DOCSLIB.ORG

Production, Purification, Properties, and Acceptor Reaction Mechanism of Dextransucrase from Leuconostoc Mesenteroides NRRL B-51

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Production, Purification, Properties, and Acceptor Reaction Mechanism of Dextransucrase from Leuconostoc Mesenteroides NRRL B-51 Iowa State University Capstones, Theses and Retrospective Theses and Dissertations Dissertations 1977 Production, purification, properties, and acceptor reaction mechanism of dextransucrase from Leuconostoc mesenteroides NRRL B-512 Timothy Francis Walseth Iowa State University Follow this and additional works at: https://lib.dr.iastate.edu/rtd Part of the Biochemistry Commons Recommended Citation Walseth, Timothy Francis, "Production, purification, properties, and acceptor reaction mechanism of dextransucrase from Leuconostoc mesenteroides NRRL B-512 " (1977). Retrospective Theses and Dissertations. 5852. https://lib.dr.iastate.edu/rtd/5852 This Dissertation is brought to you for free and open access by the Iowa State University Capstones, Theses and Dissertations at Iowa State University Digital Repository. It has been accepted for inclusion in Retrospective Theses and Dissertations by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. INFORMATION TO USERS This material was produced from a microfilm copy of the original document. While the most advanced technological means to photograph and reproduce this document have been used, the quality is heavily dependent upon the quality of the original submitted. The following explanation of techniques is provided to help you understand markings or patterns which may appear on this reproduction. 1. The sign or "target" for pages apparently lacking from the document photographed is "Missing Page(s)". If it was possible to obtain the missing page(s) or section, they are spliced into the film along with adjacent pages. This may have necessitated cutting thru an image and duplicating adjacent pages to insure you complete continuity. 2. When an image on the film is obliterated with a large round black mark, it is an indication that the photographer suspected that the copy may have moved during exposure and thus cause a blurred image. You will find a good image of the page in the adjacent frame. 3. When a map, drawing or chart, etc., was part of the material being photographed the photographer followed a definite method in "sectioning" the material, it is customary to begin photoing at the upper left hand corner of a large sheet and to continue photoing from left to right in equal sections with a small overlap. If necessary, sectioning is continued again — beginning below the first row and continuing on until complete. 4. The majority of users indicate that the textual content is of greatest value, however, a somewhat higher quality reproduction could be made from "photographs" if essential to the understanding of the dissertation. SiNer prints of "photographs" may be ordered at additional charge by writing the Order Department, giving the catalog number, title, author and specific pages you wish reproduced. 5. PLEASE NOTE: Some pages may have indistinct print. Filmed as received. University Microfilms International 300 North Zeeb Road Ann Arbor. Michigan 48106 USA St. John's Road. Tyler's Green High Wycombe. Bucks. England HP10 8HR I I 77-16,983 WALSEIH, Timothy Francis, 1950- PRODOCriON, PURIFICATION, PROPERTIES, AND ACCEPTOR REACnON MECHANISM OF DEXTRANSIOASE FROM LEUOQNOSTOC iESENTEROIDES NRRL B-512. Iowa State University, Ph.D., 1977 Chemistry, biological Xerox University Microfilms, Ann Arbor, Michigan 48106 Production, purification, properties, and acceptor reaction mechanism of dextransucrase from Leuconostoc mesenteroides NBRL B-$Ï2 by Timothy Francis Walseth A Dissertation Submitted to the Graduate Faculty in Partial Fulfillment of The Requirements for the Degree of DOCTOR OF PHILOSOPHY Department; Biochemistry and Biophysics Major: Biochemistry Approved ; Signature was redacted for privacy. Work