DOCSLIB.ORG

Metabolic Engineering of Microbial Cell Factories for Biosynthesis of Flavonoids: a Review

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Metabolic Engineering of Microbial Cell Factories for Biosynthesis of Flavonoids: a Review molecules Review Metabolic Engineering of Microbial Cell Factories for Biosynthesis of Flavonoids: A Review Hanghang Lou 1,†, Lifei Hu 2,†, Hongyun Lu 1, Tianyu Wei 1 and Qihe Chen 1,* 1 Department of Food Science and Nutrition, Zhejiang University, Hangzhou 310058, China; [email protected] (H.L.); [email protected] (H.L.); [email protected] (T.W.) 2 Hubei Key Lab of Quality and Safety of Traditional Chinese Medicine & Health Food, Huangshi 435100, China; [email protected] * Correspondence: [email protected]; Tel.: +86-0571-8698-4316 † These authors are equally to this manuscript. Abstract: Flavonoids belong to a class of plant secondary metabolites that have a polyphenol structure. Flavonoids show extensive biological activity, such as antioxidative, anti-inflammatory, anti-mutagenic, anti-cancer, and antibacterial properties, so they are widely used in the food, phar- maceutical, and nutraceutical industries. However, traditional sources of flavonoids are no longer sufficient to meet current demands. In recent years, with the clarification of the biosynthetic pathway of flavonoids and the development of synthetic biology, it has become possible to use synthetic metabolic engineering methods with microorganisms as hosts to produce flavonoids. This article mainly reviews the biosynthetic pathways of flavonoids and the development of microbial expression systems for the production of flavonoids in order to provide a useful reference for further research on synthetic metabolic engineering of flavonoids. Meanwhile, the application of co-culture systems in the biosynthesis of flavonoids is emphasized in this review. Citation: Lou, H.; Hu, L.; Lu, H.; Wei, Keywords: flavonoids; metabolic engineering; co-culture system; biosynthesis; microbial cell factories T.; Chen, Q. Metabolic Engineering of Microbial Cell Factories for Biosynthesis of Flavonoids: A Review. Molecules 2021, 26, 4522. https:// 1. Introduction doi.org/10.3390/molecules26154522 Flavonoids are a kind of secondary metabolite produced by plants in the process of long-term natural selection. They are widely found in vegetables, fruits, and medicinal Academic Editor: Luciana Mosca plants. Structurally, they possess a C6–C3–C6 carbon framework, which is formed by con- necting two aromatic rings (rings A and B) through a heterocyclic ring that contains three Received: 5 July 2021 carbon atoms (ring C) [1] (Figure1). Thus far, about 10,000 flavonoids have been discovered, Accepted: 25 July 2021 which comprise one of the largest families of natural products [2]. There are two forms of Published: 27 July 2021 flavonoids in nature, one occurs in aglycone form in plants, and the other combines with sugar to form glycosides [3]. Common sugars attached to flavonoids include D-glucose, Publisher’s Note: MDPI stays neutral D-galactose, L-rhamnose, D-xylose, D-glucuronic acid, sophorose, rutinose, tangerine peel with regard to jurisdictional claims in published maps and institutional affil- sugar, and gentianose [3,4]. Flavonoids normally occur in plants as O-glycosylated deriva- iations. tives [5], and sugars are mostly attached to the hydroxyl groups at position 3 of the C-ring and positions 5 and 7 of the A-ring. However, some plants, such as maize, wheat, and rice, also produce C-glycosylated flavonoids [5]. These flavonoids play important roles in plants, animals, and humans. In plants, flavonoids are responsible for flower color and aroma and attract pollinators to disperse fruit and help plants propagate [6]. In addition, they also help Copyright: © 2021 by the authors. plants combat various biotic and abiotic stresses, including ultraviolet protection in leaves, Licensee MDPI, Basel, Switzerland. protection from microbial infections and herbivores, and defending from physical and This article is an open access article radical damage [7,8]. As a kind of phytochemicals, flavonoids cannot be synthesized by distributed under the terms and conditions of the Creative Commons humans or animals. Therefore humans generally absorb flavonoids from fruits, vegetables, Attribution (CC BY) license (https:// legumes, and other foods, and flavonoids are thought to have health-promoting properties. creativecommons.org/licenses/by/ Most studies have found that flavonoids have multiple physiological and pharmacological 4.0/). activities, such as