DOCSLIB.ORG

Universidade Federal Do Amazonas Pró

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text

Load more

UNIVERSIDADE FEDERAL DO AMAZONAS PRÓ-REITORIA DE PESQUISA E PÓS-GRADUAÇÃO INSTITUTO DE CIÊNCIAS BIOLÓGICAS PROGRAMA MULTI-INSTITUCIONAL DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA - PPGBIOTEC CARACTERIZAÇÃO DO POTENCIAL DE DEGRADAÇÃO DE UM ISOLADO BACTERIANO ORIUNDO DA REGIÃO AMAZÔNICA DIOGO PEREIRA DE CASTRO MANAUS - AMAZONAS 2015 UNIVERSIDADE FEDERAL DO AMAZONAS PRÓ-REITORIA DE PESQUISA E PÓS-GRADUAÇÃO INSTITUTO DE CIÊNCIAS BIOLÓGICAS PROGRAMA MULTI-INSTITUCIONAL DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA - PPGBIOTEC DIOGO PEREIRA DE CASTRO CARACTERIZAÇÃO DO POTENCIAL DE DEGRADAÇÃO DE UM ISOLADO BACTERIANO ORIUNDO DA REGIÃO AMAZÔNICA Tese apresentada ao Programa Multi- Institucional de Pós-Graduação em Biotecnologia da Universidade Federal do Amazonas como parte do pré-requisito para obtenção do título de Doutor em Biotecnologia na área de concentração “Agroflorestal”. Orientadora: Dra. Patrícia Puccinelli Orlandi Co-Orientador: Dr. José Odair Pereira MANAUS - AMAZONAS 2015 ii Ficha Catalográfica Ficha catalográfica elaborada automaticamente de acordo com os dados fornecidos pelo(a) autor(a). Castro, Diogo Pereira de C355c Caracterização do potencial de degradação de um isolado bacteriano oriundo da região Amazônica / Diogo Pereira de Castro. 2015 154 f.: il. color; 31 cm. Orientador: Patrícia Puccinelli Orlandi Coorientador: José Odair Pereira Tese (Doutorado em Biotecnologia) - Universidade Federal do Amazonas. 1. pETRÓLEO. 2. Biorremediação. 3. Degradação de Hidrocarbonetos. 4. Proteômica. I. Orlandi, Patrícia Puccinelli II. Universidade Federal do Amazonas III. Título DIOGO PEREIRA DE CASTRO CARACTERIZAÇÃO DO POTENCIAL DE DEGRADAÇÃO DE UM ISOLADO BACTERIANO ORIUNDO DA REGIÃO AMAZÔNICA Tese apresentada ao Programa Multi- Institucional de Pós-Graduação em Biotecnologia da Universidade Federal do Amazonas como parte do pré-requisito para obtenção do título de Doutor APROVADA EM:______/______/______ em Biotecnologia na área de concentração “Agroflorestal”. BANCA EXAMINADORA _______________________________________________ Profa. Dra. Patrícia Puccinelli Orlandi, Presidente Centro de Pesquisas Leônidas e Maria Deane - FIOCRUZ Amazônia _______________________________________________ Profa. Dra. Ieda Hortêncio Batista Universidade do Estado do Amazonas - UEA _______________________________________________ Profa. Dra. Ana Frazão Teixeira Universidade do Estado do Amazonas - UEA _______________________________________________ Prof. Dr. Donat Alexander De Chapeaurouge Fundação Oswaldo Cruz – FIOCRUZ/IOC _______________________________________________ Prof. Dr. Jorge Luis López-Lozano Fundação Oswaldo Cruz – FIOCRUZ MANAUS – AMAZONAS 2015 iii “A minha esposa Paula Taquita, a meus familiares, a meus amigos e a todos que me ajudaram nessa jornada”. Dedico iv AGRADECIMENTOS Agradeço a Deus pela coragem e força para estar nessa jornada; Minha esposa Paula Taquita pela dedicação, companheirismo, entre outras qualidades - que são inúmeras - sem ela não conseguiria chegar até onde cheguei; A meus pais e minha sogra pelo apoio nessa etapa de vida; A minha orientadora Dra. Patrícia Orlandi pela orientação, paciência e amizade; Ao meu co-Orientador Dr. José Odair Pereira pela orientação e apoio no desenvolvimento da tese; Aos professores Dr. Paulo Afonso Nogueira, Dra. Priscila Aquino, Dra. Tainá Raiol Alencar pelas orientações e apoio cientifico; Ao Dr. Jefferson Rocha pela orientação e colaboração; Ao Dr. Fabrício Klerynon Marchini pelo apoio científico e colaboração; Ao Programa de Pós Graduação em Biotecnologia (PPGBIOTEC) pela oportunidade de estudo; A Sra. Nubiane pela amizade e apoio pedagógico; A todos os amigos do Laboratório da FIOCRUZ-ILMD pelo apoio científico, momentos alegres e ajuda nos experimentos; A Fundação Oswaldo Cruz (FIOCRUZ) ILMD pelo espaço para desenvolvimento do nosso estudo; A FAPEAM e CnPQ agradeço pelo apoio financeiro fornecido no decorrer deste projeto. v “Devemos ter fé. Não existem esforços inúteis se empregados em prol do bem comum”. Getúlio Vargas vi RESUMO A biorremediação é a utilização de microrganismos capazes de metabolizar contaminantes e transforma-los em produtos menos tóxicos. Em estudos anteriores, foi realizado o isolamento de bactérias provenientes de área contaminada por resíduos de petróleo na Amazônia. Apesar de possuírem o potencial de degradação de petróleo, a aplicação de bactérias em ambientes contaminados pode interferir em seu equilíbrio ecológico. Este estudo propôs-se a seleção de uma linhagem bacteriana para promover estudos de caracterização gênica, identificação do potencial de degradação de hidrocarbonetos e definição das proteínas envolvidas no processo de degradação. Para alcançar os objetivos, foram identificadas substâncias degradadas por cromatografia gasosa e proteínas envolvidas por