• Title/Summary/Keyword: Pyrophosphorylase

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Sucrose Synthase, UDP-glucose pyrophosphorylase and ADP-glucose Pyrophosphorylnse in Korea Ginseng Roots

  • Yelena V.Sundukova;Lee, Mi-Ja;Park, Hoon
    • Journal of Ginseng Research
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    • v.24 no.2
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    • pp.83-88
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    • 2000
  • The seasonal variation in the activity of sucrose synthase, ADP-glucose pyrophosphorylase and UDP-glucose pyrophosphorylase in roots of Panax ginseng C.A.Meyer have been studied. It was revealed that sucrose synthase and ADP-glucose pyrophosphorylase are adaptive enzymes and can serve as markers of sink strength, while UDP-glucose pyrophosphorylase is the maintenance enzyme. The average day temperature exceeded 24。C appeared to cause the disturbance in refilling process, affecting the starch synthesis. Study on the dependence of oxygen consumption in stele tissue with temperature revealed the sharp accelerating of this process after 24。C.

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Changes in the Expression of ADP-Glucose Pyrophosphorylase Genes During Fruit Ripening in Strawberry

  • Park, Jeong-Il;Kim, In-Jung
    • Food Science and Biotechnology
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    • v.16 no.3
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    • pp.343-348
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    • 2007
  • Starch contents play important roles in determining the fruit quality. Stawberry accumulates starch in the early stages and then mobilized into soluble sugars during fruit ripening. To date the molecular studies on the ADP-glucose pyrophosphorylase (AGPase), a key enzyme of starch biosynthesis, were not reported. cDNAs encoding small (FagpS) and large (FagpL1 and FaspL2) AGPase subunits were isolated from strawberry (Fragaria ${\times}$ ananassa Duch. cv. Niyobou). Both FagpS and FagpL1 cDNAs have open reading frames deriving 55-58 kDa polypeptides, where FagpL2 contains a partial fragment. Sequence analyses showed that FagpS has a glutamate-threonine-cysteine-leucine (ETCL) instead of a glutamine-threonine-cysteine-leucine (QTCL) motif found in all the dicot plants except for Citrus. In fruits, FagpS and FagpL1 were expressed in all stages with a little change in the amounts of transcripts. In the case of FagpL2, we were not able to detect any signal from all stages of fruit development and all tissues except for very a weak signal from the leaf. The results indicate that FagpL1 and FagpL2 show ubiquitous and leaf-specific expression patterns, respectively. The studies suggest that the starch contents in strawberry might be controlled by the expression of AGPase gene at both the transcriptional and post-transcriptional levels during fruit development.

Isolation and Nucleotide Sequence Analysis of ADP-glucose Pyrophosphorylase gene from Chinese cabbage (Brassica rapa L.)

  • Kim, In-Jung;Park, Jee-Young;Lee, Young-Wook;Chung, Won-Il;Lim, Yong-Pyo
    • Journal of Plant Biotechnology
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    • v.4 no.2
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    • pp.59-65
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    • 2002
  • ADP-glucose pyrophosphorylase (AGPase) catalyzes the key regulatory step in starch biosynthesis. Two cDNA clones encoding AGPase subunits were isolated from the leaf cDNA library of Chinese cabbage (Brassica campestris L. spp. pekinensis). One was designated as BCAGPS for the small subunit and the other as BCAGPL for the large subunit. Both cDNAs have uninterrupted open reading frames deriving 57 kDa and 63 kDa polypeptides for BCAGPS and BCAGPL, respectively, which showed significant similarity to those of other dicot plants. Also, However, the deduced amino acid sequence of BCAGPL has a unique feature. That is, it contains two regions (Rl and R2) lacking in all other plant enzymes. This is the first report of BCAGPL containing Rl and R2 among plant large subunits as well as small subunits. From the genomic Southern analysis and BAC library screening, we inferred the genomic status of BCAGPS and BCAGPL gene.

