@article{xi_wang_cagle_zhu_odle_xie_2023, title={Exploring the Prebiotic Activities of Proanthocyanidins on a Platform Using the Three-Dimensionally (3D)-Cultured Organoids}, volume={101}, ISSN={["1525-3163"]}, DOI={10.1093/jas/skad281.421}, abstractNote={Abstract Using antibiotics, the antimicrobial substance active against bacteria and promoting growth efficiency, in feedstuffs and food supply has been attracting great attention due to its side effects and contribution to antibiotic resistance. Therefore, exploring talternatives to antibiotics is of great significance and urgent need for human health and animal industries with a sustainable high efficiency. The objective of this study is to evaluate the biological and medicinal activities of the monomers of proanthocyanidins (PAs), such as epicatechin, epigallocatechin gallate, and flavanols such as quercetin, isoquercetrin, and rutin generated from engineered plants. The evaluations were performed in intestinal organoids isolated from ileum of neonatal piglets. The organoids after expended in vitro were incubated with or without the candidate compounds for 24 hours, and then treated with or without polyinosinic-polycytidylic acid [Poly (I:C), 10 µg/mL] for another 24 hours, a synthetic analog of double-stranded RNA (can induce a molecular pattern associated with viral infections). Cell proliferation, barrier function, toll-like receptor pathway and apoptosis as well as organic cation transporters were assessed by measuring the abundance of the corresponding genes. Concentration gradient (0 to 200 µM) measurements showed that isoquercetrin and epigallocatechin gallate might stimulate the intestinal cell proliferation, promote the uptake of L-carnitine, and increase barrier function and mucin secretion (P < 0.05). Epigallocatechin gallate and epicatechin might affect the inflammation responses via modifying expression of Interleukins but not induce apoptosis. Interactions between the examined these compounds and Poly (I: C) were not observed (P > 0.05), but the influence on the challenged pattern were tested for some of the measured genes (P < 0.05). Based on the data collected from this in vitro study, we conclude that epigallocatechin gallate is a potential PA monomer with all prebiotic characteristics and its applicable values in feedstuff and food supply should be studied in vivo studies in domestic (food) animals with and without antigen challenges. Supported by NC Biotechnology Center project (1107)2022-3082 and the North Carolina Agricultural Research Hatch Projects 02780.}, journal={JOURNAL OF ANIMAL SCIENCE}, author={Xi, Lin and Wang, Feng and Cagle, Daisy and Zhu, Yue and Odle, Jack and Xie, Deyu}, year={2023}, month={Nov}, pages={355–356} } @article{zhu_yuzuak_sun_xie_2023, title={Identification and biosynthesis of plant papanridins, a group of novel oligomeric flavonoids}, volume={16}, ISSN={["1752-9867"]}, DOI={10.1016/j.molp.2023.09.015}, abstractNote={The discovery of novel flavonoids and elucidation of their biosynthesis are fundamental to understanding their roles in plants and their benefits for human and animal health. Here, we report a new pathway for polymerization of a group of novel oligomeric flavonoids in plants. We engineered red cells for discovering genes of interest involved in the flavonoid pathway and identified a gene encoding a novel flavanol polymerase (FP) localized in the central vacuole. FP catalyzes the polymerization of flavanols, such as epicatechin and catechin, to produce yellowish dimers or oligomers. Structural elucidation shows that these compounds feature a novel oligomeric flaven–flavan (FF) skeleton linked by interflavan–flaven and interflaven bonds, distinguishing them from proanthocyanidins and dehydrodicatechins. Detailed chemical and physical characterizations further confirmed the novel FFs as flavonoids. Mechanistic investigations demonstrated that FP polymerizes flavan-3-ols and flav-2-en-3-ol carbocation, forming dimeric or oligomeric flaven-4→8-flavans, which we term “papanridins.” Data from transgenic experiments, mutant analysis, metabolic profiling, and phylogenetic analyses show that the biosynthesis of papanridins is prevalent in cacao, grape, blueberry, corn, rice, Arabidopsis, and other species in the plant kingdom. In summary, our study discoveries a group of novel oligomeric flavonoids, namely papanridins, and reveals that a novel FP-mediated polymerization mechanism for the biosynthesis of papanridins in plants.