• Title/Summary/Keyword: debranching

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Molecular Structural Properties of Legume Starches (두류 전분의 분자구조적 특성)

  • Kweon, Mee-Ra;Ahn, Seung-Yo
    • Korean Journal of Food Science and Technology
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    • v.25 no.3
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    • pp.264-269
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    • 1993
  • Molecular structural properties of legume starches were investigated. In intrinsic viscosity and degree of Polymerization of amylose and amylopectins, cow pea and mung bean were high, but kidney bean was low. Low molecular weight fractions for kidney bean starch were much eluted by gel chromatography. In the elution profiles of their amylose by Sepharose 2B-CL, molecular weight of kidney bean amylose was smaller than that of other amylose Molecular weights of cow pea and mung bean amyloses were large, but that of kidney bean amylose was small and red bean amylose was medium. The elution profiles by Sephadex G-50 after debranching amylopectins with pullulanase showed similar patterns.

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Characterization of amylopectins isolated from dry and moist type sweet potato starch (분질 및 점질 고구마 전분의 아밀로펙틴 특성)

  • Kim, Sung-Ran;Ahn, Seung-Yo
    • Applied Biological Chemistry
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    • v.34 no.1
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    • pp.26-31
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    • 1991
  • The physicochemical properties of starches from dry(Suwon 147) and moist type(Hwangmi) sweet potato were investigated and molecular structural properties of their amylopectins were also studied by gel chromatography. Suwon 147 starch bad lower swelling power and higher gelatinization temperature than Hwangmi starch. $\beta-Amylolysis\;limit(%)$ of Suwon 147 and Hwangmi amylopectin were 57.6% and 57.0%, respectively. Average unit chain length of amylopectins were 24.8 glucose units for Suwon 147 and 21.9 for Hwangmi. The elution profiles by Sephadex G-50 after debranched amylopectins of the two starches were similar but DPs of each peak were different.

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Glucose Chain Length Distribution of Starches from Endosperm Mutant Rices and Its Relationship with Adaptability in Rice Bread Processing (변이체벼 배유 전분분자의 포도당 사슬길이 분포와 쌀빵 가공성간의 상관관계)

  • Kang, Mi-Young;Han, Ji-Yeun
    • Korean Journal of Food Science and Technology
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    • v.33 no.1
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    • pp.50-54
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    • 2001
  • The amylose content, based on iodine blue value, of eight rice cultivars decreased in order of Nampungbyeo>Whachungbyeo>Punchilmi>Nampung CB243>Whachung du-1>Nampung EM90>Whachungchalbyeo>shr. The amylopectin chain length distribution was obtained by enzyme treatments followed by high-performance size-exclusion chromatographic separation. Chain length distribution profiles of the isoamylase-debranched starches showed distinct patterns according to cultivars. Based on the sensory evaluation result of the bread prepared from gluten and rice flours of eight rice cultivars, chewiness of the product was related with the presence of amylose while the short-chain amylopectin fraction was contributed to the texture and overall quality.

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Characterization of PUL Haplotypes and Its Evolutionary Analyses in Korean Rice Accessions

  • Thant Zin Maung;Yong-Jin Park
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.217-217
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    • 2022
  • Pullulanase (PUL), a debranching enzyme, has been utilized in hydrolyzing the a-1,6 glucosidic linkages in starch, amylopectin, pullulan, as well as related oligosaccharides. It has also been indicated that PUL is a novel indicator of inherent RS (Resistant Starch) formation in rice. In this study, we performed haplotype analysis on 320 bred rice accessions, and additional 54 wild accessions were added to study genetic diversity along with other population-based analyses of the PUL gene. Through these investigations, we summarized a total of 10 functional (non-synonymous) SNPs from 7 different exons on chromosome 4. There were 10 haplotypes, of which only six haplotypes were functional, implicating different subpopulations. Diversity reduction was noticed in temperate japonica (0.0005) compared to the highest one (aus, 0.0154), illustrating their higher genetic differentiation by FST-value (0.926). The highest Tajima^ D value was observed in indica (3.6613), indicating PUL gene domestication signature under balancing selection, while the lowest Tajima's D value was found in temperate japonica (-2.2191) which might have undergone under positive selection and purified due to the excess of rare alleles. PCA, population structure, and phylogenetic analyses provide information on the genetic relatedness between and or among the cultivated subpopulations and the wild based on PUL genomic region.