Impact of low temperature during ripening stage, amylose content and activities of starch biosynthesis in rice endosperm

  • Baek, Jung-Sun;Hwang, Woon-Ha;Jeong, Han-Yong;An, Sung-Hyun;Jeong, Jae-Heok;Lee, Hyeon-Seok;Yoon, Jong-Tak;Choi, Kyung-Jin;Lee, Gun-Hwi
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.229-229
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    • 2017
  • This research study was conducted to analyze the characteristics of different rice cultivars in abnormal temperature conditions (low temperature) for ripening period abnormalities, and to investigate the physiological causes behind the abnormalities. Four Korean high quality japonica-type rice cultivars, Jinbu (JB), Junamjosaeng (JJ), Geumyoung (GY), Hwawang (HW) were used in the experiment. The following day after flowering, they were then moved into two phytotrons under natural daylight with 65% RH but controlled at different temperatures - one at $19/29^{\circ}C$ (night/day) and the other at $13/23^{\circ}C$ as the low - temperature study on ripening. For the cultivars at $13/23^{\circ}C$ (low temperature study), JB and JJ had a ripening rate of 93% which is similar to the ripening rates of cultivars at $19/29^{\circ}C$ at 45 days after heading (DAH). In contrast, GY and HW recorded lower ripening rates of 86% and 57% respectively. However, when the cultivars at $13/23^{\circ}C$ were harvested at 61 DAH (when the accumulated temperature reached $1100^{\circ}C$), the difference in ripening rates compared to the 4 cultivars of $19/29^{\circ}C$ harvested at 45 DAH was not obvious (JB 94%, JJ 97%, GY 97%, HW 88%). Starch content showed little difference among the 4 cultivars at different temperature conditions while amylose content was higher for cultivars at $13/23^{\circ}C$ compared to those at $19/29^{\circ}C$. In addition, the enzyme activities of starch biosynthesis were about 5~10 days slower in cultivars at $13/23^{\circ}C$ compared to cultivars at $19/29^{\circ}C$. The grain-filling rate showed highly significant correlations with the enzyme activities of Sucrose synthase ($R^2=0.70^{***}$), ADP glucose pyrophosphorylase ($R^2=0.63^{***}$), UDP glucose pyrophosphorylase ($R^2=0.36^{***}$), Starch synthase ($R^2=0.51^{***}$), and Starch branching enzyme ($R^2=0.59^{***}$). Among the enzymes, Sucrose synthase activity had the highest correlation coefficient with grain-filling rate. In conclusion, the activity of enzymes such as Sucrose synthase, UDP glucose pyrophosphorylase, ADP glucose pyrophosphorylase, Starch synthase, Starch branching enzyme in starch biosynthesis is proven to be highly related to the grain filling process. Notably, the decrease in the activity of Sucrose synthase and Starch branching enzyme and the late increase in ADP glucose pyrophosphorylase activity at low temperature in the ripening stage are considered to be disadvantageous as they delay ripening and increased amylose content.

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Cloning and Characterization of GDP-mannose Pyrophosphorylase from Solanum Tuberosum L.

  • Hyun, Tae-Kyung;Lim, Jung-Dae;Kim, Jae-Kwang;Seong, Eun-Soo;Lee, Jae-Geun;Yoon, Byeong-Sung;Kim, Myong-Jo;Cho, Dong-Ha;Yu, Chang-Yeon
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.5
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    • pp.276-283
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    • 2005
  • Ascorbic acid is a great antioxidant and helps protect the body against pollutants. GDP-mannose pyrophosphorylase (GMPase) is a key enzyme in manufacturing GDP-mannose, a glycosyl donor for ascorbate and cell wall biosynthesis as well as for protein glycosylation. In this study, we described molecular cloning of a full-length cDNA from Potato (Solanum tuberosum L. cv. Jasim), using tuber. The cDNA isolated encoded a GDP-mannose pyrophosphrylase. The nucleotide sequence of pGMPC showed about 95%, 89% and 80% homology with S. tuberosum (AF022716), N. tabacum (AB066279) and A. thaliana (AF076484) cDNAs clone known as GMPase, respectively. We detected the expression of GMPase using RT-PCR. The highest expression of GMPase was found in stems, and the largest amount of ascorbic acid was also presented in stems. In contrast, the leaf showed minimal level of GMPase transcript and ascorbic acid content. We propose that GMPase expression patterns were similar to the changes of ascorbic acid content in the leaves treated with diverse stresses.

