DOCSLIB.ORG

1508311112.Full.Pdf

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
1508311112.Full.Pdf Discovery and molecular and biocatalytic properties of hydroxynitrile lyase from an invasive millipede, Chamberlinius hualienensis Mohammad Dadashipoura,b,1, Yuko Ishidaa,b,1, Kazunori Yamamotoa,b, and Yasuhisa Asanoa,b,2 aBiotechnology Research Center and Department of Biotechnology, Toyama Prefectural University, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan; and bAsano Active Enzyme Molecule Project, Exploratory Research for Advanced Technology, Japan Science and Technology Agency, 5180 Kurokawa, Imizu, Toyama 939-0398, Japan Edited by Jerrold Meinwald, Cornell University, Ithaca, NY, and approved July 8, 2015 (received for review April 29, 2015) Hydroxynitrile lyase (HNL) catalyzes the degradation of cyanohy- of mandelonitrile using emulsin from the almond, Prunus amyg- drins and causes the release of hydrogen cyanide (cyanogenesis). dalus, as an HNL source (PaHNL), has been demonstrated (8). HNL can enantioselectively produce cyanohydrins, which are valuable Versatile HNLs, homologous to FAD-dependent oxidoreductase, + building blocks for the synthesis of fine chemicals and pharmaceu- α/β-hydrolase, carboxypeptidase, a Zn2 -dependent alcohol de- + ticals, and is used as an important biocatalyst in industrial bio- hydrogenase, and a Mn2 -dependent cupin structure have all been technology. Currently, HNLs are isolated from plants and bacteria. identified from plants and bacteria (9–12). Because PaHNL Because industrial biotechnology requires more efficient and stable shows the highest specific activity in the synthesis of mandelonitrile enzymes for sustainable development, we must continuously explore among known HNLs (13), along with a wide substrate specificity other potential enzyme sources for the desired HNLs. Despite the and high stability, it serves as a valuable catalyst in industrial bio- abundance of cyanogenic millipedes in the world, there has been no technology. Although many scientists have identified HNLs in the precise study of the HNLs from these arthropods. Here we report the last two decades (14, 15), further screening for better enzyme(s) isolation of HNL from the cyanide-emitting invasive millipede Cham- with high reaction rates and stabilities from unexplored bio- berlinius hualienensis, along with its molecular properties and appli- resources is necessary to enrich the current tool box for sus- CHEMISTRY cation in biocatalysis. The purified enzyme displays a very high tainable industrial development. specific activity in the synthesis of mandelonitrile. It is a glycosylated The millipede Chamberlinius hualienensis (16) (Movie S1), homodimer protein and shows no apparent sequence identity or originally from Hualien, Taiwan, invaded Okinawa Island, Japan homology with proteins in the known databases. It shows biocata- in 1983 and has been expanding its habitat in Kyushu, Japan. lytic activity for the condensation of various aromatic aldehydes with Large swarms of the millipedes enter houses and sometimes potassium cyanide to produce cyanohydrins and has high stability cause train delays (17). The animal secretes mandelonitrile, over a wide range of temperatures and pH values. It catalyzes the benzaldehyde, and hazardous hydrogen cyanide as defense chem- synthesis of (R)-mandelonitrile from benzaldehyde with a 99% enan- icals, as previously reported for polydesmid millipede species (18, BIOCHEMISTRY tiomeric excess, without using any organic solvents. Arthropod fauna 19). In the millipede species Apheloria corrugate, a predicted comprise 80% of terrestrial animals. We propose that these animals enzyme is thought to play a role in the decomposition of man- can be valuable resources for exploring not only HNLs but also di- delonitrile and the release of hydrogen cyanide and benzaldehyde verse, efficient, and stable biocatalysts in industrial biotechnology. millipede hydroxynitrile lyase | bioresource exploration | biocatalysis | Significance white biotechnology | arthropod Hydroxynitrile lyase (HNL) has been isolated from plants and bacteria and is a valuable tool in the chiral-specific synthesis of nzymes are very specific and efficient catalysts. They are able cyanohydrins, which are important building blocks of fine to catalyze reactions without extreme conditions, such as high E chemicals and