}, number={11}, journal={MOLECULAR PLANT}, author={Zhu, Yue and Yuzuak, Seyit and Sun, Xiaoyan and Xie, De-Yu}, year={2023}, month={Nov}, pages={1773–1793} } @article{judd_dong_sun_zhu_li_xie_2023, title={Metabolic engineering of the anthocyanin biosynthetic pathway in Artemisia annua and relation to the expression of the artemisinin biosynthetic pathway}, volume={257}, ISSN={["1432-2048"]}, url={https://doi.org/10.1007/s00425-023-04091-6}, DOI={10.1007/s00425-023-04091-6}, number={3}, journal={PLANTA}, author={Judd, Rika and Dong, Yilun and Sun, Xiaoyan and Zhu, Yue and Li, Mingzhuo and Xie, De-Yu}, year={2023}, month={Mar} } @article{zhu_scholle_kisthardt_xie_2022, title={
Flavonols and dihydroflavonols inhibit the main protease activity of SARS-CoV-2 and the replication of human coronavirus 229E
}, volume={571}, ISSN={["1089-862X"]}, DOI={10.1016/j.virol.2022.04.005}, abstractNote={Since December 2019, the deadly novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the current COVID-19 pandemic. To date, vaccines are available in the developed countries to prevent the infection of this virus; however, medicines are necessary to help control COVID-19. Human coronavirus 229E (HCoV-229E) causes the common cold. The main protease (Mpro) is an essential enzyme required for the multiplication of these two viruses in the host cells, and thus is an appropriate candidate to screen potential medicinal compounds. Flavonols and dihydroflavonols are two groups of plant flavonoids. In this study, we report docking simulation with two Mpro enzymes and five flavonols and three dihydroflavonols, in vitro inhibition of the SARS-CoV-2 Mpro, and in vitro inhibition of the HCoV 229E replication. The docking simulation results predicted that (+)-dihydrokaempferol, (+)- dihydroquercetin, (+)-dihydromyricetin, kaempferol, quercetin, myricentin, isoquercitrin, and rutin could bind to at least two subsites (S1, S1', S2, and S4) in the binding pocket and inhibit the activity of SARS-CoV-2 Mpro. Their affinity scores ranged from -8.8 to -7.4 (kcal/mol). Likewise, these compounds were predicted to bind and inhibit the HCoV-229E Mpro activity with affinity scores ranging from -7.1 to -7.8 (kcal/mol). In vitro inhibition assays showed that seven available compounds effectively inhibited the SARS-CoV-2 Mpro activity and their IC50 values ranged from 0.125 to 12.9 μM. Five compounds inhibited the replication of HCoV-229E in Huh-7 cells. These findings indicate that these antioxidative flavonols and dihydroflavonols are promising candidates for curbing the two viruses.}, journal={VIROLOGY}, author={Zhu, Yue and Scholle, Frank and Kisthardt, Samantha C. and Xie, De-Yu}, year={2022}, month={Jun}, pages={21–33} } @article{li_he_la hovary_zhu_dong_liu_xing_liu_jie_ma_et al._2022, title={A de novo regulation design shows an effectiveness in altering plant secondary metabolism}, volume={37}, ISSN={["2090-1224"]}, url={http://dx.doi.org/10.1016/j.jare.2021.06.017}, DOI={10.1016/j.jare.2021.06.017}, abstractNote={Transcription factors (TFs) and cis-regulatory elements (CREs) control gene transcripts involved in various biological processes. We hypothesize that TFs and CREs can be effective molecular tools for De Novo regulation designs to engineer plants. We selected two Arabidopsis TF types and two tobacco CRE types to design a De Novo regulation and evaluated its effectiveness in plant engineering. G-box and MYB recognition elements (MREs) were identified in four Nicotiana tabacum JAZs (NtJAZs) promoters. MRE-like and G-box like elements were identified in one nicotine pathway gene promoter. TF