proteômica; A análise do genoma permitiu a caracterização dos genes relacionados a degradação de compostos xenobióticos através da revisão da literatura. A cepa foi identificada como Pseudomonas putida S16. Foram detectadas 51 substâncias por cromatografia gasosa, destas, 23 foram completamente degradadas, incluindo HPAs contendo naftaleno (100%), antraceno (11,49%) e fenantreno (7,41%). Foram identificados 89 genes relacionados a degradação de xenobíoticos no genoma da P. putida S16 AM, genes capazes de degradar naftaleno, antraceno e fenantreno (nah, phn, fdx, catA), produção de citocromo e quimiotaxia. Na análise proteômica, foram expressas proteínas de óxido-redução, processos metabólicos, poliaminas, aldeído desidrogenase, armazenamento de carbono, quimiotaxia e síntese de aminoácidos. Foram identificadas 355 proteínas com redundância expressas durante o contato de 7h da P. putida S16 AM (I4) com o petróleo, e 268 no contato de 75h; Para o glicerol, foram identificadas 467 na interação de 7h, e 228 no contato de 75h. A presença de proteínas relacionadas ao metabolismo e óxido- redução quando a cepa em estudo esteve em contato com o petróleo evidenciam a utilização deste como fonte de carbono para o metabolismo bacteriano. Estes estudos são importantes devido à identificação de genes e proteínas que possuem o potencial de se tornarem produtos biotecnológicos de aplicação prática. Palavras-chave: Petróleo; Biorremediação; Degradação de Hidrocarbonetos; Proteômica. vii ABSTRACT Bioremediation is the use of microorganisms capable of metabolizing contaminants and converts them into less toxic products. In previous studies, we performed bacteria isolation from contaminated area for oil waste in Amazon. Despite its oil degradation potential, the application of bacteria in contaminated sites may interfere with their ecological balance. This study aimed to promote studies of gene characterization and hydrocarbon degradation potential with a selected bacterial strain and identify and define the proteins involved in degradation process. To achieve this objectives degraded substances have been identified by gas chromatography and proteins involved by proteomics; Genome analysis allowed characterization of genes related to xenobiotics degradation. The strain was identified as Pseudomonas putida S16. 51 substances were detected by gas chromatography, of these, 23 were completely degraded, including PAH containing naphthalene (100%), anthracene (11.49%) and phenanthrene (7.41%). We identified 89 genes related to xenobiotic degradation in P. putida S16 AM genome, genes capable of degrading naphthalene, anthracene and phenanthrene (nah, phn, fdx, catA), cytochrome production and chemotaxis. In proteome analysis, proteins were expressed oxidation-reduction, metabolic processes, polyamines, aldehyde dehydrogenase, carbon storage, chemotaxis and amino acid synthesis. 355 proteins were identified with redundancy expressed during 7h contact to P. putida S16 AM (I4) with petroleum, and 268 during 75h contact; For glycerol, 467 were identified in 7h interaction, and 228 in 75h contact. The presence of proteins related to metabolism and oxidation- reduction when the strain under study was in contact to the petroleum suggest the use of this as a carbon source for bacterial metabolism. These studies are important because of identification of genes and proteins with potential to become practical application of biotechnology products. Keywords : Petroleum; Bioremediation; Degradation of Hydrocarbons; Proteomics. viii LISTA DE FIGURAS Figura 1. Biodegradação de um n-alcano.... ............................................................................ 22 Figura 2. Degradação inicial do benzeno por oxigenases como exemplo de biodegradação de substâncias aromáticas. ............................................................................................................. 22 Figura 3. Degradação do catecol pelas vias de orto e meta clivagem. .................................... 23 Figura 4. Degradação de Naftaleno. ........................................................................................ 26 Figura 5. Rota de degradação do fenantreno. .......................................................................... 27 Figura 6. Degradação de antraceno proposto para bactérias aeróbicas ................................... 28 Figura 7. Ion Torrent. Reação de incorporação de uma base pela polimerase. ....................... 31 Figura 8. Esquema do processo de Cromatografia Gasosa acoplada a Espectro de Massa .... 32 Figura 9. Esquema de um espectrômetro de massas ............................................................... 33 Figura 10. Micro gotas expelidas pelo cone de Taylor ..........................................................
Recommended publications
  • Purification, Crystallization and Preliminary X-Ray Diffraction Analysis of Exodeoxyribonuclease III from Crenarchaeon Sulfolobus Tokodaii Strain 7

