• Title/Summary/Keyword: Starch gel electrophoresis

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Evolutionary Study on the Dark Chub (Zocco temmincki) IX. Phylogeny of the Subfamily Danioninae (Pisces, Cyprinidae) (갈겨니(Zacco temmincki)의 진화에 관한 연구 IX. 피라미아과 어류의 계통진화)

  • 양서영;민미숙
    • The Korean Journal of Zoology
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    • v.32 no.4
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    • pp.393-403
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    • 1989
  • Using starch gel electrophoresis of proteins, we examined variation at 24 genetic loci in five species (three general of the Subfamily Danioninae (Class Pisces, Family Cyprinidae) inhabiting In Korea. The average degree of genic variation of the Subfamily Danioninae (A=1.8, P=16.5%, HD=0.043, HG=0.049) was far less than that of the fresh water fishes in general. But the degree of genic variation of A. chinensis (P=32.5 HD=0.100, HG=0.119) was highe, than other fishes and the value of O. biders was the lowest (P=4.2%, HD=0.008, HG=0.008). The average genetic similarities among the populations in each species of A. chinensis, Z. platypus, Z. temmincki MS type, Z. temmincki W type, and O. widens were S=0.902, 0.899, 0.990, 0.914, and 0.992, respectively. The average genetic similarity of A. chinensis to the other species was the lowest value of S=0.235, whereas that between two species of Z. temmincki was the highest of S=0.822. Genetically, Z. plypus is more similiar to O. widens than ho species of the same genus Zacco. Based on estimated divergent time of 5 species of the Subfamily Danioninae (7.6 to 0.9 million years) it is assumed that they are speciated during late Pliocene to middle Pleistocene epoch.

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Characteristics of Biochemical Markers and Whole-Wheat Flours Using Small-Scaled Sampling Methods in Korean Wheats

  • Park Chul Soo;Kim Yang-Kil;Han Ouk-Kyu;Lee Mi Ja;Park Jong-Chul;Seo Jae-Hwan;Hwang Jong-Jin;Kim Jung-Gon;Kim Tae Wan
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.50 no.5
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    • pp.346-355
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    • 2005
  • To investigate the application of biochemical markers' and small-sample methods using whole-wheat flours for screening in early generation in Korean wheat breeding system, 74 Korean wheats, including cultivars, local breeding lines and experimental lines, were analyzed. Seed storage protein and amylose contents of grains were evaluated. Biochemical makers, including granule bound starch synthase (GBSS), high molecular weigh glutenin subunits (HMW-GS) and friabilin were also evaluated by using one-dimensional sodium dodecyl sulfate-polyacryla-mide gel electrophoresis with a single kernel. The small­sample methods, including modified SDS-sedimentation test (MST), micro-alkaline water retention capacity (AWRC) and whole-wheat flour swelling volume (WSV) were also tested in this study. Protein content, MST and AWRC was $11.0 - 15.8\%$, 2.7 - 26.2 ml and $71.9 - 109.7\%$, respectively. Apparent and total amylose content and WSV was $20.6 - 25.0\%$, $26.1 - 32.4\%$ and 9.0 - 16.9 ml, respectively. There were highly significant correlations between MST and AWRC (r=0.592, P<0.001), but Korean wheats showed no significant difference in protein content, amylose content and small-sample methods. In the biochemical markers, Korean wheats contained all three GBSS encoded by Wx loci, except for Suwon 252. Korean wheats showed the high frequency ($58.1\%$) of 1Dx2.2 + 1Dy12 subunits of HMW-GS. Friabilin band was present in 46 lines ($62.2\%$) and absent in 28 lines ($37.8\%$). Friabilin-absence lines showed the higher MST (14.9 ml) and AWRC ($92.1\%$) value than friabilin-presence lines (8.5 ml and $82.4\%$, respectively).

Studies on $\alpha$-amylase of Bocillus circulans F-2 (Part II) Enzymatic characteristics of the purified $\alpha$-amylase (Bacillus circulans F-2가 생산하는 $\alpha$-amylase에 관한 연구 (제 I I 보) 정제$\alpha$-amylase의 효소적특성)

