• Title/Summary/Keyword: desaturase-1

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Expression Patterns of Genes Involved in Carotenoid Biosynthesis in Pepper

  • Ha, Sun-Hwa;Lee, Shin-Woo;Kim, Jong-Guk;Hwang, Young-Soo
    • Journal of Applied Biological Chemistry
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    • v.42 no.2
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    • pp.92-96
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    • 1999
  • To study the regulatory mechanism of isoprenoid (carotenoid) biosynthesis, we have compared the expression patterns of nine isoprenoid biosynthetic genes in Korean red pepper (Capsicum. annuum cv. NocKaung). The expression of geranylgeranyl pyrophosphate synthase gene was initially induced at early ripening stage (I1) and was rather slightly decreased during pepper fruit ripening. The ex-pression of phytoene synthase gene was strongly induced at semi-ripening stage (I2) and the phytoene desaturase transcript was maximally induced at the fully ripened stage (R). Our results suggest that genes encoding two 3-hydroxy-3-methylglutaryl-CoA reductase isozymes (HMGR1 and HMGR2) and farnesyl pyrophosphate synthase might be not so critical in pepper carotenoid biosynthesis but three genes encoding geranylgeranyl pyrophosphate synthase, phytoene synthase and phytoene desaturase were induced in a sequential manner and coordinately regulated during the ripening of pepper fruit.

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Effect of ω3-Fatty Acid Desaturase Gene Expression on Invasion and Tumorigenicity in Human Tongue Squamous Cell Carcinoma Cells (인체 혀의 편평세포암 세포에서 ω3-fatty acid desaturase 유전자 발현이 침윤 및 종양형성에 미치는 영향)

  • Hong, Tae-Hwa;Shin, Soyeon;Han, Seung-Hyeon;Hwang, Byung-Doo;Lim, Kyu
    • Journal of Life Science
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    • v.28 no.8
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    • pp.945-954
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    • 2018
  • Omega-3 polyunsaturated fatty acids (${\omega}3$-fatty acid) have been found to possess anticancer properties in a variety of cancer cell lines and animal models, but their effects in human tongue squamous cell carcinomas (SCCs) remain unclear. This study was designed to examine the effect of ${\omega}3$-fatty acid desaturase (fat-1) gene expression on invasion and tumorigenicity in human tongue SCC cells and the molecular mechanism of its action. Docosahexaenoic acid (DHA) treatment inhibited in vitro invasion in a dose-dependent manner. In zymography, matrix metalloproteinase-9 (MMP-9) and Matrix metallopeptidase-2 (MMP-2) activities were reduced, and MMP-9 and MMP-2 promoter activities were inhibited by the DHA treatment. In addition, cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) promoter reporter activities were inhibited in SCC-4 and SCC-9 cells after the DHA treatment. To investigate the effect of a high level of endogenous ${\omega}3$ fatty acids, a stable SCC-9 cell line expressing the ${\omega}3$-desaturase gene (fSCC-9sc) was generated. The growth rate and colony-forming capacity of fSCC-9sc were remarkably decreased as compared with those of fSCC-9cc. Likewise, the tumor size and volume of fSCC-9sc implanted into nude mice were significantly inhibited, with increases in the cell death index. Furthermore, a transwell chamber invasion assay showed a reduction in cell invasion of the fSCC-9sc lines when compared with that of the fSCC-9cc line. These findings suggested that fat-1 gene expression inhibited tumorigenicity, as well as invasion in human tongue SCC cells. Thus, utilization of ${\omega}3$ fatty acids may represent a promising therapeutic approach for chemoprevention and the treatment of human tongue SCCs.

Development of Perilla frutescens with Low Levels of Alpha-Linolenic Acid by Inhibition of a delta 15 desaturase Gene (Delta 15 desaturase 유전자 억제에 의해 알파리놀렌산 함량이 낮은 들깨 육성)

  • Kim, Kyung-Hwan;Lee, Kyeong-Ryeol;Kim, Jung-Bong;Lee, Myoung Hee;Lee, Eungyeong;Kim, Nyunhee;Lee, Hongseok;Kim, Song Lim;Baek, JeongHo;Choi, Inchan;Ji, Hyeonso
    • Korean Journal of Breeding Science
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    • v.50 no.4
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    • pp.463-471
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    • 2018
  • Perilla is an oilseed crop cultivated in Korea since ancient times. Due to the high ${\alpha}-linolenic$ acid content in perilla, perilla seed oil can easily become rancid. ${\alpha}-Linolenic$ acid is synthesized by two enzymes, endoplasmic reticulum-localized ${\Delta}15$ desaturase (FAD3) and chloroplast-localized ${\Delta}15$ desaturase (FAD7) in vivo. In order to lower the ${\alpha}-linolenic$ acid content of the seed oil without disturbing plant growth, we tried to suppress the expression of only the FAD3 gene using RNA interference, whilst maintaining the expression of the FAD7 gene. Seventeen transgenic plants with herbicide ($Basta^{TM}$) resistance were obtained by Agrobacterium-mediated transformation using hypocotyls of perilla plants. The transgenic plants were firstly confirmed by treatment with 0.3% (v/v) $Basta^{TM}$ herbicide, and the expression of FAD3 was measured by Northern blot analysis. The ${\alpha}-linolenic$ acid content was 10-20%, 30-40%, and 60% in two, seven, and three of the twelve $T_1$ transgenic perilla plants which had enough seeds to be analyzed for fatty acid composition, respectively. Analysis of the fatty acid composition of $T_2$ progeny seeds from $T_1$ plants with the lowest ${\alpha}-linolenic$ acid content showed that the homozygous lines had 6-10% ${\alpha}-linolenic$ acid content and the heterozygous lines had 20-26% ${\alpha}-linolenic$ acid content. It is expected that the reduction in ${\alpha}-linolenic$ acid content in perilla seed oil will prevent rancidity and can be utilized for the production of high-value functional ingredients such as high ${\gamma}-linolenic$ acid.