    Purification, Crystallization and Preliminary X-Ray Diffraction Analysis of Exodeoxyribonuclease III from Crenarchaeon Sulfolobus Tokodaii Strain 7

    Crystal Structure Theory and Applications, 2013, 2, 155-158 Published Online December 2013 (http://www.scirp.org/journal/csta) http://dx.doi.org/10.4236/csta.2013.24021 Purification, Crystallization and Preliminary X-Ray Diffraction Analysis of Exodeoxyribonuclease III from Crenarchaeon Sulfolobus tokodaii Strain 7 Shuichi Miyamoto1*, Chieko Naoe2, Masaru Tsunoda3, Kazuo T. Nakamura2 1Faculty of Pharmaceutical Sciences, Sojo University, Kumamoto, Japan 2School of Pharmacy, Showa University, Tokyo, Japan 3Faculty of Pharmacy, Iwaki Meisei University, Iwaki, Japan Email: *[email protected] Received October 13, 2013; revised November 12, 2013; accepted December 6, 2013 Copyright © 2013 Shuichi Miyamoto et al. This is an open access article distributed under the Creative Commons Attribution Li- cense, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. ABSTRACT Exodeoxyribonuclease III (EXOIII) acts as a 3’→5’ exonuclease and is homologous to purinic/apyrimidinic (AP) en- donuclease (APE), which plays an important role in the base excision repair pathway. To structurally investigate the reaction and substrate recognition mechanisms of EXOIII, a crystallographic study of EXOIII from Sulfolobus tokodaii strain 7 was carried out. The purified enzyme was crystallized by using the hanging-drop vapor-diffusion method. The crystals belonged to space group C2, with unit-cell parameters a = 154.2, b = 47.7, c = 92.4 Å, β = 125.8˚ and diffracted to 1.5 Å resolution. Keywords: Crenarchaeon; Crystallization; Exodeoxyribonuclease; Sulfolobus tokodaii; X-Ray Diffraction 1. Introduction formational change upon protein binding that permits complex formation and activation of attacking water, A variety of mechanisms exist to repair damaged DNA leading to incision, in the presence of Mg2+ [10,11].
  • Separation and Partial Purification of Collagenolytic Protease