pharmaceuticals. To discover more efficient and temperature and pressure, that are often required in chemical stable HNLs, we focused on the invasive cyanogenic millipede synthetic processes. Industrial biotechnologies using these bio- as a bioresource. The HNL identified from the millipede showed catalysts have the advantages of higher reaction rates, efficiencies, not only the highest specific activity toward benzaldehyde and enantioselectivities. This technology also reduces energy among known HNLs, including the almond HNL in industrial consumption and decreases the production of hazardous chem- use, along with wide temperature and pH stabilities, but also ical waste. Conventional industrial chemistry is gradually being high enantioselectivity in the synthesis of various cyanohy- replaced by industrial biotechnology. For example, nitrile hydra- drins. These properties make it suitable as an industrial biocat- tase is used for the synthesis of acrylamide from acrylonitrile. This alyst. Arthropods are likely to be valuable sources of potential strategy produces more than 600,000 tons of acrylamide world- biocatalysts for the next generation of industrial biotechnology. wide every year (1). Hydroxynitrile lyase (HNL) is found as a molecular component Author contributions: M.D., Y.I., K.Y., and Y.A. designed research; M.D., Y.I., and K.Y. in plant defense systems and plays a role in the decomposition of performed research; M.D., Y.I., K.Y., and Y.A. analyzed data; and M.D., Y.I., K.Y., and Y.A. stored cyanogenic glycosides into hydrogen cyanide and aldehydes wrote the paper. (2). The enzyme’s reverse activity is used industrially in the enan- The authors declare no conflict of interest. tioselective production of cyanohydrins. Cyanohydrins are valuable This article is a PNAS Direct Submission. building blocks for the synthesis of fine chemicals, pharmaceuticals, Freely available online through the PNAS open access option. and agrochemicals such as denopamine (β1-adrenergic receptor Data deposition: The cDNA sequence reported in this paper has been deposited in the agonist) (3, 4), clopidogrel (platelet aggregation inhibitor) (5), py- DNA Data Bank of Japan (accession no. LC004755). rethroids (insecticides) (6), and (R)-pantolactone [starting com- 1M.D. and Y.I. contributed equally to this work. pound for synthesis of (R)-pantothenic acid, (R)-panthenol, and 2To whom correspondence should be addressed. Email: [email protected]. (R)-pantetheine] (7). The enantioselective condensation of This article contains supporting information online at www.pnas.org/lookup/suppl/doi:10. benzaldehyde with hydrogen cyanide in the asymmetric synthesis 1073/pnas.1508311112/-/DCSupplemental. www.pnas.org/cgi/doi/10.1073/pnas.1508311112 PNAS Early Edition | 1of6 Downloaded by guest on October 1, 2021 (20). If the enzyme derived from C. hualienensis is an HNL, the properties and inhibitor susceptibility, and demonstrate its substrate large population of invasive millipedes could be used as an im- specificity, kinetic parameters, and gene expression. We describe portant resource for HNL purification. Here we identify HNL from the potential of the millipede HNL to be widely used as a bio- the invasive cyanogenic millipede, characterize its physicochemical catalyst for the enantioselective synthesis of cyanohydrins, building Fig. 1. Characterization of ChuaHNL. (A) Cleavage reaction of racemic mandelonitrile by ChuaHNL. Production of benzaldehyde in 100 mM citrate buffer, pH 5.5, at 22 °C was measured by monitoring the absorbance at 280 nm. Solid and dotted lines indicate absorbance with and without the enzyme, re- spectively. (B) Synthesis of (R)-mandelonitrile from 50 mM benzaldehyde in 400 mM citrate buffer, pH 4.2, at 22 °C. ChuaHNL specifically produces (R)-mandelonitrile, whereas the chemical reaction produces a scant amount of (R)- and (S)-mandelonitrile. Production of mandelonitrile was measured by monitoring the absorbance at 254 nm using an HPLC equipped with a chiral column. Retention times of (R)- and (S)-mandelonitrile were 11.4 min and 14.3 min, respectively. (Inset) Magnified view in the range of 10–15 min. Solid and dotted lines indicate the synthetic reaction of mandelonitrile from benzal- dehyde and potassium cyanide with and without ChuaHNL, respectively. (C) Temperature stability. Enzyme samples were incubated at each temperature for 1 h and assayed for the remaining HNL activity (as shown in B). (D) Optimum temperature. Enzyme samples were assayed at each temperature (as shown in B). (E) pH stability. Enzyme samples were incubated at each temperature for 1 h and assayed for the remaining HNL activity (as shown in B). ●, enzymatic activity in a citrate-phosphate buffer; □, enzymatic activity in a glycine-sodium hydroxide buffer. (F) Optimum pH. Enzyme samples were assayed at each pH value (as shown in B). Values are the mean ± SD; n = 3. 