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

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Isozyme electrophoresis patterns of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China

  • Park, Gab-Man;Yong, Tai-Woon;Im, Kyung-Il;Lee, Kyu-Je
    • Parasites, Hosts and Diseases
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    • v.38 no.1
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    • pp.45-48
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    • 2000
  • An enzyme analysis of the liver fluke, Clonorchis sinensis from Kimhae, Korea and from Shenyang, China was conducted using a horizontal. starch gel electrophoresis in order to elucidate their genetic relationships. A total of eight enzymes was employed from two different kinds of buffer systems. Two loci from each enzyme of aconitase and esterase (${\alpha}-Na{\;}and{\;}{\beta}-Na$) : and only one locus each from six enzymes, gluucose-6-phosphate dehydrogenase (G6PD), ${\alpha}-glycerophosphate$ dehydrogenase (GPD), 3-hydroxybutyrate dehydrogenase (HBDH), malate dehydrogenase (MDH), phosphoglucose isomerase (PGI), and phosphoglucomutase (PGM) were detected. Most of loci in two populations of C. sinensis showed homozygous monomorphic banding patterns and one of them, GPD was specific as genetic markers between two different populations. However, esterase (${\alpha}-Na$), GPD, HBDH and PGI loci showed polymorphic banding patterns. Two populations of C. sinensis were more closely clustered within the range of genetic identity value of 0.998-1.0. In summarizing the above results, two populations of C. sinensis employed in this study showed mostly monomorphic enzyme protein banding patterns, and genetic differences specific between two populations.

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Blood Glucose Lowering Effects of Mulberry Leaves and Silkworm Extracts on Mice Fed with High-Carbohydrate Diet (고탄수화물 식이 섭취 마우스에서 상엽 및 누에 추출물의 혈당강하 효과)

  • 김미선
    • Journal of Nutrition and Health
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    • v.31 no.2
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    • pp.117-125
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    • 1998
  • Mulberry leaves(Mori folium) and silkworm(Bombyx mori) are potnet inhibiters of intestinal $\alpha$-glycosidase, and inhibit the digestion of starch and sucrose in the small intestine. They are able to prevent postprandial hyperglycemia and decrease blood insulin levels. In this study , a high-carbohydrate diet(CHO ; 67.5%, protein ; 20.8%, fat : 11.7%) was received by the control group. In contrast, the experimental groups received a high-carbohydrate diet with extracts of mulberry leaves and silkwork(50mg.100g diet), and acarbose(6.7mg/100g diet). after a 10 week study period , the experimental groups had lower blood glucose and triglyceride levels. The experimental groups tended to have lwer Hb Alc levels. Also, blood insulin levels were lower than the control groups in accordance with blood glucose levels. The activities of intestinal $\alpha$-glucosidase in the middle and distal parts of small intestine were induced by the extracts of mulberry leaves and silkworm in the experimental groups. However, the activities of liver lysosomal glucosidase and the contents of glycogen in the liver were not affected by the mulberry leave and silkworm extracts nor by acarbose. Mulberry leaves and silkworm were able to prevent sudden postprandial peaks in blood glucose as a result of $\alpha$-glycosidase, inhibition, there by decreasing unnecessary insulin secretion.

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Screening of Potato Cultivars for Infestation by Selatosomus puncticollis Mot.(Coleoptera: Elasteridae) and Analysis of Factors Assiciated with Resistance (청동방아벌레(Selatosomus puncticollis Mot.) 유충에 의한 감자 품종별 피해도 검정과 저항성 관련요인 분석)

