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Functional Expression of the Sweet-Tasting Protein Brazzein In
a ISSN 0101-2061 (Print) Food Science and Technology ISSN 1678-457X (Online) DOI: https://doi.org/10.1590/fst.40521 Functional expression of the sweet-tasting protein brazzein in transgenic tobacco Hyo-Eun CHOI1, Ji-In LEE1, Seon-Yeong JO1, Yun-Cheol CHAE2, Jeong-Hwan LEE3, Hyeon-Jin SUN4, Kisung KO3, Sungguan HONG1* , Kwang-Hoon KONG1* Abstract The sweet-tasting protein, brazzein, has potential as a low-calorie sugar substitute owing to its high sweetness, stability, and water solubility. In this study, the synthetic brazzein gene was expressed in the tobacco plant, Nicotiana tabacum. Three types of expression cassettes containing the brazzein gene were constructed to examine the expression and purification efficiency of the brazzein: pBI-BZ1 containing a signal sequence and His-tag, pBI-BZ2 containing a signal sequence, and pBI-BZ3 containing only the brazzein gene. Brazzein expression confirmed by ELISA was purified using ammonium sulfate precipitation, heat treatment, and CM-sepharose chromatography. The purity and conformational state of the brazzein were confirmed using SDS-PAGE, HPLC, and circular dichroism. The identity of the brazzein was confirmed by N-terminal amino acid analysis, ESI-MS/MS, and sweetness analysis. We successfully generated brazzein overexpression tobacco plants, suggesting that this method could be used as a brazzein production platform to provide an alternative to currently produced sweeteners. Keywords: alternative sweetener; brazzein; Agrobacterium-mediated transformation; transgenic tobacco plant; protein purification. Practical Application: Our study provides an economic mass-production system for this sweet-tasting protein, brazzein, with potential use as an alternative sweetener in the food industry. 1 Introduction The appeal sweet food is undeniable, however, excessive (Assadi-Porter et al., 2000). -
Secondary Successions After Shifting Cultivation in a Dense Tropical Forest of Southern Cameroon (Central Africa)
Secondary successions after shifting cultivation in a dense tropical forest of southern Cameroon (Central Africa) Dissertation zur Erlangung des Doktorgrades der Naturwissenschaften vorgelegt beim Fachbereich 15 der Johann Wolfgang Goethe University in Frankfurt am Main von Barthélemy Tchiengué aus Penja (Cameroon) Frankfurt am Main 2012 (D30) vom Fachbereich 15 der Johann Wolfgang Goethe-Universität als Dissertation angenommen Dekan: Prof. Dr. Anna Starzinski-Powitz Gutachter: Prof. Dr. Katharina Neumann Prof. Dr. Rüdiger Wittig Datum der Disputation: 28. November 2012 Table of contents 1 INTRODUCTION ............................................................................................................ 1 2 STUDY AREA ................................................................................................................. 4 2.1. GEOGRAPHIC LOCATION AND ADMINISTRATIVE ORGANIZATION .................................................................................. 4 2.2. GEOLOGY AND RELIEF ........................................................................................................................................ 5 2.3. SOIL ............................................................................................................................................................... 5 2.4. HYDROLOGY .................................................................................................................................................... 6 2.5. CLIMATE ........................................................................................................................................................ -
Nested Whole-Genome Duplications Coincide with Diversification And
