DOCSLIB.ORG

(12) United States Patent (10) Patent No.: US 8,802,393 B2 Kim Et Al

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
(12) United States Patent (10) Patent No.: US 8,802,393 B2 Kim Et Al USOO8802393B2 (12) United States Patent (10) Patent No.: US 8,802,393 B2 Kim et al. (45) Date of Patent: Aug. 12, 2014 (54) ARABINOSE ISOMERASE EXPRESSED FOREIGN PATENT DOCUMENTS FROM CORYNEBACTERIUM GENUS AND KR 99-16118 5, 1999 TAGATOSE MANUFACTURING METHOD BY KR 10-2002-0051835 6, 2002 USING IT KR 10-044.3865 T 2004 KR 10-2006-0068505 6, 2006 (75) Inventors: Seong-bo Kim, Seoul (KR); Young-mi WO O2/052021 T 2002 Lee, Seoul (KR); Seung-won Park, WO WO-2006-065095 * 6, 2006 Gyeonggi-do (KR); Jung-hoon Kim, WO 2008/066280 6, 2008 Seoul (KR); Sang-hoon Song, Incheon OTHER PUBLICATIONS (KR); Kang-pyo Lee, Seoul (KR): Whisstocket al. Quaterly Reviews of Biophysics, 2003, “Prediction Hye-won Kim, Gyeonggi-do (KR): of protein function from protein sequence and structure', 36(3): Hye-jin Choi, Seoul (KR) 307-340. Chica et al. Semi-rational approaches to engineering enzyme activ (73) Assignee: CJ Cheiljedang Corporation, Seoul ity: combining the benefits of directed evolution and rational design, (KR) Curr Opin Biotechnol. Aug. 2005:16(4):378-84. Review.* Witkowski et al. Conversion of a beta-ketoacyl synthase to a malonyl decarboxylase by replacement of the active-site cysteine with (*) Notice: Subject to any disclaimer, the term of this glutamine, Biochemistry. Sep. 7, 1999;38(36): 11643-50.* patent is extended or adjusted under 35 Srivastava et al. Gene expression systems in Corynebacteria, Protein U.S.C. 154(b) by 271 days. Expr Purif. Apr. 2005:40(2):221-9, Review.* Accession No. AAK18729 GI: 19879240 of NCBI database (pub (21) Appl. No.: 12/253.454 lished Apr. 1, 2000). Kim et al. FEMS Microbiology Letters, 2002, 212: 121-126. (22) Filed: Oct. 17, 2008 Branden et al. Introduction to protein structure, Gerald Publishing Inc., New York, p. 247, 1991. (65) Prior Publication Data Witkowski et al., Conversion of a beta-ketoacylsynthase to a malonyl decarboxylase by replacement of the active-site cysteine with US 2009/O246828A1 Oct. 1, 2009 glutamine, Biochemistry, Sep. 7, 1999;38(36): 11643-50. Seffernicket al., Melamine deaminase and atrazine chlorohydrolase: 98 percent identical but functionally different, J Bacteriol. Apr. 2001; Related U.S. Application Data 183 (8): 2405-10. Lee, S.J., et al. Characterization of a Thermoacidophilic L-Arabinose (62) Division of application No. 11/564,934, filed on Nov. Isomerase from Alicyclobacillus acidocaldarius: Role of Lys-269 in 30, 2006, now abandoned. pH Optimum. Office Action in connection with New Zealand application No. (30) Foreign Application Priority Data 576797 dated Jul. 23, 2010, 7 pages. Kim, Pil. “Current studies on biological tagatose production using Nov. 27, 2006 (KR) ........................ 10-2006-01 17795 L-arabinose isomerase: a review and future perspective.' Applied Microbiol Biotechnol (2004) 65: pp. 243-249. (51) Int. Cl. Chinese Office Action issued in Application No. 20078004400.X CI2P2/06 (2006.01) dated Nov. 3, 2010, 4 pages. CI2P 19/00 (2006.01) * cited by examiner CI2P 19/20 (2006.01) CI2N 9/00 (2006.01) Primary Examiner — Iqbal H Chowdhury CI2N 9/92 (2006.01) (74) Attorney, Agent, or Firm — Occhiuti & Rohlicek LLP CI2N L/20 (2006.01) CI2N IS/00 (2006.01) (57) ABSTRACT C7H 2L/04 (2006.01) The present invention relates to a thermophilic arabinose (52) U.S. Cl. isomerase and a method of manufacturing tagatose using the USPC .............. 435/69.1; 435/72:435/94; 435/183; same, and more precisely, a gene encoding arabinose 435/234:435/252.32:435/320.1536/23.2 isomerase originating from the thermophile Thermotoga nea (58) Field of Classification Search politana DSM 5068, a recombinant expression vector con None taining the gene, a method of preparing a food grade thermo See application file for complete search history. philic arabinose isomerase from the recombinant GRAS (Generally Recognized As Safe) strain transformed with the (56) References Cited said expression vector, and a method of preparing tagatose U.S. PATENT DOCUMENTS from galactose using the said enzyme. 