    Separation and Partial Purification of Collagenolytic Protease

    Biocatalysis and Agricultural Biotechnology 24 (2020) 101509 Contents lists available at ScienceDirect Biocatalysis and Agricultural Biotechnology journal homepage: http://www.elsevier.com/locate/bab Separation and partial purificationof collagenolytic protease from peacock bass (Cichla ocellaris) using different protocol: Precipitation and partitioning approaches Vagne de Melo Oliveira a, Marcia� Nieves Carneiro da Cunha a, Caio Rodrigo Dias de Assis b,f, Juanize Matias da Silva Batista a, Thiago Pajeú Nascimento a, Juliana Ferreira dos Santos c, Carolina de Albuquerque Lima d, Daniela de Araújo Viana Marques e, Ranilson de Souza Bezerra b, Ana Lúcia Figueiredo Porto a,* a Laboratorio� de Tecnologia de Produtos Bioativos, Departamento de Morfologia e Fisiologia Animal, DMFA, Universidade Federal Rural de Pernambuco, Av. Dom Manoel, de Medeiros, s/n, 52171-900, Recife, PE, Brazil b Laboratorio� de Enzimologia, Departamento de Bioquímica, Universidade Federal de Pernambuco, Cidade Universitaria,� 50670-420, Recife, PE, Brazil c Laboratorio� de Aquicultura e Biotecnologia, LABITEC, Unidade Acad^emica de Serra Talhada, Universidade Federal Rural de Pernambuco, Serra Talhada, PE, Brazil d Universidade de Pernambuco, UPE, Rua Capitao~ Pedro Rodrigues, 105, 55294-902, Garanhuns, PE, Brazil e Universidade de Pernambuco, UPE, Instituto de Ci^encias Biologicas� - ICB, Departmento de Parasitologia, Rua Arnobio� Marques, 310 - Santo Amaro, Recife, PE, 50100- 130, Brazil f Laboratorio� de Compostos Organicos^ em Ecossistemas Costeiros e Marinhos, OrganoMAR, Departamento de Oceanografia,Universidade Federal de Pernambuco, Cidade Universitaria,� 50670-420, Recife, PE, Brazil ARTICLE INFO ABSTRACT Keywords: This comparative study provides a useful protocol for the separation and partial purification of collagenolytic ATPS proteases obtained from peacock bass, by using precipitation and partitioning.
  • The Rnase H-Like Superfamily: New Members, Comparative Structural Analysis and Evolutionary Classification Karolina A

    The Rnase H-Like Superfamily: New Members, Comparative Structural Analysis and Evolutionary Classification Karolina A

    4160–4179 Nucleic Acids Research, 2014, Vol. 42, No. 7 Published online 23 January 2014 doi:10.1093/nar/gkt1414 The RNase H-like superfamily: new members, comparative structural analysis and evolutionary classification Karolina A. Majorek1,2,3,y, Stanislaw Dunin-Horkawicz1,y, Kamil Steczkiewicz4, Anna Muszewska4,5, Marcin Nowotny6, Krzysztof Ginalski4 and Janusz M. Bujnicki1,3,* 1Laboratory of Bioinformatics and Protein Engineering, International Institute of Molecular and Cell Biology, ul. Ks. Trojdena 4, PL-02-109 Warsaw, Poland, 2Department of Molecular Physiology and Biological Physics, University of Virginia, 1340 Jefferson Park Avenue, Charlottesville, VA USA-22908, USA, 3Bioinformatics Laboratory, Institute of Molecular Biology and Biotechnology, Adam Mickiewicz University, Umultowska 89, PL-61-614 Poznan, Poland, 4Laboratory of Bioinformatics and Systems Biology, Centre of New Technologies, University of Warsaw, Zwirki i Wigury 93, PL-02-089 Warsaw, Poland, 5Institute of Biochemistry and Biophysics PAS, Pawinskiego 5A, PL-02-106 Warsaw, Poland and 6Laboratory of Protein Structure, International Institute of Molecular and Cell Biology, ul. Ks. Trojdena 4, PL-02-109 Warsaw, Poland Received September 23, 2013; Revised December 12, 2013; Accepted December 26, 2013 ABSTRACT revealed a correlation between the orientation of Ribonuclease H-like (RNHL) superfamily, also called the C-terminal helix with the exonuclease/endo- the retroviral integrase superfamily, groups together nuclease function and the architecture of the numerous enzymes involved in nucleic acid metab- active site. Our analysis provides a comprehensive olism and implicated in many biological processes, picture of sequence-structure-function relation- including replication, homologous recombination, ships in the RNHL superfamily that may guide func- DNA repair, transposition and RNA interference.
  • CYP2A6) by P53

