DOCSLIB.ORG

In Vitro and in Vivo Exploration of the Cellobiose and Cellodextrin Phosphorylases Panel in Ruminiclostridium Cellulolyticum: Im

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
In Vitro and in Vivo Exploration of the Cellobiose and Cellodextrin Phosphorylases Panel in Ruminiclostridium Cellulolyticum: Im Liu et al. Biotechnol Biofuels (2019) 12:208 https://doi.org/10.1186/s13068-019-1549-x Biotechnology for Biofuels RESEARCH Open Access In vitro and in vivo exploration of the cellobiose and cellodextrin phosphorylases panel in Ruminiclostridium cellulolyticum: implication for cellulose catabolism Nian Liu1, Aurélie Fosses1, Clara Kampik1, Goetz Parsiegla2, Yann Denis3, Nicolas Vita1, Henri‑Pierre Fierobe1 and Stéphanie Perret1* Abstract Background: In anaerobic cellulolytic micro‑organisms, cellulolysis results in the action of several cellulases gathered in extracellular multi‑enzyme complexes called cellulosomes. Their action releases cellobiose and longer cellodex‑ trins which are imported and further degraded in the cytosol to fuel the cells. In Ruminiclostridium cellulolyticum, an anaerobic and cellulolytic mesophilic bacteria, three cellodextrin phosphorylases named CdpA, CdpB, and CdpC, were identifed in addition to the cellobiose phosphorylase (CbpA) previously characterized. The present study aimed at characterizing them, exploring their implication during growth on cellulose to better understand the life‑style of cellulolytic bacteria on such substrate. Results: The three cellodextrin phosphorylases from R. cellulolyticum displayed marked diferent enzymatic char‑ acteristics. They are specifc for cellodextrins of diferent lengths and present diferent kcat values. CdpC is the most active enzyme before CdpA, and CdpB is weakly active. Modeling studies revealed that a mutation of a conserved histidine residue in the phosphate ion‑binding pocket in CdpB and CdpC might explain their activity‑level diferences. The genes encoding these enzymes are scattered over the chromosome of R. cellulolyticum and only the expression of the gene encoding the cellobiose phosphorylase and the gene cdpA is induced during cellulose growth. Char‑ acterization of four independent mutants constructed in R. cellulolyticum for each of the cellobiose and cellodextrin phosphorylases encoding genes indicated that only the cellobiose phosphorylase is essential for growth on cellulose. Conclusions: Unexpectedly, the cellobiose phosphorylase but not the cellodextrin phosphorylases is essential for the growth of the model bacterium on cellulose. This suggests that the bacterium adopts a “short” dextrin strategy to grow on cellulose, even though the use of long cellodextrins might be more energy‑saving. Our results suggest marked diferences in the cellulose catabolism developed among cellulolytic bacteria, which is a result that might impact the design of future engineered strains for biomass‑to‑biofuel conversion. Keywords: Cellobiose, Cellodextrins, Phosphorylase, Cellulolysis, Cellulose *Correspondence: [email protected] 1 Aix‑Marseille Univ, CNRS, LCB UMR 7283, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France Full list of author information is available at the end of the article © The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creat iveco mmons .org/licen ses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/ publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Liu et al. Biotechnol Biofuels (2019) 12:208 Page 2 of 14 Background located upstream of cuaABC-cbpA. In our previous Cellulose is the most abundant polysaccharide produced study, we constructed an R. cellulolyticum cuaD mutant on Earth and is constituted of linear chains of β-1,4- strain (MTLcuaD) in which a type II intron inactivates linked glucose units. It represents a large reservoir of cuaD. Te modifcation