2of6 | www.pnas.org/cgi/doi/10.1073/pnas.1508311112 Dadashipour et al. Downloaded by guest on October 1, 2021 Table 1. Purification of ChuaHNL from the millipede C. hualienensis Specific Yield, Purification − Purification step Activity, U* Protein, mg activity, U·mg 1 % fold Body crude homogenate 26,600 4,390 6.10 100 1.00 Saturated ammonium sulfate 19,600 1,000 19.6 73.7
Load more

Recommended publications
  • Bioprospecting for Hydroxynitrile Lyases by Blue Native PAGE Coupled HCN Detection
    Send Orders for Reprints to [email protected] Current Biotechnology, 2015, 4, 111-117 111 Bioprospecting for Hydroxynitrile Lyases by Blue Native PAGE Coupled HCN Detection Elisa Lanfranchi1, Eva-Maria Köhler1, Barbara Darnhofer1,2,3, Kerstin Steiner1, Ruth Birner-Gruenberger1,2,3, Anton Glieder1,4 and Margit Winkler*,1 1ACIB GmbH, Graz, Austria; 2Institute for Pathology, Medical University of Graz, Graz, Austria; 3Omics Center Graz, BioTechMed, Graz, Austria; 4Institute of Molecular Biotechnology, Graz University of Technology, NAWI Graz, Graz, Austria Abstract: Hydroxynitrile lyase enzymes (HNLs) catalyze the stereoselective addition of HCN to carbonyl compounds to give valuable chiral hydroxynitriles. The discovery of new sources of HNL activity has been reported several times as the result of extensive screening of diverse plants for cyanogenic activity. Herein we report a two step-method that allows estimation of not only the native size of the active HNL enzyme but also its substrate specificity. Specifically, crude protein extracts from plant tissue are first subjected to blue native-PAGE. The resulting gel is then directly used for an activity assay in which the formation of hydrocyanic acid (HCN) is detected upon the cyanogenesis reaction of any cyanohydrin catalyzed by the enzyme of interest. The same gel may be used with different substrates, thus exploring the enzyme’s substrate scope already on the screening level. In combination with mass spectrometry, sequence information can be retrieved, which is demonstrated
    [Show full text]
  • Argininosuccinate Lyase Deficiency
    ©American College of Medical Genetics and Genomics GENETEST REVIEW Argininosuccinate lyase deficiency Sandesh C.S. Nagamani, MD1, Ayelet Erez, MD, PhD1 and Brendan Lee, MD, PhD1,2 The urea cycle consists of six consecutive enzymatic reactions that citrulline together with elevated argininosuccinic acid in the plasma convert waste nitrogen into urea. Deficiencies of any of these enzymes or urine. Molecular genetic testing of ASL and assay of ASL enzyme of the cycle result in urea cycle disorders (UCDs), a group of inborn activity are helpful when the biochemical findings are equivocal. errors of hepatic metabolism that often result in life-threatening However, there is no correlation between the genotype or enzyme hyperammonemia. Argininosuccinate lyase (ASL) catalyzes the activity and clinical outcome. Treatment of acute metabolic decom- fourth reaction in this cycle, resulting in the breakdown of arginino- pensations with hyperammonemia involves discontinuing oral pro- succinic acid to arginine and fumarate. ASL deficiency (ASLD) is the tein intake, supplementing oral intake with intravenous lipids and/ second most common UCD, with a prevalence of ~1 in 70,000 live or glucose, and use of intravenous arginine and nitrogen-scavenging births. ASLD can manifest as either a severe neonatal-onset form therapy. Dietary restriction of protein and dietary supplementation with hyperammonemia within the first few days after birth or as a with arginine are the mainstays in long-term management. Ortho- late-onset form with episodic hyperammonemia and/or long-term topic liver transplantation (OLT) is best considered only in patients complications that include liver dysfunction, neurocognitive deficits, with recurrent hyperammonemia or metabolic decompensations and hypertension.
