysing enzyme activities during germination, respectively. ${\alpha}-Amylase$ activity of rices germinated for 3 days found to be higher than that of malt. Especially, Whachung-giant embryonic rice and Shinsunchal-giant embryonic rice were greater in activity than other rice cultivars and possessed the activities double that of malt. In contrast, ${\beta}-amylase$ of germinated rice found to be considerably less active than malt, although the giant embryonic rice group showed prevalent activity as compared o the normal rice group. With the starch granules, the amount of long glucose chains from amylose molecules were reduced in the non-waxy type giant embryonic rices, while the chain length increase was found in the waxy type giant embryonic rices. For the distribution profile of the glucose chain length from amylopectin molecules, we could observed that the chain length with DP (degree of polymerization) ranged 33 to 66 and 14 to 32 increased with the decreasing rate of that above 67 and below 13 regardless of starch waxiness. With non-waxy type of giant embryonic rices, susceptibility for glucoamylase were found to reduce along with germination, however, increase in susceptibility was observed with waxy rice types. In addition, we found the reduction in both initiation and termination temperature, and enthalpy for gelatinization.