Signature was redacted for privacy. Signature was redacted for privacy. Iowa State University Ames, Iowa 1977 ii TABLE OF CONTENTS Page ABBREVIATIONS USED V NOTES ON THE NOMENCLATURE OF SUGARS vii ABSTRACT ix I. INTRODUCTION 1 II. LITERATURE REVIEW 11 A. Production 11 B. Purification 1^ C. Properties l8 D. Acceptor Reaction Mechanism 22 III. MATERIALS AND METHODS 30 A. Materials 30 1. Organisms 30 2. Enzymes 30 3. Polysaccharides 31 4. Radioactive sugars 32 5. Chromatographic standards 32 6. Chromatographic supplies 33 7. Chemicals 33 8. Miscellaneous 33 B. Methods 3^ 1. Culturing of L. mesenteroides NRRL B-512 3^ 2. Determination"of radioactivity 35 3. Radioautography 35 4. Paper chromatography 35 5. Paper electrophoresis 38 6. Total protein analysis 38 7. Total carbohydrate analysis 38 8. Reducing value determinations 38 9. Enzyme assays 39 10. Concentration of dextransucrase solu­ tions 4-2 iii 11. Concentration of carbohydrate solutions 4-3 12. NaB% reduction of l^C-oligosac- charides and -polysaccharides ^3 13. Complete acid hydrolysis of poly­ saccharides and oligosaccharides 44 14-. Partial acid hydrolysis of poly­ saccharides and oligosaccharides 4-5 15. Agar gel double diffusion precipitin test 46 16. Preparation of gel and ion-exchange chromatography materials 4-7 17. Detection of contaminating enzymatic activities 4-7 18. Conditions for the purification of dextransucrase 4-9 EXPERIMENTAL 50 A. Production 50 1. Production of dextransucrase from cul­ tures of L. mesenteroides NEîRL B-512 50 2. The effect of calcium on the production of dextransucrase 50 3. The effect of various calcium levels on the production of dextransucrase 53 4-. Structural analysis of the fructan pro­ duced by L. mesenteroides B-512 53 B. Purification 60 1. Dialysis and concentration of the cul­ ture supernatant 60 2. Ammonium sulfate precipitation of CSC 63 3. Hydroxylapatite chromatography of CSC 64 4. Ion-exchange chromatography of CSC 65 5. Chromatography of CSC on Bio-Gel P-I50 65 6. Chromatography of the culture super­ natant and CSC on agarose gel columns 66 7. Chromatography of endo-dextranase treated CSC on Bio-Gel A-5m 66 C. Properties 69 1. Contaminating activities analysis of the A-5m concentrate 69 2. The action of the A-5m concentrate on l4-c~(u)-sucrose 69 iv 3. The effect of chelating agents on the A-^m concentrate 76 4-. The effect of metal ions on untreated and EDTA treated concentrate 78 5. The effect of dextran on the A-^m concentrate 80 6. The stability of the A-5m concentrate to storage 80 7. The stability of the A-5m concentrate stored in varying amounts of dextran 83 8. Chromatography of the A-^m concentrate stored in the presence and absence of dextran on Bio-Gel A-l^m 83 9. Examination of the carbohydrate content of the A-5m concentrate 88 10. The effect of concanavalin A on the A-5m concentrate 89 11. ConA-Sepharose chromatography of the A-5m concentrate 89 12. Examination of the polysaccharide produced by the A-5m concentrate 95 D. Acceptor Reactions 99 1. Preparation of charged enzyme 99 2. Acceptor reaction conditions 100 3. Analysis of the acceptor reaction products 104- 4. Structural determination of Fraction II 109 5. Studies on Fraction I 126 V. DISCUSSION 133 A. Production of Dextransucrase 133 B. Purification of Dextransucrase I36 C. Properties of Dextransucrase 142 D. Acceptor Reaction Mechanism of Dextran­ sucrase 150 VI. SUMMARY 165 VII. BIBLIOGRAPHY 168 VIII. ACKNOWLEDGMENTS I8I V ABBREVIATIONS USED AE aminoethyl aMDG a-methyl-D-glucoside ara arabinose atm atmosphere CM carboxymethyl ConA concanavalin A DEAE diethylaminoethyl DP degree of polymerization DPg disaccharide DP^ trisaccharide DPi^ tetrasaccharide DPj pentasaccharide DP^ hexasaccharide ECTEOLA epichlorohydrintr i ethanolamine EDTA ethylenediami netetraacetic acid EGTA ethylenegilycol-bis(t3 -aminoethyl ether)-N,N' - tetraacetic acid fuc fucose fru fructose gal galactose glc glucose glcUA glucuronic acid IM isomaltose IM-, isomaltotriose vi IMi^ isomaltotetraose isomaltop