2003/012971.0 A1 7/2003 Hansen et al. 3 Claims, 4 Drawing Sheets U.S. Patent Aug. 12, 2014 Sheet 1 of 4 US 8,802,393 B2 Figure 1 Genomic DNA of Tineapolitana DSM5063 TNA PCR (1509bp: SEQ ID NO. 1 and 2) MCS Etely. Enzyme cutting pCJ-1 EcoRVIPst!) kanamycin EggRf U.S. Patent Aug. 12, 2014 Sheet 2 of 4 US 8,802,393 B2 Figure 2 CJ(1)-TNAI (2.5% sucrose) 16 - OO 85,088 - 80 12 - 60 - cS 8 5o R 40 E O 4. - 20 O l --- -- O O 5 O 15 20 25 30 Time (hour) -O-OD600 -O-pH -O-Activity U.S. Patent Aug. 12, 2014 Sheet 3 of 4 US 8,802,393 B2 FIGURE 3 U.S. Patent Aug. 12, 2014 Sheet 4 of 4 US 8,802,393 B2 Figure 4 35O.O E. Corynebacterium 3000 E. coli 2 5 O. O - 2OO. O - 150.0 - 100.0 sists 50.0 O.O – PBS -- 1096 PBS -- 2006 PBS - 30 96 PBS -- 40% Gu Gu Gu Gu Reaction Conditions US 8,802,393 B2 1. 2 ARABINOSE SOMERASE EXPRESSED was 25%, indicating that both thermostability and conversion FROM CORYNEBACTERIUM GENUS AND yield were very low (Korean Patent Application No. TAGATOSE MANUFACTURING METHOD BY 99-16118). Professor Oh, Deok-keun and his colleagues (Se USING IT jong University) Succeeded in heterogeneous expression of 5 arabinose isomerase originating from Geobacillus Stearo This application is a divisional of U.S. application Ser. No. thermophilus in E. coli and based on that they proposed an isomerization procedure at high temperature to convert galac 11/564,934, filed Nov. 30, 2006, which claims priority to tose into tagatose. The present inventors succeeded in cloning Korean Application No. 10-2006-01 17795, filed Nov. 27, a novel arabinose isomerase gene from Thermotoga neapoli 2006. The content of the application is incorporated by ref tana and producing it in E. coli, and further developed a erence in its entirety. 10 method of manufacturing tagatose at hyper-thermophilic TECHNICAL FIELD condition using the enzyme (Korean Patent No. 10-0443865). The production of tagatose using a derived arabinose The present invention relates to an arabinose isomerase isomerase derived from thermophiles still depends on the gene expressed in a Corynebacterium sp. strain and a method 15 method of using a recombinant enzyme mass-expressed in of preparing tagatose using the same, and more particularly, recombinant E. coli or the isomerization of galactose to taga the present invention provides a gene encoding arabinose tose using a host containing the recombinant enzyme. How isomerase originating from Thermotoga neapolitana DSM ever, this biological production oftagatose using recombinant 5068, a recombinant expression vector containing the gene, a E. coli is not appropriate for the production of tagatose as a method of preparing a thermophilic arabinose isomerase by food ingredient. To produce tagatose as a food additive, ara expressing the gene in a food grade GRAS Corynebacterium binose isomerase expressed in a host which is a GRAS micro sp. Strain transfected with the recombinant expression vector, organism appropriate for the mass-production of the same is and a method of preparing tagatose from galactose using the essential. Corynebacterium is an industrial microorganism that has SaC. been used for the production of chemical compounds appli BACKGROUND ART 25 cable to a variety offields including L-lysine, L-threonine and various nucleic acid containing feeds, medical Supplies and With the increasing interest in well-being or a healthy life, foods. The Corynebacterium is a GRAS (Generally Recog tagatose has been proposed as an alternative to Sugar as it has nized AS Safe) strain, which favors gene manipulation and less side effects and Sugar is one of the major factors causing mass production. In addition, this strain is highly stable under various adult diseases. Tagatose is the isomer of galactose