Identification of a Potential Gene for Elevation ω-3 Concentration and its Efficiency for Improving ω-6/ω-3 Ratio in Soybean

  • Hyun Jo;Jeong-Dong Lee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2022.10a
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    • pp.24-24
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    • 2022
  • This present study was to identify a novel candidate gene that contribute to the elevated α-linolenic acid (ALA, ω-3) concentration in PE2166 from mutagenesis of Pungsannamul. Major loci qALA5_1 and qALA5_2 were detected on chromosome 5 of soybean through quantitative trait loci mapping analyses of recombinant inbred lines. With next generation sequencing of parental lines and Pungsannamul, and recombinant analyses, a potential gene, Glyma. 05g221500 (HD) controlling elevated ALA concentration was identified. HD is a homeodomain-like transcriptional regulator that may regulate the expression level of microsomal ω-3 fatty acid desaturase (FAD3) genes responsible for the conversion of linoleic acid into ALA in the fatty acid biosynthetic pathway. In addition, we hypothesized that combination of mutant alleles, HD and either of microsomal delta-12 fatty acid desaturase 2-1 (FAD2-1\ could reduce the ω-6/ω-3 ratio. In populations where HD, and FAD2-1A and FAD2-1B genes were segregated, combination of a hd allele from PE2166 and either of the variant FAD2-1 alleles were sufficient to reduce the ω-6/ω-3 ratio in seeds.

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Mating Disruption of Grapholita molesta by RNA Interference of a Fatty Acid Desaturase Expressed in Adult Abdomen (복숭아순나방 성충 복부에서 발현하는 불포화효소의 RNA 간섭과 교미교란)

  • Kim, Kyusoon;Jung, Chung Ryul;Yang, Chang Yeol;Kwon, Gimyeon;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.56 no.1
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    • pp.61-67
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    • 2017
  • Two major sex pheromone components (Z-8-dodecenyl acetate and E-8-dodecenyl acetate) are known in the peach fruit moth, Grapholita molesta. From a putative biosynthetic pathway of these sex pheromone components, delta 10 desaturase ($10{\Delta}$ DES) has been proposed to play a crucial role in synthesizing a species-specific stereoisomer of the double bond. However, its molecular identity was not known. This study determined a putative desaturase (Gm-comp1575) as a $10{\Delta}$ DES candidate from G. molesta transcriptome constructed from the sex pheromone gland. Its open reading frame encodes 370 amino acid sequence with a predicted molecular weight at 43.2 kDa and isoelectric point at 8.77. It was predicted to have four transmembrane domains and six glycosylation sites at N-terminal or cytosolic domains. A phylogenetic analysis with its predicted amino acid sequence indicated that Gm-comp1575 is closely related with known $10{\Delta}$ DES genes of other insects. Gm-comp1575 transcript was detected in female adults at sex pheromone gland and other abdominal tissues. RNA interference of Gm-comp1575 significantly reduced attractiveness of virgin females in apple orchard compared to control females. These results suggest that Gm-comp1575 is associated with sex pheromone biosynthesis of G. molesta.

Stearoyl-CoA desaturase induces lipogenic gene expression in prostate cancer cells and inhibits ceramide-induced cell death

  • Kim, Seung-Jin;Kim, Eung-Seok
    • Animal cells and systems
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    • v.15 no.1
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    • pp.1-8
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    • 2011
  • Perturbation of metabolism with increased expression of lipogenic enzymes is a common characteristic of human cancers, including prostate cancer. In the present work the overexpression of stearoyl-CoA desaturase (SCD) in LNCaP cells led to increased mRNA levels of fatty acid synthase (FAS) and acetyl-CoA-carboxylase-a, whereas micro RNA-mediated silencing of SCD inhibited the expression of these lipogenic genes in LNCaP cells. Treatment with the FAS-specific inhibitor cerulenin inhibited SCD induction of LNCaP cell proliferation. In addition, a transient transfection assay revealed the capability of cerulenin to suppress SCD and dihydrotestosterone induction of androgen receptor transcriptional activity. Furthermore, overexpression of SCD in LNCaP cells produced marked resistance to ceramide-induced cell death with reduced poly(ADP-ribose) polymerase (PARP) cleavage. In contrast, silencing of SCD expression increased Bax protein in LNCaP cells. Furthermore, addition of ceramide to SCD knockdown LNCaP cells increased cell death and caspase-3 activity with drastic increase of PARP cleavage. Together, the data indicate that SCD may provide resistance of prostate cancer cells to ceramide-induced cell death.