induced a polar efect on the glucose and an attractive renewable energy source. Nev- expression of the downstream genes cuaS (sensor) and ertheless, glucose molecules are scarcely available from cuaR (regulator), thus preventing the upregulation of cellulose because of the tight crystalline packing of the the expression of cuaABC-cbpA operon encoding the cellulosic chains which makes this material recalcitrant cellodextrins ABC transporter and the cellobiose phos- to enzymatic degradation. Its biological deconstruction phorylase A. In consequence, the MTLcuaD strain is, therefore, a limiting step in the carbon cycle on Earth was unable to grow on either cellobiose or cellulose. and also a bottleneck in the process of biofuel or bio- Te transformation of the strain with a vector contain- based chemicals production [1]. ing the ABC transporter genes but not the cellobiose Nonetheless, several anaerobic bacteria are able to phosphorylase-encoding gene cbpA restored growth on use this recalcitrant substrate as the sole carbon and cellulose but not on cellobiose. Tis observation sug- energy source [2]. Among them, Ruminiclostridium gests that cellodextrins of degree of polymerization cellulolyticum, a mesophilic, anaerobic model bacte- (DP) greater than 2 might be imported in the cytosol, rium raises special interest for years due to its ability thus ensuring growth on cellulose of this strain. Simi- to efciently degrade and use plant cell wall polysac- larly, another cellulolytic strain (Hungatei) Clostridium charides including cellulose and hemicellulose, and the thermocellum was reported to assimilate long cellodex- availability of genetic tools [3–8]. To achieve the enzy- trins of 5 and 6 glucose residues when grown on cel- matic degradation of plant cell wall polysaccharides, it lulose [11]. In general, the import of long cellodextrins produces multi-enzyme complexes called cellulosomes is believed to be more cost-efective compared to the by assembling on a scafolding protein diverse enzymes import of short ones, since for the same ATP transport belonging to families of glycoside hydrolase (GH), car- cost, long cellodextrins carry more glucose units and, bohydrate esterase (CE), or polysaccharide lyase (PL) therefore, generate more energy than short ones [11]. [6, 9]. Te released mono- and oligosaccharides are In anaerobic cellulolytic bacteria, the cytosolic degra- subsequently imported by the bacteria and catabolized. dation of cellodextrins is usually ensured by cellobiose/ For example, the uptake of xyloglucan and cellodextrins cellodextrin phosphorylases [12, 13]. Te cellodextrin was shown to be ensured by specifc ABC transporters, phosphorylases catalyze reversible phosphorolysis reac- the imported dextrins being further degraded into sim- tion in which a β-1,4-glycosidic bond of a cellodextrin ple monosaccharides by cytosolic GHs. Two distinct of n glucose units (called Gn with n ≥ 2) is cleaved in clusters of genes dedicated to either the catabolism the presence of inorganic phosphate, releasing one G-1P of xyloglucan or cellodextrins were shown to encode from the non-reducing end and one G n-1 molecule. Te ABC transporter components (including a solute- phosphorylated glucose can directly enter the glyco- binding protein collecting the solute to be imported lysis pathway after conversion into glucose 6-phosphate and two transmembrane domains forming a channel), (G-6P), without consumption of an ATP molecule for intracellular GH(s), and a signal transduction system its phosphorylation, in contrast to the unphosphoryl- [8, 10]. Te ABC transporter called CuaABC (for cel- ated glucoses generated by hydrolysis of cellodextrins. lulose utilization associated) has a solute-binding pro- Tis pathway, therefore, represents an energetically more tein which binds to cellodextrins with