and 30 the various process conditions and has a comparatively stable is known to have fructose-like physiochemical properties. cell membrane, compared with other bacteria. Thus, this Tagatose is a natural low-calorie Sugar, and has recently been strain remains stable even under high osmotic pressure gen approved by the FDA in the USA as GRAS (Generally Rec erated by a high concentration of galactose. ognized AS Safe), so it is now allowed to be added as a The present inventors found that thermophilic arabinose Sweetener to foods, beverages, health foods, diet additives, as isomerase originating from the hyperthermophile Thermo etc. toga neapolitana is overexpressed as an active form in a GRAS indicates a Substance that is generally recognized as GRAS Corynebacterium sp. strain, and further completed safe, which is judged by specialized people having enough this invention by developing a novel art to induce tagatose experience and skills through Scientific procedure and exami isomerization from the high concentration of galactose by nation under the indicated conditions and purpose of use. using the expressed recombinant enzymes from Corynebac GRAS is a unique system used only in the USA to evaluate the 40 terium species. safety of foods and food chemical Substances (under certain conditions) but it is recognized world-wide. DISCLOSURE OF THE INVENTION Tagatose is produced by either isomerization, which is a chemical method using a catalyst to produce an isomer, of It is an object of the present invention to provide a method galactose or a biological method using isomerase to convert 45 of preparing GRAS strains expressing arabinose isomerase galactose enzymatically. stably in their cells under the conditions of high temperature One of the biological methods well-known to those in the and high concentration of galactose by taking advantage of art is to convert aldose or aldose derivatives into ketose or mass-production of the recombinant enzyme in the Coryne ketose derivatives by using an enzyme.
Load more

Recommended publications
  • Comparison of Strand-Specific Transcriptomes Of
    Landstorfer et al. BMC Genomics 2014, 15:353 http://www.biomedcentral.com/1471-2164/15/353 RESEARCH ARTICLE Open Access Comparison of strand-specific transcriptomes of enterohemorrhagic Escherichia coli O157:H7 EDL933 (EHEC) under eleven different environmental conditions including radish sprouts and cattle feces Richard Landstorfer1, Svenja Simon2, Steffen Schober3, Daniel Keim2, Siegfried Scherer1 and Klaus Neuhaus1* Abstract Background: Multiple infection sources for enterohemorrhagic Escherichia coli O157:H7 (EHEC) are known, including animal products, fruit and vegetables. The ecology of this pathogen outside its human host is largely unknown and one third of its annotated genes are still hypothetical. To identify genetic determinants expressed under a variety of environmental factors, we applied strand-specific RNA-sequencing, comparing the SOLiD and Illumina systems. Results: Transcriptomes of EHEC were sequenced under 11 different biotic and abiotic conditions: LB medium at pH4, pH7, pH9, or at 15°C; LB with nitrite or trimethoprim-sulfamethoxazole; LB-agar surface, M9 minimal medium, spinach leaf juice, surface of living radish sprouts, and cattle feces. Of 5379 annotated genes in strain EDL933 (genome and plasmid), a surprising minority of only 144 had null sequencing reads under all conditions. We therefore developed a statistical method to distinguish weakly transcribed genes from background transcription. We find that 96% of all genes and 91.5% of the hypothetical genes exhibit a significant transcriptional signal under at least one condition. Comparing SOLiD and Illumina systems, we find a high correlation between both approaches for fold-changes of the induced or repressed genes. The pathogenicity island LEE showed highest transcriptional activity in LB medium, minimal medium, and after treatment with antibiotics.