  • Kwon, Min;Park, Cheon-Soo;Hahm, Young-Il;Ahn, Yong-Joon
    • Korean journal of applied entomology
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    • v.39 no.3
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    • pp.157-163
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    • 2000
  • In a previous report, 50 potato cultivars were screened for infestation levels by the coppery click beetle (Selatosomus puncticollis Motschulsky) in the field. Subsequently, we selected 10 potato cultivars (Anco, Atlantic, Bintje, Dejima, Denali, Jopung, Irish Cobbler, Namsuh, Shepody, and Superior) to evaluate feeding preferences by wireworms, and to analyze some factors associated with resistance. The injury rates and number of holes in potato tubers damaged by larvae of S. puncticollis were checked in the field and laboratory. Additionally, some of their chemical characters (contents of glycoalkaloids, total-nitrogen, Ca, K, Mg, sugars, and starch) were quantified. And finally correlation analysis was conducted to see whether there is a possible relationship between these characteristics and the damage level. The tuber injury rates by S. puncticollis larvae were generally high showing 19% to 73% of damage level. The highest number of tuber hole damaged by S. puncticollis larvae was found on cv. Namsuh, but generally fewer on cvs. Anco, Atlantic, Bintje, Denali and Superior. No activities for $\alpha$-chaconine, $\alpha$-solanine and $\alpha$-tomatine at a concentration of 2,500 ppm were found to S. puncticollis larvae. The contents of glycoalkaloids in tuber were different depending on cultivars. In tubers, cv. Superior contained the highest level of 18.8 mg%, but cv. Irish Cobbler had the lowest level of 6.39 mg%. Concentrations of reducing sugars and total free sugars in tubers of cv. Namsuh were 0.71 % and 2.95%, but 0.26% and 1.77% in those of cv. Dejima, respectively. For the content of potato starch, cvs. Bintje, Dejima and Irish Cobbler showed higher level, but cvs. Jopung and Shepody lower. The highest contents of total nitrogen, Ca, K and Mg in tuber were found on cvs. Anco, Atlantic, Anco and Jopung, respectively. From correlation analyses, injury rate by S. puncticollis larvae was correlated with total nitrogen content (r = -0.71435*) and total sugar content in tuber (r = 0.78018*). Such information will become essential in developing integrated pest management programs and also in breeding new potato cultivars resistant to the wireworms.

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Changes in Microflora, Enzyme Activities and Microscopic Structure of Waxy Rice and Steeping Water in Response to Different Steeping Conditions During Preparation of Gangjung (강정 제조시 찹쌀의 수침조건에 따른 찹쌀과 수침액의 미생물상, 효소활성 및 미세구조 변화)

  • Kim, Haeng-Ran;Kim, Kyung-Mi;Kim, Kwang-Ok
    • Korean Journal of Food Science and Technology
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    • v.41 no.6
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    • pp.644-651
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    • 2009
  • This study was conducted to investigate the effects of steeping periods and temperatures of waxy rice on the microflora and enzyme activities of steeping water and waxy rice, as well as on the microscopic structures of waxy rice during the preparation of gangjung. When the steeping water was 15 and $35^{\circ}C$, yeast and lactic acid bacteria were the major microflora after 1 day. After 11 days, Lactobacillus spp. were the major bacteria, while Bacillus spp. were the primary microflora after 21 days. At $25^{\circ}C$ the primary species were Bacillus spp., Leuconostoc spp., and Bacillus spp. after 1, 11 and 21 days, respectively. The $\alpha$-amylase activity of steeping water increased proportionally with the increase of temperature and period, while the protease activity tended to increase for 11 days, but decreased at 21 days. The enzyme activities of the steeped waxy rice, especially the $\alpha$-amylase activity, decreased with higher steeping temperature, while the $\beta$-amylase and glucoamylase activity remained constant for up to 21 days. Observation using scanning electron microscopy indicated that intracellular cleavage of steeped waxy rice endosperm resulted in a rough surface morphology due to the exposure of polygonal starch granules. Tiny pinholes on the surface of waxy rice starch granules were also observed after steeping for 11 days.

Effect of Germination Condition and Drying Methods on Physicochemical Properties of Sprouted Brown Rice (발아조건 및 건조방법이 발아현미의 이화학적 특성에 미치는 영향)

  • 김선림;손영구;손종록;허한순
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.3
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    • pp.221-228
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    • 2001
  • This study was carried out to investigate the effect of germination condition and drying temperature on growth and physicochemical properties of brown rice. Three brown rice seeds of Ilpumbyeo, Dasanbyeo and Heugjinjubyeo were stored at room temperature for six weeks to test the time-sequence germination viability. Relatively stable germination ratio was maintained until 2 weeks after storage. However, 3 weeks after storage, germination ratio of brown rice seeds started to decrease rapidly and their germination ratio was lower than 80%. For this reason, brown rice was recommended for seeding within 2 weeks after hulling. During the initial 5 days, germination ratio of 24 hours pre-soaking brown rice was higher about 2-3% than that of non-soaking brown rice. The $25^{\circ}C$ was considered as the most favorable temperature for brown rice germination, because of the high germination ratio and desirable coleoptile growth of the brown rice, and little seed rotting symptoms. The scanning electron micrographs showed the structural differences between hot-air dried and freeze dried germinated-brown rice kernel. In the freeze dried germinated-brown rice, seed coat (pericarp, tegmen and aleurone layer) was mechanically disrupted from the endosperm, and many cleavages were observed among starch storing cells and starch granules. The endosperm of freeze-dried brown rice kernels formed the sponge-like structures and showed the fragile traits. For this reason, hot-air drying is considered as more suitable method than freeze drying for germinated-brown rice. The crude protein and amylose contents were slightly changed, but there were no significant differences during the germination period. Crude fiber content was decreased, but crude Int and total amino acid contents were increased as seeding days increased. A rapid increase in $\alpha$-amylase activities of germinating brown rice was observed at S days after seeding, and $\alpha$-amylase activities were decreased from 8 days after seeding. Total free sugar contents were decreased during the germination period. There was continuous decline in the contents of sucrose and glucose until 8 days after seeding, but fructose and maltose content were gradually increased from the 5 days after seeding.

