• Title/Summary/Keyword: $C_4$ pathway

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The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.167-167
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_ Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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The overexpression of Arachis hypogaea resveratrol synthase 3 (AhRS3) modified the expression pattern of phenylpropanoid pathway genes in developing rice seeds

  • Lee, Choonseok;Jeong, Namhee;Kim, Dool-Yi;Ok, Hyun-Choong;Choi, Man-Soo;Park, Ki-Do;Kim, Jaehyun
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.105-105
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    • 2017
  • Our previous study for developing seeds of Iksan 526 (I.526), an inbred line of resveratrol-producing transgenic rice line, showed that, in 20 days after heading (DAH) seeds, resveratrol was almost saturated and accumulation of piceid was highest though the expression of Arachis hypogaea resveratrol synthase 3 (AhRS3, GenBank DQ124938) was highest in 31 DAH seeds. In this study, it was investigated how the overexpression of AhRS3 affects phenylpropanoid pathway genes. p-Coumaroyl-CoA is derived from phenylpropanoid pathway and used as a substrate of AhRS3 reaction for resveratrol production. In 6, 13, 20, 31 and 41 (45 for Dongjin) DAH seeds of I526 and Dongjin, a wild type of I.526, respectively, the expression pattern of phenylpropanoid pathway genes, including phenylalanine ammonia-lyase (PAL: LOC_Os02g41630.2, LOC_Os04g43760.1), cinnamate 4-hydroxylase (C4H: LOC_Os05g25640.1), 4-coumarate-CoA ligase (4CL: LOC_Os02g08100.1), cinnamoyl-CoA reductase (CCR: LOC_Os09g25150.1, LOC_Os08g34280.1), hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl transferase (HCT: LOC_Os04g42250.2, LOC_Os02g39850.1) and cinnamyl alcohol dehydrogenase (CAD: LOC_Os02g09490.1), was examined using real time (RT)-PCR. Compared to developing seeds of Dongjin, RT-PCR results showed that the expression pattern of phenylpropanoid pathway genes was modified in developing seeds of I.526. In most genes, except for CAD, of I.526 developing seeds, the gene expression was highest in 20 DAH corresponding to biosynthesis of resveratrol and piceid, i.e. the expression of phenylpropanoid pathway genes was gradually increased by 20 DAH and decreased as seeds develop. Especially, in Dongjin, the highest expression of PALs and 4CL was in 6 DAH and their expression was gradually decreased as seeds develop. These genes expression data also exhibited that, in developing seeds of I.526, phenylpropanoid pathway genes were slightly or significantly (in some genes) upregulated compared to Dongjin. Therefore, the overexpression of AhRS3 changed the expression pattern of phenylpropanoid pathway genes in I.526 developing seeds and this modification for gene expression is closely related to biosynthesis of resveratrol and piceid.

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Application of the Extended Grunwald-Winstein Equation to the Solvolyses of 4-(Chlorosulfonyl)biphenyl

  • Kang, Suk Jin;Koh, Han Joong
    • Journal of the Korean Chemical Society
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    • v.61 no.1
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    • pp.25-28
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    • 2017
  • Solvolyses with the reaction center being the sulfur of 4-(chlorosulfonyl)biphenyl ($C_6H_5C_6H_4SO_2Cl$, 1) was studied under solvolytic conditions and the extended Grunwald-Winstein equation was applied. The thirty five kinds of solvents gave a reasonable extended Grunwald-Winstein plot with a correlation coefficient (R) of 0.940. The sensitivity values (l = 0.60 and m = 0.47) of 1 were smaller than those obtained for benzenesulfonyl chloride ($C_6H_5SO_2Cl$, 2; l = 1.10 and m = 0.61) proposed to undergo dissociative $S_N2$ mechanism. These l and m values for the solvolyses of 1 can be considered to support a $S_N2$ pathway with some ionization reaction. The activation parameters, ${\Delta}H^{\neq}$ and ${\Delta}S^{\neq}$, were determined and they are also in line with values expected for a bimolecular reaction. The kinetic solvent isotope effect (KSIE) of 1.26 is also in accord with a bimolecular mechanism, probably assisted by general-base catalysis.