antioxidative, anti-inflammatory, anti-mutagenic, anti-cancer, antiviral, Molecules 2021, 26, 4522. https://doi.org/10.3390/molecules26154522 https://www.mdpi.com/journal/molecules Molecules 2021, 26, x FOR PEER REVIEW 2 of 18 and flavonoids are thought to have health-promoting properties. Most studies have found that flavonoids have multiple physiological and pharmacological activities, such Molecules 2021, 26, 4522 as antioxidative, anti-inflammatory, anti-mutagenic, anti-cancer, antiviral, antibacterial,2 of 17 and protecting the heart and cerebrovascular vessels [6,9,10]. Therefore, flavonoids show extensive applications in functional foods and some medicine. In fact, some flavonoid extracts,antibacterial, such as andMasquelier’s protecting grape the heartseed extr andact, cerebrovascular have been sold vessels as dietary [6,9 ,10supplements]. Therefore, [11].flavonoids show extensive applications in functional foods and some medicine. In fact, some flavonoid extracts, such as Masquelier’s grape seed extract, have been sold as dietary supplements [11]. Figure 1. General structure of flavonoids. Figure 1. General structure of flavonoids. The wide application of flavonoids has led to their increasing demand. At present, flavonoids are generally extracted from plants. However, plants have a long culture period, The wide application of flavonoids has led to their increasing demand. At present, and their growth is also limited by the temperature and season. Additionally, the process flavonoids are generally extracted from plants. However, plants have a long culture pe- of extraction and purification could increase the cost of production and lead to the loss of riod, and their growth is also limited by the temperature and season. Additionally, the the product and the reduction of product activity [12]. Therefore, it is a huge challenge process of extraction and purification could increase the cost of production and lead to to produce these compounds on a large scale through plant extraction to meet medical the loss of the product and the reduction of product activity [12]. Therefore, it is a huge and nutritional needs [13]. The total synthesis of some flavonoids has been realized by challenge to produce these compounds on a large scale through plant extraction to meet chemical approaches; however, this method required some extreme reaction conditions, medical and nutritional needs [13]. The total synthesis of some flavonoids has been real- such as high temperature, strong acid, strong base, and the use of heavy metal, which is izedunfriendly by chemical to the approaches; environment however, and hindered this method its scale-up required and commercial some extreme application. reaction In conditions,recent years, such the as productionhigh temperature, of flavonoids strong by acid, microbial strong cell base, factories and the has use attracted of heavy wide metal,attention which with is unfriendly the development to the environm of metabolicent engineeringand hindered tools, its scale-up the in-depth and commercial studies on the application.synthesis pathwayIn recent ofyears, flavonoids the production and available of flavonoids substrates by [7, 12microbial,14]. Engineering cell factories the wholehas attractedbiosynthetic wide pathway attention to producewith the flavonoids developm inent microorganisms of metabolic hasengineering many advantages, tools, the such in-depthas a short studies production on the cycle, synthesis low waste pathway production, of flavonoids low energy and requirements, available substrates and mass [7,12,14].production. Engineering Some microorganisms, the whole biosynthetic such as Escherichia pathway to coli produce(E. coli), flavonoidsSaccharomyces in micro- cerevisiae organisms(S. cerevisiae has), manyYarrowia advantages, lipolytica (Y. such lipolytica) as a sh,ort grow production faster, and cycle, various low genetic waste manipulationsproduction, lowof energy this species requirements, and metabolic and modificationsmass production. have Some been microorganisms, studied in detail, such so it hasas Esche- become richiapossible coli (E. to coli use), theseSaccharomyces microorganisms cerevisiae as ( hostsS. cerevisiae for the), productionYarrowia lipolytica of flavonoids (Y. lipolytica) [2,15–17, ]. growIn a faster, previous and study, various the genetic production manipulations of flavonoids of this by geneticallyspecies and engineered metabolic bacteriamodifica- had tionsnot have been been achieved, studied and in one detail, of the so barriersit has become to produce possible these to compounds use these microorganisms was the difficulty as inhosts the for expression the production of active of C4H.flavonoids In 2003, [2,15– a 4CL17]. (4-coumarin-CoA In a previous study, ligase) the production was discovered of flavonoidsin the gram-positive by genetically filamentous engineered bacterium bacteria hadStreptomyces not been achieved, coelicolor [and18]. one The of enzyme the bar- not riersonly to activatedproduce these cinnamic compounds acid to was cinnamoyl-CoA the difficulty but in the also expression activated of 4-coumaric active C4H. acid In to 2003,4-coumaroyl-CoA. a 4CL (4-coumarin-CoA The use of ligase) the enzyme was woulddiscovered