entaos e IM6 isomaltohexaose ma milliampere man mannose nm nanometer NRRL Northern Regional Research Laboratory PAB p-aminobenzyl REL reducing end labeled rha rhamnose rib ribose SDS sodium dodecyl sulfate sue sucrose V volt xyl xylose TFA trifluoroacetic acid vii NOTES ON THE NOMENCLATURE OF SUGARS For disaccharides, the following system is used; a-O-b-D-substituent-D-glycose a: The numeral indicates the position of the oxygen through which the substituent attaches to the glycose unit, b: a or p indicates the type of linkage. For example: ^-0-a-D-glucopyranosyl-D-glucose is the nomenclature for maltose. For oligosaccharides larger than a disaccharide and containing more than one type of sugar, the following system is used: V a -0-c-D-substituent-remainder of the oligosac­ charide a: The numeral indicates the position of the oxygen through which the substituent is attached to the remainder of the oligosaccharide. b: The superscript is a three letter abbreviation of the sugar to which the substituent is attached. c; a or P indicates the type of linkage by which the substituent is attached to the remainder of the mole­ cule. For example: 6^^^-0-a-D-galactopyranosylsucrose is the nomenclature for raffinose. vili For isomaltooligosaccharides possessing other types of linkages besides a-i,6 linkages, the following system is used: a^-0-c-D-( substituent) isomaltooligosaccharide a: The numeral indicates the position of the oxygen through which the substituent attaches to the isomaltooligo­ saccharide. b: The superscript is a numeral vdaich identifies the glu­ cose unit to which the substituent is attached. The numbering starts with 1 at the reducing end. c; a or P indicate the type of linkage between the sub­ stituent and remainder of the molecule. For example: 3^-0-a-D-glucopyranosyl isomaltotriose is the nomenclature for a tetrasaccharide consisting of a glucose unit linked to the C^-OH of the glucose unit at the nonreducing end of isomaltotriose. ix ABSTRACT The production of dextransucrase from Leuconostoc mes enter oides NRRLB-512 was found to be stimulated 1.5-2 fold by the addition of 0,00^% (w/v) CaClg to the media. Stimulation of levansucrase occurred only at CaClg concen­ trations of 0.5-1/^ (w/v). Calcium appears to stabilize dextransucrase from loss of activity during dialysis and column chromatography.
Load more

Recommended publications
  • 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]
  • Ijms-20-05263-V2
    Delft University of Technology Leloir Glycosyltransferases in Applied Biocatalysis A Multidisciplinary Approach Mestrom, Luuk; Przypis, Marta; Kowalczykiewicz, Daria; Pollender, André; Kumpf, Antje; Marsden, Stefan R.; Szymańska, Katarzyna; Hanefeld, Ulf; Hagedoorn, Peter Leon; More Authors DOI 10.3390/ijms20215263 Publication date 2019 Document Version Final published version Published in International Journal of Molecular Sciences Citation (APA) Mestrom, L., Przypis, M., Kowalczykiewicz, D., Pollender, A., Kumpf, A., Marsden, S. R., Szymańska, K., Hanefeld, U., Hagedoorn, P. L., & More Authors (2019). Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach. International Journal of Molecular Sciences, 20(21), [5263]. https://doi.org/10.3390/ijms20215263 Important note To cite this publication, please use the final published version (if applicable). Please check the document version above. Copyright Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons. Takedown policy Please contact us and provide details if you believe this document breaches copyrights. We will remove access to the work immediately and investigate your claim. This work is downloaded from Delft University of Technology. For technical reasons the number of authors shown on this cover page is limited to a maximum of 10. International Journal of Molecular Sciences Review Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach Luuk Mestrom 1, Marta Przypis 2,3 , Daria Kowalczykiewicz 2,3, André Pollender 4 , Antje Kumpf 4,5, Stefan R. Marsden 1, Isabel Bento 6, Andrzej B.
