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Keywords = short chain dehydrogenase/reductase (SDR) genes

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13 pages, 2778 KiB  
Article
Tissue-Specific Expression of the Porcine DHRS3 Gene and Its Impact on the Proliferation and Differentiation of Myogenic Cells
by Jifeng Li, Yong Ruan, Chuanmei Jiang, Jinkui Sun, Dongwei An, Bo Zhou, Huan Liu, Ziyang Li and Houqiang Xu
Animals 2025, 15(8), 1101; https://doi.org/10.3390/ani15081101 - 10 Apr 2025
Viewed by 467
Abstract
The DHRS3 gene, a member of the short-chain dehydrogenase/reductase (SDR) family, is involved in critical metabolic processes in animals. This study investigated the expression patterns of DHRS3 across various tissues of developmental stages in pigs and preliminarily evaluated its effects on myoblast proliferation, [...] Read more.
The DHRS3 gene, a member of the short-chain dehydrogenase/reductase (SDR) family, is involved in critical metabolic processes in animals. This study investigated the expression patterns of DHRS3 across various tissues of developmental stages in pigs and preliminarily evaluated its effects on myoblast proliferation, apoptosis, and differentiation. RT-qPCR (real-time quantitative PCR) was employed to analyze DHRS3 expression in the heart, liver, spleen, lungs, kidneys, longissimus dorsi, foreleg, and hind leg of pigs at 3 days, 6 months, and 12 months of age. Cell proliferation was analyzed using EdU (5-ethynyl-2′-deoxyuridine) assays, RT-qPCR, and flow cytometry, while the expression changes of proliferation-, apoptosis-, and differentiation-related genes were assessed via RT-qPCR. The results indicated that DHRS3 was expressed in all eight tissues at all three developmental stages. At 3 days, DHRS3 expression was the highest in the kidneys; at 6 months, it peaked in the liver; and at 12 months, it was again the highest in the kidneys. Across all stages, the liver and kidneys exhibited the highest DHRS3 expression levels. Functional studies revealed that DHRS3 overexpression suppressed myoblast proliferation and differentiation while promoting apoptosis. In contrast, DHRS3 inhibition enhanced myoblast proliferation and differentiation and reduced apoptosis. These findings underscore the regulatory role of DHRS3 in myogenesis and provide insights into its metabolic and developmental functions in pigs. Full article
(This article belongs to the Special Issue Better Fattening Performance, Better Pig Meat Quality)
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18 pages, 6578 KiB  
Article
Genome-Wide Analysis and Characterization of the SDR Gene Superfamily in Cinnamomum camphora and Identification of Synthase for Eugenol Biosynthesis
by Yueting Zhang, Chao Fu, Shifang Wen, Ting Zhang and Xindong Wang
Int. J. Mol. Sci. 2024, 25(18), 10084; https://doi.org/10.3390/ijms251810084 - 19 Sep 2024
Cited by 1 | Viewed by 1557
Abstract
Short-chain dehydrogenase/reductases (SDRs) are the largest NAD(H)-dependent oxidoreductase superfamilies and are involved in diverse metabolisms. This study presents a comprehensive genomic analysis of the SDR superfamily in Cinnamomum camphora, a species that is one of the most significant woody essential oil plants in [...] Read more.