    [Show full text]
  • Synthesis of Tryptophan from Indole, Pyruvate, and Ammonia (E
    Proc. Nat. Acad. S&i. USA Vol. 69, No. 5, pp. 1086-1090, May 1972 Reversibility of the Tryptophanase Reaction: Synthesis of Tryptophan from Indole, Pyruvate, and Ammonia (E. coli/a-aminoacrylate/Michaelis-Menten kinetics/pyridoxal 5'-phosphate) TAKEHIKO WATANABE AND ESMOND E. SNELL Department of Biochemistry, University of California, Berkeley, Calif. 94720 Contributed by Esmond E. Snell, February 14, 1972 ABSTRACT Degradation of tryptophan to indole, tain substituted indoles. Reactions 1-3 were shown (4)t to pro- pyruvate, and ammonia by tryptophanase (EC 4 ....) from ceed through a common intermediate, probably an enzyme- Escherichia coli, previously thought to be an irreversible reaction, is readily reversible at high concentrations of bound a-aminoacrylic acid, which could either decompose to pyruvate and ammonia. Tryptophan and certain of its pyruvate and ammonia (in reactions 1 and 2) or add indole to analogues, e.g., 5-hydroxytryptophan, can be synthesized form tryptophan (in reaction 3). At concentrations previously by this reaction from pyruvate, ammonia, and indole or an tested, reactions 1 and 2 were irreversible (4). appropriate derivative at maximum velocities approaching to Yamada et al. those of the degradative reactions. Concentrations of Subsequent these investigations, (5-7) ammonia required for the synthetic reactions produce showed that 0-tyrosinase from Escherichia intermedia cata- specific changes in the spectrum of tryptophanase that lyzes reaction 4 but not reaction 1, and is similar in many differ from those produced by K+ and indicate that am- respects to tryptophanase. monia interacts with bound pyridoxal 5'-phosphate to form an imine. Kinetic results indicate that pyruvate is Tyrosine + H20 Phenol + Pyruvate + NH3 (4) the second substrate bound, hence indole must be the too, catalyzes degradation of serine, cysteine, etc.