  • ;Hajime Taniguchi;Yoshiharu Maruyama
    • Microbiology and Biotechnology Letters
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    • v.10 no.2
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    • pp.123-132
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    • 1982
  • These experiments were conducted to investigate the enzymatic characteristics of the purified $\alpha$-amylase (F-2A) of Bacillus circulans F-2 and the digestion rate of various starches. 1. The molecular weight was estimated to be 93000 by SDS-polyacrylamide disc gel electrophoresis. The isoelectric point was about pH 5.0. The optimum pH for the enzyme action was 6.0-6.5 and the stable pH ranged pH 5.5-12.0. The optimum temperature was 6$0^{\circ}C$, and the purified $\alpha$-amylase was stable below 4$0^{\circ}C$. 2. The purified $\alpha$-amylase was activated by Mn$^{++}$ and Co$^{++}$, whereas it was inhibited by Ag$^{+}$, HT$^{++}$, Cu$^{++}$ and Pb$^{++}$. 3. The purified $\alpha$-amylase is considered to have no sulfhydryl residue essential for its catalytic activity. 4. Michaelis constant (Km) was 1.704 mg/$m\ell$. Activation energy between 25-4$0^{\circ}C$ was 12.297 Kcal/mole, and between 40-6$0^{\circ}C$, it was 7.831 Kcal/mole. 5. The hydrolysis product from soluble starch, amylose and amylopectin in the early stage of hydrolysis was G$_{6}$, and as hydrolysis proceeds, G$_4$and G$_2$appeared. 6. Products from each oligosaccarides are as follows: G$_4$longrightarrow G$_2$+ G$_2$,G$_3$ +G$_1$,G$_{5}$longrightarrow G$_4$+G$_1$,G$_{6}$longrightarrowG$_4$+ G$_2$,G$_{7}$ G$_4$,G$_{8}$longrightarrow G$_4$+G$_4$, 7. On raw potato starch, raw sago starch and raw yam starch, the purified enzyme exhibited a remarkably high digestion rate than Porcine pancreatic amylase and Streptococcus bovis amylase.

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Characterization of Sporulation-Specific Glucoamylase of Saccharomyces diastaticus (Saccharomyces diastaticus의 포자형성 특이 글루코아밀라제의 특성)

  • Kim, Eun-Ju;Ahn, Jong-Seog;Kang, Dae-Ook
    • Journal of Life Science
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    • v.20 no.5
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    • pp.683-690
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    • 2010
  • The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

Studies on the Effects of Copper on the Lactate Dehydrogenase and Esterase Isozymes in Various Tissues of Carassius carassius (붕어(Carassius carassius)의 조직내 젖산수소이탈효소와 에스테라아제 아이소자임에 미치는 동의 영향에 관한 연구)

  • Lee, Choon-Koo;Choo, Il-Young
    • The Korean Journal of Zoology
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    • v.16 no.2
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    • pp.79-96
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    • 1973
  • In order to elucidate the effects of copper on Corassius carassius, the following were studied: 1) lactate dehydrogenase isozyme patterns by cellulose acetate electrophoresis, 2) LDH activity and copper effect on LDH enzyme system y spectrophotometry, 3) esterase isozyme patterns by agar thin layer electrophoresis, 4) hemoglobin patterns by starch gel electrophoresis, and 5) histological study. 1. There were two bands of LDH isozymes (LDH-3 and LDH-5) in the gill, three bands (LDH-2, LDH-4, and LDH-5) in the liver, and two bands (LDH-3 and LDH-4) in the muscle of the normal fish. The LDH-1 bond was not found in the above three tissues. When the fish were exposed to copper, LDH-3 appeared in the liver, LDH-5 in the muscle, but no new LDH band appeared in the gill. 2. The sepcific activities of the LDH were lowest in the gill and highest in the muscle of the normal fish, and they were gradually decreassed in the gill and highest in the muscle of the normal fish, and they were gradually decreased in the liver and mucle except in the gill from 1-day to 10-day exposure to copper. It indicates that LDH activities in the liver and muscle of the fish were inhibited by copper. 3. Through in vitro experiment, it is clear that the decrease of the LDH activities of the liver and muscle of the fish exposed to copper is mainly caused by the inhibition on the M-LDH in the fish. 4. The numbers of the esterase isozyme bands of the gill, liver, muscle, blood, brain, and kidney of the normal fish were 3, 6, 2, 2, 2, and 2 respectively, and these numbers were the same as those exposed to copper. The relative mobilities of the esterase bands in the gill, liver, blood, and kidney of the exposed group were different from those of the control. 5. There was one hemoglobin band on the anode in the normal fish. It seems that the nobility of hemoglobin band of the fish exposed to copper was slightly faster than that of the normal fish. 6. The normal gill lamellae of the fish consisted of centrally located pillar cells and a number of mucus cells. When the fish were exposed to copper, the epithelial layer was divorced first, disintegrated, and then destroyed completely. 7. The liver of the normal fish had prominent central veins, cords of hepatic cells, and sinusoids. When the fish were exposed to copper, numerous droplets of fat appeared in the cells around the central vein of the liver. It is assumed that the fatty droplets were accumulated by the lesion due to fatty metamorphosis of the liver caused by copper. 8. There was no histological difference between the muscle of the normal fish and that of the fish exposed to copper. 9. In the normal fish, the tubules of the kidney were surrounded by hemopoetic tissues. However, the kidney tissue of the fish exposed to copper received some damage on the proximal tubules. Since the tubule cells were reduced in height, the lumens of the tubules were enlarged. Consequently many proximal tubules exhibited some pink-stained granular casts and various stages of degeneration.