lengths ranging advantageous way of degrading oligosaccharides com- from cellobiose (G2) to cellopentaose (G5), suggest- pared to hydrolysis, which is especially benefcial for ing that at least these cellodextrins might be imported anaerobic organisms [12–14]. Te cellobiose phosphory- in the cytosol. CuaABC was shown to be essential for lase A from R. cellulolyticum belongs to the GH94 Fam- growth of the bacterium on both cellobiose and cel- ily. It is specifc for cellobiose as well as other cellobiose lulose growth substrate [10]. We have formerly shown phosphorylases described so far, with the exception of that imported cellobiose is subsequently converted into the cellobiose phosphorylase from Termosipho afri- glucose and α-glucose 1-phosphate (G-1P) by the cello- canus which is active on both cellobiose and long cello- biose phosphorylase A (CbpA) [10]. Te gene encoding dextrins [10, 15–19]. If R. cellulolyticum is able to import this enzyme forms with the genes cuaABC an operon long cellodextrins, enzyme(s) other than the cellobiose named (cuaABC-cbpA). Tis operon is regulated by a phosphorylase A might be implicated in their degrada- predicted three-component system CuaDSR encom- tion. We analyzed the genome of R. cellulolyticum and passing a binding protein CuaD, a sensor (CuaS), and identifed three genes encoding putative cytosolic cel- a regulator (CuaR) encoded by the operon cuaDSR lodextrin phosphorylases belonging to the GH94 family. Liu et al. Biotechnol Biofuels (2019) 12:208 Page 3 of 14 We characterized the three cellodextrin phosphorylases tree was generated based on the amino acid sequence and addressed the question of their role in the cellulose from characterized bacterial GH94 phosphorylases. It catabolism achieved by R. cellulolyticum. encompasses phosphorylases
Load more

Recommended publications
  • Bacteria Belonging to Pseudomonas Typographi Sp. Nov. from the Bark Beetle Ips Typographus Have Genomic Potential to Aid in the Host Ecology
    insects Article Bacteria Belonging to Pseudomonas typographi sp. nov. from the Bark Beetle Ips typographus Have Genomic Potential to Aid in the Host Ecology Ezequiel Peral-Aranega 1,2 , Zaki Saati-Santamaría 1,2 , Miroslav Kolaˇrik 3,4, Raúl Rivas 1,2,5 and Paula García-Fraile 1,2,4,5,* 1 Microbiology and Genetics Department, University of Salamanca, 37007 Salamanca, Spain; [email protected] (E.P.-A.); [email protected] (Z.S.-S.); [email protected] (R.R.) 2 Spanish-Portuguese Institute for Agricultural Research (CIALE), 37185 Salamanca, Spain 3 Department of Botany, Faculty of Science, Charles University, Benátská 2, 128 01 Prague, Czech Republic; [email protected] 4 Laboratory of Fungal Genetics and Metabolism, Institute of Microbiology of the Academy of Sciences of the Czech Republic, 142 20 Prague, Czech Republic 5 Associated Research Unit of Plant-Microorganism Interaction, University of Salamanca-IRNASA-CSIC, 37008 Salamanca, Spain * Correspondence: [email protected] Received: 4 July 2020; Accepted: 1 September 2020; Published: 3 September 2020 Simple Summary: European Bark Beetle (Ips typographus) is a pest that affects dead and weakened spruce trees. Under certain environmental conditions, it has massive outbreaks, resulting in attacks of healthy trees, becoming a forest pest. It has been proposed that the bark beetle’s microbiome plays a key role in the insect’s ecology, providing nutrients, inhibiting pathogens, and degrading tree defense compounds, among other probable traits. During a study of bacterial associates from I. typographus, we isolated three strains identified as Pseudomonas from different beetle life stages. In this work, we aimed to reveal the taxonomic status of these bacterial strains and to sequence and annotate their genomes to mine possible traits related to a role within the bark beetle holobiont.