    CYP2A6) by P53

    Transcriptional Regulation of Human Stress Responsive Cytochrome P450 2A6 (CYP2A6) by p53 Hao Hu M.Biotech. (Biotechnology) 2012 The University of Queensland B.B.A. 2009 University of Electronic Science and Technology of China B.Sc. (Pharmacy) 2009 University of Electronic Science and Technology of China A thesis submitted for the degree of Doctor of Philosophy at The University of Queensland in 2016 School of Medicine ABSTRACT Human cytochrome P450 (CYP) 2A6 is highly expressed in the liver and the encoding gene is regulated by various stress activated transcription factors, such as the nuclear factor (erythroid-derived 2)-like 2 (Nrf-2). Unlike the other xenobiotic metabolising CYP enzymes (XMEs), CYP2A6 only plays a minor role in xenobiotic metabolism. The CYP2A6 is highly induced by multiple forms of cellular stress conditions, where XMEs expression is normally inhibited. Recent findings suggest that the CYP2A6 plays an important role in regulating BR homeostasis. A computer based sequence analysis on the 3 kb proximate CYP2A6 promoter revealed several putative binding sites for p53, a protein that mediates regulation of antioxidant and apoptosis pathways. In this study, the role of p53 in CYP2A6 gene regulation is demonstrated. The site closest to transcription start site (TSS) is highly homologous with the p53 consensus sequence. The p53 responsiveness of this site was confirmed by transfections with various stepwise deleted of CYP2A6-5’-Luc constructs containing the putative p53RE. Deletion of the putative p53RE resulted in a total abolishment of p53 responsiveness of CYP2A6 promoter. Specific binding of p53 to the putative p53RE was detected by electrophoresis mobility shift assay.
  • Characterisation of Bilirubin Metabolic Pathway in Hepatic Mitochondria Siti Nur Fadzilah Muhsain M.Sc

    Characterisation of Bilirubin Metabolic Pathway in Hepatic Mitochondria Siti Nur Fadzilah Muhsain M.Sc

    Characterisation of Bilirubin Metabolic Pathway in Hepatic Mitochondria Siti Nur Fadzilah Muhsain M.Sc. (Medical Research) 2005 Universiti Sains Malaysia Postgrad. Dip. (Toxicology) 2003 University of Surrey B.Sc.(Biomed. Sc.) 2000 Universiti Putra Malaysia A thesis submitted for the degree of Doctor of Philosophy at The University of Queensland in 2014 School of Medicine ABSTRACT Bilirubin (BR), a toxic waste product of degraded haem, is a potent antioxidant at physiological concentrations. To achieve the maximum benefit of BR, its intracellular level needs to be carefully regulated. A system comprising of two enzymes, haem oxygenase-1 (HMOX1) and cytochrome P450 2A5 (CYP2A5) exists in the endoplasmic reticulum (ER), responsible for regulating BR homeostasis. This system is induced in response to oxidative stress. In this thesis, oxidative stress caused accumulation of these enzymes in mitochondria — major producers and targets of reactive oxygen species (ROS) — is demonstrated. To understand the significance of this intracellular targeting, properties of microsomal and mitochondrial BR metabolising enzymes were compared and the capacity of mitochondrial CYP2A5 to oxidise BR in response to oxidative stress is reported. Microsomes and mitochondrial fractions were isolated from liver homogenates of DBA/2J mice, administered with sub-toxic dose of pyrazole, an oxidant stressor. The purity of extracted organelles was determined by analysing the expressions and activities of their respective marker enzymes. HMOX1 and CYP2A5 were significantly increased in microsomes and even more so in mitochondria in response to pyrazole-induced oxidative stress. By contrast, the treatment did not increase either microsomes or mitochondrial Uridine-diphosphate-glucuronosyltransferase 1A1 (UGT1A1), the sole enzyme that catalyses BR elimination through glucuronidation.
  • Dissertation