Load more

Recommended publications
  • Key Enzymes Involved in the Synthesis of Hops Phytochemical Compounds: from Structure, Functions to Applications
    International Journal of Molecular Sciences Review Key Enzymes Involved in the Synthesis of Hops Phytochemical Compounds: From Structure, Functions to Applications Kai Hong , Limin Wang, Agbaka Johnpaul , Chenyan Lv * and Changwei Ma * College of Food Science and Nutritional Engineering, China Agricultural University, 17 Qinghua Donglu Road, Haidian District, Beijing 100083, China; [email protected] (K.H.); [email protected] (L.W.); [email protected] (A.J.) * Correspondence: [email protected] (C.L.); [email protected] (C.M.); Tel./Fax: +86-10-62737643 (C.M.) Abstract: Humulus lupulus L. is an essential source of aroma compounds, hop bitter acids, and xanthohumol derivatives mainly exploited as flavourings in beer brewing and with demonstrated potential for the treatment of certain diseases. To acquire a comprehensive understanding of the biosynthesis of these compounds, the primary enzymes involved in the three major pathways of hops’ phytochemical composition are herein critically summarized. Hops’ phytochemical components impart bitterness, aroma, and antioxidant activity to beers. The biosynthesis pathways have been extensively studied and enzymes play essential roles in the processes. Here, we introduced the enzymes involved in the biosynthesis of hop bitter acids, monoterpenes and xanthohumol deriva- tives, including the branched-chain aminotransferase (BCAT), branched-chain keto-acid dehydroge- nase (BCKDH), carboxyl CoA ligase (CCL), valerophenone synthase (VPS), prenyltransferase (PT), 1-deoxyxylulose-5-phosphate synthase (DXS), 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (HDR), Geranyl diphosphate synthase (GPPS), monoterpene synthase enzymes (MTS), cinnamate Citation: Hong, K.; Wang, L.; 4-hydroxylase (C4H), chalcone synthase (CHS_H1), chalcone isomerase (CHI)-like proteins (CHIL), Johnpaul, A.; Lv, C.; Ma, C.
    [Show full text]
  • Building Blue E. Coli: Engineering the Anthocyanin Biosynthetic Pathway
    Building blue E. coli: Engineering the anthocyanin biosynthetic pathway Leah Johnston1, Lucas Jarche1, Alexander Porter1, Julie Ryu1, Emma Price1, Emma Finlayson-Trick1, Khaled Aly1, Paul Bissonnette2, Eric Pace3, Dr. John Rohde1 1Department of Microbiology and Immunology, 2Department of Chemistry, 3Department of Agriculture Dalhousie University, Halifax, Canada, B3H 1X5 THE ANTHOCYANIN PATHWAY METHODS RESULTS 1kb (B) Method of Template DNA for each of the genes required for the production of the Anthocyanin biosynthetic Gene Size (bp) Function Epitope ladder (B) Acquisition of pathway in E. coli were acquired from the following sources outlined in Figure 1 (B). (A) Name Tag Template DNA 10 Phenylalanine Lyase involved in Amplified from A. (A) 2762 polyphenol S-Tag thaliana cDNA, Ammonia . synthesis. insert in Phenylalanine Ammonia Lyase and 4-Coumaroyl CoA Ligase were acquired as inserts in a 3.0. Lyase pACYCDuet-1 pACYCDuet-1 plasmid from Sinyuan Wang et. al (2005) at Utah State University. Cinnamate 4- 2.0 Phenylalanine from Utah State Hydroxylase-HA fusion protein was inserted into this plasmid at the end of the PAL coding region 1.5 Phenylalanine ammonia lyase University using restriction enzymes and transformed into BL21 CaCl2 competent E. coli with T7 RNA Cinnamate 4- Oxidoreductase Amplified from A. Cinnamic Acid 1518 involved in HA-Tag thaliana cDNA, Polymerase activity. A western blot with anti-HA antibody was used to detect the presence of the 1.0 Hydroxylase phenylalanine purchased from C4H fusion protein in the resultant cells. Cinnamate 4-hydroxylase metabolism. TAIR database P-Coumaric Acid 4-Coumaroyl Ligase involved in Insert in 2546 phenylpropanoid His-Tag pACYCDuet-1 Chalcone Synthase, Chalcone Isomerase and Flavanone 3-Hydroxylase were amplified from the CoA Ligase 0.5 4-coumaryl-coenzyme A ligase biosynthesis.