    [Show full text]
  • Enzymatic Synthesis of L-Fucose from L-Fuculose Using a Fucose Isomerase
    Kim et al. Biotechnol Biofuels (2019) 12:282 https://doi.org/10.1186/s13068-019-1619-0 Biotechnology for Biofuels RESEARCH Open Access Enzymatic synthesis of L-fucose from L-fuculose using a fucose isomerase from Raoultella sp. and the biochemical and structural analyses of the enzyme In Jung Kim1†, Do Hyoung Kim1†, Ki Hyun Nam1,2 and Kyoung Heon Kim1* Abstract Background: L-Fucose is a rare sugar with potential uses in the pharmaceutical, cosmetic, and food industries. The enzymatic approach using L-fucose isomerase, which interconverts L-fucose and L-fuculose, can be an efcient way of producing L-fucose for industrial applications. Here, we performed biochemical and structural analyses of L-fucose isomerase identifed from a novel species of Raoultella (RdFucI). Results: RdFucI exhibited higher enzymatic activity for L-fuculose than for L-fucose, and the rate for the reverse reaction of converting L-fuculose to L-fucose was higher than that for the forward reaction of converting L-fucose to L-fuculose. In the equilibrium mixture, a much higher proportion of L-fucose (~ ninefold) was achieved at 30 °C and pH 7, indicating that the enzyme-catalyzed reaction favors the formation of L-fucose from L-fuculose. When biochemical analysis was conducted using L-fuculose as the substrate, the optimal conditions for RdFucI activity were determined to be 40 °C and pH 10. However, the equilibrium composition was not afected by reaction temperature in the range 2 of 30 to 50 °C. Furthermore, RdFucI was found to be a metalloenzyme requiring Mn + as a cofactor. The comparative crystal structural analysis of RdFucI revealed the distinct conformation of α7–α8 loop of RdFucI.
    [Show full text]
  • Table 6. Putative Genes Involved in the Utilization of Carbohydrates in G
    Table 6. Putative genes involved in the utilization of carbohydrates in G. thermodenitrificans NG80-2 genome Carbohydrates* Enzymes Gene ID Glycerol Glycerol Kinase GT1216 Glycerol-3-phosphate dehydrogenase, aerobic GT2089 NAD(P)H-dependent glycerol-3-phosphate dehydrogenase GT2153 Enolase GT3003 2,3-bisphosphoglycerate-independentphosphoglycerate mutase GT3004 Triosephosphate isomerase GT3005 3-phosphoglycerate kinase GT3006 Glyceraldehyde-3-phosphate dehydrogenase GT3007 Phosphoglycerate mutase GT1326 Pyruvate kinase GT2663 L-Arabinose L-arabinose isomerase GT1795 L-ribulokinase GT1796 L-ribulose 5-phosphate 4-epimerase GT1797 D-Ribose Ribokinase GT3174 Transketolase GT1187 Ribose 5-phosphate epimerase GT3316 D-Xylose Xylose kinase GT1756 Xylose isomerase GT1757 D-Galactose Galactokinase GT2086 Galactose-1-phosphate uridyltransferase GT2084 UDP-glucose 4-epimerase GT2085 Carbohydrates* Enzymes Gene ID D-Fructose 1-phosphofructokinase GT1727 Fructose-1,6-bisphosphate aldolase GT1805 Fructose-1,6-bisphosphate aldolase type II GT3331 Triosephosphate isomerase GT3005 D-Mannose Mannnose-6 phospate isomelase GT3398 6-phospho-1-fructokinase GT2664 D-Mannitol Mannitol-1-phosphate dehydrogenase GT1844 N-Acetylglucosamine N-acetylglucosamine-6-phosphate deacetylase GT2205 N-acetylglucosamine-6-phosphate isomerase GT2204 D-Maltose Alpha-1,4-glucosidase GT0528, GT1643 Sucrose Sucrose phosphorylase GT3215 D-Trehalose Alpha-glucosidase GT1643 Glucose kinase GT2381 Inositol Myo-inositol catabolism protein iolC;5-dehydro-2- GT1807 deoxygluconokinase
    [Show full text]
  • SUPPY Liglucosexlmtdh
    US 20100314248A1 (19) United States (12) Patent Application Publication (10) Pub. No.: US 2010/0314248 A1 Worden et al. (43) Pub. Date: Dec. 16, 2010 (54) RENEWABLE BOELECTRONIC INTERFACE Publication Classification FOR ELECTROBOCATALYTC REACTOR (51) Int. Cl. (76) Inventors: Robert M. Worden, Holt, MI (US); C25B II/06 (2006.01) Brian L. Hassler, Lake Orion, MI C25B II/2 (2006.01) (US); Lawrence T. Drzal, Okemos, GOIN 27/327 (2006.01) MI (US); Ilsoon Lee, Okemo s, MI BSD L/04 (2006.01) (US) C25B 9/00 (2006.01) (52) U.S. Cl. ............... 204/403.14; 204/290.11; 204/400; Correspondence Address: 204/290.07; 427/458; 204/252: 977/734; PRICE HENEVELD COOPER DEWITT & LIT 977/742 TON, LLP 695 KENMOOR, S.E., PO BOX 2567 (57) ABSTRACT GRAND RAPIDS, MI 495.01 (US) An inexpensive, easily renewable bioelectronic device useful for bioreactors, biosensors, and biofuel cells includes an elec (21) Appl. No.: 12/766,169 trically conductive carbon electrode and a bioelectronic inter face bonded to a surface of the electrically conductive carbon (22) Filed: Apr. 23, 2010 electrode, wherein the bioelectronic interface includes cata lytically active material that is electrostatically bound directly Related U.S. Application Data or indirectly to the electrically conductive carbon electrode to (60) Provisional application No. 61/172,337, filed on Apr. facilitate easy removal upon a change in pH, thereby allowing 24, 2009. easy regeneration of the bioelectronic interface. 7\ POWER 1 - SUPPY|- LIGLUCOSEXLMtDH?till pi 6.0 - esses&aaaas-exx-xx-xx-xx-xxxxixax-e- Patent Application Publication Dec. 16, 2010 Sheet 1 of 18 US 2010/0314248 A1 Potential (nV) Patent Application Publication Dec.