Isolation and functional analysis of three microsomal delta-12 fatty acid desaturase genes from Camelina sativa (L.) cv. CAME (카멜리나 (Camelina sativa L. cv. CAME)로부터 3 microsomal delta-12 fatty acid desaturase 유전자들의 분리 및 기능 분석)

  • Kim, Hyojin;Go, Young Sam;Kim, Augustine Yonghwi;Lee, Sanghyeob;Kim, Kyung-Nam;Lee, Geung-Joo;Kim, Gi-Jun;Suh, Mi Chung
    • Journal of Plant Biotechnology
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    • v.41 no.3
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    • pp.146-158
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    • 2014
  • Camelina sativa that belongs to Brassicaceae family is an emerging oilseed crop. Camelina seeds contain approximately 40% storage oils per seed dry weight, which are useful for human and animal diets and industrial applications. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. Microsomal delta-12 fatty acid desaturase2 (FAD2) catalyzes the conversion of oleic acid to linoleic acid. The polymorphisms of FAD2 genes are correlated with the levels of oleic acids in seed oils. In this study, three CsFAD2 genes (CsFAD2-1, CsFAD2-2 and CsFAD2-3.1) were isolated from developing seeds of Camelina sativa (L.) cv. CAME. The nucleotide and deduced amino acid sequences of three CsFAD2 genes were compared with those from dicotyledon and monocotyledon plants including Camelina cultivars Sunesone and SRS933. Three histidine motifs (HECGH, HRRHH, and HVAHH) required for FAD activity and a hydrophobic valine or isoleucine residue, which is a SNP (single nucleotide polymorphism) marker related with enzyme activity are well conserved in three CsFAD2s. The expressions of CsFAD2-1 and CsFAD2-3.1 were ubiquitously detected in various Camelina organs, whereas the CsFAD2-2 transcripts were predominantly detected in flowers and developing seeds. The contents of oleic acids decreased, whereas the amounts of linoleic acid increased in dry seeds of transgenic fad2-2 lines expressing each CsFAD2 gene compared with fad2-2 mutant, indicating that three CsFAD2 genes are functionally active. The isolated CsFAD2 genes might be applicable in metabolic engineering of storage oils with high oleic acids in oilseed crops.

Linseed oil supplementation affects fatty acid desaturase 2, peroxisome proliferator activated receptor gamma, and insulin-like growth factor 1 gene expression in turkeys (Meleagris gallopavo)

  • Szalai, Klaudia;Tempfli, Karoly;Zsedely, Eszter;Lakatos, Erika;Gaspardy, Andras;Papp, Agnes Bali
    • Animal Bioscience
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    • v.34 no.4
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    • pp.662-669
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    • 2021
  • Objective: Effects of linseed oil (LO) supplementation on the fat content and fatty acid profile of breast meat, and the expression of three genes in the liver, breast muscle and fat tissues of commercial 154-day-old hybrid male turkeys were investigated. Methods: The animals in the control group were fed a commercially available feed and received no LO supplementation (n = 70), whereas animals in the LO group (n = 70) were fed the same basic diet supplemented with LO (day 15 to 21, 0.5%; day 22 to 112, 1%). The effect of dietary LO supplementation on fatty acid composition of breast muscle was examined by gas chromatography, and the expression of fatty acid desaturase 2 (FADS2), peroxisome proliferator activated receptor gamma (PPARγ), and insulin-like growth factor 1 (IGF1) genes was analysed by means of quantitative reverse transcription polymerase chain reaction. Results: The LO supplementation affected the fatty acid composition of breast muscle. Hepatic FADS2 levels were considerably lower (p<0.001), while adipose tissue expression was higher (p<0.05) in the control compared to the LO group. The PPARγ expression was lower (p<0.05), whereas IGF1 was higher (p<0.05) in the fat of control animals. There were no significant (p>0.05) differences in FADS2, PPARγ, and IGF1 gene expressions of breast muscle; however, omega-6/omega-3 ratio of breast muscle substantially decreased (p<0.001) in the LO group compared to control. Conclusion: Fatty acid composition of breast meat was positively influenced by LO supplementation without deterioration of fattening parameters. Remarkably, increased FADS2 expression in the liver of LO supplemented animals was associated with a significantly decreased omega-6/omega-3 ratio, providing a potentially healthier meat product for human consumption. Increased PPARγ expression in fat tissue of the LO group was not associated with fat content of muscle, whereas a decreased IGF1 expression in fat tissue was associated with a trend of decreasing fat content in muscle of the experimental LO group.