    [Show full text]
  • Interpreting Sources and Endocrine Active Components of Trace Organic Contaminant Mixtures in Minnesota Lakes
    INTERPRETING SOURCES AND ENDOCRINE ACTIVE COMPONENTS OF TRACE ORGANIC CONTAMINANT MIXTURES IN MINNESOTA LAKES by Meaghan E. Guyader © Copyright by Meaghan E. Guyader, 2018 All Rights Reserved A thesis submitted to the Faculty and the Board of Trustees of the Colorado School of Mines in partial fulfillment of the requirements for the degree of Doctor of Philosophy (Civil and Environmental Engineering). Golden, Colorado Date _____________________________ Signed: _____________________________ Meaghan E. Guyader Signed: _____________________________ Dr. Christopher P. Higgins Thesis Advisor Golden, Colorado Date _____________________________ Signed: _____________________________ Dr. Terri S. Hogue Professor and Department Head Department of Civil and Environmental Engineering ii ABSTRACT On-site wastewater treatment systems (OWTSs) are a suspected source of widespread trace organic contaminant (TOrC) occurrence in Minnesota lakes. TOrCs are a diverse set of synthetic and natural chemicals regularly used as cleaning agents, personal care products, medicinal substances, herbicides and pesticides, and foods or flavorings. Wastewater streams are known to concentrate TOrC discharges to the environment, particularly accumulating these chemicals at outfalls from centralized wastewater treatment plants. Fish inhabiting these effluent dominated environments are also known to display intersex qualities. Concurrent evidence of this phenomenon, known as endocrine disruption, in Minnesota lake fish drives hypotheses that OWTSs, the primary form of wastewater treatment in shoreline residences, may contribute to TOrC occurrence and the endocrine activity in these water bodies. The causative agents specific to fish in this region remain poorly understood. The objective of this dissertation was to investigate OWTSs as sources of TOrCs in Minnesota lakes, and TOrCs as potential causative agents for endocrine disruption in resident fish.
    [Show full text]
  • L.) Leaves Tao Jiang1,3, Kunyuan Guo2,3, Lingdi Liu1, Wei Tian1, Xiaoliang Xie1, Saiqun Wen1 & Chunxiu Wen1*
    www.nature.com/scientificreports OPEN Integrated transcriptomic and metabolomic data reveal the favonoid biosynthesis metabolic pathway in Perilla frutescens (L.) leaves Tao Jiang1,3, Kunyuan Guo2,3, Lingdi Liu1, Wei Tian1, Xiaoliang Xie1, Saiqun Wen1 & Chunxiu Wen1* Perilla frutescens (L.) is an important medicinal and edible plant in China with nutritional and medical uses. The extract from leaves of Perilla frutescens contains favonoids and volatile oils, which are mainly used in traditional Chinese medicine. In this study, we analyzed the transcriptomic and metabolomic data of the leaves of two Perilla frutescens varieties: JIZI 1 and JIZI 2. A total of 9277 diferentially expressed genes and 223 favonoid metabolites were identifed in these varieties. Chrysoeriol, apigenin, malvidin, cyanidin, kaempferol, and their derivatives were abundant in the leaves of Perilla frutescens, which were more than 70% of total favonoid contents. A total of 77 unigenes encoding 15 enzymes were identifed as candidate genes involved in favonoid biosynthesis in the leaves of Perilla frutescens. High expression of the CHS gene enhances the accumulation of favonoids in the leaves of Perilla frutescens. Our results provide valuable information on the favonoid metabolites and candidate genes involved in the favonoid biosynthesis pathways in the leaves of Perilla frutescens. Perilla frutescens (L.), which is a self-compatible annual herb, belongs to the family Lamiaceae. Tis species has been widely cultivated in China, Japan, and Korea for centuries. Perilla frutescens is an important medicinal and edible plant in China with medical and nutritional uses 1. Its leaves can be utilized as a transitional medicinal herb, as a vegetable, and as a spice, and its seeds can be processed into foods and nutritional edible oils 2.