    [Show full text]
  • Taming the Wild Rubisco: Explorations in Functional Metagenomics
    Taming the Wild RubisCO: Explorations in Functional Metagenomics DISSERTATION Presented in Partial Fulfillment of the Requirements for the Degree Doctor of Philosophy in the Graduate School of The Ohio State University By Brian Hurin Witte, M.S. Graduate Program in Microbiology The Ohio State University 2012 Dissertation Committee : F. Robert Tabita, Advisor Joseph Krzycki Birgit E. Alber Paul Fuerst Copyright by Brian Hurin Witte 2012 Abstract Ribulose bisphosphate carboxylase/oxygenase (E.C. 4.1.1.39) (RubisCO) is the most abundant protein on Earth and the mechanism by which the vast majority of carbon enters the planet’s biosphere. Despite decades of study, many significant questions about this enzyme remain unanswered. As anthropogenic CO2 levels continue to rise, understanding this key component of the carbon cycle is crucial to forecasting feedback circuits, as well as to engineering food and fuel crops to produce more biomass with few inputs of increasingly scarce resources. This study demonstrates three means of investigating the natural diversity of RubisCO. Chapter 1 builds on existing DNA sequence-based techniques of gene discovery and shows that RubisCO from uncultured organisms can be used to complement growth in a RubisCO-deletion strain of autotrophic bacteria. In a few short steps, the time-consuming work of bringing an autotrophic organism in to pure culture can be circumvented. Chapter 2 details a means of entirely bypassing the bias inherent in sequence-based gene discovery by using selection of RubisCO genes from a metagenomic library. Chapter 3 provides a more in-depth study of the RubisCO from the methanogenic archaeon Methanococcoides burtonii.
    [Show full text]
  • Purification of Uroporphyrinogen Decarboxylase from Human Erythrocytes
    Biochem. J. (1983) 215,45-55 45 Printed in Great Britain Purification of uroporphyrinogen decarboxylase from human erythrocytes Immunochemical evidence for a single protein with decarboxylase activity in human erythrocytes and liver George H. ELDER, John A. TOVEY and Diane M. SHEPPARD Department ofMedical Biochemistry, Welsh National School ofMedicine, Heath Park, CardiffCF 4XN, Wales, U.K. (Received 21 March 1983/Accepted I June 1983) Uroporphyrinogen decarboxylase (EC 4.1.1.37) has been purified 4419-fold to a specific activity of 58.3 nmol of coproporphyrinogen III formed/min per mg of protein (with pentacarboxyporphyrinogen III as substrate) from human erythrocytes by adsorption to DEAE-cellulose, (NH4)2SO4 fractionation, gel filtration, phenyl- Sepharose chromatography and polyacrylamide-gel electrophoresis. Progressive loss of activity towards uroporphyrinogens I and III occurred during purification. Experiments employing immunoprecipitation, immunoelectrophoresis and titration with solid-phase antibody indicated that all the uroporphyrinogen decarboxylase activity of human erythrocytes resides in one protein, and that the substrate specificity of this protein had changed during purification. The purified enzyme had a minimum mol.wt. of 39 500 on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Gel filtration gave a mol.wt. of 58 000 for the native enzyme. Isoelectric focusing showed a single band with a pl of 4.60. Reaction with N-ethylmaleimide abofished both catalytic activity and immunoreactivity. Incubation with substrates or porphyrins prevented inactivation by N-ethylmaleimide. An antiserum raised against purified erythrocyte enzyme pre- cipitated more than 90% of the uroporphyrinogen decarboxylase activity from human liver. Quantitative immunoprecipitation and crossed immunoelectrophoresis showed that the erythrocyte and liver enzymes are very similar but not identical.
    [Show full text]
  • Evidence for Lack of DNA Photoreactivating Enzyme in Humans
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 4389-4393, May 1993 Biochemistry Evidence for lack of DNA photoreactivating enzyme in humans (thymine dimer/skin cancer/mammais/reptiles) YWAN FENG Li, SANG-TAE KIM, AND AZIZ SANCAR Department of Biochemistry and Biophysics, University of North Carolina, School of Medicine, Chapel Hill, NC 27599 Communicated by Mary Ellen Jones, January 27, 1993 (receivedfor review November 13, 1992) ABSTRACT Photoreactivating enzyme (DNA photolyase; fore, we have reexamined this question, employing new deoxyribocyclobutadipyrimidine pyrimidine-lyase, EC methodology that has become available within the last few 4.1.99.3) repairs UV damage to DNA by utilizing the energy of years-specifically, the ability to make large quantities of near-UV/visible light to split pyrimidine dimers into mono- substrate with a single TOT at a defined position (13). Such mers. The enzyme is widespread in nature but is absent in a substrate makes it possible to detect very low levels of certain species in a seemingly unpredictable manner. Its pres- photolyase. Further, since the photoproduct is chemically ence in humans has been a source of considerable controversy. incorporated into DNA the ambiguity arising from the nature To help resolve the issue we used a very specific and sensitive of the photoproduct affected by photoreactivation treatment assay to compare photoreactivation activity in human, rattle- is eliminated. Our results with this substrate system and snake, yeast, and Escherichia coli cells. Photolyase was easily extracts from cultured human cells and from white blood cells detectable in E. coli, yeast, and rattlesnake cell-free extracts (WBCs) taken from volunteers reveal that humans do not but none was detected in cell-free extracts from HeLa cells or have a photoreactivating enzyme.