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Bacterial Community Dynamics during Swine In vitro Fermentation Using Starch as a Substrate with Different Feed Additives for Odor Reduction

  • Alam, Md.J.;Jeong, C.D.;Mamuad, L.L.;Sung, H.G.;Kim, D.W.;Cho, S.B.;Lee, K.;Jeon, C.O.;Lee, Sang-S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.5
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    • pp.690-700
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    • 2012
  • The experiment was conducted by in vitro fermentation and bacterial community analysis to investigate the reduction of odorous compounds in response to the use of feed additives (FA) during carbohydrate overload in growing pigs. Soluble starch at 1% (control) and various FA at 0.1% Ginseng meal (FA1); Persimmon leaf (FA2); Gingko nut (FA3) and Oregano lippia (FA4) were added to fecal slurry and incubated anaerobically for 12 and 24 h. In vitro parameters and microbial diversity of the dominant bacteria following fermentation were analyzed using Denaturing Gradient Gel Electrophoresis (DGGE), band cloning and sequencing of the V3 region. Results showed that total gas production increased with the advancement of incubation (p<0.05). pH values of FAs and control groups were decreased except the FA4 group which increased somewhat from 12 to 24 h (p<0.05). Ammonia nitrogen ($NH_3$-N) and $H_2S$ gas concentrations were comparatively lower in both stages in FA4 treatment than in the other groups (p<0.05). Hence, $NH_3$-N concentrations in liquid phases were increased (p<0.05) from 12 to 24 h, but the trend was lowest in FA4 than in the other groups at both stages. The total VFA production was comparatively lower and butyrate levels were moderate in FA4 group than in the the other groups during both stages (p<0.05). Indirect odor-reducing compounds such as $NO_2$, $NO_3$ and $SO_4$ concentrations were higher in the FA4 and FA3 than in the other groups at 24 h (p<0.05). After fermentation, ten dominant bands appeared, six of which appeared in all samples and four in only the FA4 treated group. The total number of DGGE bands and diversity was higher in the FA4-group compared to other groups. Additionally, similarity indices were lowest (71%) in the FA4, which represented a different bacterial community compared with the other groups. These findings indicate that $NH_3$-N, $H_2S$ and VFA production was minimal, and pH was also better in the FA4 group than in the other groups. Furthermore, the conversion of odor-reducing indirect compounds or their intermediates was higher in the FA4 group in compared to the other groups. FA4 group generated less odorous products and more indirect products by in vitro fermentation at 24 h, and their microbial pattern appeared to differ from that of the other groups. These findings suggest that this particular FA could change the microbial population, which may have a beneficial effect on odor reduction. It is recommended that the oregano lippia may be supplied to growing pigs as FA along with excess carbohydrate sources to reduce the production of odorous compounds.

Genetic Diversity and Population Structure of Codium fragile (SURINGAR) HARlOT in Korea Using Allozymes (알로자임을 이용한 청각의 유전적 다양성과 집단구조)

  • Lee Bok-Kyu;Park So-Hye;Heo Youn-Seong;Ju Mu-Teol;Choi Joo-Soo;Huh Man-Kyu
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.213-218
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    • 2006
  • The study of genetic diversity and population structure was carried out in the Codium fragile using allozyme analysis. Although this species has been regarded as a ecologically and economically important source, there is no report on population structure in Korea. Starch gel electrophoresis was used to investigate the allozyme variation and genetic structure of four Korean populations of this species. Of the 15 genetic loci surveyed, nine (60.0%) was polymorphic in at least one population. Genetic diversity was high at the species level ($H_{ES}$=0.144), and, that of the population level was relatively low ($H_{EP}$=0.128). Nearly 87% of the total genetic diversity in C. fragile was apportioned within populations. The predominant asexual reproduction, population fragmentation, low fecundity, geographic isolation and colonization process are proposed as possible factors contributing to low genetic diversity in this species. The indirect estimated of gene flow based on $G_{ST}$ was 1.69. The moderate level of gene flow in C. fragile populations is mainly caused by thallus developed from isolated utricles dispersal via sea current.