    [Show full text]
  • Analysis of the Chromosomal Clustering of Fusarium-Responsive
    www.nature.com/scientificreports OPEN Analysis of the chromosomal clustering of Fusarium‑responsive wheat genes uncovers new players in the defence against head blight disease Alexandre Perochon1,2, Harriet R. Benbow1,2, Katarzyna Ślęczka‑Brady1, Keshav B. Malla1 & Fiona M. Doohan1* There is increasing evidence that some functionally related, co‑expressed genes cluster within eukaryotic genomes. We present a novel pipeline that delineates such eukaryotic gene clusters. Using this tool for bread wheat, we uncovered 44 clusters of genes that are responsive to the fungal pathogen Fusarium graminearum. As expected, these Fusarium‑responsive gene clusters (FRGCs) included metabolic gene clusters, many of which are associated with disease resistance, but hitherto not described for wheat. However, the majority of the FRGCs are non‑metabolic, many of which contain clusters of paralogues, including those implicated in plant disease responses, such as glutathione transferases, MAP kinases, and germin‑like proteins. 20 of the FRGCs encode nonhomologous, non‑metabolic genes (including defence‑related genes). One of these clusters includes the characterised Fusarium resistance orphan gene, TaFROG. Eight of the FRGCs map within 6 FHB resistance loci. One small QTL on chromosome 7D (4.7 Mb) encodes eight Fusarium‑ responsive genes, fve of which are within a FRGC. This study provides a new tool to identify genomic regions enriched in genes responsive to specifc traits of interest and applied herein it highlighted gene families, genetic loci and biological pathways of importance in the response of wheat to disease. Prokaryote genomes encode co-transcribed genes with related functions that cluster together within operons. Clusters of functionally related genes also exist in eukaryote genomes, including fungi, nematodes, mammals and plants1.
    [Show full text]
  • Synthesis of Ochnaflavone and Its Inhibitory Activity on PGE 2
    Synthesis and Biological Activity of Ochnaflavone Bull. Korean Chem. Soc. 2014, Vol. 35, No. 11 3219 http://dx.doi.org/10.5012/bkcs.2014.35.11.3219 Synthesis of Ochnaflavone and Its Inhibitory Activity on PGE2 Production Sung Soo Kim,† Van Anh Vo,† and Haeil Park* College of Pharmacy and †Medicine of Kangwon National University, Chuncheon 200-701, Korea *E-mail: [email protected] Received June 5, 2014, Accepted July 11, 2014 Ochnaflavone, a naturally occurring biflavonoid composed of two units of apigenin (5,7,4'-trihydroxyflavone) joined via a C-O-C linkage, was first synthesized and evaluated its inhibitory activity on PGE2 production. Total synthesis was accomplished through modified Ullmann diaryl ether formation as a key step. Coupling reactions of 4'-halogenoflavones and 3'-hydroxy-5,7,4'-trimethoxyflavone were explored in diverse reaction conditions. The reaction of 4'-fluoro-5,7-dimethoxyflavone (2c) and 3'-hydroxy-5,7,4'-trimethoxyflavone (2d) in N,N-dimethylacetamide gave the coupled compound 3 in 58% yield. Synthetic ochnaflavone strongly inhibited PGE2 production (IC50 = 1.08 µM) from LPS-activated RAW 264.7 cells, which was due to reduced expression of COX-2. On the contrary, the inhibition mechanism of wogonin was somewhat different from that of ochnaflavone although wogonin, a natural occurring anti-inflammatory flavonoid, showed strong inhibitory activity of PGE2 production (IC50 = 0.52 µM), and seems to be COX-2 enzyme inhibition. Our concise total synthesis of ochnaflavone enable us to provide sufficient quantities of material for advanced biological studies as well as to efficiently prepare derivatives for structure-activity relationship study.