    Dissertation

    Dissertation Submitted to the Combined Faculties for the Natural Sciences and for Mathematics of the Ruperto-Carola University of Heidelberg, Germany for the Degree of Doctor of Natural Sciences Presented by Ann-Cathrin Hofer (M.Sc.) Born in Heidelberg, Germany Oral Examination: 12th of September 2016 Regulatory T cells protect the neonatal liver and secure the hepatic circadian rhythm Referees 1st Referee: Prof. Dr. Peter Angel 2nd Referee: Dr. Markus Feuerer This dissertation was performed and written during the period from November 2012 to May 2016 in the German Cancer Research Center (DKFZ) under the supervision of Prof. Dr. Peter Angel and direct supervision of Dr. Markus Feuerer. The dissertation was submitted to the Combined Faculties for the Natural Sciences and for Mathematics of the Ruperto-Carola University of Heidelberg, Germany in June 2016. German Cancer Research Center (DKFZ) Immune Tolerance (D100) Im Neuenheimer Feld 280 69120 Heidelberg, Germany I II Confirmation Hereby, I confirm that I have written this thesis independently, using only the results of my investigation unless otherwise stated. Furthermore, I declare that I have not submitted this thesis for a degree to any other academic or similar institution. Parts of this dissertation have been submitted for publishing: Regulatory T cells protect the liver early in life and safeguard the hepatic circadian rhythm Ann-Cathrin Hofer, Thomas Hielscher, David M. Richards, Michael Delacher, Ulrike Träger, Sophia Föhr, Artyom Vlasov, Marvin Wäsch, Marieke Esser, Annette Kopp-Schneider, Achim Breiling, Frank Lyko, Ursula Klingmüller, Peter Angel, Jakub Abramson, Jeroen Krijgsveld & Markus Feuerer Parts of the experiments in this dissertation were performed in collaboration with other research groups as follows: CG methylation analysis with the 454 pyrosequencing technology: Division of Epigenetics, DKFZ, Heidelberg Dr.
  • Transcriptome Analysis of Differential Gene Expression in Dichomitus Squalens

    Transcriptome Analysis of Differential Gene Expression in Dichomitus Squalens

    bioRxiv preprint doi: https://doi.org/10.1101/359646; this version posted June 30, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. 1 Transcriptome analysis of differential gene expression in Dichomitus squalens 2 during interspecific mycelial interactions and the potential link with laccase 3 induction 4 Zixuan Zhong1, Nannan Li1, Binghui He, Yasuo Igarashi, Feng Luo* 5 6 Research Center of Bioenergy and Bioremediation, College of Resources and Environment, 7 Southwest University, Beibei, Chongqing 400715, People’s Republic of China 8 9 *corresponding author: Feng Luo (FL) [email protected] 10 11 1These authors contributed equally to the paper 12 13 ABSTRACT 14 Interspecific mycelial interactions between white rot fungi are always accompanied by increased 15 production of laccase. In this study, the potential of white rot fungi Dichomitus squalens for 16 enhancing laccase production during interaction with two other white rot fungi Trametes 17 versicolor or Pleurotus ostreatus was identified. To probe the mechanism of laccase induction and 18 the role of laccase played during the combative interaction, we analyzed the laccase induction 19 response to stressful conditions during fungal interaction related to the differential gene expression 20 profile. We further confirmed the expression patterns of 16 selected genes by qRT-PCR analysis. 21 We noted that many differential expression genes (DEGs) encoding proteins were involved in 22 xenobiotics detoxification and ROS generation or reduction, including aldo/keto reductase, 23 glutathione S-transferases, cytochrome P450 enzymes, alcohol oxidases and dehydrogenase, 24 manganese peroxidase and laccase.
  • Supplementary Materials

    Supplementary Materials

    Supplementary Materials COMPARATIVE ANALYSIS OF THE TRANSCRIPTOME, PROTEOME AND miRNA PROFILE OF KUPFFER CELLS AND MONOCYTES Andrey Elchaninov1,3*, Anastasiya Lokhonina1,3, Maria Nikitina2, Polina Vishnyakova1,3, Andrey Makarov1, Irina Arutyunyan1, Anastasiya Poltavets1, Evgeniya Kananykhina2, Sergey Kovalchuk4, Evgeny Karpulevich5,6, Galina Bolshakova2, Gennady Sukhikh1, Timur Fatkhudinov2,3 1 Laboratory of Regenerative Medicine, National Medical Research Center for Obstetrics, Gynecology and Perinatology Named after Academician V.I. Kulakov of Ministry of Healthcare of Russian Federation, Moscow, Russia 2 Laboratory of Growth and Development, Scientific Research Institute of Human Morphology, Moscow, Russia 3 Histology Department, Medical Institute, Peoples' Friendship University of Russia, Moscow, Russia 4 Laboratory of Bioinformatic methods for Combinatorial Chemistry and Biology, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow, Russia 5 Information Systems Department, Ivannikov Institute for System Programming of the Russian Academy of Sciences, Moscow, Russia 6 Genome Engineering Laboratory, Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, Russia Figure S1. Flow cytometry analysis of unsorted blood sample. Representative forward, side scattering and histogram are shown. The proportions of negative cells were determined in relation to the isotype controls. The percentages of positive cells are indicated. The blue curve corresponds to the isotype control. Figure S2. Flow cytometry analysis of unsorted liver stromal cells. Representative forward, side scattering and histogram are shown. The proportions of negative cells were determined in relation to the isotype controls. The percentages of positive cells are indicated. The blue curve corresponds to the isotype control. Figure S3. MiRNAs expression analysis in monocytes and Kupffer cells. Full-length of heatmaps are presented.
  • (51) International Patent Classification: A61K 8/66 (2006.01) A61Q 11/00