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Molecular Cloning and Characterization of Trehalose Biosynthesis Genes from Hyperthermophilic Archaebacterium Metallosphaera hakonesis

  • Seo, Ju-Seok;An, Ju-Hee;Baik, Moo-Yeol;Park, Cheon-Seok;Cheong, Jong-Joo;Moon, Tae-Wha;Park, Kwan-Hwa;Choi, Yang-Do;Kim, Chung-Ho
    • Journal of Microbiology and Biotechnology
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    • v.17 no.1
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    • pp.123-129
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    • 2007
  • The trehalose $({\alpha}-D-glucopyranosyl-[1,1]-{\alpha}-D-glucopyranose)$ biosynthesis genes MhMTS and MhMTH, encoding a maltooligosyltrehalose synthase (MhMTS) and a maltooligosyltrehalose trehalohydrolase (MhMTH), respectively, have been cloned from the hyperthermophilic archaebacterium Metallosphaera hakonesis. The ORF of MhMTS is 2,142 bp long, and encodes 713 amino acid residues constituting a 83.8 kDa protein. MhMTH is 1,677 bp long, and encodes 558 amino acid residues constituting a 63.7 kDa protein. The deduced amino acid sequences of MhMTS and MhMTH contain four regions highly conserved for MTSs and three for MTHs that are known to constitute substrate-binding sites of starch-hydrolyzing enzymes. Recombinant proteins obtained by expressing the MhMTS and MhMTH genes in E. coli catalyzed a sequential reaction converting maltooligosaccharides to produce trehalose. Optimum pH of the MhMTS/MhMTH enzyme reaction was around 5.0 and optimum temperature was around 70 C. Trehalose-producing activity of the MhMTS/ MhMTH was notably stable, retaining 80% of the activity after preincubation of the enzyme mixture at $70^{\circ}C$ for 48 h, but was gradually abolished by incubating at above $85^{\circ}C$. Addition of thermostable $4-{\alpha}-glucanotransferase$ increased the yield of trehalose production from maltopentaose by 10%. The substrate specificity of the MhMTS/MhMTH-catalyzed reaction was extended to soluble starch, the most abundant maltodextrin in nature.

Immobilization of Cyclodextrin Glucanotransferase for Production of 2-O-\alpha-D-Glucopyranosyl L-Ascorbic Acid. (2-O-\alpha-D-Glucopyranosyl L-Ascorbic acid 생산을 위한 Cyclodextrin glucanotransferase의 고정화)

  • 성경혜;김성구;장경립;전홍기
    • Microbiology and Biotechnology Letters
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    • v.31 no.4
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    • pp.368-376
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    • 2003
  • Cyclodextrin glucanotransferase (CGTase) from Paenibacillus sp. JB-13 was immobilized on various carriers by several immobilization methods such as ionic binding, covalent linkage and ultrafiltration to improve the process performance. The ultrafiltration and covalent linkage with CNBr-activated sepharose 4B were found as the best method for immobilization of CGTase. The ability of CGTase immobilization onto CNBr-activated sepharose 4B was as high as 18,000 units/g resin when the conditions was as follows: contact time 9 hrs at $37^{\circ}C$, pH 6.0, 100 nm and enzyme loading 24,000 units/g resin. The optimum conditions for production of 2-O-$\alpha$-D-Glucopyranosyl L-Ascorbic acid by immobilized CGTase turned out to be: pH 5.0, temperature $37^{\circ}C$, 20% substrate solution containing 8% (w/v) of soluble starch and 12% (w/v) of L-ascorbic acid sodium salt, 100 rpm, far 25 hrs and with 800 units of immobilized CGTase/ml substrate solution. Moreover the CGTase activity could be stably maintained for 8 times of repetitive reactions after removing products by ultrafiltration through YM 10 membrane.