    [Show full text]
  • 1 Termite Feeding Deterrent from Japanese Larch Wood K. Chen A, W
    Termite feeding deterrent from Japanese larch wood K. Chen a, W. Ohmurab, S. Doic, M. Aoyamad* a Kunming University of Science and Technology, Kunming, People’s Republic of China b Forestry and Forest Products Research Institute, Tsukuba 305-8687, Japan c Institute of Wood Technology, Akita Prefectural University, Noshiro 016-0876, Japan d Laboratory of Bioresource Science, Department of Applied Chemistry, Kitami Institute of Technology, Kitami 090-8507, Japan Abstract Extraction of flavonoids from Japanese larch (Larix leptolepis) wood with water was carried out to prepare a termite feeding deterrent. A two-stage procedure for the extraction was adopted. The first extraction step was performed at ambient temperature (22C) and the second at elevated temperatures ranging 50-100C. The first step mainly gave a mixture of polysaccharides together with small amount of flavonoids. At the *Corresponding author. I will move to new laboratory (kitami Institute of Technology) in January, 2004. I can not, at present, supply my facsimile number and e-mail address. They will be available at the time of proof-reading. 1 second step, the yield of extract and its chemical composition were greatly affected by the temperature. The yield of solubilised carbohydrates steadily increased with a rise in the temperature, while the overall yield of flavonoids reached its optimum at 70C. An additional increase in the temperature resulted in a decrease in the yield. Model experiments using dihydroflavonols confirmed the occurrence of oxidative dehydrogenation and/or intramolecular rearrangement during the hydrothermal treatment at higher temperatures. The crude water extracts showed strong feeding deterrent activities against the subterranean termite, Coptotermes formosanus, in a choice paper disc assay.
    [Show full text]
  • Combinatorial Biosynthesis of Non-Bacterial and Unnatural Flavonoids, Stilbenoids and Curcuminoids by Microorganisms Sueharu Horinouchi
    J. Antibiot. 61(12): 709–728, 2008 THE JOURNAL OF REVIEW ARTICLE ANTIBIOTICS Combinatorial Biosynthesis of Non-bacterial and Unnatural Flavonoids, Stilbenoids and Curcuminoids by Microorganisms Sueharu Horinouchi Received: August 1, 2008 / Accepted: October 14, 2008 © Japan Antibiotics Research Association Abstract One of the approaches of combinatorial biosynthesis is combining genes from different organisms and designing a new set of gene clusters to produce bioactive compounds, leading to diversification of both chemical and natural product libraries. This makes efficient use of the potential of the host organisms, especially when microorganisms are used. An Escherichia coli system, in which artificial biosynthetic pathways for production of plant-specific medicinal polyketides, such as flavonoids, stilbenoids, isoflavonoids, and curcuminoids, are assembled, has been designed and expressed. Starting with amino acids tyrosine and phenylalanine as substrates, this system yields naringenin, resveratrol, genistein, and curcumin, for example, all of which are beneficial to human health because of their wide variety of biological activities. Supplementation of unnatural carboxylic acids to the recombinant E. coli cells carrying the artificial pathways by precursor-directed biosynthesis results in production of unnatural compounds. Addition of decorating or modification enzymes to the artificial pathway leads to production of natural and unnatural flavonols, flavones, and methylated resveratrols. This microbial system is promising for construction of larger libraries by employing other polyketide synthases and decorating enzymes of various origins. In addition, the concept of building and expressing artificial biosynthetic pathways for production of non-bacterial and unnatural compounds in microorganisms should be successfully applied to production of not only plant-specific polyketides but also many other useful compound classes.