    [Show full text]
  • WO 2014/207087 Al 31 December 2014 (31.12.2014) P O P C T
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2014/207087 Al 31 December 2014 (31.12.2014) P O P C T (51) International Patent Classification: Seville (ES). GUTIERREZ GOMEZ, Pablo; Campus C12N 9/00 (2006.01) C12N 9/90 (2006.01) Palmas Altas, Calle Energia Solar 1, E-41014 Seville (ES). C12N 9/02 (2006.01) C12P 5/02 (2006.01) (74) Agent: NEDERLANDSCH OCTROOIBUREAU; J.W. C12N 9/10 (2006.01) C12P 7/76 (2006.01) Frisolaan 13, NL-2517 JS The Hague (NL). C12N 9/12 (2006.01) C12P 7/64 (2006.01) C12N 9/88 (2006.01) C12N 1/1 9 (2006.01) (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, (21) International Application Number: AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, PCT/EP2014/063484 BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DK, DM, (22) International Filing Date: DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, 26 June 2014 (26.06.2014) HN, HR, HU, ID, IL, IN, IR, IS, JP, KE, KG, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, ME, (25) Filing Language: English MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, (26) Publication Language: English OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, (30) Priority Data: TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, 13382241 .1 26 June 2013 (26.06.2013) EP ZW.
    [Show full text]
  • Supplementary Informations SI2. Supplementary Table 1
    Supplementary Informations SI2. Supplementary Table 1. M9, soil, and rhizosphere media composition. LB in Compound Name Exchange Reaction LB in soil LBin M9 rhizosphere H2O EX_cpd00001_e0 -15 -15 -10 O2 EX_cpd00007_e0 -15 -15 -10 Phosphate EX_cpd00009_e0 -15 -15 -10 CO2 EX_cpd00011_e0 -15 -15 0 Ammonia EX_cpd00013_e0 -7.5 -7.5 -10 L-glutamate EX_cpd00023_e0 0 -0.0283302 0 D-glucose EX_cpd00027_e0 -0.61972444 -0.04098397 0 Mn2 EX_cpd00030_e0 -15 -15 -10 Glycine EX_cpd00033_e0 -0.0068175 -0.00693094 0 Zn2 EX_cpd00034_e0 -15 -15 -10 L-alanine EX_cpd00035_e0 -0.02780553 -0.00823049 0 Succinate EX_cpd00036_e0 -0.0056245 -0.12240603 0 L-lysine EX_cpd00039_e0 0 -10 0 L-aspartate EX_cpd00041_e0 0 -0.03205557 0 Sulfate EX_cpd00048_e0 -15 -15 -10 L-arginine EX_cpd00051_e0 -0.0068175 -0.00948672 0 L-serine EX_cpd00054_e0 0 -0.01004986 0 Cu2+ EX_cpd00058_e0 -15 -15 -10 Ca2+ EX_cpd00063_e0 -15 -100 -10 L-ornithine EX_cpd00064_e0 -0.0068175 -0.00831712 0 H+ EX_cpd00067_e0 -15 -15 -10 L-tyrosine EX_cpd00069_e0 -0.0068175 -0.00233919 0 Sucrose EX_cpd00076_e0 0 -0.02049199 0 L-cysteine EX_cpd00084_e0 -0.0068175 0 0 Cl- EX_cpd00099_e0 -15 -15 -10 Glycerol EX_cpd00100_e0 0 0 -10 Biotin EX_cpd00104_e0 -15 -15 0 D-ribose EX_cpd00105_e0 -0.01862144 0 0 L-leucine EX_cpd00107_e0 -0.03596182 -0.00303228 0 D-galactose EX_cpd00108_e0 -0.25290619 -0.18317325 0 L-histidine EX_cpd00119_e0 -0.0068175 -0.00506825 0 L-proline EX_cpd00129_e0 -0.01102953 0 0 L-malate EX_cpd00130_e0 -0.03649016 -0.79413596 0 D-mannose EX_cpd00138_e0 -0.2540567 -0.05436649 0 Co2 EX_cpd00149_e0
    [Show full text]
  • The University of Chicago the Metaproteomic Analysis Of