    (51) International Patent Classification: A61K 8/66 (2006.01) A61Q 11/00

    ( (51) International Patent Classification: A61K 8/66 (2006.01) A61Q 11/00 (2006.01) (21) International Application Number: PCT/EP20 19/08 1186 (22) International Filing Date: 13 November 2019 (13. 11.2019) (25) Filing Language: English (26) Publication Language: English (30) Priority Data: 18206133.3 14 November 2018 (14. 11.2018) EP (71) Applicant: NOVOZYMES A/S [DK/DK]; Krogshoejvej 36, 2880 Bagsvaerd (DK). (72) Inventors: DURHUUS, Thomas, Thomasen; Krogshoe¬ jvej 36, 2880 Bagsvaerd (DK). PALMEN, Lorena, Gonzalez,; Krogshoejvej 36, 2880 Bagsvaerd (DK). REISER, Anna, Verena,; Krogshoejvej 36, 2880 Bagsvaerd (DK). STREICHER, Werner, W,; Krogshoe¬ jvej 36, 2880 Bagsvaerd (DK). (81) Designated States (unless otherwise indicated, for every kind of national protection available) : AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, 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, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available) : ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, Cl, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG).
  • Letters to Nature

    Letters to Nature

    letters to nature Received 7 July; accepted 21 September 1998. 26. Tronrud, D. E. Conjugate-direction minimization: an improved method for the re®nement of macromolecules. Acta Crystallogr. A 48, 912±916 (1992). 1. Dalbey, R. E., Lively, M. O., Bron, S. & van Dijl, J. M. The chemistry and enzymology of the type 1 27. Wolfe, P. B., Wickner, W. & Goodman, J. M. Sequence of the leader peptidase gene of Escherichia coli signal peptidases. Protein Sci. 6, 1129±1138 (1997). and the orientation of leader peptidase in the bacterial envelope. J. Biol. Chem. 258, 12073±12080 2. Kuo, D. W. et al. Escherichia coli leader peptidase: production of an active form lacking a requirement (1983). for detergent and development of peptide substrates. Arch. Biochem. Biophys. 303, 274±280 (1993). 28. Kraulis, P.G. Molscript: a program to produce both detailed and schematic plots of protein structures. 3. Tschantz, W. R. et al. Characterization of a soluble, catalytically active form of Escherichia coli leader J. Appl. Crystallogr. 24, 946±950 (1991). peptidase: requirement of detergent or phospholipid for optimal activity. Biochemistry 34, 3935±3941 29. Nicholls, A., Sharp, K. A. & Honig, B. Protein folding and association: insights from the interfacial and (1995). the thermodynamic properties of hydrocarbons. Proteins Struct. Funct. Genet. 11, 281±296 (1991). 4. Allsop, A. E. et al.inAnti-Infectives, Recent Advances in Chemistry and Structure-Activity Relationships 30. Meritt, E. A. & Bacon, D. J. Raster3D: photorealistic molecular graphics. Methods Enzymol. 277, 505± (eds Bently, P. H. & O'Hanlon, P. J.) 61±72 (R. Soc. Chem., Cambridge, 1997).
  • The Microbiota-Produced N-Formyl Peptide Fmlf Promotes Obesity-Induced Glucose