Short-chain dehydrogenase/reductases (SDRs) are the largest NAD(H)-dependent oxidoreductase superfamilies and are involved in diverse metabolisms. This study presents a comprehensive genomic analysis of the SDR superfamily in Cinnamomum camphora, a species that is one of the most significant woody essential oil plants in southern China. We identify a total of 222 CcSDR proteins and classify them into five types based on their cofactor-binding and active sites: ‘atypical’, ‘classic’, ‘divergent’, ‘extended’, and ‘unknown’. Phylogenetic analysis reveals three evolutionary branches within the CcSDR proteins, and further categorization using the SDR-initiative Hidden Markov model resulted in 46 families, with the CcSDR110C, CcSDR108E, and CcSDR460A families being the most populous. Collinearity analysis identified 34 pairs of CcSDR paralogs in C. camphora, 141 pairs of SDR orthologs between C. camphora and Populus trichocarpa, and 59 pairs between C. camphora and Oryza sativa. Expression profile analysis indicates a preference for the expression of 77 CcSDR genes in specific organs such as flowers, bark, twigs, roots, leaves, or fruits. Moreover, 77 genes exhibit differential expression patterns during the four developmental stages of leaves, while 130 genes show variance across the five developmental stages of fruits. Additionally, to explore the biosynthetic mechanism of methyl eugenol, a key component of the leaf essential oil in the methyl eugenol chemotype, this study also identifies eugenol synthase (EGS) within the CcSDR460A family through an integrated strategy. Real-time quantitative PCR analysis demonstrates that the expression of CcEGS in the leaves of the methyl eugenol chemotype is more than fourfold higher compared to other chemotypes. When heterologously expressed in Escherichia coli, it catalyzes the conversion of coniferyl acetate into a mixture predominantly composed of eugenol (71.44%) and isoeugenol (21.35%). These insights pave the way for future research into the functional diversity of CcSDR genes, with a focus on secondary metabolism. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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17 pages, 4026 KiB  
Article
Rv0687 a Putative Short-Chain Dehydrogenase Is Required for In Vitro and In Vivo Survival of Mycobacterium tuberculosis
by Gunapati Bhargavi, Mohan Krishna Mallakuntla, Deepa Kale and Sangeeta Tiwari
Int. J. Mol. Sci. 2024, 25(14), 7862; https://doi.org/10.3390/ijms25147862 - 18 Jul 2024
Cited by 2 | Viewed by 1585
Abstract
Mycobacterium tuberculosis (Mtb), a successful human pathogen, resides in host sentinel cells and combats the stressful intracellular environment induced by reactive oxygen and nitrogen species during infection. Mtb employs several evasion mechanisms in the face of the host as a survival [...] Read more.
Mycobacterium tuberculosis (Mtb), a successful human pathogen, resides in host sentinel cells and combats the stressful intracellular environment induced by reactive oxygen and nitrogen species during infection. Mtb employs several evasion mechanisms in the face of the host as a survival strategy, including detoxifying enzymes as short-chain dehydrogenases/reductases (SDRs) to withstand host-generated insults. In this study, using specialized transduction, we have generated a Rv0687 deletion mutant and its complemented strain and investigated the functional role of Rv0687, a member of SDRs family genes in Mtb pathogenesis. A wildtype (WT) and a mutant Mtb strain lacking Rv0687 (RvΔ0687) were tested for the in vitro stress response and in vivo survival in macrophages and mice models of infection. The study demonstrates that the deletion of Rv0687 elevated the sensitivity of Mtb to oxidative and nitrosative stress-inducing agents. Furthermore, the lack of Rv0687 compromised the survival of Mtb in primary bone marrow macrophages and led to an increase in the levels of the secreted proinflammatory cytokines TNF-α and MIP-1α. Interestingly, the growth of WT and RvΔ0687 was similar in the lungs of infected immunocompromised mice; however, a significant reduction in RvΔ0687 growth was observed in the spleen of immunocompromised Rag−/− mice at 4 weeks post-infection. Moreover, Rag−/− mice infected with RvΔ0687 survived longer compared to those infected with the WT Mtb strain. Additionally, we observed a significant reduction in the bacterial burden in the spleens and lungs of immunocompetent C57BL/6 mice infected with RvΔ0687 compared to those infected with complemented and WT Mtb strains. Collectively, this study reveals that Rv0687 plays a role in Mtb pathogenesis. Full article
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26 pages, 10815 KiB  
Article
Silicon-Induced Morphological, Biochemical and Molecular Regulation in Phoenix dactylifera L. under Low-Temperature Stress
by Saqib Bilal, Taimoor Khan, Sajjad Asaf, Nasir Ali Khan, Syed Saad Jan, Muhammad Imran, Ahmed Al-Rawahi, Abdul Latif Khan, In-Jung Lee and Ahmed Al-Harrasi
Int. J. Mol. Sci. 2023, 24(7), 6036; https://doi.org/10.3390/ijms24076036 - 23 Mar 2023
Cited by 8 | Viewed by 2640
Abstract
Climate changes abruptly affect optimum growth temperatures, leading to a negative influence on plant physiology and productivity. The present study aimed to investigate the extent of low-temperature stress effects on date palm growth and physiological indicators under the exogenous application of silicon (Si). [...] Read more.