    [Show full text]
  • Dependent Enzymes. a Hypothesis Philipp Christen*, Patrik Kasper, Heinz Gehring, Michael Sterk Bioehemisches Institut, Universitgit Ziirich, Winterthurerstr
    FEBS Letters 389 (1996) 12-14 FEBS 16909 Minireview Stereochemical constraint in the evolution of pyridoxal-5'-phosphate- dependent enzymes. A hypothesis Philipp Christen*, Patrik Kasper, Heinz Gehring, Michael Sterk Bioehemisches Institut, Universitgit Ziirich, Winterthurerstr. 190, CH-8057 Ziirieh, Switzerland Received 11 January 1996 cular evolution of B6 enzymes. Alanine aminotransferase, as- Abstract In the transamination reactions undergone by pyri- doxal-5'-phosphate-dependent enzymes that act on L-amino partate aminotransferase, 2,2-dialkylglycine decarboxylase, acids, the C4' atom of the cofactor is without exception glutamate decarboxylase, and serine hydroxymethyltransfer- protonated from the si side. This invariant absolute stereo- ase indeed belong to the large c~ family of homologous B6 chemistry of enzymes not all of which are evolutionarily related enzymes [1-3]. However, the pyridoxal-5'-phosphate-depen- to each other and the inverse stereochemistry in the case of D- dent 13 subunit of tryptophan synthase which shows the alanine aminotransferase might reflect a stereochemical con- same stereochemistry is a member of the [3 family of B6 en- straint in the evolution of these enzymes rather than an zymes which is not homologous with the e~ family [1,4]. accidental historical trait passed on from a common ancestor (About the seventh enzyme, pyridoxamine pyruvate amino- enzyme. Conceivably, the coenzyme and substrate binding sites transferase, no information on primary or tertiary structure of primordial pyridoxal-5'-phosphate-dependent
    [Show full text]
  • Identification, Cloning, and Nucleotide Sequencing of the Ornithine Decarboxylase Antizyme Gene of Escherichia Coli E
    Proc. Natl. Acad. Sci. USA Vol. 90, pp. 7129-7133, August 1993 Biochemistry Identification, cloning, and nucleotide sequencing of the ornithine decarboxylase antizyme gene of Escherichia coli E. S. CANELLAKIS*, A. A. PATERAKISt, S.-C. HUANG*, C. A. PANAGIOTIDIS*t, AND D. A. KYRIAKIDISt *Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06510; and tLaboratory of Biochemistry, School of Chemistry, Aristotelian University of Thessaloniki, 540 06 Thessaloniki, Greece Communicated by Philip Siekevitz, April 5, 1993 ABSTRACT The ornithine decarboxylase antizyme gene (11). The study of the Az has been very difficult because of of Escherichia coli was identified by immunological screening its low abundance and because its N terminus is blocked of an E. coli genomic library. A 6.4-kilobase fragment con- (unpublished results). taining the antizyme gene was subdoned and sequenced. The We now report the isolation of the E. coli Az gene, its open reading frame encoding the antizyme was identified on the location on the E. coli chromosome, as well as the identifi- basis of its ability to direct the synthesis of immunoreactive cation and sequence of the open reading frame (ORF) en- antizyme. Antizyme shares significant homology with bacterial coding the Az.§ transcriptional activators of the two-component regulatory system family; these systems consist of a "sensor" kinase and a transcriptional regulator. The open reading frame next to MATERIALS AND METHODS antizyme is homologous to sensor kinases. Antizyme overpro- Bacterial Strains, Plasmids, and Phages. E. coli MG1655 duction inhibits the activities of both ornithine and arginine (A-, F-) (13); KL527 [MG1655 gyrA, A (speA-speB)97, decarboxylases without affecting their protein levels.