    [Show full text]
  • Leloir Glycosyltransferases in Applied Biocatalysis: a Multidisciplinary Approach
    International Journal of Molecular Sciences Review Leloir Glycosyltransferases in Applied Biocatalysis: A Multidisciplinary Approach Luuk Mestrom 1, Marta Przypis 2,3 , Daria Kowalczykiewicz 2,3, André Pollender 4 , Antje Kumpf 4,5, Stefan R. Marsden 1, Isabel Bento 6, Andrzej B. Jarz˛ebski 7, Katarzyna Szyma ´nska 8, Arkadiusz Chru´sciel 9, Dirk Tischler 4,5 , Rob Schoevaart 10, Ulf Hanefeld 1 and Peter-Leon Hagedoorn 1,* 1 Department of Biotechnology, Delft University of Technology, Section Biocatalysis, Van der Maasweg 9, 2629 HZ Delft, The Netherlands; [email protected] (L.M.); [email protected] (S.R.M.); [email protected] (U.H.) 2 Department of Organic Chemistry, Bioorganic Chemistry and Biotechnology, Silesian University of Technology, B. Krzywoustego 4, 44-100 Gliwice, Poland; [email protected] (M.P.); [email protected] (D.K.) 3 Biotechnology Center, Silesian University of Technology, B. Krzywoustego 8, 44-100 Gliwice, Poland 4 Environmental Microbiology, Institute of Biosciences, TU Bergakademie Freiberg, Leipziger Str. 29, 09599 Freiberg, Germany; [email protected] (A.P.); [email protected] (A.K.); [email protected] (D.T.) 5 Microbial Biotechnology, Faculty of Biology & Biotechnology, Ruhr-Universität Bochum, Universitätsstr. 150, 44780 Bochum, Germany 6 EMBL Hamburg, Notkestraβe 85, 22607 Hamburg, Germany; [email protected] 7 Institute of Chemical Engineering, Polish Academy of Sciences, Bałtycka 5, 44-100 Gliwice, Poland; [email protected] 8 Department of Chemical and Process Engineering, Silesian University of Technology, Ks. M. Strzody 7, 44-100 Gliwice Poland.; [email protected] 9 MEXEO Wiesław Hreczuch, ul.
    [Show full text]
  • The Microbiota-Produced N-Formyl Peptide Fmlf Promotes Obesity-Induced Glucose
    Page 1 of 230 Diabetes Title: The microbiota-produced N-formyl peptide fMLF promotes obesity-induced glucose intolerance Joshua Wollam1, Matthew Riopel1, Yong-Jiang Xu1,2, Andrew M. F. Johnson1, Jachelle M. Ofrecio1, Wei Ying1, Dalila El Ouarrat1, Luisa S. Chan3, Andrew W. Han3, Nadir A. Mahmood3, Caitlin N. Ryan3, Yun Sok Lee1, Jeramie D. Watrous1,2, Mahendra D. Chordia4, Dongfeng Pan4, Mohit Jain1,2, Jerrold M. Olefsky1 * Affiliations: 1 Division of Endocrinology & Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California, USA. 2 Department of Pharmacology, University of California, San Diego, La Jolla, California, USA. 3 Second Genome, Inc., South San Francisco, California, USA. 4 Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA. * Correspondence to: 858-534-2230, [email protected] Word Count: 4749 Figures: 6 Supplemental Figures: 11 Supplemental Tables: 5 1 Diabetes Publish Ahead of Print, published online April 22, 2019 Diabetes Page 2 of 230 ABSTRACT The composition of the gastrointestinal (GI) microbiota and associated metabolites changes dramatically with diet and the development of obesity. Although many correlations have been described, specific mechanistic links between these changes and glucose homeostasis remain to be defined. Here we show that blood and intestinal levels of the microbiota-produced N-formyl peptide, formyl-methionyl-leucyl-phenylalanine (fMLF), are elevated in high fat diet (HFD)- induced obese mice. Genetic or pharmacological inhibition of the N-formyl peptide receptor Fpr1 leads to increased insulin levels and improved glucose tolerance, dependent upon glucagon- like peptide-1 (GLP-1). Obese Fpr1-knockout (Fpr1-KO) mice also display an altered microbiome, exemplifying the dynamic relationship between host metabolism and microbiota.