Climate changes abruptly affect optimum growth temperatures, leading to a negative influence on plant physiology and productivity. The present study aimed to investigate the extent of low-temperature stress effects on date palm growth and physiological indicators under the exogenous application of silicon (Si). Date palm seedlings were treated with Si (1.0 mM) and exposed to different temperature regimes (5, 15, and 30 °C). It was observed that the application of Si markedly improved fresh and dry biomass, photosynthetic pigments (chlorophyll and carotenoids), plant morphology, and relative water content by ameliorating low-temperature-induced oxidative stress. Low-temperature stress (5 and 15 °C), led to a substantial upregulation of ABA-signaling-related genes (NCED-1 and PyL-4) in non Si treated plants, while Si treated plants revealed an antagonistic trend. However, jasmonic acid and salicylic acid accumulation were markedly elevated in Si treated plants under stress conditions (5 and 15 °C) in comparison with non Si treated plants. Interestingly, the upregulation of low temperature stress related plant plasma membrane ATPase (PPMA3 and PPMA4) and short-chain dehydrogenases/reductases (SDR), responsible for cellular physiology, stomatal conductance and nutrient translocation under silicon applications, was observed in Si plants under stress conditions in comparison with non Si treated plants. Furthermore, a significant expression of LSi-2 was detected in Si plants under stress, leading to the significant accumulation of Si in roots and shoots. In contrast, non Si plants demonstrated a low expression of LSi-2 under stress conditions, and thereby, reduced level of Si accumulation were observed. Less accumulation of oxidative stress was evident from the expression of superoxide dismutase (SOD) and catalase (CAT). Additionally, Si plants revealed a significant exudation of organic acids (succinic acid and citric acid) and nutrient accumulation (K and Mg) in roots and shoots. Furthermore, the application of Si led to substantial upregulation of the low temperature stress related soybean cold regulated gene (SRC-2) and ICE-1 (inducer of CBF expression 1), involved in the expression of CBF/DREB (C-repeat binding factor/dehydration responsive element binding factor) gene family under stress conditions in comparison with non Si plants. The current research findings are crucial for exploring the impact on morpho-physio-biochemical attributes of date palms under low temperature and Si supplementation, which may provide an efficient strategy for growing plants in low-temperature fields. Full article
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14 pages, 672 KiB  
Article
Gene Structure Evolution of the Short-Chain Dehydrogenase/Reductase (SDR) Family
by Franco Gabrielli, Marco Antinucci and Sergio Tofanelli
Genes 2023, 14(1), 110; https://doi.org/10.3390/genes14010110 - 30 Dec 2022
Cited by 10 | Viewed by 3926
Abstract
SDR (Short-chain Dehydrogenases/Reductases) are one of the oldest and heterogeneous superfamily of proteins, whose classification is problematic because of the low percent identity, even within families. To get clearer insights into SDR molecular evolution, we explored the splicing site organization of the 75 [...] Read more.
SDR (Short-chain Dehydrogenases/Reductases) are one of the oldest and heterogeneous superfamily of proteins, whose classification is problematic because of the low percent identity, even within families. To get clearer insights into SDR molecular evolution, we explored the splicing site organization of the 75 human SDR genes across their vertebrate and invertebrate orthologs. We found anomalous gene structures in members of the human SDR7C and SDR42E families that provide clues of retrogene properties and independent evolutionary trajectories from a common invertebrate ancestor. The same analyses revealed that the identity value between human and invertebrate non-allelic variants is not necessarily associated with the homologous gene structure. Accordingly, a revision of the SDR nomenclature is proposed by including the human SDR40C1 and SDR7C gene in the same family. Full article
(This article belongs to the Section Population and Evolutionary Genetics and Genomics)
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28 pages, 5097 KiB  
Article
Silicon-Induced Tolerance against Arsenic Toxicity by Activating Physiological, Anatomical and Biochemical Regulation in Phoenix dactylifera (Date Palm)
by Taimoor Khan, Saqib Bilal, Sajjad Asaf, Safiya Salim Alamri, Muhammad Imran, Abdul Latif Khan, Ahmed Al-Rawahi, In-Jung Lee and Ahmed Al-Harrasi
Plants 2022, 11(17), 2263; https://doi.org/10.3390/plants11172263 - 31 Aug 2022
Cited by 15 | Viewed by 3331
Abstract
Arsenic is a toxic metal abundantly present in agricultural, industrial, and pesticide effluents. To overcome arsenic toxicity and ensure safety for plant growth, silicon (Si) can play a significant role in its mitigation. Here, we aim to investigate the influence of silicon on [...] Read more.