    [Show full text]
  • A Hierarchical Classification of Polysaccharide Lyases for Glycogenomics Vincent Lombard, Thomas Bernard, Corinne Rancurel, Harry Brumer, Pedro M
    A hierarchical classification of polysaccharide lyases for glycogenomics Vincent Lombard, Thomas Bernard, Corinne Rancurel, Harry Brumer, Pedro M. Coutinho, Bernard Henrissat To cite this version: Vincent Lombard, Thomas Bernard, Corinne Rancurel, Harry Brumer, Pedro M. Coutinho, et al.. A hierarchical classification of polysaccharide lyases for glycogenomics. Biochemical Journal, Portland Press, 2010, 432 (3), pp.437-444. 10.1042/BJ20101185. hal-00539724 HAL Id: hal-00539724 https://hal.archives-ouvertes.fr/hal-00539724 Submitted on 25 Nov 2010 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. Biochemical Journal Immediate Publication. Published on 07 Oct 2010 as manuscript BJ20101185 A hierarchical classification of polysaccharide lyases for glycogenomics V. Lombard*, T. Bernard*†, C. Rancurel*, H Brumer‡, P.M. Coutinho* & B. Henrissat*1 *Architecture et Fonction des Macromolécules Biologiques, UMR6098, CNRS, Université de la Méditerranée, Université de Provence, Case 932, 163 Avenue de Luminy, 13288 Marseille cedex 9, France ‡School of Biotechnology, Royal Institute of Technology (KTH), AlbaNova University Centre, 106 91 Stockholm, Sweden † Present address: Biométrie et Biologie Évolutive, UMR CNRS 5558, UCB Lyon 1, Bât. Grégor Mendel, 43 bd du 11 novembre 1918, 69622 Villeurbanne cedex, France 1To whom correspondence should be addressed: [email protected]‐mrs.fr Abstract: Carbohydrate‐active enzymes face huge substrate diversity in a highly selective manner with only a limited number of available folds.
    [Show full text]
  • Structure and Expression of Spinach Leaf Cdna Encoding
    Proc. Nati. Acad. Sci. USA Vol. 85, pp. 787-791, February 1988 Botany Structure and expression of spinach leaf cDNA encoding ribulosebisphosphate carboxylase/oxygenase activase (photosynthesis/Arabidopsis thaliana/nucleotide binding site/mRNA processing) JEFFREY M. WERNEKE*, RAYMOND E. ZIELINSKI*, AND WILLIAM L. OGRENtt *Department of Plant Biology, University of Illinois, Urbana, IL 61801; and tAgricultural Research Service, U.S. Department of Agriculture, 1102 South Goodwin Avenue, Urbana, IL 61801 Contributed by William L. Ogren, October 12, 1987 ABSTRACT Ribulosebisphosphate carboxylase/oxygen- gene, expressed the cDNAs in Escherichia coli, and used the ase activase is a recently discovered enzyme that catalyzes the clones as hybridization probes to address the specific nature activation of ribulose-1,5-bisphosphate carboxylase/oxygenase of the rca mutation.§ ["rubisco"; ribulose-bisphosphate carboxylase; 3-phospho-D- glycerate carboxy-lyase (dimerizing), EC 4.1.1.39] in vivo. MATERIALS AND METHODS Clones of rubisco activase cDNA were isolated immunologi- Purification of Rubisco Activase. Intact spinach chloro- cally from spinach (Spinacea oleracea L.) and Arabidopsis plasts were lysed by 1:10 dilution into 20 mM Tris HCI, pH thaliana libraries. Sequence analysis of the spinach and Ara- 8/4 mM 2-mercaptoethanol (8). After centrifugation at bidopsis cDNAs identified consensus nucleotide binding sites, 10,000 x g for 10 min, the supernatant was passed through consistent with an ATP requirement for rubisco activase a 22-ptm Milex filter. Forty milligrams of soluble protein was activity. A derived amino acid sequence common to chloro- then loaded onto a Mono Q column (Pharmacia) equilibrated plast transit peptides was also identified. After synthesis of in the same buffer.