screening led to select Arabidopsis Production of Anthocyanin Pigment 1 (PAP1/MYB) and Transparent Testa 8 (TT8/bHLH). Two NtJAZ and two nicotine pathway gene promoters were cloned from commercial Narrow Leaf Madole (NL) and KY171 (KY) tobacco cultivars. Electrophoretic mobility shift assay (EMSA), cross-linked chromatin immunoprecipitation (ChIP), and dual-luciferase assays were performed to test the promoter binding and activation by PAP1 (P), TT8 (T), PAP1/TT8 together, and the PAP1/TT8/Transparent Testa Glabra 1 (TTG1) complex. A DNA cassette was designed and then synthesized for stacking and expressing PAP1 and TT8 together. Three years of field trials were performed by following industrial and GMO protocols. Gene expression and metabolic profiling were completed to characterize plant secondary metabolism. PAP1, TT8, PAP1/TT8, and the PAP1/TT8/TTG1 complex bound to and activated NtJAZ promoters but did not bind to nicotine pathway gene promoters. The engineered red P + T plants significantly upregulated four NtJAZs but downregulated the tobacco alkaloid biosynthesis. Field trials showed significant reduction of five tobacco alkaloids and four carcinogenic tobacco specific nitrosamines in most or all cured leaves of engineered P + T and PAP1 genotypes. G-boxes, MREs, and two TF types are appropriate molecular tools for a De Novo regulation design to create a novel distant-pathway cross regulation for altering plant secondary metabolism.}, journal={JOURNAL OF ADVANCED RESEARCH}, publisher={Elsevier BV}, author={Li, Mingzhuo and He, Xianzhi and La Hovary, Christophe and Zhu, Yue and Dong, Yilun and Liu, Shibiao and Xing, Hucheng and Liu, Yajun and Jie, Yucheng and Ma, Dongming and et al.}, year={2022}, month={Mar}, pages={43–60} } @article{zhu_xie_2020, title={Docking Characterization and in vitro Inhibitory Activity of Flavan-3-ols and Dimeric Proanthocyanidins Against the Main Protease Activity of SARS-Cov-2}, volume={11}, ISSN={1664-462X}, url={http://dx.doi.org/10.3389/fpls.2020.601316}, DOI={10.3389/fpls.2020.601316}, abstractNote={We report to use the main protease (M pro ) of SARS-Cov-2 to screen plant flavan-3-ols and proanthocyanidins. Twelve compounds, (–)-afzelechin (AF), (–)-epiafzelechin (EAF), (+)-catechin (CA), (–)-epicatechin (EC), (+)-gallocatechin (GC), (–)-epigallocatechin (EGC), (+)-catechin-3-O-gallate (CAG), (–)-epicatechin-3-O-gallate (ECG), (–)-gallocatechin-3-O-gallate (GCG), (–)-epigallocatechin-3-O-gallate (EGCG), procyanidin A2 (PA2), and procyanidin B2 (PB2), were selected for docking simulation. The resulting data predicted that all 12 metabolites could bind to M pro . The affinity scores of PA2 and PB2 were predicted to be −9.2, followed by ECG, GCG, EGCG, and CAG, −8.3 to −8.7, and then six flavan-3-ol aglycones, −7.0 to −7.7. Docking characterization predicted that these compounds bound to three or four subsites (S1, S1′, S2, and S4) in the binding pocket of M pro via different spatial ways and various formation of one to four hydrogen bonds. In vitro analysis with 10 available compounds showed that CAG, ECG, GCG, EGCG, and PB2 inhibited the M pro activity with an IC 50 value, 2.98 ± 0.21, 5.21 ± 0.5, 6.38 ± 0.5, 7.51 ± 0.21, and 75.3 ± 1.29 μM, respectively, while CA, EC, EGC, GC, and PA2 did not have inhibitory activities. To further substantiate the inhibitory activities, extracts prepared from green tea (GT), two muscadine grapes (MG), cacao, and dark chocolate (DC), which are rich in CAG, ECG, GAG, EGCG, or/and PB2, were used for inhibitory assay. The resulting data showed that GT, two MG, cacao, and DC extracts inhibited the M pro activity with an IC 50 value, 2.84 ± 0.25, 29.54 ± 0.41, 29.93 ± 0.83, 153.3 ± 47.3, and 256.39 ± 66.3 μg/ml, respectively. These findings indicate that on the one hand, the structural features of flavan-3-ols are closely associated with the affinity scores; on the other hand, the galloylation and oligomeric types of flavan-3-ols are critical in creating the inhibitory activity against the M pro activity.