    [Show full text]
  • Isolation and Characterization of Achromobacter Sp. CX2 From
    Ann Microbiol (2015) 65:1699–1707 DOI 10.1007/s13213-014-1009-6 ORIGINAL ARTICLE Isolation and characterization of Achromobacter sp. CX2 from symbiotic Cytophagales, a non-cellulolytic bacterium showing synergism with cellulolytic microbes by producing β-glucosidase Xiaoyi Chen & Ying Wang & Fan Yang & Yinbo Qu & Xianzhen Li Received: 27 August 2014 /Accepted: 24 November 2014 /Published online: 10 December 2014 # Springer-Verlag Berlin Heidelberg and the University of Milan 2014 Abstract A Gram-negative, obligately aerobic, non- degradation by cellulase (Carpita and Gibeaut 1993). There- cellulolytic bacterium was isolated from the cellulolytic asso- fore, efficient degradation is the result of multiple activities ciation of Cytophagales. It exhibits biochemical properties working synergistically to efficiently solubilize crystalline cel- that are consistent with its classification in the genus lulose (Sánchez et al. 2004;Lietal.2009). Most known Achromobacter. Phylogenetic analysis together with the phe- cellulolytic organisms produce multiple cellulases that act syn- notypic characteristics suggest that the isolate could be a novel ergistically on native cellulose (Wilson 2008)aswellaspro- species of the genus Achromobacter and designated as CX2 (= duce some other proteins that enhance cellulose hydrolysis CGMCC 1.12675=CICC 23807). The strain CX2 is the sym- (Wang et al. 2011a, b). Synergistic cooperation of different biotic microbe of Cytophagales and produces β-glucosidase. enzymes is a prerequisite for the efficient degradation of cellu- The results showed that the non-cellulolytic Achromobacter lose (Jalak et al. 2012). Both Trichoderma reesi and Aspergillus sp. CX2 has synergistic activity with cellulolytic microbes by niger were co-cultured to increase the levels of different enzy- producing β-glucosidase.
    [Show full text]
  • Insights Into the Oxidative Degradation of Cellulose by a Copper Metalloenzyme That Exploits Biomass Components
    Insights into the oxidative degradation of cellulose by a copper metalloenzyme that exploits biomass components R. Jason Quinlana,1, Matt D. Sweeneya,1, Leila Lo Leggiob, Harm Ottenb, Jens-Christian N. Poulsenb, Katja Salomon Johansenc,2, Kristian B. R. M. Kroghc, Christian Isak Jørgensenc, Morten Tovborgc, Annika Anthonsenc, Theodora Tryfonad, Clive P. Walterc, Paul Dupreed, Feng Xua, Gideon J. Daviese, and Paul H. Waltone aNovozymes, Inc., Davis, CA 95618; bDepartment of Chemistry, University of Copenhagen, 2100 Copenhagen Ø, Denmark; cNovozymes A/S, DK-2880 Bagsværd, Denmark; dDepartment of Biochemistry, School of Biological Sciences, University of Cambridge, Cambridge CB2 1QW, United Kingdom; and eDepartment of Chemistry, University of York, Heslington, York YO10 5DD, United Kingdom Edited* by Diter von Wettstein, Washington State University, Pullman, WA, and approved August 2, 2011 (received for review April 13, 2011) The enzymatic degradation of recalcitrant plant biomass is one lysis. From this work, it was suggested that GH61s act directly on of the key industrial challenges of the 21st century. Accordingly, cellulose rendering it more accessible to traditional cellulase there is a continuing drive to discover new routes to promote action (11). Moreover, recent genomic sequencing of the brown polysaccharide degradation. Perhaps the most promising approach rot fungi Postia placenta showed a number of GH61 genes in this involves the application of “cellulase-enhancing factors,” such as organism (13–15), indicating the widespread nature of this those from the glycoside hydrolase (CAZy) GH61 family. Here we family of enzymes in cellulose degradation. As such, GH61s show that GH61 enzymes are a unique family of copper-depen- likely hold major potential for industrial decomposition of dent oxidases.