Anti-complement Activity of Triterpenoids from the Whole Plant of Patrinia saniculaefolia

  • An, Ren-Bo;Na, Min-Kyun;Min, Byung-Sun;Lee, Hyeong-Kyu;Bae, Ki-Hwan
    • Natural Product Sciences
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    • v.14 no.4
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    • pp.249-253
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    • 2008
  • Two oleanane-type triterpenes (1, 2) and their glycosides (4-6), and one ursane-type triterpene (3) have been isolated from a methanolic extract of Patrinia saniculaefolia Hemsley (Valerianaceae) through repeated silica gel and reversed-phase C-18 column chromatography. Their chemical structures were determined as oleanolic acid (1), oleanonic acid (2), 23-hydroxyursolic acid (3), 3-O-${\alpha}$-L-arabinopyranosyl-oleanolic acid (4), 3-O-${\beta}$-D-glucopyranosyl-oleanolic acid (5), and oleanolic acid 3-O-[${\alpha}$-D-xylopyranosyl-($1{\rightarrow}3$)-${\beta}$-D-glucuronopyranoside-6-O-butyl-ester] (6) on the basis of their MS, $^1H$-, and $^{13}C$-NMR spectral data. All compounds were isolated from the whole plant of the P. saniculaefolia for the first time. These compounds were examined for their anti-complement activity against the classical pathway of the complement system. Among them, compounds 1 - 3 exhibited anti-complement activity with $IC_{50}$ values of 470.1, 212.2, and 121.0 ${\mu}M$, respectively, whereas compounds 4 - 6 were inactive. These results suggest that the carbonyl or hydroxy group at C-3 in the oleananeand/or ursane-triterpenes are important for the anti-complement activity against the classical pathway.

Complete genome sequence of Pantoea intestinalis SRCM103226, a microbial C40 carotenoid zeaxanthin producer (식용곤충 갈색거저리에서 분리한 카로테노이드 생성균주인 Pantoea intestinalis SRCM103226 균주의 유전체 해독)

  • Kim, Jin Won;Ha, Gwangsu;Jeong, Seong-Yeop;Jeong, Do-Youn
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.167-170
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    • 2019
  • Pantoea intestinalis SRCM103226, isolated from edible insect mealworm overproduces zeaxanthin as a main carotenoid. The complete genome of P. intestinalis SRCM103226 was sequenced using the Pacific Biosciences (PacBio) RS II platform. The genome of P. intestinalis SRCM103226 comprises a 4,784,919 bp circular chromosome (53.41% G+C content), and is devoid of any extrachromosomal plasmids. Annotation using the RAST server reveals 4,332 coding sequences and 107 RNAs (22 rRNA genes, 85 tRNA genes). Genome annotation analysis revealed that it has five genes involved in the carotenoid pathway. The genome information provides fundamental knowledge for comparative genomics studies of the zeaxanthin pathway.

Co-Localization of GABA Shunt Enzymes for the Efficient Production of Gamma-Aminobutyric Acid via GABA Shunt Pathway in Escherichia coli

  • Pham, Van Dung;Somasundaram, Sivachandiran;Park, Si Jae;Lee, Seung Hwan;Hong, Soon Ho
    • Journal of Microbiology and Biotechnology
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    • v.26 no.4
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    • pp.710-716
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    • 2016
  • Gamma-aminobutyric acid (GABA) is a non-protein amino acid, which is an important inhibitor of neurotransmission in the human brain. GABA is also used as the precursor of biopolymer Nylon-4 production. In this study, the carbon flux from the tricarboxylic acid cycle was directed to the GABA shunt pathway for the production of GABA from glucose. The GABA shunt enzymes succinate-semialdehyde dehydrogenase (GabD) and GABA aminotransferase (GabT) were co-localized along with the GABA transporter (GadC) by using a synthetic scaffold complex. The co-localized enzyme scaffold complex produced 0.71 g/l of GABA from 10 g/l of glucose. Inactivation of competing metabolic pathways in mutant E. coli strains XBM1 and XBM6 increased GABA production 13% to reach 0.80 g/l GABA by the enzymes co-localized and expressed in the mutant strains. The recombinant E. coli system developed in this study demonstrated the possibility of the pathway of the GABA shunt as a novel GABA production pathway.

Fungal and Plant Phenylalanine Ammonia-lyase

  • Hyun, Min-Woo;Yun, Yeo-Hong;Kim, Jun-Young;Kim, Seong-Hwan
    • Mycobiology
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    • v.39 no.4
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    • pp.257-265
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    • 2011
  • L-Phenylalanine is one of the essential amino acids that cannot be synthesized in mammals in adequate amounts to meet the requirements for protein synthesis. Fungi and plants are able to synthesize phenylalanine via the shikimic acid pathway. L-Phenylalanine, derived from the shikimic acid pathway, is used directly for protein synthesis in plants or metabolized through the phenylpropanoid pathway. This phenylpropanoid metabolism leads to the biosynthesis of a wide array of phenylpropanoid secondary products. The first step in this metabolic sequence involves the action of phenylalanine ammonialyase (PAL). The discovery of PAL enzyme in fungi and the detection of $^{14}CO_2$ production from $^{14}C$-ring-labeled phenylalanine and cinnamic acid demonstrated that certain fungi can degrade phenylalanine by a pathway involving an initial deamination to cinnamic acid, as happens in plants. In this review, we provide background information on PAL and a recent update on the presence of PAL genes in fungi.