ARTICLE https://doi.org/10.1038/s41467-020-17605-7 OPEN Nested whole-genome duplications coincide with diversification and high morphological disparity in Brassicaceae Nora Walden 1,7, Dmitry A. German 1,5,7, Eva M. Wolf 1,7, Markus Kiefer 1, Philippe Rigault 1,2, Xiao-Chen Huang 1,6, Christiane Kiefer 1, Roswitha Schmickl3, Andreas Franzke 1, Barbara Neuffer4, ✉ Klaus Mummenhoff4 & Marcus A. Koch 1 1234567890():,; Angiosperms have become the dominant terrestrial plant group by diversifying for ~145 million years into a broad range of environments. During the course of evolution, numerous morphological innovations arose, often preceded by whole genome duplications (WGD). The mustard family (Brassicaceae), a successful angiosperm clade with ~4000 species, has been diversifying into many evolutionary lineages for more than 30 million years. Here we develop a species inventory, analyze morphological variation, and present a maternal, plastome-based genus-level phylogeny. We show that increased morphological disparity, despite an apparent absence of clade-specific morphological innovations, is found in tribes with WGDs or diversification rate shifts. Both are important processes in Brassicaceae, resulting in an overall high net diversification rate. Character states show frequent and independent gain and loss, and form varying combinations. Therefore, Brassicaceae pave the way to concepts of phy- logenetic genome-wide association studies to analyze the evolution of morphological form and function. 1 Centre for Organismal Studies, University of Heidelberg, Im Neuenheimer Feld 345, 69120 Heidelberg, Germany. 2 GYDLE, 1135 Grande Allée Ouest, Québec, QC G1S 1E7, Canada. 3 Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01, Prague, Czech Republic. -
Enhancing Sweetness and Stability of the Single Chain Monellin MNEI
www.nature.com/scientificreports OPEN Sweeter and stronger: enhancing sweetness and stability of the single chain monellin MNEI through Received: 08 July 2016 Accepted: 07 September 2016 molecular design Published: 23 September 2016 Serena Leone1, Andrea Pica1, Antonello Merlino1, Filomena Sannino1, Piero Andrea Temussi1,2 & Delia Picone1 Sweet proteins are a family of proteins with no structure or sequence homology, able to elicit a sweet sensation in humans through their interaction with the dimeric T1R2-T1R3 sweet receptor. In particular, monellin and its single chain derivative (MNEI) are among the sweetest proteins known to men. Starting from a careful analysis of the surface electrostatic potentials, we have designed new mutants of MNEI with enhanced sweetness. Then, we have included in the most promising variant the stabilising mutation E23Q, obtaining a construct with enhanced performances, which combines extreme sweetness to high, pH-independent, thermal stability. The resulting mutant, with a sweetness threshold of only 0.28 mg/L (25 nM) is the strongest sweetener known to date. All the new proteins have been produced and purified and the structures of the most powerful mutants have been solved by X-ray crystallography. Docking studies have then confirmed the rationale of their interaction with the human sweet receptor, hinting at a previously unpredicted role of plasticity in said interaction. Sweet proteins are a family of structurally unrelated proteins that can elicit a sweet sensation in humans. To date, eight sweet and sweet taste-modifying proteins have been identified: monellin1, thaumatin2, brazzein3, pentadin4, mabinlin5, miraculin6, neoculin7 and lysozyme8. With the sole exception of lysozyme, all sweet proteins have been purified from plants, but, besides this common feature, they share no structure or sequence homology9. -
Sweet Sensations by Judie Bizzozero | Senior Editor
[Confections] July 2015 Sweet Sensations By Judie Bizzozero | Senior Editor By R.J. Foster, Contributing Editor For many, terms like “reduced-sugar” or “sugar-free” do not go with the word “candy.” And yet, the confectionery industry is facing growing demand for treats that offer the taste people have grown to love without the adverse health effects they’re looking to avoid. Thankfully, there is a growing palette of ingredients from which candy makers can paint a new picture of sweetness that will be appreciated by the even most discerning of confectionery critics. SUGAR ALCOHOLS Also referred to as polyols, sugar alcohols are a common ingredient in reduced-sugar and sugar-free applications, especially confections. Funny thing, they’re not sugars or alcohols. Carbohydrate chains composed of monomeric, dimeric and polymeric units, polyols resemble both sugars and alcohols, but do not contain an ethanol molecule. All but two sugar alcohols are less sweet than sugar. Being only partially digestible, though, replacing a portion of a formulation’s sugar with a sugar alcohol reduces total calories without losing bulk (which can occur when replacing sugar with high-intensity sweeteners). Unique flavoring, texturizing and moisture-controlling effects also make polyols well-suited for confectionery products. Two very common and very similar monomeric polyols are sorbitol and mannitol. Present in a variety of fruits and vegetables, both are derived from products of cornstarch hydrolysis. Sorbitol is made via hydrogenation of glucose, which is why sorbitol is sometimes referred to as glucitol. Mannitol is created when fructose hydrogenation converts fructose into mannose, for which the final product, mannitol, is named. -