    [Show full text]
  • Enzymatic Glycosylation of Small Molecules
    View metadata, citation and similar papers at core.ac.uk brought to you by CORE provided by University of Groningen University of Groningen Enzymatic Glycosylation of Small Molecules Desmet, Tom; Soetaert, Wim; Bojarova, Pavla; Kren, Vladimir; Dijkhuizen, Lubbert; Eastwick- Field, Vanessa; Schiller, Alexander; Křen, Vladimir Published in: Chemistry : a European Journal DOI: 10.1002/chem.201103069 IMPORTANT NOTE: You are advised to consult the publisher's version (publisher's PDF) if you wish to cite from it. Please check the document version below. Document Version Publisher's PDF, also known as Version of record Publication date: 2012 Link to publication in University of Groningen/UMCG research database Citation for published version (APA): Desmet, T., Soetaert, W., Bojarova, P., Kren, V., Dijkhuizen, L., Eastwick-Field, V., ... Křen, V. (2012). Enzymatic Glycosylation of Small Molecules: Challenging Substrates Require Tailored Catalysts. Chemistry : a European Journal, 18(35), 10786-10801. https://doi.org/10.1002/chem.201103069 Copyright Other than for strictly personal use, it is not permitted to download or to forward/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 (like Creative Commons). Take-down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Downloaded from the University of Groningen/UMCG research database (Pure): http://www.rug.nl/research/portal. For technical reasons the number of authors shown on this cover page is limited to 10 maximum.
    [Show full text]
  • New Computational Approaches for Investigating the Impact of Mutations on the Transglucosylation Activity of Sucrose Phosphorylase Enzyme
    Thesis submitted to the Université de la Réunion for award of Doctor of Philosophy in Sciences speciality in bioinformatics New computational approaches for investigating the impact of mutations on the transglucosylation activity of sucrose phosphorylase enzyme Mahesh VELUSAMY Thesis to be presented on 18th December 2018 in front of the jury composed of Mme Isabelle ANDRE, Directeur de Recherche CNRS, INSA de TOULOUSE, Rapporteur M. Manuel DAUCHEZ, Professeur, Université de Reims Champagne Ardennes, Rapporteur M. Richard DANIELLOU, Professeur, Université d'Orléans, Examinateur M. Yves-Henri SANEJOUAND, Directeur de Recherche CNRS, Université de Nantes, Examinateur Mme Irène MAFFUCCI, Maître de Conférences, Université de Technologie de Compiègne, Examinateur Mme Corinne MIRAL, Maître de Conférences HDR, Université de Nantes, Examinateur M. Frédéric CADET, Professeur, Université de La Réunion, Co-directeur de thèse M. Bernard OFFMANN, Professeur, Université de Nantes, Directeur de thèse ெ்்ந்ி ிைு்ூுத் ெ்யாம் ெ்த உதி்ு ையகு் ானகு் ஆ்ற் அிு. -ிு்ு் ுதி் அ்ப் ுுக், எனு த்ைத ேுாி, தா் க்ூி, அ்ண் ு்ு்ுமா், த்ைக பா்பா, ஆ்தா ுு்மா், ீனா, அ்ண் ்ுக், ெபிய்பா, ெபிய்மா ம்ு் உுுைணயா் இு்த அைண்ு ந்ப்கு்ு் எனு மனமா்்த ந்ி. இ்த ஆ்ி்ைக ுுைம அை3த்ு ுுுத்்காரண், எனு ஆ்ி்ைக இய்ுன் ேபராிிய் ெப்னா்் ஆஃ்ேம். ப்ேு துண்கி் நா் மனதாு், ெபாுளாதார அளிு் க்3்ி் இு்தேபாு, என்ு இ்ெனாு த்ைதயாகே இு்ு எ்ைன பா்்ு்ெகா்3ா். ுி்பாக, எனு ூ்றா் ஆ்ு இுிி், அ்்ு எ்ளோ தி்ப்3 க3ைமக் ம்ு் ிர்ிைனக் இு்தாு், அைத்ெபாு்பு்தாு, அ் என்ு ெ்த ெபாுளாதார உதி, ப்கைB்கழக பிு ம்ு் இதர ி்ாக ்ப்த்ப்3 உதிகு்ு எ்ன ைகமா்ு ெகாு்தாு் ஈ3ாகாு.