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The New Neurobiology of Depression (우울증의 새로운 신경생물학)

  • Kim, Yong Ku
    • Korean Journal of Biological Psychiatry
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    • v.8 no.1
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    • pp.3-19
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    • 2001
  • Recent basic and clinical studies demonstrate a major role for neural plasticity in the etiology and treatment of depression and stress-related illness. The neural plasticity is reflected both in the birth of new cell in the adult brain(neurogenesis) and the death of genetically healthy cells(apoptosis) in the response to the individual's interaction with the environment. The neural plasticity includes adaptations of intracellular signal transduction pathway and gene expression, as well as alterations in neuronal morphology and cell survival. At the cellular level, repeated stress causes shortening and debranching of dendrite in the CA3 region of hippocampus and suppress neurogenesis of dentate gyrus granule neurons. At the molecular level, both form of structural remodeling appear to be mediated by glucocorticoid hormone working in concert with glutamate and N-methyl-D-aspartate(NMDA) receptor, along with transmitters such as serotonin and GABA-benzodiazepine system. In addition, the decreased expression and reduced level of brain-derived neurotrophic factor(BDNF) could contribute the atrophy and decreased function of stress-vulnerable hippocampal neurons. It is also suggested that atrophy and death of neurons in the hippocampus, as well as prefrontal cortex and possibly other regions, could contribute to the pathophysiology of depression. Antidepressant treatment could oppose these adverse cellular effects, which may be regarded as a loss of neural plasticity, by blocking or reversing the atrophy of hippocampal neurons and by increasing cell survival and function via up-regulation of cyclic adenosine monophosphate response element-binding proteins(CREB) and BDNF. In this article, the molecular and cellular mechanisms that underlie stress, depression, and action of antidepressant are precisely discussed.

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Genetically Engineered Yeast by Heterologous Transformation and Intergeneric Two-Step Protoplast Fusion for Ethanol Fermentation

  • Kim, Young-Ho;Lee, Jae-Ran;Seu, Jung-Hwn
    • Journal of Microbiology and Biotechnology
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    • v.3 no.4
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    • pp.232-237
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    • 1993
  • A strain of yeast which can convert starch directly to ethanol was developed by the intergeneric protoplast fusion between Schwanniomyces alluvius possessing $\alpha$ amylase as well as glucoamylase with debranching activity and FSC-14-75 which previously had been formed from a heterologous transformation and subsequent intergeneric protoplast fusion. Fusants were selected on minimal medium after protoplasts of auxotrophic mutant of S. alluvius fused with heat-treated protoplasts of FSC-14-75 in the presence of 30%(w/v) PEG and 20 mM $CaCl_2$. The fusion frequency was in the range of $10^{-6}$ order. All fusants tested were intermediate types of parental strains for carbon compound assimilation, and their cell volumes were approximately 1.1 times larger than FSC-14-75 and 1.8 times larger than S. alluvius. The fusants were unable to sporulate like FSC-14-75, while S. alluvius could sporulate. In flask scale the most promising fusant, FSCSa-R10-6, produced 7.83%(v/v) and 10.17%(v/v) ethanol from 15% and 20% of liquefied potato starch, respectively, indicating that the fermetation efficiency of each case increased 1.2 times and 1.6 times than that of FSC-14-75. The elution pattern on DEAE-cellulose chromatography showed that FSCSa-R10-6 has four distinct amylase peaks of which two peaks originated from S. alluvius and the other two from FSC-14-75. These results suggest that the enhanced fermentation efficiency of the fusant might be due to almost-complemented parental amylases.

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The Fine Structure of Amylopectin and Physicochemical Properties of Starch Granules from Endosperm Varieties in Glutinous Rice (찹쌀 전분의 미세구조 및 이화학적 특성 비교)

  • Sung, You-Me;Nam, Seok-Hyun;Kang, Mi-Young
    • Applied Biological Chemistry
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    • v.43 no.1
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    • pp.18-23
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    • 2000
  • stract : Fourteen varieties of glutinous rices were examined on amylopectin fine structure and physicochemical properties of starch granules. The amylopectin chain length distribution and short chain/long chain ratio were investigated by enzymatic treatments followed by high-performance size-exclusion chromatographic separation. Chain length distribution profiles of the isoamylase-debranched amylopectins showed distinct patterns according to varieties. Beongok showed the highest short chain/long chain ratio, while TP2579A1 showed the lowest one. Sharebyeo-152-1-B showed the highest hydrolysis rate to 15% $H_2SO_4$, while Sandong 47 showed the lowest one. Fourteen varieties of rice starch granules showed A-type pattern on X-ray diffractograms. Non-gelitinized starch granules from Keochang 1 and Beongok had almost 100% hydrolysed by glucoamylase for 3 hrs at $370^{\circ}C$.