Reports of the Scientific Committee for Food
Commission of the European Communities food - science and techniques Reports of the Scientific Committee for Food (Sixteenth series) Commission of the European Communities food - science and techniques Reports of the Scientific Committee for Food (Sixteenth series) Directorate-General Internal Market and Industrial Affairs 1985 EUR 10210 EN Published by the COMMISSION OF THE EUROPEAN COMMUNITIES Directorate-General Information Market and Innovation Bâtiment Jean Monnet LUXEMBOURG LEGAL NOTICE Neither the Commission of the European Communities nor any person acting on behalf of the Commission is responsible for the use which might be made of the following information This publication is also available in the following languages : DA ISBN 92-825-5770-7 DE ISBN 92-825-5771-5 GR ISBN 92-825-5772-3 FR ISBN 92-825-5774-X IT ISBN 92-825-5775-8 NL ISBN 92-825-5776-6 Cataloguing data can be found at the end of this publication Luxembourg, Office for Official Publications of the European Communities, 1985 ISBN 92-825-5773-1 Catalogue number: © ECSC-EEC-EAEC, Brussels · Luxembourg, 1985 Printed in Luxembourg CONTENTS Page Reports of the Scientific Committee for Food concerning - Sweeteners (Opinion expressed 14 September 1984) III Composition of the Scientific Committee for Food P.S. Elias A.G. Hildebrandt (vice-chairman) F. Hill A. Hubbard A. Lafontaine Mne B.H. MacGibbon A. Mariani-Costantini K.J. Netter E. Poulsen (chairman) J. Rey V. Silano (vice-chairman) Mne A. Trichopoulou R. Truhaut G.J. Van Esch R. Wemig IV REPORT OF THE SCIENTIFIC COMMITTEE FOR FOOD ON SWEETENERS (Opinion expressed 14 September 1984) TERMS OF REFERENCE To review the safety in use of certain sweeteners. -
Biogeography and Diversification of Brassicales
Molecular Phylogenetics and Evolution 99 (2016) 204–224 Contents lists available at ScienceDirect Molecular Phylogenetics and Evolution journal homepage: www.elsevier.com/locate/ympev Biogeography and diversification of Brassicales: A 103 million year tale ⇑ Warren M. Cardinal-McTeague a,1, Kenneth J. Sytsma b, Jocelyn C. Hall a, a Department of Biological Sciences, University of Alberta, Edmonton, Alberta T6G 2E9, Canada b Department of Botany, University of Wisconsin, Madison, WI 53706, USA article info abstract Article history: Brassicales is a diverse order perhaps most famous because it houses Brassicaceae and, its premier mem- Received 22 July 2015 ber, Arabidopsis thaliana. This widely distributed and species-rich lineage has been overlooked as a Revised 24 February 2016 promising system to investigate patterns of disjunct distributions and diversification rates. We analyzed Accepted 25 February 2016 plastid and mitochondrial sequence data from five gene regions (>8000 bp) across 151 taxa to: (1) Available online 15 March 2016 produce a chronogram for major lineages in Brassicales, including Brassicaceae and Arabidopsis, based on greater taxon sampling across the order and previously overlooked fossil evidence, (2) examine Keywords: biogeographical ancestral range estimations and disjunct distributions in BioGeoBEARS, and (3) determine Arabidopsis thaliana where shifts in species diversification occur using BAMM. The evolution and radiation of the Brassicales BAMM BEAST began 103 Mya and was linked to a series of inter-continental vicariant, long-distance dispersal, and land BioGeoBEARS bridge migration events. North America appears to be a significant area for early stem lineages in the Brassicaceae order. Shifts to Australia then African are evident at nodes near the core Brassicales, which diverged Cleomaceae 68.5 Mya (HPD = 75.6–62.0). -
Highly Sweet Compounds of Plant Origin T