    [Show full text]
  • Taxonomic Binning Approaches and Functional Characteristics of the Microbial Community During the Anaerobic Digestion of Hydrolyzed Corncob
    energies Article Taxonomic Binning Approaches and Functional Characteristics of the Microbial Community during the Anaerobic Digestion of Hydrolyzed Corncob Luz Breton-Deval 1 , Ilse Salinas-Peralta 1 , Jaime Santiago Alarcón Aguirre 2, Belkis Sulbarán-Rangel 2,* and Kelly Joel Gurubel Tun 2,* 1 Catedras Conacyt, Instituto de Biotecnología, Universidad Nacional Autónoma de México, Cuernavaca 62210, Mexico; [email protected] (L.B.-D.); [email protected] (I.S.-P.) 2 Department of Water and Energy, University of Guadalajara Campus Tonalá, Tonalá 45425, Mexico; [email protected] * Correspondence: [email protected] (B.S.-R.); [email protected] (K.J.G.T.); Tel.: +52-33-2000-2300 (K.J.G.T.) Abstract: Maize forms the basis of Mexican food. As a result, approximately six million tons of corncob are produced each year, which represents an environmental issue, as well as a potential feedstock for biogas production. This research aimed to analyze the taxonomic and functional shift in the microbiome of the fermenters using a whole metagenome shotgun approach. Two strategies were used to understand the microbial community at the beginning and the end of anaerobic digestion: (i) phylogenetic analysis to infer the presence and coverage of clade-specific markers to assign taxonomy and (ii) the recovery of the individual genomes from the samples using the binning of the assembled scaffolds. The results showed that anaerobic digestion brought some noticeable changes and the main microbial community was composed of Corynebacterium variable, Desulfovibrio desulfuricans, Vibrio furnissii, Shewanella spp., Actinoplanes spp., Pseudoxanthomonas spp., Saccharomonospora azurea, Citation: Breton-Deval, L.; Agromyces spp., Serinicoccus spp., Cellulomonas spp., Pseudonocardia spp., Rhodococcus rhodochrous, Salinas-Peralta, I.; Alarcón Aguirre, Sphingobacterium spp.
    [Show full text]
  • Protein & Peptide Letters
    696 Send Orders for Reprints to [email protected] Protein & Peptide Letters, 2017, 24, 696-709 REVIEW ARTICLE ISSN: 0929-8665 eISSN: 1875-5305 Impact Factor: 1.068 Glycan Phosphorylases in Multi-Enzyme Synthetic Processes BENTHAM Editor-in-Chief: SCIENCE Ben M. Dunn Giulia Pergolizzia, Sakonwan Kuhaudomlarpa, Eeshan Kalitaa,b and Robert A. Fielda,* aDepartment of Biological Chemistry, John Innes Centre, Norwich Research Park, Norwich NR4 7UH, UK; bDepartment of Molecular Biology and Biotechnology, Tezpur University, Napaam, Tezpur, Assam -784028, India Abstract: Glycoside phosphorylases catalyse the reversible synthesis of glycosidic bonds by glyco- A R T I C L E H I S T O R Y sylation with concomitant release of inorganic phosphate. The equilibrium position of such reac- tions can render them of limited synthetic utility, unless coupled with a secondary enzymatic step Received: January 17, 2017 Revised: May 24, 2017 where the reaction lies heavily in favour of product. This article surveys recent works on the com- Accepted: June 20, 2017 bined use of glycan phosphorylases with other enzymes to achieve synthetically useful processes. DOI: 10.2174/0929866524666170811125109 Keywords: Phosphorylase, disaccharide, α-glucan, cellodextrin, high-value products, biofuel. O O 1. INTRODUCTION + HO OH Glycoside phosphorylases (GPs) are carbohydrate-active GH enzymes (CAZymes) (URL: http://www.cazy.org/) [1] in- H2O O GP volved in the formation/cleavage of glycosidic bond together O O GT O O + HO O + HO with glycosyltransferase (GT) and glycoside hydrolase (GH) O -- NDP OPO3 NDP -- families (Figure 1) [2-6]. GT reactions favour the thermody- HPO4 namically more stable glycoside product [7]; however, these GS R enzymes can be challenging to work with because of their O O + HO current limited availability and relative instability, along R with the expense of sugar nucleotide substrates [7].