    [Show full text]
  • Supplementary Materials Evodiamine Inhibits Both Stem Cell and Non-Stem
    Supplementary materials Evodiamine inhibits both stem cell and non-stem-cell populations in human cancer cells by targeting heat shock protein 70 Seung Yeob Hyun, Huong Thuy Le, Hye-Young Min, Honglan Pei, Yijae Lim, Injae Song, Yen T. K. Nguyen, Suckchang Hong, Byung Woo Han, Ho-Young Lee - 1 - Table S1. Short tandem repeat (STR) DNA profiles for human cancer cell lines used in this study. MDA-MB-231 Marker H1299 H460 A549 HCT116 (MDA231) Amelogenin XX XY XY XX XX D8S1179 10, 13 12 13, 14 10, 14, 15 13 D21S11 32.2 30 29 29, 30 30, 33.2 D7S820 10 9, 12 8, 11 11, 12 8 CSF1PO 12 11, 12 10, 12 7, 10 12, 13 D3S1358 17 15, 18 16 12, 16, 17 16 TH01 6, 9.3 9.3 8, 9.3 8, 9 7, 9.3 D13S317 12 13 11 10, 12 13 D16S539 12, 13 9 11, 12 11, 13 12 D2S1338 23, 24 17, 25 24 16 21 D19S433 14 14 13 11, 12 11, 14 vWA 16, 18 17 14 17, 22 15 TPOX 8 8 8, 11 8, 9 8, 9 D18S51 16 13, 15 14, 17 15, 17 11, 16 D5S818 11 9, 10 11 10, 11 12 FGA 20 21, 23 23 18, 23 22, 23 - 2 - Table S2. Antibodies used in this study. Catalogue Target Vendor Clone Dilution ratio Application1) Number 1:1000 (WB) ADI-SPA- 1:50 (IHC) HSP70 Enzo C92F3A-5 WB, IHC, IF, IP 810-F 1:50 (IF) 1 :1000 (IP) ADI-SPA- HSP90 Enzo 9D2 1:1000 WB 840-F 1:1000 (WB) Oct4 Abcam ab19857 WB, IF 1:100 (IF) Nanog Cell Signaling 4903S D73G4 1:1000 WB Sox2 Abcam ab97959 1:1000 WB ADI-SRA- Hop Enzo DS14F5 1:1000 WB 1500-F HIF-1α BD 610958 54/HIF-1α 1:1000 WB pAkt (S473) Cell Signaling 4060S D9E 1:1000 WB Akt Cell Signaling 9272S 1:1000 WB pMEK Cell Signaling 9121S 1:1000 WB (S217/221) MEK Cell Signaling 9122S 1:1000
    [Show full text]
  • BIOSYNTHESIS of PROANTHOCYANIDINS in BARLEY: GENETIC CONTROL of the CONVERSION of DIHYDROQUERCETIN to CATECHIN and PROCYANIDINS by KLAUS NYEGAARD KRISTIANSEN
    Carlsberg Res. Commun. Vol. 49, p. 503-524, 1984 BIOSYNTHESIS OF PROANTHOCYANIDINS IN BARLEY: GENETIC CONTROL OF THE CONVERSION OF DIHYDROQUERCETIN TO CATECHIN AND PROCYANIDINS by KLAUS NYEGAARD KRISTIANSEN Department of Physiology, Carlsberg Laboratory, Gamle Carlsberg Vej 10, DK-2500 Copenhagen Valby and Institute of Genetics, University of Copenhagen Oster Farimagsgade 2A, DK-1353 Copenhagen K Keywords: Flavonoid biosynthesis, leucocyanidin isomers, ant mutants, genetic control, high pressure liquid chromatography, 'H NMR, mass spectroscopy The conversion of dihydroquercetin to catechin and procyanidin was studied in maturing wild type barley (Hordeum vulgare L., cv. Nordal) seeds and proanthocyanidin free mutants blocked in four different genes, ant 13, ant 17. ant 18 and ant 19. In the wild type barley grown under controlled conditions, maximal rate of synthesis of catechin, procyanidin B3 and procyanidin C2 occurred 8-16 days after flowering. Dihydroquercetin was radioactively labelled by feeding ( 1-'4C)-acetate and (2-'4C)-acetate to flowerbuds of a petunia mutant accumulating this flavonoid. When fed to pericarp-testa tissue of wild type barley labelled catechin, procyanidin B3 and procyanidin C2 were synthesized establishing dihydroquercetin as a precursor of these compounds. In addition labelled 2,3-trans-3,4-cis-leucocyanidin was synthesized indicating that this compound is an intermediate. The leucocyanidin was identified by co-chromatography with an authentic standard prepared chemically by reduction ofdihydroquercetin with NaBH,. The major product of this reduction, however, was the 2,3-trans-3,4-trans-leuco- cyanidin. Only mutant ant 18-102 accumulated dihydroquercetin in the seeds. Feeding ('4C)-dihydroquercetin to pericarp-testa tissue from the mutants revealed that ant 17-139 was capable of synthesizing significant amounts of labelled catechin and procyanidin, whereas ant 13-101, ant 13-152, ant 18-102 and ant 19-109 synthesized none or only very small amounts of these compounds.