glrthi~eS of ~armata[ ge~earth Arch Pharm Res Vol 25, No 6, 725-746, 2002 http://apr.psk.or.kr Highly Sweet Compounds of Plant Origin t Nam-Cheol Kim I and A. Douglas Kinghorn 2 1Chemistry and Life Sciences, Research Triangle Institute, Research Triangle Park, North Carolina, U.S.A. and 2Program for Collaborative Research in the Pharmaceutical Sciences and Department of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of Illinois at Chicago, Chicago, Illinois, U.S.A. (Received October 18, 2002) The demand for new alternative "low calorie" sweeteners for dietetic and diabetic purposes has increased worldwide. Although the currently developed and commercially used highly sweet sucrose substitutes are mostly synthetic compounds, the search for such compounds from natural sources is continuing. As of mid-2002, over 100 plant-derived sweet compounds of 20 major structural types had been reported, and were isolated from more than 25 different families of green plants. Several of these highly sweet natural products are marketed as sweeteners or flavoring agents in some countries as pure compounds, compound mixtures, or refined extracts. These highly sweet natural substances are reviewed herein. Key words: Low-Calorie Natural Sweeteners, Plants, Glycyrrhizin, Mogroside V, Rebaudio- side A, Stevioside, Thaumatin, Terpenoids, Steroids, Flavonoids, Proteins INTRODUCTION Most of the currently available potently sweet, low calorie sucrose substitutes in the world market are synthetic com- The consumption of sucrose as a sweetener has been pounds, inclusive of acesulfame-K, alitame, aspartame, associated with several nutritional and medical problems, cyclamate, saccharin, and sucralose (Duffy and Anderson, with dental caries being the most widely described (Grenby, 1998). -
The Butterfly Plant Arms-Race Escalated by Gene and Genome Duplications
The butterfly plant arms-race escalated by gene and genome duplications Patrick P. Edgera,b,c,1, Hanna M. Heidel-Fischerd,1, Michaël Bekaerte, Jadranka Rotaf, Gernot Glöcknerg,h, Adrian E. Plattsi, David G. Heckeld, Joshua P. Derj,k, Eric K. Wafulaj, Michelle Tanga, Johannes A. Hofbergerl, Ann Smithsonm,n, Jocelyn C. Hallo, Matthieu Blanchettei, Thomas E. Bureaup, Stephen I. Wrightq, Claude W. dePamphilisj, M. Eric Schranzl, Michael S. Barkerb, Gavin C. Conantr,s, Niklas Wahlbergf, Heiko Vogeld, J. Chris Piresa,s,2, and Christopher W. Wheatt,2 aDivision of Biological Sciences, University of Missouri, Columbia, MO 65211; bDepartment of Ecology and Evolutionary Biology, University of Arizona, Tucson, AZ 85721; cDepartment of Plant and Microbial Biology, University of California, Berkeley, CA 94720; dDepartment of Entomology, Max Planck Institute for Chemical Ecology, 07745 Jena, Germany; eInstitute of Aquaculture, University of Stirling, Stirling FK9 4LA, Scotland, United Kingdom; fDepartment of Biology, University of Turku, FI-20014 Turku, Finland; gLeibniz Institute for Age Research, Fritz Lipmann Institute, 07745 Jena, Germany; hInstitute for Biochemistry I, University of Cologne, 50931 Koeln, Germany; iMcGill Centre for Bioinformatics, McGill University, Montreal, QC, Canada H3A 0E9; jDepartment of Biology, Pennsylvania State University, University Park, PA 16803; kDepartment of Biological Science, California State University Fullerton, Fullerton, CA 92831; lBiosystematics Group, Plant Sciences, Wageningen University, Wageningen 6700 -
Stevia Leaf Reb M” (I, 2018) Suppliers: • 2017: I • 2018 C, D
9/27/2018 1 Answer Today’s High Sugar and Clean Label Concerns with 3rd Generation Stevia Alex Woo, PhD Chief Innovation Officer Nascent SoPure Stevia 9/27/2018 2 We love it! Nascent Innovation Core Competencies • Taste • Plant-based High • Smell potency sweeteners • Sight • Non/low caloric • Sound bulk sweeteners • Touch • Natural flavors Sweeteners Neuroscience and Flavors Taste Formulation Modulation • Sweetness • Stacking modulators • Matrix • Bitterness • Beverages & Foods modulators • Enhancement without ingredients 9/27/2018 3 We love it! Executive Summary • 2nd generation stevia extracts were all about high purity RA, the higher the purity the better the taste. • Farm-based 3rd generation stevia extracts are the newer 2-way and 3-way blends of RABCDM for even more sugar like taste but at higher cost. Alternatively, fermentation and enzymology-based stevia already co-exist with farm-based stevia in 2018. • Enzymatically modified stevia extracts are sweet taste enhancers