    [Show full text]
  • Recent Advances in Enzymatic Synthesis of β-Glucan and Cellulose
    Scotland's Rural College Recent advances in enzymatic synthesis of -glucan and cellulose Bulmer, Gregory S.; de Andrade, Peterson; Field, Robert A.; van Munster, Jolanda M. Published in: Carbohydrate Research DOI: 10.1016/j.carres.2021.108411 Print publication: 01/10/2021 Document Version Publisher's PDF, also known as Version of record Link to publication Citation for pulished version (APA): Bulmer, G. S., de Andrade, P., Field, R. A., & van Munster, J. M. (2021). Recent advances in enzymatic synthesis of -glucan and cellulose. Carbohydrate Research, 508, [108411]. https://doi.org/10.1016/j.carres.2021.108411 General rights Copyright and moral rights for the publications made accessible in the public portal are retained by the authors and/or other copyright owners and it is a condition of accessing publications that users recognise and abide by the legal requirements associated with these rights. • Users may download and print one copy of any publication from the public portal for the purpose of private study or research. • You may not further distribute the material or use it for any profit-making activity or commercial gain • You may freely distribute the URL identifying the publication in the public portal ? Take down policy If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim. Download date: 04. Oct. 2021 Carbohydrate Research 508 (2021) 108411 Contents lists available at ScienceDirect Carbohydrate Research journal homepage: www.elsevier.com/locate/carres Recent advances in enzymatic synthesis of β-glucan and cellulose Gregory S.
    [Show full text]
  • アミロース製造に利用する酵素の開発と改良 Phosphorylase and Muscle Phosphorylase B
    125 J. Appl. Glycosci., 54, 125―131 (2007) !C 2007 The Japanese Society of Applied Glycoscience Proceedings of the Symposium on Amylases and Related Enzymes, 2006 Developing and Engineering Enzymes for Manufacturing Amylose (Received December 5, 2006; Accepted January 5, 2007) Michiyo Yanase,1,＊ Takeshi Takaha1 and Takashi Kuriki1 1 Biochemical Research Laboratory, Ezaki Glico Co., Ltd. (4 ―6 ―5, Utajima, Nishiyodogawa-ku, Osaka 555-8502, Japan) Abstract: Amylose is a functional biomaterial and is expected to be used for various industries. However at present, manufacturing of amylose is not done, since the purification of amylose from starch is very difficult. It has been known that amylose can be produced in vitro by using α-glucan phosphorylases. In order to ob- tain α-glucan phosphorylase suitable for manufacturing amylose, we isolated an α-glucan phosphorylase gene from Thermus aquaticus and expressed it in Escherichia coli. We also obtained thermostable α-glucan phos- phorylase by introducing amino acid replacement onto potato enzyme. α-Glucan phosphorylase is suitable for the synthesis of amylose; the only problem is that it requires an expensive substrate, glucose 1-phosphate. We have avoided this problem by using α-glucan phosphorylase either with sucrose phosphorylase or cellobiose phosphorylase, where inexpensive raw material, sucrose or cellobiose, can be used instead. In these combined enzymatic systems, α-glucan phosphorylase is a key enzyme. This paper summarizes our work on engineering practical α-glucan phosphorylase for industrial processes and its use in the enzymatic synthesis of essentially linear amylose and other glucose polymers. Key words: amylose, glucose polymer, α-glucan phosphorylase, glycogen debranching enzyme, protein engi- neering α-1,4 glucan is the major form of energy reserve from Enzymes for amylose synthesis.
