Bioethanol Production

Roberto Ruller

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Research Biography

Ruller develops research for CTBE since September of 2009, working in the study, development and Biotechnological Applications for feasibility of microorganisms and enzymes on the industrial context. For the development of enzymes, he has used Microbiology, Molecular Biology, Biochemistry and Biophysics techniques for Protein Engineering, highlighting a vast experience using Directed Evolution of enzymes, SMD (site directed mutagenesis), Chimeragenesis (creation of multifunctional enzymes) and production of recombinant enzymes. He uses bioprospecting techniques for studies of microorganism and enzymes from extremophilic environments and protocols of adaptive evolution to improve bacteria, yeast and fungi for better fermentation of sugars (pentoses and hexoses) from pretreated plant biomasses.


2007 – 2009

MBA (Master Business Administration).
University of São Paulo (USP) – Foundation for Research and Development of Administration, Accounting and Economics (Fundace) – Ribeirão Preto-SP, Brazil.

2008 – 2009

Post-doctorate in Protein Biochemistry.
University of São Paulo (USP) – Medicine College of Ribeirão Preto (FMRP) – Ribeirão Preto-SP, Brazil.

2001 – 2006

Ph.D. in Cellular and Molecular Biology.
University of São Paulo (USP) – Medicine College of Ribeirão Preto (FMRP) – Ribeirão Preto-SP, Brazil.

1999 – 2001

M.Sc. in Biochemistry.
University of São Paulo (USP) – Medicine College of Ribeirão Preto (FMRP) – Ribeirão Preto-SP, Brazil.

1995 – 1999

B.Sc. in Biology.
Paulista State University (Unesp) – São Paulo-SP, Brazil.

Areas of Expertise
  1. Bioprospecting and Biochemical Studies of extremophile enzymes (extremozymes)
    Prospecting of microorganism enzymes from extremophile environments for studies and biotechnological applications;
  2. Development for enzymes to customization of enzymatic cocktails
    Application of Directed Evolution and chimeragenisis of proteins for development of enzyme cocktails for lignocellulose hydrolysis;
  3. Adaptive Engineering of microorganism
    Accelerated evolution (directed evolution) of microorganism for biotechnological applications in the biofuels production and value-added bio-products.
Selected Publications and Patents

Selected Publications

  • Zanphorlin LM, de Giuseppe PO, Honorato RV, Tonoli CC, Fattori J, Crespim E, de Oliveira PS, Ruller R, Murakami MT. Oligomerization as a strategy for cold adaptation: Structure and dynamics of the GH1 β-glucosidase from Exiguobacterium antarcticum B7. Sci Rep. 2016 Mar 31;6:23776.
  • Santos CA, Zanphorlin LM, Crucello A, Tonoli CC, Ruller R, Horta MA, Murakami MT, de Souza AP. Crystal structure and biochemical characterization of the recombinant ThBgl, a GH1 β-glucosidase overexpressed in Trichoderma harzianum under biomass degradation conditions. Biotechnol Biofuels. 2016 Mar 22;9:71.
  • Hoffmam ZB, Zanphorlin LM, Cota J, Diogo JA, Almeida GB, Damásio AR, Squina F, Murakami MT, Ruller R. Xylan-specific carbohydrate-binding module belonging to family 6 enhances the catalytic performance of a GH11 endo-xylanase. N.Biotechnol. 2016 Feb 23. pii: S1871-6784(16)00019-4.
  • de Lima EA, Machado CB, Zanphorlin LM, Ward RJ, Sato HH, Ruller R. GH53 Endo-Beta-1,4-Galactanase from a Newly Isolated Bacillus licheniformis CBMAI 1609 as an Enzymatic Cocktail Supplement for Biomass Saccharification. Appl Biochem Biotechnol. 2016 Feb 15.
  • Crespim E, Zanphorlin LM, de Souza FH, Diogo JA, Gazolla AC, Machado CB, Figueiredo F, Sousa AS, Nóbrega F, Pellizari VH, Murakami MT, Ruller R. A novel cold-adapted and glucose-tolerant GH1 β-glucosidase from Exiguobacterium antarcticum B7. Int J Biol Macromol. 2016 Jan;82:375-80.
  • Diogo JA, Zanphorlin LM, Sato HH, Murakami MT, Ruller R. Molecular cloning, overexpression, purification and crystallographic analysis of a GH43 β-xylosidase from Bacillus licheniformis. Acta Crystallogr F Struct Biol Commun. 2015 Aug;71(Pt 8):962-
  • Diogo JA, Hoffmam ZB, Zanphorlin LM, Cota J, Machado CB, Wolf LD, Squina F, Damásio AR, Murakami MT, Ruller R. Development of a chimeric hemicellulase to enhance the xylose production and thermotolerance. Enzyme Microb Technol. 2015 Feb;69:31-7.
  • Santos CR, Hoffmam ZB, de Matos Martins VP, Zanphorlin LM, de Paula Assis LH, Honorato RV, Lopes de Oliveira PS, Ruller R, Murakami MT. Molecular mechanisms associated with xylan degradation by Xanthomonas plant pathogens. J Biol Chem. 2014 Nov 14;289(46):32186-200.
  • Ruller R, Alponti J, Deliberto LA, Zanphorlin LM, Machado CB, Ward RJ. Concommitant adaptation of a GH11 xylanase by directed evolution to create an alkali-tolerant/thermophilic enzyme. Protein Eng Des Sel. 2014 Aug;27(8):255-62.
  • Ruller R, Deliberto L, Ferreira TL, Ward RJ. Thermostable variants of the recombinant xylanase A from Bacillus subtilis produced by directed evolution show reduced heat capacity changes. Proteins. 2008 Mar;70(4):1280-93. PubMed PMID: 17876824.
  • Goldbeck R, Damásio AR, Gonçalves TA, Machado CB, Paixão DA, Wolf LD, Mandelli F, Rocha GJ, Ruller R, Squina FM. Development of hemicellulolytic enzyme mixtures for plant biomass deconstruction on target biotechnological applications. Appl. Microbiol Biotechnol. 2014 Oct;98(20):8513-25.
  • Delabona Pda S, Cota J, Hoffmam ZB, Paixão DA, Farinas CS, Cairo JP, Lima DJ,  Squina FM, Ruller R, Pradella JG. Understanding the cellulolytic system of Trichoderma harzianum P49P11 and enhancing saccharification of pretreated sugarcane bagasse by supplementation with pectinase and α-L-arabinofuranosidase.  Bioresour Technol. 2013 Mar;131:500-7.