}, journal={Frontiers in Plant Science}, publisher={Frontiers Media SA}, author={Zhu, Yue and Xie, De-Yu}, year={2020}, month={Nov} } @article{ma_li_zhu_xie_2017, title={Overexpression and Suppression of Artemisia annua 4-Hydroxy-3-Methylbut-2-enyl Diphosphate Reductase 1 Gene (AaHDR1) Differentially Regulate Artemisinin and Terpenoid Biosynthesis}, volume={8}, ISSN={["1664-462X"]}, DOI={10.3389/fpls.2017.00077}, abstractNote={4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR) catalyzes the last step of the 2-C-methyl-D-erythritol 4- phosphate (MEP) pathway to synthesize isopentenyl pyrophosphate (IPP) and dimethylallyl diphosphate (DMAPP). To date, little is known regarding effects of an increase or a decrease of a HDR expression on terpenoid and other metabolite profiles in plants. In our study, an Artemisia annua HDR cDNA (namely AaHDR1) was cloned from leaves. Expression profiling showed it was highly expressed in leaves, roots, stems, and flowers with different levels. Green florescence protein fusion and confocal microscope analyses showed that AaHDR1 was localized in chloroplasts. The overexpression of AaHDR1 increased contents of artemisinin, arteannuin B and other sesquiterpenes, and multiple monoterpenes. By contrast, the suppression of AaHDR1 by anti-sense of led to opposite results. In addition, an untargeted metabolic profiling showed that the overexpression and suppression altered non-polar metabolite profiles. In conclusion, the overexpression and suppression of AaHDR1 protein level in plastids differentially affect artemisinin and other terpenoid biosynthesis, and alter non-polar metabolite profiles of A. annua. Particularly, its overexpression leading to the increase of artemisinin production is informative to future metabolic engineering of this antimalarial medicine.}, journal={FRONTIERS IN PLANT SCIENCE}, author={Ma, Dongming and Li, Gui and Zhu, Yue and Xie, De-Yu}, year={2017}, month={Jan} } @article{ma_li_alejos-gonzalez_zhu_xue_wang_zhang_li_ye_wang_et al._2017, title={Overexpression of a type-I isopentenyl pyrophosphate isomerase of Artemisia annua in the cytosol leads to high arteannuinB production and artemisinin increase}, volume={91}, DOI={10.1111/tpj.13583}, abstractNote={Summary We recently characterized a gene–terpene network that is associated with artemisinin biosynthesis in self‐pollinated ( SP ) Artemisia annua , an effective antimalarial plant. We hypothesize that an alteration of gene expression in the network may improve the production of artemisinin and its precursors. In this study, we cloned an isopentenyl pyrophosphate isomerase ( IPPI ) cDNA , Aa IPPI 1 , from Artemisia annua (Aa). The full‐length cDNA encodes a type‐I IPPI containing a plastid transit peptide ( PTP ) at its amino terminus. After the removal of the PTP , the recombinant truncated Aa IPPI 1 isomerized isopentenyl pyrophosphate ( IPP ) to dimethyl allyl pyrophosphate ( DMAPP ) and vice versa . The steady‐state equilibrium ratio of IPP / DMAPP in the enzymatic reactions was approximately 1:7. The truncated Aa IPPI 1 was overexpressed in the cytosol of the SP A. annua variety. The leaves of transgenic plants produced approximately 4% arteannuin B (g g −1 , dry weight, dw) and 0.17–0.25% artemisinin (g g −1 , dw), the levels of which were significantly higher than those in the leaves of wild‐type plants. In addition, transgenic plants showed an increase in artemisinic acid production of more than 1% (g g −1 , dw). In contrast, isoprene formation was significantly reduced in transgenic plants. These results provide evidence that overexpression of AaIPPI1 in the cytosol can lead to metabolic alterations of terpenoid biosynthesis, and show that these transgenic plants have the potential to yield high production levels of arteannuin B as a new precursor source for artemisinin.}, number={3}, journal={Plant Journal}, author={Ma, D. M. and Li, G. and Alejos-Gonzalez, F. and Zhu, Y. and Xue, Z. and Wang, A. M. and Zhang, H. and Li, X. and Ye, H. C. and Wang, H. and et al.}, year={2017}, pages={466–479} }