    [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]
  • Downloaded from the Ribosomal Database Project Website [22], and Aligned Using MUSCLE [23]
    Diversity 2013, 5, 627-640; doi:10.3390/d5030627 OPEN ACCESS diversity ISSN 1424-2818 www.mdpi.com/journal/diversity Article Untangling the Genetic Basis of Fibrolytic Specialization by Lachnospiraceae and Ruminococcaceae in Diverse Gut Communities Amy Biddle 1, Lucy Stewart 1, Jeffrey Blanchard 2 and Susan Leschine 3,* 1 Department of Microbiology, University of Massachusetts, Amherst, MA 01003, USA; E-Mails: [email protected] (A.B.); [email protected] (L.S.) 2 Department of Biology, University of Massachusetts, Amherst, MA 01003, USA; E-Mail: [email protected] 3 Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +1-413-545-0673; Fax: +1-413-545-6326. Received: 17 May 2013; in revised form: 21 June 2013 / Accepted: 26 June 2013 / Published: 9 August 2013 Abstract: The Lachnospiraceae and Ruminococcaceae are two of the most abundant families from the order Clostridiales found in the mammalian gut environment, and have been associated with the maintenance of gut health. While they are both diverse groups, they share a common role as active plant degraders. By comparing the genomes of the Lachnospiraceae and Ruminococcaceae with the Clostridiaceae, a more commonly free-living group, we identify key carbohydrate-active enzymes, sugar transport mechanisms, and metabolic pathways that distinguish these two commensal groups as specialists for the degradation of complex plant material. Keywords: Clostridiales; Ruminococcaceae; Lachnospiraceae; carbohydrate-active enzymes; comparative genomics; plant degradation Diversity 2013, 5 628 1. Introduction 1.1. Taxonomic Revision of the Clostridiales Is a Work in Progress Classically, the genus Clostridium was described as comprising spore-forming, non-sulfate reducing obligate anaerobic bacteria with a gram-positive cell wall.
    [Show full text]
  • 55856192.Pdf
    Members of the jury: Prof. dr. ir. Frank Devlieghere (chairman) Prof. dr. ir. Wim Soetaert (promotor) Prof. dr. ir. Erick Vandamme (promotor) Prof. dr. Els Vandamme Prof. dr. Savvas Savvides Dr. Tom Desmet Dr. Henk-Jan Joosten Promotors: Prof. dr. ir. Wim SOETAERT (promotor) Prof. dr. ir. Erick VANDAMME (promotor) Centre of expertise – Industrial Biotechnology and Biocatalysis Department of Biochemical and Microbial Technology Ghent University, Belgium Dean: Prof. dr. ir. Guido Van Huylenbroeck Rector: Prof. dr. Paul Van Cauwenberge The research was conducted at the Centre of expertise - Industrial Biotechnology and Biocatalysis, Department of Biochemical and Microbial Technology, Faculty of Bioscience Engineering, Ghent University (Ghent, Belgium) ir. An CERDOBBEL ENGINEERING THE THERMOSTABILITY OF SUCROSE PHOSPHORYLASE FOR INDUSTRIAL APPLICATIONS Thesis submitted in fulfilment of the requirements for the degree of Doctor (PhD) in Applied Biological Sciences Dutch translation of the title: Engineering van de thermostabiliteit van sucrose phosphorylase voor industriële toepassingen Cover illustration: “Three-dimensional structure of sucrose phosphorylase colored by B-factor.” Printed by University Press, Zelzate To refer to this thesis: Cerdobbel, A. (2011). Engineering the thermostability of sucrose phosphorylase for industrial applications. PhD thesis, Faculty of Bioscience Engineering, Ghent University, Ghent, 200 p. ISBN-number: 978-90-5989-414-3 The author and the promoter give the authorization to consult and to copy parts of this work for personal use only. Every other use is subject to the copyright laws. Permission to reproduce any material contained in this work should be obtained from the author. WOORD VOORAF Er wordt wel eens gezegd dat het woord vooraf het meest gelezen stukje is van een proefschrift.
    [Show full text]