Genetic Analysis of Some Polymorphic Isozymes in Pinus densiflora(II) - Inheritance of acid phosphatase, alcohol dehydrogenase and catalase isozymes - (소나무의 몇가지 다형적(多形的) 동위효소(同位酵素)의 유전분석(遺傳分析)(II) - Acid phosphatase, alcohol dehydrogenase와 catalase 동위효소(同位酵素)의 유전양식(遺傳樣式) -)

  • Kim, Z.S.;Hong, Y.P.
    • Journal of Korean Society of Forest Science
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    • v.68 no.1
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    • pp.32-36
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    • 1985
  • Megagametophyte tissues of Pinus densiflora were subjected to study the inheritance of acid phosphatase (ACP), alcohol dehydrogenase (ADH) and catalase (CAT) isozymes by starch gel zone-electrophoresis. At least three or four zones were segregated for ACP isozyme. However, as one isozyme of ACP-A zone was separated clearly, only that isozyme was analysed. Five isozyme phenotypes (A1-A5), observed in ACP-A zone, were segregated to a simple Mendelian ratio, suggesting that these are controlled by five codominant alleles existed at ACP-A locus. Two zones of activity were segregated in the gels after staining for ADH, the more anodal zone (ADH-A) of the two was invariant in our materials. Three isozyme phenotypes (B1-B3) were observed in ADH-B zone and these variants showed a 1:1 segregation pattern, suggesting that each variant is controlled by three codominant alleles at ADH-B locus. A total of five isozyme phenotypes, composed of multiple bands, were observed in CAT isozyme. The segregation of these phenotypes in heterozygous trees did not show any significant deviation from a 1:1 segregation. Therefore, the genetic control of CAT isozyme in Pinus densiflora seeds seems to be based on a single locus (CAT-A) with Five codominant alleles ($A_1-A_5$).

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Genotypic Variation of Esterase Isozyme in Breeding Lines of Two-rowed Barley by Electrophoretic Banding Pattern (전기영동 패턴에 의한 2조보리 계통의 Esterase 동위효소 유전자형 변이분석)

  • 박광근;최홍집;이종호;서세정;김재철;남중현;김상효
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.47 no.6
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    • pp.465-470
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    • 2002
  • This study presents results of electro-phoretically detectable isozyme variation in Crossing Block (CB) lines of two-rowed barley maintained by the National Crop Experiment Station. The specific objectives were to determine allelic frequencies at the four Est loci(Est1, Est2, Est4, and Est5) and their distribution over 380CB lines of two-rowed barley. A total of 17 alleles were detected over the four Est loci in these lines. There were 4 alleles (Pr, Al, Ca, and Af at the Est1 locus and their frequencies were 69.7, 1.1, 28.4, and 0.8%, respectively. At the Est2 locus, 5 different alleles (Dr, Fr, Sp, Un and a recessive null allele) were detected and their frequencies were 2.9,84.5,0.5,2.1, and 10%, respectively. four alleles (Nz, Su, At, and null were detected at the Est4 locus and the allelic frequency of Su was about 84%. Four alleles(Mi, Pi, Te, and a null allele(od)) were detected at the Est5 locus and their frequencies were 34.2, 61.0, 2.4, and 2.4%, respectively. Based on the allelic frequencies over the four Est loci, 380 CB lines were classified into 25 genotypes. The most frequent genotypes were G1(Pr-Fr-Su-Mi) and G2(Pr-Fr-Su-Pi), and their frequencies were 28.1 and 39.5%, respectively. The frequencies of other genotypes were less than 10%.

The Roles of Lipid Supplements in Ethanol Production Using a Continuous Immobilized and Suspended Cell Bioreactor (연속식 고정화 및 현탁 세포 생물 반응기에 의한 에탄을 생성중 지질 첨가 영향)

  • Gil, Gwang-Hoon
    • Applied Biological Chemistry
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    • v.39 no.1
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    • pp.1-8
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    • 1996
  • A one-stage, continuous-flow bioreactor with both immobilized and suspended cells was used to investigate the roles of lipid supplements in ethanol production by Saccharomyces cerevisiae. The reactor performance and the level of alcohol dehydrogenase(ADH) activities of the suspended cells, grown under various conditions, were measured. When ergosterol and/or oleic acid were added with surfactants to the yeast culture grown under non-aerated conditions, remarkable increases in ethanol production and cell growth was achieved, but specific ADH activities were not affected. Especially, no difference of specific ADH activities of the suspended cells grown under aerated and non-aerated condition was observed. The addition of the surfactant as a supplement also resulted in significant increases in ethanol production, cell growth, and specific ADH activity. When ergosterol and oleic acid were added to the yeast culture exposed to higher ethanol concentration($>40\;g/{\ell}$) level, ethanol production, cell growth, and specific ADH activity were increased, but the addition of surfactant was as effective as at lower ethanol concentration level. The results indicated that lipid supplements were more effective at higher ethanol concentration level than at lower ethanol concentration level during ethanol production. ADH isozyme patterns of the yeast cultures grown under various conditions on starch gel electrophoresis showed only one major band, probably ADH I. The migrating distance of the major isozyme, however, varied slightly according to the culture conditions of the cells. No apparent correlation was found between specific ADH activity and amount of ethanol produced. Cell mass was more important factor for ethanol production than specific ADH activity of the cells.

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