Selected Patents

  • PRADELLA, J. G. ; PRADELLA, J. G. C. ; RULLER, R. ; DELABONA, P. ; SQUINA F. M. ; DILLON, A. ; PIMENTA, M. T. B.. PROCESSO DE PRODUÇÃO DE ENZIMAS COM FUNGO FILAMENTOSO PENICILLIUM ECHINULATUN PARA USO NA HIDRÓLISE ENZIMÁTICA DE BIOMASSA LIGNOCELULOSICA. 2012, Brasil. Patent: Privilégio de Inovação. Registration number: BR1020120268728, Registration institution: INPI (Instituto Nacional da Propriedade Industrial)
  • RULLER, R. ; DIOGO, JOSÉ A. ; Hoffmam, Zaira ; Cota, Junio ; Squina, Fabio Marcio ; PRADELLA, J. G. C. . ENZIMA BIFUNCIONAL UTILIZADA NA DEGRADAÇÃO DE BIOMASSA NA PRODUÇÃO DE XILOSE EM UMA ÚNICA OPERAÇÃO. 2013, Brasil. Patent: Privilégio de Inovação. Registration number: BR1020130180513, data de depósito: 15/07/2013, Registration institution: INPI  (Instituto Nacional da Propriedade Industrial).
  • PRADELLA, J. G. C. ; PRADELLA, J. G. C. ; Ienczak J.L. ; RABELO, S. ; RULLER, R. . PROCESSO DE PRODUÇÃO DE SUPORTE INERTE A PARTIR DE MATERIAL LIGNOCELULÓSICO, IMOBILIZAÇÃO DE MICRO-ORGANISMOS EM DITO SUPORTE EM UM BIORREATOR DE LEITO EMPACOTADO E SEUS USOS. 2013, Brasil. Patent: Privilégio de Inovação. Registration number: BR102013019252, deposit date: 29/07/2013, Registration instituition: INPI (Instituto Nacional da Propriedade Industrial).
  • RULLER, R. ; HOFFMAM, ZAIRA B. ; Lima E A ; Sato H H . Composição de meio de cultura, processo de obtenção de um coquetel enzimático para hidrólise de polissacarídeos e seus usos. 2013, Brasil. Patent: Privilégio de Inovação. Registration number: BR1020130280682, data de depósito: 31/10/2013, Registration instituiton: INPI – Instituto Nacional da Propriedade Industrial.
  • PRADELLA, J.G.C. ; DELABONA, PRISCILA DA SILVA ; Hoffmam, Zaira ; SQUINA F. M. ; CAIRO, JOÃO PAULO LOURENÇO FRANCO ; RULLER, R. . COQUETEL ENZIMÁTICO DE Trichoderma harzianum SUPLEMENTADO COM ENZIMAS AUXILIARES, PROCESSO DE OBTENÇÃO E SEUS USOS. 2013, Brasil. Patent: Privilégio de Inovação. Registration number: BR1020130299472, deposit date: 21/11/2013, Registration institution: INPI (Instituto Nacional da Propriedade Industrial)