Arsenic is a toxic metal abundantly present in agricultural, industrial, and pesticide effluents. To overcome arsenic toxicity and ensure safety for plant growth, silicon (Si) can play a significant role in its mitigation. Here, we aim to investigate the influence of silicon on date palm under arsenic toxicity by screening antioxidants accumulation, hormonal modulation, and the expression profile of abiotic stress-related genes. The results showed that arsenic exposure (As: 1.0 mM) significantly retarded growth attributes (shoot length, root length, fresh weight), reduced photosynthetic pigments, and raised reactive species levels. Contrarily, exogenous application of Si (Na2SiO3) to date palm roots strongly influenced stress mitigation by limiting the translocation of arsenic into roots and shoots as compared with the arsenic sole application. Furthermore, an enhanced accumulation of polyphenols (48%) and increased antioxidant activities (POD: 50%, PPO: 75%, GSH: 26.1%, CAT: 51%) resulted in a significant decrease in superoxide anion (O2•−: 58%) and lipid peroxidation (MDA: 1.7-fold), in silicon-treated plants, compared with control and arsenic-treated plants. The Si application also reduced the endogenous abscisic acid (ABA: 38%) under normal conditions, and salicylic acid (SA: 52%) and jasmonic acid levels (JA: 62%) under stress conditions as compared with control and arsenic. Interestingly, the genes; zeaxanthin epoxidase (ZEP) and 9-cis-epoxycarotenoid dioxygenase (NCED-1) involved in ABA biosynthesis were upregulated by silicon under arsenic stress. Likewise, Si application also upregulated gene expression of plant plasma membrane ATPase (PMMA-4), aluminum-activated malate transporter (ALMT) responsible for maintaining cellular physiology, stomatal conductance, and short-chain dehydrogenases/reductases (SDR) involved in nutrients translocation. Hence, the study demonstrates the remarkable role of silicon in supporting growth and inducing arsenic tolerance by increasing antioxidant activities and endogenous hormones in date palm. The outcomes of our study can be employed in further studies to better understand arsenic tolerance and decode mechanism. Full article
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11 pages, 1706 KiB  
Article
An NmrA-Like Protein, Lws1, Is Important for Pathogenesis in the Woody Plant Pathogen Lasiodiplodia theobromae
by Junbo Peng, Janith V. S. Aluthmuhandiram, K. W. Thilini Chethana, Qi Zhang, Qikai Xing, Hui Wang, Mei Liu, Wei Zhang, Xinghong Li and Jiye Yan
Plants 2022, 11(17), 2197; https://doi.org/10.3390/plants11172197 - 24 Aug 2022
Cited by 8 | Viewed by 2514
Abstract
The NmrA-like proteins have been reported to be important nitrogen metabolism regulators and virulence factors in herbaceous plant pathogens. However, their role in the woody plant pathogen Lasiodiplodia theobromae is less clear. In the current study, we identified a putative NmrA-like protein, Lws1, [...] Read more.