Development of Detection Method of Unapproved Genetically Modified Potato (EH92-527-1) in Korea using Duplex Polymerase Chain Reaction (Duplex PCR을 이용한 국내 미승인 유전자변형 감자(EH92-527-1)의 검사법 개발)

  • Yoo, Myung-Ryul;Kim, Jae-Hwan;Yea, Mi-Chi;Kim, Hae-Yeong
    • Korean Journal of Food Science and Technology
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    • v.45 no.2
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    • pp.156-160
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    • 2013
  • A duplex polymerase chain reaction (PCR) method was developed to detect unapproved genetically modified (GM) potato (EH92-527-1) in Korea. The UDP-glucose pyrophosphorylase (UGP) gene was selected as an endogenous reference gene for potato and used to validate the specificity for 14 different crops. The primer pair EH92-F/R was designed to amplify the junction sequence between the genome and transgenic region introduced in GM potato. Its specificity was also validated using several different GM events. The detection limit of the duplex PCR method is approximately 0.05%. This duplex PCR method could be useful for monitoring cultivation of unauthorized GM potato in Korea.

Cloning, Expression, and Characterization of UDP-glucose Pyrophosphorylase from Sphingomonas chungbukensis DJ77

  • Yoon, Moon-Young;Lee, Kyoung-Jin;Park, Hea-Chul;Park, Sung-Ha;Kim, Sang-Gon;Kim, Sung-Kun;Choi, Jung-Do
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1360-1364
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    • 2009
  • The bacterium Sphingomonas chungbukensis DJ77 produces the extracellular polysaccharide gellan in high yield. Gellan produced by this bacterium is widely used as a gelling agent, and the enzyme UDP-glucose pyrophosphorylase (UGP) is thought to play a key role in the gellan biosynthetic pathway. The UGP gene has been successfully cloned and over-expressed in E. coli. The expressed enzyme was purified with a molecular weight of approximately 32 kDa, as determined by a SDS-polyacrylamide gel, but the enzyme appears as ca. 63 kDa on a native gel, suggesting that the enzyme is present in a homodimer. Kinetic analysis of UDP-glucose for UGP indicates $K_m$ = 1.14 mM and $V_{max}$ = 10.09 mM/min/mg at pH 8.0, which was determined to be the optimal pH for UGP catalytic activity. Amino acid sequence alignment against other bacteria suggests that the UGP contains two conserved domains: An activator binding site and a glucose-1-phosphate binding site. Site-directed mutagenesis of Lys194, located within the glucose-1-phosphate binding site, indicates that substitution of the charge-reversible residue Asp for Lys194 dramatically impairs the UGP activity, supporting the hypothesis that Lys194 plays a critical role in the catalysis.

Comparison of Agronomic Characteristics and Activity Variation of ADP-Glucose Pyrophosphorylase at Different Growth Stages in Soybean Cultivars (콩 품종의 생육특성 및 생육단계별 ADP-Glucose Pyrophosphorylase의 활성변화 비교)

  • Kim, Young-Jin;Lee, Si-Myeong;Cho, Sang-Kyun;Oh, Young-Jin;Kim, Hag-Sin
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.55 no.2
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    • pp.139-143
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    • 2010
  • The relationship between ADP-glucose pyrophosphorylase (AGP) activity and the characteristics of related pod setting in developing seed of soybean cv. Pungsannamulkong, Iksannamulkong, Geumjeongkong #1 and Danpaheuk was studied. AGP activity during the accumulate of the majority of dry matter in all cultivars suggested that this enzyme might be associated with this process. At the Vn and R1 stages, AGP activity of full-grown leaves of Pungsannamulkong, Iksannamulkong, Geumjeongkong #1 was the highest and then decreased progressively. However AGP activity of Danpaheuk was the lowest and also had lower seed weight. So regulation of matter accumulation in developing soybean seeds may also depend on AGP activity. AGP capacities as expressed by AGP activity seem to have a good predicting value for the dry matter of leaf and seed at R1 to R5 stages in our series of R3 stage genotypes. Western blots probed with antibody specific to the subunit of potato AGP revealed a single 60KD immunoreactive band that changed in intensity during the growth cycle in association with changes in total AGP activity.