    THE UNIVERSITY OF CHICAGO THE METAPROTEOMIC ANALYSIS OF ARCTIC SOILS WITH NOVEL BIOINFORMATIC METHODS A DISSERTATION SUBMITTED TO THE FACULTY OF THE DIVISION OF THE PHYSICAL SCIENCES IN CANDIDACY FOR THE DEGREE OF DOCTOR OF PHILOSOPHY DEPARTMENT OF THE GEOPHYSICAL SCIENCES BY SAMUEL MILLER CHICAGO, ILLINOIS DECEMBER 2018 COPYRIGHT © 2018 Samuel Edward Miller All Rights Reserved TABLE OF CONTENTS LIST OF FIGURES ................................................................................................................. vii LIST OF TABLES .....................................................................................................................ix ACKNOWLEDGMENTS ........................................................................................................... x ABSTRACT ..............................................................................................................................xi I. INTRODUCTION ................................................................................................................... 1 I.A. IMPETUS FOR RESEARCH ...........................................................................................1 I.B. PROTEOMICS BACKGROUND ....................................................................................2 I.B.1. TANDEM MASS SPECTROMETRY .......................................................................3 I.B.2. METHODS OF AUTOMATIC SEQUENCE ASSIGNMENT ...................................9 I.B.2.i. DE NOVO SEQUENCING BY PEPNOVO+ ...................................................
    [Show full text]
  • The Secreted L-Arabinose Isomerase Displays Anti-Hyperglycemic Effects
    Rhimi et al. Microb Cell Fact (2015) 14:204 DOI 10.1186/s12934-015-0391-5 Microbial Cell Factories RESEARCH Open Access The secreted L‑arabinose isomerase displays anti‑hyperglycemic effects in mice Moez Rhimi1,2*, Luis G. Bermudez‑Humaran3,4, Yuan Huang1,2, Samira Boudebbouze1,2, Nadia Gaci1,2, Alexandrine Garnier1,2, Jean‑Jacques Gratadoux3,4, Héla Mkaouar1,2, Philippe Langella3,4 and Emmanuelle Maguin1,2* Abstract Background: The L-arabinose isomerase is an intracellular enzyme which converts L-arabinose into L-ribulose in living systems and D-galactose into D-tagatose in industrial processes and at industrial scales. D-tagatose is a natural ketohex‑ ose with potential uses in pharmaceutical and food industries. The D-galactose isomerization reaction is thermody‑ namically equilibrated, and leads to secondary subproducts at high pH. Therefore, an attractive L-arabinose isomerase should be thermoactive and acidotolerant with high catalytic efficiency. While many reports focused on the set out of a low cost process for the industrial production of D-tagatose, these procedures remain costly. When compared to intracellular enzymes, the production of extracellular ones constitutes an interesting strategy to increase the suitabil‑ ity of the biocatalysts. Results: The L-arabinose isomerase (L-AI) from Lactobacillus sakei was expressed in Lactococcus lactis in fusion with the signal peptide of usp45 (SPUsp45). The L-AI protein and activity were detected only in the supernatant of the induced cultures of the recombinant L. lactis demonstrating the secretion in the medium of the intracellular L. sakei L-AI in an active form. Moreover, we showed an improvement in the enzyme secretion using either (1) L.