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Metagenomic Insight into Lignocellulose Degradation of the Thermophilic Microbial Consortium TMC7

  • Wang, Yi;Wang, Chen;Chen, Yonglun;Chen, Beibei;Guo, Peng;Cui, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.31 no.8
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    • pp.1123-1133
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    • 2021
  • Biodegradation is the key process involved in natural lignocellulose biotransformation and utilization. Microbial consortia represent promising candidates for applications in lignocellulose conversion strategies for biofuel production; however, cooperation among the enzymes and the labor division of microbes in the microbial consortia remains unclear. In this study, metagenomic analysis was performed to reveal the community structure and extremozyme systems of a lignocellulolytic microbial consortium, TMC7. The taxonomic affiliation of TMC7 metagenome included members of the genera Ruminiclostridium (42.85%), Thermoanaerobacterium (18.41%), Geobacillus (10.44%), unclassified_f__Bacillaceae (7.48%), Aeribacillus (2.65%), Symbiobacterium (2.47%), Desulfotomaculum (2.33%), Caldibacillus (1.56%), Clostridium (1.26%), and others (10.55%). The carbohydrate-active enzyme annotation revealed that TMC7 encoded a broad array of enzymes responsible for cellulose and hemicellulose degradation. Ten glycoside hydrolases (GHs) endoglucanase, 4 GHs exoglucanase, and 6 GHs β-glucosidase were identified for cellulose degradation; 6 GHs endo-β-1,4-xylanase, 9 GHs β-xylosidase, and 3 GHs β-mannanase were identified for degradation of the hemicellulose main chain; 6 GHs arabinofuranosidase, 2 GHs α-mannosidase, 11 GHs galactosidase, 3 GHs α-rhamnosidase, and 4 GHs α-fucosidase were identified as xylan debranching enzymes. Furthermore, by introducing a factor named as the contribution coefficient, we found that Ruminiclostridium and Thermoanaerobacterium may be the dominant contributors, whereas Symbiobacterium and Desulfotomaculum may serve as "sugar cheaters" in lignocellulose degradation by TMC7. Our findings provide mechanistic profiles of an array of enzymes that degrade complex lignocellulosic biomass in the microbial consortium TMC7 and provide a promising approach for studying the potential contribution of microbes in microbial consortia.

Characterization of the Transglycosylation Reaction of 4-α-Glucanotransferase (MalQ) and Its Role in Glycogen Breakdown in Escherichia coli

  • Nguyen, Dang Hai Dang;Park, Sung-Hoon;Tran, Phuong Lan;Kim, Jung-Wan;Le, Quang Tri;Boos, Winfried;Park, Jong-Tae
    • Journal of Microbiology and Biotechnology
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    • v.29 no.3
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    • pp.357-366
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    • 2019
  • We first confirmed the involvement of MalQ (4-${\alpha}$-glucanotransferase) in Escherichia coli glycogen breakdown by both in vitro and in vivo assays. In vivo tests of the knock-out mutant, ${\Delta}malQ$, showed that glycogen slowly decreased after the stationary phase compared to the wild-type strain, indicating the involvement of MalQ in glycogen degradation. In vitro assays incubated glycogen-mimic substrate, branched cyclodextrin (maltotetraosyl-${\beta}$-CD: G4-${\beta}$-CD) and glycogen phosphorylase (GlgP)-limit dextrin with a set of variable combinations of E. coli enzymes, including GlgX (debranching enzyme), MalP (maltodextrin phosphorylase), GlgP and MalQ. In the absence of GlgP, the reaction of MalP, GlgX and MalQ on substrates produced glucose-1-P (glc-1-P) 3-fold faster than without MalQ. The results revealed that MalQ led to disproportionate G4 released from GlgP-limit dextrin to another acceptor, G4, which is phosphorylated by MalP. In contrast, in the absence of MalP, the reaction of GlgX, GlgP and MalQ resulted in a 1.6-fold increased production of glc-1-P than without MalQ. The result indicated that the G4-branch chains of GlgP-limit dextrin are released by GlgX hydrolysis, and then MalQ transfers the resultant G4 either to another branch chain or another G4 that can immediately be phosphorylated into glc-1-P by GlgP. Thus, we propose a model of two possible MalQ-involved pathways in glycogen degradation. The operon structure of MalP-defecting enterobacteria strongly supports the involvement of MalQ and GlgP as alternative pathways in glycogen degradation.

Heterologous Expression of a Thermostable α-Galactosidase from Parageobacillus thermoglucosidasius Isolated from the Lignocellulolytic Microbial Consortium TMC7

  • Wang, Yi;Wang, Chen;Chen, Yonglun;Cui, MingYu;Wang, Qiong;Guo, Peng
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.749-760
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    • 2022
  • α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca2+ and Mg2+ promoted the activity of T26GAL while Zn2+ and Cu2+ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.