The NmrA-like proteins have been reported to be important nitrogen metabolism regulators and virulence factors in herbaceous plant pathogens. However, their role in the woody plant pathogen Lasiodiplodia theobromae is less clear. In the current study, we identified a putative NmrA-like protein, Lws1, in L. theobromae and investigated its pathogenic role via gene silencing and overexpression experiments. We also evaluated the effects of external carbon and nitrogen sources on Lws1 gene expression via qRT-PCR assays. Moreover, we analyzed the molecular interaction between Lws1 and its target protein via the yeast two-hybrid system. The results show that Lws1 contained a canonical glycine-rich motif shared by the short-chain dehydrogenase/reductase (SDR) superfamily proteins and functioned as a negative regulator during disease development. Transcription profiling revealed that the transcription of Lws1 was affected by external nitrogen and carbon sources. Interaction analyses demonstrated that Lws1 interacted with a putative GATA family transcription factor, LtAreA. In conclusion, these results suggest that Lws1 serves as a critical regulator in nutrition metabolism and disease development during infection. Full article
(This article belongs to the Special Issue The Research of Plant Fungal Disease)
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23 pages, 5220 KiB  
Article
Plastidial Expression of 3β-Hydroxysteroid Dehydrogenase and Progesterone 5β-Reductase Genes Confer Enhanced Salt Tolerance in Tobacco
by Muhammad Sameeullah, Muhammet Yildirim, Noreen Aslam, Mehmet Cengiz Baloğlu, Buhara Yucesan, Andreas G. Lössl, Kiran Saba, Mohammad Tahir Waheed and Ekrem Gurel
Int. J. Mol. Sci. 2021, 22(21), 11736; https://doi.org/10.3390/ijms222111736 - 29 Oct 2021
Cited by 9 | Viewed by 4137
Abstract
The short-chain dehydrogenase/reductase (SDR) gene family is widely distributed in all kingdoms of life. The SDR genes, 3β-hydroxysteroid dehydrogenase (3β-HSD) and progesterone 5-β-reductases (P5βR1, P5βR2) play a crucial role in cardenolide biosynthesis pathway in the Digitalis species. However, [...] Read more.
The short-chain dehydrogenase/reductase (SDR) gene family is widely distributed in all kingdoms of life. The SDR genes, 3β-hydroxysteroid dehydrogenase (3β-HSD) and progesterone 5-β-reductases (P5βR1, P5βR2) play a crucial role in cardenolide biosynthesis pathway in the Digitalis species. However, their role in plant stress, especially in salinity stress management, remains unexplored. In the present study, transplastomic tobacco plants were developed by inserting the 3β-HSD, P5βR1 and P5βR2 genes. The integration of transgenes in plastomes, copy number and transgene expression at transcript and protein level in transplastomic plants were confirmed by PCR, end-to-end PCR, qRT-PCR and Western blot analysis, respectively. Subcellular localization analysis showed that 3β-HSD and P5βR1 are cytoplasmic, and P5βR2 is tonoplast-localized. Transplastomic lines showed enhanced growth in terms of biomass and chlorophyll content compared to wild type (WT) under 300 mM salt stress. Under salt stress, transplastomic lines remained greener without negative impact on shoot or root growth compared to the WT. The salt-tolerant transplastomic lines exhibited enhanced levels of a series of metabolites (sucrose, glutamate, glutamine and proline) under control and NaCl stress. Furthermore, a lower Na+/K+ ratio in transplastomic lines was also observed. The salt tolerance, mediated by plastidial expression of the 3β-HSD, P5βR1 and P5βR2 genes, could be due to the involvement in the upregulation of nitrogen assimilation, osmolytes as well as lower Na+/K+ ratio. Taken together, the plastid-based expression of the SDR genes leading to enhanced salt tolerance, which opens a window for developing saline-tolerant plants via plastid genetic engineering. Full article
(This article belongs to the Special Issue Plant Genomics and Genome Editing)
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16 pages, 5634 KiB  
Article
Genome-Wide Identification and Characterization of Short-Chain Dehydrogenase/Reductase (SDR) Gene Family in Medicago truncatula
by Shuhan Yu, Qiguo Sun, Jiaxuan Wu, Pengcheng Zhao, Yanmei Sun and Zhenfei Guo
Int. J. Mol. Sci. 2021, 22(17), 9498; https://doi.org/10.3390/ijms22179498 - 31 Aug 2021
Cited by 25 | Viewed by 4807
Abstract
Short-chain dehydrogenase/reductase (SDR) belongs to the NAD(P)(H)-dependent oxidoreductase superfamily. Limited investigations reveal that SDRs participate in diverse metabolisms. A genome-wide identification of the SDR gene family in M. truncatula was conducted. A total of 213 MtSDR genes were identified, and they were distributed [...] Read more.