    [Show full text]
  • Dynamic Determination of the Functional State in Photolyase and the Implication for Cryptochrome
    Dynamic determination of the functional state in photolyase and the implication for cryptochrome Zheyun Liu a,b, Meng Zhanga,b,c, Xunmin Guo a,b, Chuang Tan a,b,d,JiangLia,b,LijuanWanga,b, Aziz Sancare,1, and Dongping Zhong a,b,c,d,f,1 Departments of aPhysics and bChemistry and Biochemistry, and Programs of cBiophysics, dChemical Physics, and fBiochemistry, The Ohio State University, Columbus, OH 43210; and eDepartment of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill, NC 27599 Contributed by Aziz Sancar, June 28, 2013 (sent for review May 26, 2013) – The flavin adenine dinucleotide cofactor has an unusual bent of photolyase by donating an electron from its anionic form (FAD – configuration in photolyase and cryptochrome, and such a folded in insect or FADH in plant) to a putative substrate that induces structure may have a functional role in initial photochemistry. Using a local electrostatic variation to cause conformation changes for femtosecond spectroscopy, we report here our systematic charac- signaling. Both models require electron transfer (ET) at the active terization of cyclic intramolecular electron transfer (ET) dynamics site to induce electrostatic changes for signaling. Similar to the between the flavin and adenine moieties of flavin adenine di- pyrimidine dimer, the Ade moiety near the Lf ring could also be nucleotide in four redox forms of the oxidized, neutral, and anionic an oxidant or a reductant. Thus, it is necessary to know the role of semiquinone, and anionic hydroquinone states. By comparing wild- the Ade moiety in initial photochemistry of FAD in cryptochrome type and mutant enzymes, we have determined that the excited to understand the mechanism of cryptochrome signaling.
    [Show full text]
  • Redalyc.Characterization of OAZ1 and Its Potential Functions in Goose
    Electronic Journal of Biotechnology E-ISSN: 0717-3458 [email protected] Pontificia Universidad Católica de Valparaíso Chile Kang, Bo; Jiang, Dongmei; He, Hui; Ma, Rong; Yi, Zhixin; Chen, Ziyu Characterization of OAZ1 and its potential functions in goose follicular development Electronic Journal of Biotechnology, vol. 26, 2017, pp. 1-6 Pontificia Universidad Católica de Valparaíso Valparaíso, Chile Available in: http://www.redalyc.org/articulo.oa?id=173350071001 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Electronic Journal of Biotechnology 26 (2017) 1–6 Contents lists available at ScienceDirect Electronic Journal of Biotechnology Research Article Characterization of OAZ1 and its potential functions in goose follicular development Bo Kang a,b,⁎,1, Dongmei Jiang a,b,⁎,1,HuiHea,RongMaa, Zhixin Yi a,ZiyuChena a College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, Sichuan 611130, People's Republic of China b Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, Sichuan 611130, People's Republic of China article info abstract Article history: Background: Ornithine decarboxylase antizyme 1 (OAZ1) is an important regulator of polyamine synthesis and Received 6 October 2016 uptake. Our previous studies indicated that high OAZ1 expression in the ovaries of laying geese is responsible Accepted 2 December 2016 for poor egg production. In the present study, the molecular characterization of goose OAZ1 gene was Available online 19 December 2016 analyzed, as well as the expression profile in various follicular tissues.
    [Show full text]