    [Show full text]
  • Characterization of Dextransucrase from Leuconostoc Mesenteroides T3, Water Kefir Grains Isolate
    Characterization of dextransucrase from Leuconostoc mesenteroides T3, water kefir grains isolate Miona G. Miljković1, Slađana Z. Davidović1, Slavko Kralj2, Slavica S. Šiler-Marinković1, Mirjana D. Rajilić-Stojanović1, Suzana I. Dimitrijević-Branković1 1University of Belgrade, Faculty of Technology and Metallurgy, Department for Biochemical Engineering and Biotechnology, Belgrade, Serbia 2Department for Materials Synthesis, Jožef Stefan Institute, Ljubljana, Slovenia Abstract The production of dextransucrase (DS) by Leuconostoc mesenteroides T3, novel isolate SCIENTIFIC PAPER from water kefir grain, was studied and optimized. Bacterial supernatant reached activity of 3.1 U/ml when the culture was grown at 23 °C and under static culture condition using UDC 63(497.113):632.954:66.061:543 classical Tsuchiya medium for DS production. The increase of sucrose concentration to 7% led to an increase of DS activity by 52% compared to the control. Medium with 2% beef extract and 1% yeast extract resulted in 4.52 U/ml, which was 47% higher than in the Hem. Ind. 71 (4) 351–360 (2017) control (with 2% yeast extract). Finally, the increase of K2HPO4 concentration from 2 to 3% resulted in the increased enzyme activity by 28%. Enzyme purified by polyethylene glycol 400 fractionation displayed maximum activity at 30 °C and pH 5.4. Zymogram analysis confirmed the presence of DS of approximately 180 kDa. The addition of divalent cations Ca2+, Mg2+, Fe2+ and Co2+ led to a minor increase of DS activity, while the addition of Mn2+ was the most prominent with 73% increase. These findings classify dextransucrase from Leuconostoc mesenteroides T3 as promising candidate for production of dextran, which has numerous applications in various industries.
    [Show full text]
  • Uni International 300 N
    INFORMATION TO USERS This reproduction was made from a copy of a document sent to us for microfilming. While the most advanced technology has been used to photograph and reproduce this document, the quality of the reproduction is heavily dependent upon the quality of the material submitted. The following explanation o f techniques is provided to help clarify markings or notations which may appear on this reproduction. 1.The sign or “ target” for pages apparently lacking from the document photographed is “Missing Page(s)” . I f it was possible to obtain the missing page(s) or section, they are spliced into the film along with adjacent pages. This may have necessitated cutting through an image and duplicating adjacent pages to assure complete continuity. 2. When an image on the film is obliterated with a round black mark, it is an indication of either blurred copy because of movement during exposure, duplicate copy, or copyrighted materials that should not have been filmed. For blurred pages, a good image of the page can be found in the adjacent frame. I f copyrighted materials were deleted, a target note will appear listing the pages in the adjacent frame. 3. When a map, drawing or chart, etc., is part of the material being photographed, a definite method of “sectioning” the material has been followed. It is customary to begin filming at the upper left hand comer of a large sheet and to continue from left to right in equal sections with small overlaps. I f necessary, sectioning is continued again—beginning below the first row and continuing on until complete.