Short-chain dehydrogenase/reductase (SDR) belongs to the NAD(P)(H)-dependent oxidoreductase superfamily. Limited investigations reveal that SDRs participate in diverse metabolisms. A genome-wide identification of the SDR gene family in M. truncatula was conducted. A total of 213 MtSDR genes were identified, and they were distributed on all chromosomes unevenly. MtSDR proteins were categorized into seven subgroups based on phylogenetic analysis and three types including ‘classic’, ‘extended’, and ‘atypical’, depending on the cofactor-binding site and active site. Analysis of the data from M. truncatula Gene Expression Atlas (MtGEA) showed that above half of MtSDRs were expressed in at least one organ, and lots of MtSDRs had a preference in a tissue-specific expression. The cis-acting element responsive to plant hormones (salicylic acid, ABA, auxin, MeJA, and gibberellin) and stresses were found in the promoter of some MtSDRs. Many genes of MtSDR7C,MtSDR65C, MtSDR110C, MtSDR114C, and MtSDR108E families were responsive to drought, salt, and cold. The study provides useful information for further investigation on biological functions of MtSDRs, especially in abiotic stress adaptation, in the future. Full article
(This article belongs to the Section Molecular Plant Sciences)
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17 pages, 3717 KiB  
Article
Identification, Expression and Evolution of Short-Chain Dehydrogenases/Reductases in Nile Tilapia (Oreochromis niloticus)
by Shuai Zhang, Lang Xie, Shuqing Zheng, Baoyue Lu, Wenjing Tao, Xiaoshuang Wang, Thomas D Kocher, Linyan Zhou and Deshou Wang
Int. J. Mol. Sci. 2021, 22(8), 4201; https://doi.org/10.3390/ijms22084201 - 18 Apr 2021
Cited by 11 | Viewed by 3359
Abstract
The short-chain dehydrogenases/reductases (SDR) superfamily is involved in multiple physiological processes. In this study, genome-wide identification and comprehensive analysis of SDR superfamily were carried out in 29 animal species based on the latest genome databases. Overall, the number of SDR genes in animals [...] Read more.
The short-chain dehydrogenases/reductases (SDR) superfamily is involved in multiple physiological processes. In this study, genome-wide identification and comprehensive analysis of SDR superfamily were carried out in 29 animal species based on the latest genome databases. Overall, the number of SDR genes in animals increased with whole genome duplication (WGD), suggesting the expansion of SDRs during evolution, especially in 3R-WGD and polyploidization of teleosts. Phylogenetic analysis indicated that vertebrates SDRs were clustered into five categories: classical, extended, undefined, atypical, and complex. Moreover, tandem duplication of hpgd-a, rdh8b and dhrs13 was observed in teleosts analyzed. Additionally, tandem duplications of dhrs11-a, dhrs7a, hsd11b1b, and cbr1-a were observed in all cichlids analyzed, and tandem duplication of rdh10-b was observed in tilapiines. Transcriptome analysis of adult fish revealed that 93 SDRs were expressed in more than one tissue and 5 in one tissue only. Transcriptome analysis of gonads from different developmental stages showed that expression of 17 SDRs were sexually dimorphic with 11 higher in ovary and 6 higher in testis. The sexually dimorphic expressions of these SDRs were confirmed by in situ hybridization (ISH) and qPCR, indicating their possible roles in steroidogenesis and gonadal differentiation. Taken together, the identification and the expression data obtained in this study contribute to a better understanding of SDR superfamily evolution and functions in teleosts. Full article
(This article belongs to the Section Molecular Biology)
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20 pages, 2374 KiB  
Article
Representational Difference Analysis of Transcripts Involved in Jervine Biosynthesis
by Magdalena Szeliga, Joanna Ciura and Mirosław Tyrka
Life 2020, 10(6), 88; https://doi.org/10.3390/life10060088 - 19 Jun 2020
Cited by 11 | Viewed by 3397
Abstract
Veratrum-type steroidal alkaloids (VSA) are the major bioactive ingredients that strongly determine the pharmacological activities of Veratrum nigrum. Biosynthesis of VSA at the molecular and genetic levels is not well understood. Next-generation sequencing of representational difference analysis (RDA) products after elicitation and [...] Read more.