that can be used as part of the stacking strategy for sugar reduction. • Stacking is a sugar reduction strategy for building up to the required sweetness intensity and profile while staying below the off flavor thresholds for all the plant-based ingredients used 9/27/2018 4 We love it! Agenda • Sweetness neuroscience • Stevia as sweetener • Stevia as flavor • Stacking 9/27/2018 5 We love it! Re-Defining “Flavor” = Taste + Smell + More Taste (5+ primary) Smell (aroma) Somatosensation (Touch): • Mechanoreception: Touch, Pressure and Vibration (Prescott, 2015), • Thermoception: Temperature, • Nociception: Pain (Youseff, 2015), and • Up to total 30 senses? (Smith, 2016) can they all be part of somatosensation? Vision (“Seeing the flavor”. -
A Biobrick Compatible Strategy for Genetic Modification of Plants Boyle Et Al
A BioBrick compatible strategy for genetic modification of plants Boyle et al. Boyle et al. Journal of Biological Engineering 2012, 6:8 http://www.jbioleng.org/content/6/1/8 Boyle et al. Journal of Biological Engineering 2012, 6:8 http://www.jbioleng.org/content/6/1/8 METHODOLOGY Open Access A BioBrick compatible strategy for genetic modification of plants Patrick M Boyle1†, Devin R Burrill1†, Mara C Inniss1†, Christina M Agapakis1,7†, Aaron Deardon2, Jonathan G DeWerd2, Michael A Gedeon2, Jacqueline Y Quinn2, Morgan L Paull2, Anugraha M Raman2, Mark R Theilmann2, Lu Wang2, Julia C Winn2, Oliver Medvedik3, Kurt Schellenberg4, Karmella A Haynes1,8, Alain Viel3, Tamara J Brenner3, George M Church5,6, Jagesh V Shah1* and Pamela A Silver1,5* Abstract Background: Plant biotechnology can be leveraged to produce food, fuel, medicine, and materials. Standardized methods advocated by the synthetic biology community can accelerate the plant design cycle, ultimately making plant engineering more widely accessible to bioengineers who can contribute diverse creative input to the design process. Results: This paper presents work done largely by undergraduate students participating in the 2010 International Genetically Engineered Machines (iGEM) competition. Described here is a framework for engineering the model plant Arabidopsis thaliana with standardized, BioBrick compatible vectors and parts available through the Registry of Standard Biological Parts (www.partsregistry.org). This system was used to engineer a proof-of-concept plant that exogenously expresses the taste-inverting protein miraculin. Conclusions: Our work is intended to encourage future iGEM teams and other synthetic biologists to use plants as a genetic chassis. -
Taste Responsiveness to Two Steviol Glycosides in Three Species of Nonhuman Primates
Current Zoology, 2018, 64(1), 63–68 doi: 10.1093/cz/zox012 Advance Access Publication Date: 27 February 2017 Article Article Taste responsiveness to two steviol glycosides in three species of nonhuman primates a a a,b c Sandra NICKLASSON , Desire´eSJO¨ STRO¨ M , Mats AMUNDIN , Daniel ROTH , d a, Laura Teresa HERNANDEZ SALAZAR , and Matthias LASKA * aIFM Biology, Linko¨ping University, Linko¨ping, SE-581 83, bKolma˚rden Wildlife Park, Kolma˚rden, SE-681 92, cBora˚s Zoo, Bora˚s, SE-501 13, Sweden, and dInstituto de Neuro-Etologia, Universidad Veracruzana, Xalapa, Veracruz, C.P. 91000, Mexico *Address correspondence to Matthias Laska. E-mail: [email protected]. Received on 23 December 2016; accepted on 21 February 2017 Abstract Primates have been found to differ widely in their taste perception and studies suggest that a co- evolution between plant species bearing a certain taste substance and primate species feeding on these plants may contribute to such between-species differences. Considering that only platyrrhine primates, but not catarrhine or prosimian primates, share an evolutionary history with the neotrop- ical plant Stevia rebaudiana, we assessed whether members of these three primate taxa differ in their ability to perceive and/or in their sensitivity to its two quantitatively predominant sweet- tasting substances. We found that not only neotropical black-handed spider monkeys, but also paleotropical black-and-white ruffed lemurs and Western chimpanzees are clearly able to perceive stevioside and rebaudioside A. Using a two-bottle preference test of short duration, we found that Ateles geoffroyi preferred concentrations as low as 0.05 mM stevioside and 0.01 mM rebaudioside A over tap water.