    [Show full text]
  • Ep 1 117 822 B1
    Europäisches Patentamt (19) European Patent Office & Office européen des brevets (11) EP 1 117 822 B1 (12) EUROPÄISCHE PATENTSCHRIFT (45) Veröffentlichungstag und Bekanntmachung des (51) Int Cl.: Hinweises auf die Patenterteilung: C12P 19/18 (2006.01) C12N 9/10 (2006.01) 03.05.2006 Patentblatt 2006/18 C12N 15/54 (2006.01) C08B 30/00 (2006.01) A61K 47/36 (2006.01) (21) Anmeldenummer: 99950660.3 (86) Internationale Anmeldenummer: (22) Anmeldetag: 07.10.1999 PCT/EP1999/007518 (87) Internationale Veröffentlichungsnummer: WO 2000/022155 (20.04.2000 Gazette 2000/16) (54) HERSTELLUNG VON POLYGLUCANEN DURCH AMYLOSUCRASE IN GEGENWART EINER TRANSFERASE PREPARATION OF POLYGLUCANS BY AMYLOSUCRASE IN THE PRESENCE OF A TRANSFERASE PREPARATION DES POLYGLUCANES PAR AMYLOSUCRASE EN PRESENCE D’UNE TRANSFERASE (84) Benannte Vertragsstaaten: (56) Entgegenhaltungen: AT BE CH CY DE DK ES FI FR GB GR IE IT LI LU WO-A-00/14249 WO-A-00/22140 MC NL PT SE WO-A-95/31553 (30) Priorität: 09.10.1998 DE 19846492 • OKADA, GENTARO ET AL: "New studies on amylosucrase, a bacterial.alpha.-D-glucosylase (43) Veröffentlichungstag der Anmeldung: that directly converts sucrose to a glycogen- 25.07.2001 Patentblatt 2001/30 like.alpha.-glucan" J. BIOL. CHEM. (1974), 249(1), 126-35, XP000867741 (73) Patentinhaber: Südzucker AG Mannheim/ • BUTTCHER, VOLKER ET AL: "Cloning and Ochsenfurt characterization of the gene for amylosucrase 68165 Mannheim (DE) from Neisseria polysaccharea: production of a linear.alpha.-1,4-glucan" J. BACTERIOL. (1997), (72) Erfinder: 179(10), 3324-3330, XP002129879 • GALLERT, Karl-Christian • DE MONTALK, G. POTOCKI ET AL: "Sequence D-61184 Karben (DE) analysis of the gene encoding amylosucrase • BENGS, Holger from Neisseria polysaccharea and D-60598 Frankfurt am Main (DE) characterization of the recombinant enzyme" J.
    [Show full text]
  • In Vitro Synthesis of Ten Starches by Potato Starch-Synthase and Starch
    y: C istr urr m en e t h R Mukerjea et al., Organic Chem Curr Res 2015, 4:3 C e c s i e n a a r c g DOI: 10.4172/2161-0401.1000142 r h Organic Chemistry O ISSN: 2161-0401 Current Research ResearchResearch Article Article OpenOpen Access Access In Vitro Synthesis of Ten Starches by Potato Starch-Synthase and Starch- Branching-Enzyme Giving Different Ratios of Amylopectin and Amylose Mukerjea R, Sheets RL, Gray AN and Robyt JF* Laboratory of Carbohydrate Chemistry and Enzymology, Department of Biochemistry, Biophysics and Molecular Biology, Iowa State University, Ames, IA 50011, USA Abstract The objectives of this study was the use of two highly purified enzymes, potato Starch-Synthase (SS) and Starch- Branching-Enzyme (SBE) to obtain in vitro the syntheses of ten starches, with different ratios of amylopectin and amylose. The amylose was first synthesized by the reaction of SS with ADP [14C] Glc to give ten identical amyloses; the amylopectin was then synthesized by the action of SBE to give ten starches with different amounts of amylopectin. Two of the 14C-amyloses were reacted with 1.0 mIU of purified Starch-Branching-Enzyme (SBE) for 75 and 130 sec, respectively. In a second synthesis, 3 of the amyloses were autoclaved and reacted with 1.0 mIU for 15, 45, and 75 sec. In the third synthesis, 5 amyloses were reacted with different amounts of SBE from 0.50 to 0.01 mIU for 130 sec each. The 10 synthesized starches were separated into amylopectin and amylose fractions.
    [Show full text]
  • NAPRALERT Classification Codes
    NAPRALERT Classification Codes June 1993 STN International® Copyright © 1993 American Chemical Society Quoting or copying of material from this publication for educational purposes is encouraged, providing acknowledgement is made of the source of such material. Classification Codes in NAPRALERT The NAPRALERT File contains classification codes that designate pharmacological activities. The code and a corresponding textual description are searchable in the /CC field. To be comprehensive, both the code and the text should be searched. Either may be posted, but not both. The following tables list the code and the text for the various categories. The first two digits of the code describe the categories. Each table lists the category described by codes. The last table (starting on page 56) lists the Classification Codes alphabetically. The text is followed by the code that also describes the category. General types of pharmacological activities may encompass several different categories of effect. You may want to search several classification codes, depending upon how general or specific you want the retrievals to be. By reading through the list, you may find several categories related to the information of interest to you. For example, if you are looking for information on diabetes, you might want to included both HYPOGLYCEMIC ACTIVITY/CC and ANTIHYPERGLYCEMIC ACTIVITY/CC and their codes in the search profile. Use the EXPAND command to verify search terms. => S HYPOGLYCEMIC ACTIVITY/CC OR 17006/CC OR ANTIHYPERGLYCEMIC ACTIVITY/CC OR 17007/CC 490 “HYPOGLYCEMIC”/CC 26131 “ACTIVITY”/CC 490 HYPOGLYCEMIC ACTIVITY/CC ((“HYPOGLYCEMIC”(S)”ACTIVITY”)/CC) 6 17006/CC 776 “ANTIHYPERGLYCEMIC”/CC 26131 “ACTIVITY”/CC 776 ANTIHYPERGLYCEMIC ACTIVITY/CC ((“ANTIHYPERGLYCEMIC”(S)”ACTIVITY”)/CC) 3 17007/CC L1 1038 HYPOGLYCEMIC ACTIVITY/CC OR 17006/CC OR ANTIHYPERGLYCEMIC ACTIVITY/CC OR 17007/CC 2 This search retrieves records with the searched classification codes such as the ones shown here.