Veratrum-type steroidal alkaloids (VSA) are the major bioactive ingredients that strongly determine the pharmacological activities of Veratrum nigrum. Biosynthesis of VSA at the molecular and genetic levels is not well understood. Next-generation sequencing of representational difference analysis (RDA) products after elicitation and precursor feeding was applied to identify candidate genes involved in VSA biosynthesis. A total of 12,048 contigs with a median length of 280 bases were received in three RDA libraries obtained after application of methyl jasmonate, squalene and cholesterol. The comparative analysis of annotated sequences was effective in identifying candidate genes. GABAT2 transaminase and hydroxylases active at C-22, C-26, C-11, and C-16 positions in late stages of jervine biosynthesis were selected. Moreover, genes coding pyrroline-5-carboxylate reductase and enzymes from the short-chain dehydrogenases/reductases family (SDR) associated with the reduction reactions of the VSA biosynthesis process were proposed. The data collected contribute to better understanding of jervine biosynthesis and may accelerate implementation of biotechnological methods of VSA biosynthesis. Full article
(This article belongs to the Section Plant Science)
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14 pages, 9573 KiB  
Article
Cloning and Characterization of Two Iridoid Synthase Homologs from Swertia Mussotii
by Beibei Xiang, Xiaoxue Li, Yan Wang, Xiaoxuan Tian, Zhen Yang, Lin Ma, Xia Liu and Yong Wang
Molecules 2017, 22(8), 1387; https://doi.org/10.3390/molecules22081387 - 22 Aug 2017
Cited by 18 | Viewed by 6143
Abstract
Swertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, [...] Read more.
Swertia mussotii is an important medicinal plant found on the Qinghai Tibetan Plateau that has great economic and medicinal value. This plant has enjoyed a long history of use as a curative for hepatitis. The biological activity of secoiridoids, including gentiopicroside and swertiamarin, has been mainly tested for its anti-hepatitis effects. Here, we identify two candidate genes (SmIS1 and SmIS2) that are homologues of iridoid synthase and that are components of the secoiridoid pathway in S. mussotii. Using sequencing and phylogenetic analyses, we confirm that SmIS1 and SmIS2 contain six conserved short-chain dehydrogenases/reductase (SDR) motifs and thus belong to the P5βRs group. The two purified Escherichia coli-expressed proteins reduced 8-oxogeranial to both nepetalactol and iridodials. A comparison of the kinetic parameters of SmIS1 and SmIS2 recombinant proteins revealed that SmIS2 has a lower affinity than SmIS1 for 8-oxogeranial. Transcript levels of the two genes were analysed in three different tissues of S. mussotii using semi-quantitative RT-PCR and RT-qPCR. SmIS1 and SmIS2 expression levels were more abundant in leaves and stems. This investigation adds to our knowledge of P5βRs genes in the secoiridoid synthesis pathway and provides candidate genes for genetically improving S. mussotii by enhancing secondary metabolite production. Full article
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16 pages, 4016 KiB  
Article
cDNA Isolation and Functional Characterization of UDP-d-glucuronic Acid 4-Epimerase Family from Ornithogalum caudatum
by Sen Yin, Yu-Jia Sun, Ming Liu, Li-Na Li and Jian-Qiang Kong
Molecules 2016, 21(11), 1505; https://doi.org/10.3390/molecules21111505 - 9 Nov 2016
Cited by 9 | Viewed by 5851
Abstract
d-Galacturonic acid (GalA) is an important component of GalA-containing polysaccharides in Ornithogalum caudatum. The incorporation of GalA into these polysaccharides from UDP-d-galacturonic acid (UDP-GalA) was reasonably known. However, the cDNAs involved in the biosynthesis of UDP-GalA were still unknown. [...] Read more.