Characterization of Carbohydrate Metabolism during Dark-Induced Senescence

  • Kim, Young-Sang;Chang, Ji-Young;Lee, Dong-Hee
    • Journal of Life Science
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    • v.11 no.1
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    • pp.14-21
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    • 2001
  • To investigate the changes of carbohydrate metabolism in the senescing leaves of Zea mays during dark-induced senescence, the changes in the contents of reducing sugar, sucrose and starch as well as the activities of sucrose synthase, three isozymes of invertase, and ${\alpha}$-amylase were measured. In the senescing leaves, the content of reducing sugars temporarily increased at 4 d and rapidly decreased thereafter, whereas sucrose contents gradually decreased thereafter, whereas sucrose contents gradually decreased until 3 d of senscence and significantly decreased thereafter. The activities of intracellular invertases such as soluble acid and alkaline formed gradually enhanced until 4 d of leaf senescence and significantly declined thereafter. The extracellular invertase activity showed no significant changes during leaf senescence. The deactivation of sucrose synthase was observed within 3 d of leaf senscence. On the other hand, the starch contents gradually declined during 2 d of leaf senescence, and showed a temporary increase at 3 d, which is similar to the pattern of sucrose synthase activity., These results imply that sucrose in the senescing leaves. The major enzymes which correlated to the breakdown of sucrose during dark-induced senescence were soluble acid and alkaline invertases, not sucrose and ABA accelerated leaf senescence by inducing the accumulation of reducing sugar. These result, therefore, that leaf senescence may be mediated by the temporary quantitative changes of reducing sugar induced by the activation of intracellualr inveertases.

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Evaluation of Immobilization Methods for Cyclodextrin Glucanotransferase and Characterization of its Enzymatic Properties

  • Lee, Sang-Ho;Shin, Hyun-Dong;Lee, Yong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.54-62
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    • 1991
  • Cyclodextrin glucanotransferase(CGTase) derived from Bacillus macerans was immobilized by (1) covalent linkage on chitosan and chitin with glutaraldehyde, (2) adsorption on DEAE-cellulose and Amberite IRA 900 after succinylation, and (3) entrapment on alginate and polyacrylamide by cross linking. Adsorption on Amberite IRA 900 and covalent linking on chitosan were identified to be the most suitable immobilization methods considering the yield of activity and stability of immobilized CGTase. The enzymatic properties of immobilized CGTase were investigated and compared with those of the soluble CGTase. Thermal stability of CGTase immobilized on chitosan was increased from 50 to $55^{\circ}C$, and the optimum temperature of CGTase immobilized on Amberite IRA 900 was shifted from 55 to $50^{\circ}C$. The effect of molecular size of soluble starch (substrate) on immobilized CGTase investigated using partially liquefied substrates with different dextrose equivalent(DE). Cyclodextrin(CD) conversion yield augmented according to the increase of DE level for immobilized CGTase on Amberite IRA 900. CD conversion yield of partially cyclized starch with soluble CGTase was higher compared with liquefied one with ${\alpha}-amylase$.

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Expression of Bacillus macerans Cyclodextrin Glucanotransferase in Bacillus subtilis

  • Kim, Chang-Sup;Han, Nam-Soo;Kweon, Dae-Hyuk;Seo, Jin-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.2
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    • pp.230-233
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    • 1999
  • A plasmid vector was constructed for the expression and secretion of Bacillus macerans cyclodextrin glucanotransferase (CGTase) in Bacillus subtilis. The vector, pUBACGT, was composed of the ribosome-binding sequence, signal sequence, and cgt gene from B. macerans under the control of amyR2, the promoter of amyE gene coding for $\alpha$-amylase from B. subtilis var. natto. Bacillus subtilis LKS88, a mutant strain lacking genes for an amylase and two proteases, was used as a host for the transformation of the plasmid vector. The transformants were selected on kanamycin-containing Luria-Bertani plates. The starch hydrolyzing activity was observed on the starch-containing plates by the iodine method and cyclodextrin-forming activity was detected in the culture medium. A SDS-PAGE analysis showed that most of the expressed CGTase in the recombinant B. subtilis was secreted into the medium at a high expression level.

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