    [Show full text]
  • Oil 96 20061219 150649
    0. Schamburg · 0. Stephan (Eds.) GBF- Gesellschaft für Biotechnologische Forschung Enzyme Handbock Class 2.3.2 - 2.4 Transferases 12 Springer Index (Aiphabetical order of Enzyme names) EC-No. Name EC-No. Name 2.4.1.60 Abequosyltransferase 2.4.99.7 (alpha-N-Acetylneuraminyl- 2.4.99 .3 alpha-N-Acetylgalactos­ 2,3-beta-galactosyl-1 ,3)­ aminide alpha-2,6-sialyl­ N-acetylgalactosaminide transferase alpha-2,6-sialyltransferase 2.4.1.147 Acetylgalactosaminyl-ü-gly• 2.4.1.92 (N-Acetylneuraminyl)-galac­ cosyl-glycoprotein beta- tosylglucosylceramide 1,3-N-acetylglucosaminyl­ N-acetylgalactosaminyl­ transferase transferase 2.4.1.148 Acetylgalactosaminyl-0-gly­ 2.4.1.165 N-Acetylneuraminylgalacto­ cosyl-glycoprotein beta- sylglucosylceramide beta- 1,6-N-acetylglucosaminyl 1,4-N-acetylgalactosaminyl­ transferase transferase 2.4.1.141 N-Acetylglucosaminyl­ 2.4.1.47 N-Acylsphingosine galacto­ diphosphodolichol N-ace­ syltransferase tylglucosaminyltransferase 2.4.2.7 Adenine phosphoribosyl­ 2.4.1.187 N-Acetylglucosaminyl­ transferase diphosphoundecaprenol 2.3.2.9 Agaritine gamma-glutamyl­ N-acetyl-beta-0-mannos­ transferase aminyltransferase 2.3.2.14 0-Aianine gamma-glutamyl­ 2.4.1.188 N-Acetylglucosaminyl­ transferase diphosphoundecaprenol 2.3.2.11 Alanylphosphatidylglycerol glucosyltransferase synthase 2.4.1.38 beta-N-Acetylglucosaminyl­ 2.4.1.162 Aldose beta-D-fructosyl­ glycopeptide beta-1 ,4-ga­ transferase lactosyltransferase 2.4.1.33 Alginate synthase 2.4.1.124 N-Acetyllactosamine 2.4.1.103 Alizarin 2-beta-glucosyl­ 3-alpha-galactosyltrans­ transferase ferase 2. 4.1.140 Alternansucrase 2.4.1.90 N-Acetyllactosamine syn­ 2.4.2.14 Amidophosphoribosyl­ thase transferase 2.4.1.149 N-Acetyllactosaminide beta- 2.4.1.4 Amylosucrase 1, 3-N-acetylg lucosaminyl­ 2.4.1.