    [Show full text]
  • O O2 Enzymes Available from Sigma Enzymes Available from Sigma
    COO 2.7.1.15 Ribokinase OXIDOREDUCTASES CONH2 COO 2.7.1.16 Ribulokinase 1.1.1.1 Alcohol dehydrogenase BLOOD GROUP + O O + O O 1.1.1.3 Homoserine dehydrogenase HYALURONIC ACID DERMATAN ALGINATES O-ANTIGENS STARCH GLYCOGEN CH COO N COO 2.7.1.17 Xylulokinase P GLYCOPROTEINS SUBSTANCES 2 OH N + COO 1.1.1.8 Glycerol-3-phosphate dehydrogenase Ribose -O - P - O - P - O- Adenosine(P) Ribose - O - P - O - P - O -Adenosine NICOTINATE 2.7.1.19 Phosphoribulokinase GANGLIOSIDES PEPTIDO- CH OH CH OH N 1 + COO 1.1.1.9 D-Xylulose reductase 2 2 NH .2.1 2.7.1.24 Dephospho-CoA kinase O CHITIN CHONDROITIN PECTIN INULIN CELLULOSE O O NH O O O O Ribose- P 2.4 N N RP 1.1.1.10 l-Xylulose reductase MUCINS GLYCAN 6.3.5.1 2.7.7.18 2.7.1.25 Adenylylsulfate kinase CH2OH HO Indoleacetate Indoxyl + 1.1.1.14 l-Iditol dehydrogenase L O O O Desamino-NAD Nicotinate- Quinolinate- A 2.7.1.28 Triokinase O O 1.1.1.132 HO (Auxin) NAD(P) 6.3.1.5 2.4.2.19 1.1.1.19 Glucuronate reductase CHOH - 2.4.1.68 CH3 OH OH OH nucleotide 2.7.1.30 Glycerol kinase Y - COO nucleotide 2.7.1.31 Glycerate kinase 1.1.1.21 Aldehyde reductase AcNH CHOH COO 6.3.2.7-10 2.4.1.69 O 1.2.3.7 2.4.2.19 R OPPT OH OH + 1.1.1.22 UDPglucose dehydrogenase 2.4.99.7 HO O OPPU HO 2.7.1.32 Choline kinase S CH2OH 6.3.2.13 OH OPPU CH HO CH2CH(NH3)COO HO CH CH NH HO CH2CH2NHCOCH3 CH O CH CH NHCOCH COO 1.1.1.23 Histidinol dehydrogenase OPC 2.4.1.17 3 2.4.1.29 CH CHO 2 2 2 3 2 2 3 O 2.7.1.33 Pantothenate kinase CH3CH NHAC OH OH OH LACTOSE 2 COO 1.1.1.25 Shikimate dehydrogenase A HO HO OPPG CH OH 2.7.1.34 Pantetheine kinase UDP- TDP-Rhamnose 2 NH NH NH NH N M 2.7.1.36 Mevalonate kinase 1.1.1.27 Lactate dehydrogenase HO COO- GDP- 2.4.1.21 O NH NH 4.1.1.28 2.3.1.5 2.1.1.4 1.1.1.29 Glycerate dehydrogenase C UDP-N-Ac-Muramate Iduronate OH 2.4.1.1 2.4.1.11 HO 5-Hydroxy- 5-Hydroxytryptamine N-Acetyl-serotonin N-Acetyl-5-O-methyl-serotonin Quinolinate 2.7.1.39 Homoserine kinase Mannuronate CH3 etc.
    [Show full text]
  • 12) United States Patent (10
    US007635572B2 (12) UnitedO States Patent (10) Patent No.: US 7,635,572 B2 Zhou et al. (45) Date of Patent: Dec. 22, 2009 (54) METHODS FOR CONDUCTING ASSAYS FOR 5,506,121 A 4/1996 Skerra et al. ENZYME ACTIVITY ON PROTEIN 5,510,270 A 4/1996 Fodor et al. MICROARRAYS 5,512,492 A 4/1996 Herron et al. 5,516,635 A 5/1996 Ekins et al. (75) Inventors: Fang X. Zhou, New Haven, CT (US); 5,532,128 A 7/1996 Eggers Barry Schweitzer, Cheshire, CT (US) 5,538,897 A 7/1996 Yates, III et al. s s 5,541,070 A 7/1996 Kauvar (73) Assignee: Life Technologies Corporation, .. S.E. al Carlsbad, CA (US) 5,585,069 A 12/1996 Zanzucchi et al. 5,585,639 A 12/1996 Dorsel et al. (*) Notice: Subject to any disclaimer, the term of this 5,593,838 A 1/1997 Zanzucchi et al. patent is extended or adjusted under 35 5,605,662 A 2f1997 Heller et al. U.S.C. 154(b) by 0 days. 5,620,850 A 4/1997 Bamdad et al. 5,624,711 A 4/1997 Sundberg et al. (21) Appl. No.: 10/865,431 5,627,369 A 5/1997 Vestal et al. 5,629,213 A 5/1997 Kornguth et al. (22) Filed: Jun. 9, 2004 (Continued) (65) Prior Publication Data FOREIGN PATENT DOCUMENTS US 2005/O118665 A1 Jun. 2, 2005 EP 596421 10, 1993 EP 0619321 12/1994 (51) Int. Cl. EP O664452 7, 1995 CI2O 1/50 (2006.01) EP O818467 1, 1998 (52) U.S.