Shikonin ameliorates salivary gland damage and inflammation in a mouse model of Sjögren's syndrome by modulating MAPK signaling pathway

  • Wenjing Guo;Xin Wang;Chao Sun;Jian Wang;Tao Wang
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.4
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    • pp.357-364
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    • 2023
  • Sjögren syndrome (SS) is a systemic inflammatory autoimmune disease that involves exocrine glands. Shikonin is extracted from comfrey, which is conventionally used as an anti-tumor, antibacterial, and antiviral drug in China. However, the application of Shikonin in SS remains unreported. This study aimed to verify the potential functions of Shikonin in SS progression. Firstly, non-obese diabetic mice were used as the SS mouse model, with C57BL/6 mice serving as the healthy control. It was demonstrated that the salivary gland damage and inflammation were aggravated in the SS mouse model. Shikonin improved salivary gland function decline and injury in the SS mouse model. Moreover, Shikonin reduced inflammatory cytokines and immune infiltration in the SS mouse model. Further experiments discovered that Shikonin attenuated the MAPK signaling pathway in the SS mouse model. Lastly, inhibition of the MAPK signaling pathway combined with Shikonin treatment further alleviated the symptoms of SS. In conclusion, Shikonin ameliorated salivary gland damage and inflammation in a mouse model of SS by modulating the MAPK signaling pathway. Our findings indicate that Shikonin may be a useful drug for SS treatment.

Identification of Genetic Variations in CBL, SORBS1, CRK, and RHOQ, Key Modulators in the CAP/TC10 Pathway of Insulin Signal Transduction, and Their Association with Type 2 Diabetes Mellitus in the Korean Population

  • Hong, Kyung-Won;Jin, Hyun-Seok;Lim, Ji-Eun;Go, Min-Jin;Lee, Jong-Young;Hwang, Sue-Yun;Park, Hun-Kuk;Oh, Berm-Seok
    • Genomics & Informatics
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    • v.7 no.2
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    • pp.53-56
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    • 2009
  • Recent evidence has strongly suggested that the CAP/TC10 pathway is involved in the trafficking, docking, and fusion of vesicles containing the insulin-responsive glucose transporter Glut4 to the plasma membrane. However, little is known about how the genes employed in the CAP/TC10 pathway are associated with the development of type 2 diabetes mellitus. In this study, we sequenced 4 genes of the CAP/TC10 pathway [SORBS1, CBL, CRK, and RHOQ] in 24 individuals to identify genetic variations in these loci. A total of 48 sequence variants were identified, including 23 novel variations. To investigate the possible association with type 2 diabetes mellitus, 3 single nucleotide polymorphisms from SORBS1, 3 from CBL, and 4 from RHOQ were genotyped in 1122 Korean type 2 diabetic patients and 1138 nondiabetic controls. Using logistic regression analysis, 1 significant association between SNP rs1376405 in RHOQ and type 2 diabetes mellitus [OR = 8.714 (C.I. 1.714-44.29), p = 0.009] was found in the recessive model. Our data demonstrate a positive association of the RHOQ gene in the CAP/TC10 pathway with T2DM in the Korean population.

Molecular and functional characterization of a Brmecp gene encoding 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase from Brassica rapa (배추 유래 2-C-methyl-D-erythritol 2,4-cyclodiphosphate synthase 관련 Brmecp 유전자의 발현 및 분자적 특성)

  • Jung, Yu-Jin;Choi, Jang-Sun;Sun, Ju-Nam;Nou, Ill-Sup;Cho, Yong-Gu;Kang, Kwon-Kyoo
    • Journal of Plant Biotechnology
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    • v.39 no.3
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    • pp.189-195
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    • 2012
  • In plants, the fifth step of the plastidial 2-Cmethyl-D-erythritol 4-phosphate (MEP) pathway is catalyzed by 2-C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (MECP; EC: 4. 6. 1. 12), an enzyme proposed to play a key role in the regulation of isoprenoid biosynthesis. Here we report the isolation and functional characterization of a 823 bp Brassica rapa MECP (Brmecp) cDNA encoding a deduced polypeptide of 230 amino acid residues. Transcription levels of Brmecp were two-fold higher in petal compared to leaves. In addition, Brmecp expression in cabbage seedlings treated with ABA, $H_2O_2$ and drought was higher than control seedlings. These results were consistent with changes in chlorophyll contents in transgenic Arabidopsis. Thus, the Brmecp may contribute to the production of primary (chlorophylls and carotenoids) isoprenoid end-products in chloroplasts.