    [Show full text]
  • Flavonoid Glucodiversification with Engineered Sucrose-Active Enzymes Yannick Malbert
    Flavonoid glucodiversification with engineered sucrose-active enzymes Yannick Malbert To cite this version: Yannick Malbert. Flavonoid glucodiversification with engineered sucrose-active enzymes. Biotechnol- ogy. INSA de Toulouse, 2014. English. NNT : 2014ISAT0038. tel-01219406 HAL Id: tel-01219406 https://tel.archives-ouvertes.fr/tel-01219406 Submitted on 22 Oct 2015 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Last name: MALBERT First name: Yannick Title: Flavonoid glucodiversification with engineered sucrose-active enzymes Speciality: Ecological, Veterinary, Agronomic Sciences and Bioengineering, Field: Enzymatic and microbial engineering. Year: 2014 Number of pages: 257 Flavonoid glycosides are natural plant secondary metabolites exhibiting many physicochemical and biological properties. Glycosylation usually improves flavonoid solubility but access to flavonoid glycosides is limited by their low production levels in plants. In this thesis work, the focus was placed on the development of new glucodiversification routes of natural flavonoids by taking advantage of protein engineering. Two biochemically and structurally characterized recombinant transglucosylases, the amylosucrase from Neisseria polysaccharea and the α-(1→2) branching sucrase, a truncated form of the dextransucrase from L. Mesenteroides NRRL B-1299, were selected to attempt glucosylation of different flavonoids, synthesize new α-glucoside derivatives with original patterns of glucosylation and hopefully improved their water-solubility.
    [Show full text]
  • Chondroprotective Agents
    Europaisches Patentamt J European Patent Office © Publication number: 0 633 022 A2 Office europeen des brevets EUROPEAN PATENT APPLICATION © Application number: 94109872.5 © Int. CI.6: A61K 31/365, A61 K 31/70 @ Date of filing: 27.06.94 © Priority: 09.07.93 JP 194182/93 Saitama 350-02 (JP) Inventor: Niimura, Koichi @ Date of publication of application: Rune Warabi 1-718, 11.01.95 Bulletin 95/02 1-17-30, Chuo Warabi-shi, 0 Designated Contracting States: Saitama 335 (JP) CH DE FR GB IT LI SE Inventor: Umekawa, Kiyonori 5-4-309, Mihama © Applicant: KUREHA CHEMICAL INDUSTRY CO., Urayasu-shi, LTD. Chiba 279 (JP) 9-11, Horidome-cho, 1-chome Nihonbashi Chuo-ku © Representative: Minderop, Ralph H. Dr. rer.nat. Tokyo 103 (JP) et al Cohausz & Florack @ Inventor: Watanabe, Koju Patentanwalte 2-5-7, Tsurumai Bergiusstrasse 2 b Sakado-shi, D-30655 Hannover (DE) © Chondroprotective agents. © A chondroprotective agent comprising a flavonoid compound of the general formula (I): (I) CM < CM CM wherein R1 to R9 are, independently, a hydrogen atom, hydroxyl group, or methoxyl group and X is a single bond or a double bond, or a stereoisomer thereof, or a naturally occurring glycoside thereof is disclosed. The 00 00 above compound strongly inhibits proteoglycan depletion from the chondrocyte matrix and exhibits a function to (Q protect cartilage, and thus, is extremely effective for the treatment of arthropathy. Rank Xerox (UK) Business Services (3. 10/3.09/3.3.4) EP 0 633 022 A2 BACKGROUND OF THE INVENTION 1 . Field of the Invention 5 The present invention relates to an agent for protecting cartilage, i.e., a chondroprotective agent, more particularly, a chondroprotective agent containing a flavonoid compound or a stereoisomer thereof, or a naturally occurring glycoside thereof.
    [Show full text]