    [Show full text]
  • Genomic, Transcriptomic and Enzymatic Insight Into Lignocellulolytic System of a Plant Pathogen Dickeya Sp
    biomolecules Article Genomic, Transcriptomic and Enzymatic Insight into Lignocellulolytic System of a Plant Pathogen Dickeya sp. WS52 to Digest Sweet Pepper and Tomato Stalk 1, 2, 3, 2 2 Ying-Jie Yang y, Wei Lin y, Raghvendra Pratap Singh * , Qian Xu , Zhihou Chen , Yuan Yuan 1 , Ping Zou 1, Yiqiang Li 1 and Chengsheng Zhang 1,* 1 Marine Agriculture Research Center, Tobacco Research Institute of Chinese Academy of Agricultural Sciences, Qingdao 266101, China; [email protected] (Y.-J.Y.); [email protected] (Y.Y.); [email protected] (P.Z.); [email protected] (Y.L.) 2 Tobacco Research Institute of Nanping, Nanping, Fujian 353000, China; [email protected] (W.L.); [email protected] (Q.X.); [email protected] (Z.C.) 3 Department of Research & Development, Biotechnology, Uttaranchal University, Dehradun 248007, India * Correspondence: [email protected] (R.P.S.); [email protected] (C.Z.) These authors contributed equally to this work. y Received: 31 October 2019; Accepted: 18 November 2019; Published: 20 November 2019 Abstract: Dickeya sp., a plant pathogen, causing soft rot with strong pectin degradation capacity was taken for the comprehensive analysis of its corresponding biomass degradative system, which has not been analyzed yet. Whole genome sequence analysis of the isolated soft-rotten plant pathogen Dickeya sp. WS52, revealed various coding genes which involved in vegetable stalk degradation-related properties. A total of 122 genes were found to be encoded for putative carbohydrate-active enzymes (CAZy) in Dickeya sp. WS52. The number of pectin degradation-related genes, was higher than that of cellulolytic bacteria as well as other Dickeya spp.
    [Show full text]
  • The Chemistry and Evolution of Enzyme Function
    The Chemistry and Evolution of Enzyme Function: Isomerases as a Case Study Sergio Mart´ınez Cuesta EMBL - European Bioinformatics Institute Gonville and Caius College University of Cambridge A thesis submitted for the degree of Doctor of Philosophy 31st July 2014 This dissertation is the result of my own work and contains nothing which is the outcome of work done in collaboration except where specifically indicated in the text. No part of this dissertation has been submitted or is currently being submitted for any other degree or diploma or other qualification. This thesis does not exceed the specified length limit of 60.000 words as defined by the Biology Degree Committee. This thesis has been typeset in 12pt font using LATEX according to the specifications de- fined by the Board of Graduate Studies and the Biology Degree Committee. Cambridge, 31st July 2014 Sergio Mart´ınezCuesta To my parents and my sister Contents Abstract ix Acknowledgements xi List of Figures xiii List of Tables xv List of Publications xvi 1 Introduction 1 1.1 Chemistry of enzymes . .2 1.1.1 Catalytic sites, mechanisms and cofactors . .3 1.1.2 Enzyme classification . .5 1.2 Evolution of enzyme function . .6 1.3 Similarity between enzymes . .8 1.3.1 Comparing sequences and structures . .8 1.3.2 Comparing genomic context . .9 1.3.3 Comparing biochemical reactions and mechanisms . 10 1.4 Isomerases . 12 1.4.1 Metabolism . 13 1.4.2 Genome . 14 1.4.3 EC classification . 15 1.4.4 Applications . 18 1.5 Structure of the thesis . 20 2 Data Resources and Methods 21 2.1 Introduction .
    [Show full text]