    The Microbiota-Produced N-Formyl Peptide Fmlf Promotes Obesity-Induced Glucose

    Page 1 of 230 Diabetes Title: The microbiota-produced N-formyl peptide fMLF promotes obesity-induced glucose intolerance Joshua Wollam1, Matthew Riopel1, Yong-Jiang Xu1,2, Andrew M. F. Johnson1, Jachelle M. Ofrecio1, Wei Ying1, Dalila El Ouarrat1, Luisa S. Chan3, Andrew W. Han3, Nadir A. Mahmood3, Caitlin N. Ryan3, Yun Sok Lee1, Jeramie D. Watrous1,2, Mahendra D. Chordia4, Dongfeng Pan4, Mohit Jain1,2, Jerrold M. Olefsky1 * Affiliations: 1 Division of Endocrinology & Metabolism, Department of Medicine, University of California, San Diego, La Jolla, California, USA. 2 Department of Pharmacology, University of California, San Diego, La Jolla, California, USA. 3 Second Genome, Inc., South San Francisco, California, USA. 4 Department of Radiology and Medical Imaging, University of Virginia, Charlottesville, VA, USA. * Correspondence to: 858-534-2230, [email protected] Word Count: 4749 Figures: 6 Supplemental Figures: 11 Supplemental Tables: 5 1 Diabetes Publish Ahead of Print, published online April 22, 2019 Diabetes Page 2 of 230 ABSTRACT The composition of the gastrointestinal (GI) microbiota and associated metabolites changes dramatically with diet and the development of obesity. Although many correlations have been described, specific mechanistic links between these changes and glucose homeostasis remain to be defined. Here we show that blood and intestinal levels of the microbiota-produced N-formyl peptide, formyl-methionyl-leucyl-phenylalanine (fMLF), are elevated in high fat diet (HFD)- induced obese mice. Genetic or pharmacological inhibition of the N-formyl peptide receptor Fpr1 leads to increased insulin levels and improved glucose tolerance, dependent upon glucagon- like peptide-1 (GLP-1). Obese Fpr1-knockout (Fpr1-KO) mice also display an altered microbiome, exemplifying the dynamic relationship between host metabolism and microbiota.
  • Solarbio Catalogue with PRICES

    Solarbio Catalogue with PRICES

    CAS Name Grade Purity Biochemical Reagent Biochemical Reagent 75621-03-3 C8390-1 3-((3-Cholamidopropyl)dimethylammonium)-1-propanesulfonateCHAPS Ultra Pure Grade 1g 75621-03-3 C8390-5 3-((3-Cholamidopropyl)dimethylammonium)-1-propanesulfonateCHAPS 5g 57-09-0 C8440-25 Cetyl-trimethyl Ammonium Bromide CTAB High Pure Grade ≥99.0% 25g 57-09-0 C8440-100 Cetyl-trimethyl Ammonium Bromide CTAB High Pure Grade ≥99.0% 100g 57-09-0 C8440-500 Cetyl-trimethyl Ammonium Bromide CTAB High Pure Grade ≥99.0% 500g E1170-100 0.5M EDTA (PH8.0) 100ml E1170-500 0.5M EDTA (PH8.0) 500ml 6381-92-6 E8030-100 EDTA disodium salt dihydrate EDTA Na2 Biotechnology Grade ≥99.0% 100g 6381-92-6 E8030-500 EDTA disodium salt dihydrate EDTA Na2 Biotechnology Grade ≥99.0% 500g 6381-92-6 E8030-1000 EDTA disodium salt dihydrate EDTA Na2 Biotechnology Grade ≥99.0% 1kg 6381-92-6 E8030-5000 EDTA disodium salt dihydrate EDTA Na2 Biotechnology Grade ≥99.0% 5kg 60-00-4 E8040-100 Ethylenediaminetetraacetic acid EDTA Ultra Pure Grade ≥99.5% 100g 60-00-4 E8040-500 Ethylenediaminetetraacetic acid EDTA Ultra Pure Grade ≥99.5% 500g 60-00-4 E8040-1000 Ethylenediaminetetraacetic acid EDTA Ultra Pure Grade ≥99.5% 1kg 67-42-5 E8050-5 Ethylene glycol-bis(2-aminoethylether)-N,N,NEGTA′,N′-tetraacetic acid Ultra Pure Grade ≥97.0% 5g 67-42-5 E8050-10 Ethylene glycol-bis(2-aminoethylether)-N,N,NEGTA′,N′-tetraacetic acid Ultra Pure Grade ≥97.0% 10g 50-01-1 G8070-100 Guanidine Hydrochloride Guanidine HCl ≥98.0%(AT) 100g 50-01-1 G8070-500 Guanidine Hydrochloride Guanidine HCl ≥98.0%(AT) 500g 56-81-5