d-Galacturonic acid (GalA) is an important component of GalA-containing polysaccharides in Ornithogalum caudatum. The incorporation of GalA into these polysaccharides from UDP-d-galacturonic acid (UDP-GalA) was reasonably known. However, the cDNAs involved in the biosynthesis of UDP-GalA were still unknown. In the present investigation, one candidate UDP-d-glucuronic acid 4-epimerase (UGlcAE) family with three members was isolated from O. caudatum based on RNA-Seq data. Bioinformatics analyses indicated all of the three isoforms, designated as OcUGlcAE1~3, were members of short-chain dehydrogenases/reductases (SDRs) and shared two conserved motifs. The three full-length cDNAs were then transformed to Pichia pastoris GS115 for heterologous expression. Data revealed both the supernatant and microsomal fractions from the recombinant P. pastoris expressing OcUGlcAE3 can interconvert UDP-GalA and UDP-d-glucuronic acid (UDP-GlcA), while the other two OcUGlcAEs had no activity on UDP-GlcA and UDP-GalA. Furthermore, expression analyses of the three epimerases in varied tissues of O. caudatum were performed by real-time quantitative PCR (RT-qPCR). Results indicated OcUGlcAE3, together with the other two OcUGlcAE-like genes, was root-specific, displaying highest expression in roots. OcUGlcAE3 was UDP-d-glucuronic acid 4-epimerase and thus deemed to be involved in the biosynthesis of root polysaccharides. Moreover, OcUGlcAE3 was proposed to be environmentally induced. Full article
(This article belongs to the Section Natural Products Chemistry)
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16 pages, 1484 KiB  
Article
Identification of a 4-Deoxy-l-erythro-5-hexoseulose Uronic Acid Reductase, FlRed, in an Alginolytic Bacterium Flavobacterium sp. Strain UMI-01
by Akira Inoue, Ryuji Nishiyama, Shogo Mochizuki and Takao Ojima
Mar. Drugs 2015, 13(1), 493-508; https://doi.org/10.3390/md13010493 - 16 Jan 2015
Cited by 24 | Viewed by 7990
Abstract
In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11). The monosaccharide is non-enzymatically converted to 4-deoxy-l-ery thro-5-hexoseulose uronic acid (DEH), then reduced to 2-keto-3-deoxy-d-gluconate (KDG) by a specific reductase, [...] Read more.
In alginate-assimilating bacteria, alginate is depolymerized to unsaturated monosaccharide by the actions of endolytic and exolytic alginate lyases (EC 4.2.2.3 and EC 4.2.2.11). The monosaccharide is non-enzymatically converted to 4-deoxy-l-ery thro-5-hexoseulose uronic acid (DEH), then reduced to 2-keto-3-deoxy-d-gluconate (KDG) by a specific reductase, and metabolized through the Entner–Doudoroff pathway. Recently, the NADPH-dependent reductase A1-R that belongs to short-chain dehydrogenases/reductases (SDR) superfamily was identified as the DEH-reductase in Sphingomonas sp. A1. We have subsequently noticed that an SDR-like enzyme gene, flred, occurred in the genome of an alginolytic bacterium Flavobacterium sp. strain UMI-01. In the present study, we report on the deduced amino-acid sequence of flred and DEH-reducing activity of recombinant FlRed. The deduced amino-acid sequence of flred comprised 254 residues and showed 34% amino-acid identities to that of A1-R from Sphingomonas sp. A1 and 80%–88% to those of SDR-like enzymes from several alginolytic bacteria. Common sequence motifs of SDR-superfamily enzymes, e.g., the catalytic tetrad Asn-Lys-Tyr-Ser and the cofactor-binding sequence Thr-Gly-x-x-x-Gly-x-Gly in Rossmann fold, were completely conserved in FlRed. On the other hand, an Arg residue that determined the NADPH-specificity of Sphingomonas A1-R was replaced by Glu in FlRed. Thus, we investigated cofactor-preference of FlRed using a recombinant enzyme. As a result, the recombinant FlRed (recFlRed) was found to show high specificity to NADH. recFlRed exhibited practically no activity toward variety of aldehyde, ketone, keto ester, keto acid and aldose substrates except for DEH. On the basis of these results, we conclude that FlRed is the NADH-dependent DEH-specific SDR of Flavobacterium sp. strain UMI-01. Full article
(This article belongs to the Special Issue Green Chemistry Approach to Marine Products)
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