    [Show full text]
  • Pan-Genomic and Transcriptomic Analyses of Leuconostoc
    www.nature.com/scientificreports OPEN Pan-genomic and transcriptomic analyses of Leuconostoc mesenteroides provide insights into Received: 8 June 2017 Accepted: 30 August 2017 its genomic and metabolic features Published: xx xx xxxx and roles in kimchi fermentation Byung Hee Chun1, Kyung Hyun Kim1, Hye Hee Jeon1, Se Hee Lee2 & Che Ok Jeon 1 The genomic and metabolic features of Leuconostoc (Leu) mesenteroides were investigated through pan-genomic and transcriptomic analyses. Relatedness analysis of 17 Leu. mesenteroides strains available in GenBank based on 16S rRNA gene sequence, average nucleotide identity, in silico DNA- DNA hybridization, molecular phenotype, and core-genome indicated that Leu. mesenteroides has been separated into diferent phylogenetic lineages. Pan-genome of Leu. mesenteroides strains, consisting of 999 genes in core-genome, 1,432 genes in accessory-genome, and 754 genes in unique genome, and their COG and KEGG analyses showed that Leu. mesenteroides harbors strain-specifcally diverse metabolisms, probably representing high evolutionary genome changes. The reconstruction of fermentative metabolic pathways for Leu. mesenteroides strains showed that Leu. mesenteroides produces various metabolites such as lactate, ethanol, acetate, CO2, mannitol, diacetyl, acetoin, and 2,3-butanediol through an obligate heterolactic fermentation from various carbohydrates. Fermentative metabolic features of Leu. mesenteroides during kimchi fermentation were investigated through transcriptional analyses for the KEGG pathways and reconstructed metabolic pathways of Leu. mesenteroides using kimchi metatranscriptomic data. This was the frst study to investigate the genomic and metabolic features of Leu. mesenteroides through pan-genomic and metatranscriptomic analyses, and may provide insights into its genomic and metabolic features and a better understanding of kimchi fermentations by Leu.
    [Show full text]
  • All Enzymes in BRENDA™ the Comprehensive Enzyme Information System
    All enzymes in BRENDA™ The Comprehensive Enzyme Information System http://www.brenda-enzymes.org/index.php4?page=information/all_enzymes.php4 1.1.1.1 alcohol dehydrogenase 1.1.1.B1 D-arabitol-phosphate dehydrogenase 1.1.1.2 alcohol dehydrogenase (NADP+) 1.1.1.B3 (S)-specific secondary alcohol dehydrogenase 1.1.1.3 homoserine dehydrogenase 1.1.1.B4 (R)-specific secondary alcohol dehydrogenase 1.1.1.4 (R,R)-butanediol dehydrogenase 1.1.1.5 acetoin dehydrogenase 1.1.1.B5 NADP-retinol dehydrogenase 1.1.1.6 glycerol dehydrogenase 1.1.1.7 propanediol-phosphate dehydrogenase 1.1.1.8 glycerol-3-phosphate dehydrogenase (NAD+) 1.1.1.9 D-xylulose reductase 1.1.1.10 L-xylulose reductase 1.1.1.11 D-arabinitol 4-dehydrogenase 1.1.1.12 L-arabinitol 4-dehydrogenase 1.1.1.13 L-arabinitol 2-dehydrogenase 1.1.1.14 L-iditol 2-dehydrogenase 1.1.1.15 D-iditol 2-dehydrogenase 1.1.1.16 galactitol 2-dehydrogenase 1.1.1.17 mannitol-1-phosphate 5-dehydrogenase 1.1.1.18 inositol 2-dehydrogenase 1.1.1.19 glucuronate reductase 1.1.1.20 glucuronolactone reductase 1.1.1.21 aldehyde reductase 1.1.1.22 UDP-glucose 6-dehydrogenase 1.1.1.23 histidinol dehydrogenase 1.1.1.24 quinate dehydrogenase 1.1.1.25 shikimate dehydrogenase 1.1.1.26 glyoxylate reductase 1.1.1.27 L-lactate dehydrogenase 1.1.1.28 D-lactate dehydrogenase 1.1.1.29 glycerate dehydrogenase 1.1.1.30 3-hydroxybutyrate dehydrogenase 1.1.1.31 3-hydroxyisobutyrate dehydrogenase 1.1.1.32 mevaldate reductase 1.1.1.33 mevaldate reductase (NADPH) 1.1.1.34 hydroxymethylglutaryl-CoA reductase (NADPH) 1.1.1.35 3-hydroxyacyl-CoA
    [Show full text]