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Targeted analysis of microbial lignocellulolytic secretomes: a new approach to enzyme discovery

Grant number: 10/52362-5
Support type:Regular Research Grants
Duration: November 01, 2011 - August 31, 2015
Field of knowledge:Biological Sciences - Biochemistry
Cooperation agreement: BBSRC, UKRI
Principal Investigator:Igor Polikarpov
Grantee:Igor Polikarpov
Principal investigator abroad: Neil Bruce
Institution abroad: University of York, England
Home Institution: Instituto de Física de São Carlos (IFSC). Universidade de São Paulo (USP). São Carlos , SP, Brazil
Associated scholarship(s):13/19236-4 - Targeted analysis of microbial lignocellulolytic secretomes: a new approach to enzyme discovery, BP.TT
13/05842-0 - Targeted analysis of microbial lignocellulolytic secretomes: a new approach to enzyme discovery, BP.TT
12/19674-9 - Targeted analysis of microbial lignocellulolytic secretomes: a new approach to enzyme discovery, BP.TT

Abstract

From both a fundamental and industrial biotech viewpoint understanding the deconstruction of lignocellulose in soil and compost is of central importance. In the natural environments microbial communities can efficiently degrade or modify lignin to enable the effective enzymatic hydrolysis of the polysaccharides present in plant cell walls. The aim of this proposal is to use metatranscriptomics and proteomics to determine gene - and protein - centred details to determine new mechanisms and improved methods of lignocellulose deconstruction in mixed microbial communities from composting wheat straw and sugar cane bagasse. Secreted proteins will be tagged, affinity purified and analyzed by LC-ESI-MS. In order to have a picture of the overall community dynamics in terms of species composition during the composting process DNA will be extracted for SSU rRNA profiling: Saccharification of the lignocellulose will be monitored and the lignin content of the straw or bagasse analyzed using FTIR spectroscopy and solid state NMR. Metatranscriptome analysis will be performed by preparing cDNA from samples taken at various time points from the Iignocellulose enriched cultures, the cDNA will be sequenced using the Roche 454 GS FLX Titanium platform. The peptide sequences from the proteomics analysis will allow the identification of full and partial coding sequences in the library. These coding sequences will be cloned and expressed in established recombinant expression systems and the recombinant proteins screened for activity. (AU)

Articles published in Agência FAPESP about the research grant
Perfecting enzyme cocktails is the path to cellulosic ethanol 

Scientific publications (18)
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
PUHL, ANA C.; PRATES, ERICA T.; ROSSETO, FLAVIO R.; MANZINE, LIVIA R.; STANKOVIC, IVANA; DE ARAUJO, SIMARA S.; ALVAREZ, THABATA M.; SQUINA, FABIO M.; SKAF, MUNIR S.; POLIKARPOV, IGOR. Crystallographic structure and molecular dynamics simulations of the major endoglucanase from Xanthomonas campestris pv. campestris shed light on its oligosaccharide products release pattern. International Journal of Biological Macromolecules, v. 136, p. 493-502, SEP 1 2019. Web of Science Citations: 0.
ALESSI, ANNA M.; BIRD, SUSANNAH M.; OATES, NICOLA C.; LI, YI; DOWLE, ADAM A.; NOVOTNY, ETELVINO H.; DEAZEVEDO, EDUARDO R.; BENNETT, JOSEPH P.; POLIKARPOV, IGOR; YOUNG, J. PETER W.; MCQUEEN-MASON, SIMON J.; BRUCE, NEIL C. Defining functional diversity for lignocellulose degradation in a microbial community using multi-omics studies. BIOTECHNOLOGY FOR BIOFUELS, v. 11, JUN 18 2018. Web of Science Citations: 4.
DE ARAUJO, EVANDRO ARES; MANZINE, LIVIA REGINA; PIIADOV, VASSILI; SEIKI KADOWAKI, MARCO ANTONIO; POLIKARPOV, IGOR. Biochemical characterization, low-resolution SAXS structure and an enzymatic cleavage pattern of BlCel48 from Bacillus licheniformis. International Journal of Biological Macromolecules, v. 111, p. 302-310, MAY 2018. Web of Science Citations: 1.
EVANGELISTA, DANILO ELTON; SEIKI KADOWAKI, MARCO ANTONIO; MELLO, BRUNO LUAN; POLIKARPOV, IGOR. Biochemical and biophysical characterization of novel GH10 xylanase prospected from a sugar cane bagasse compost-derived microbial consortia. International Journal of Biological Macromolecules, v. 109, p. 560-568, APR 2018. Web of Science Citations: 3.
SANTO, MELISSA ESPIRITO; REZENDE, CAMILA A.; BERNARDINELLI, OIGRES D.; PEREIRA, JR., NEI; CURVELO, ANTONIO A. S.; DEAZEVEDO, EDUARDO R.; GUIMARAES, FRANSCISCO E. G.; POLIKARPOV, IGOR. Structural and compositional changes in sugarcane bagasse subjected to hydrothermal and organosolv pretreatments and their impacts on enzymatic hydrolysis. INDUSTRIAL CROPS AND PRODUCTS, v. 113, p. 64-74, MAR 2018. Web of Science Citations: 8.
GODOY, ANDRE S.; PEREIRA, CAROLINE S.; RAMIA, MARINA PAGLIONE; SILVEIRA, RODRIGO L.; CAMILO, CESAR M.; KADOWAKI, MARCO A.; LANGE, LENE; BUSK, PETER K.; NASCIMENTO, ALESSANDRO S.; SKAF, MUNIR S.; POLIKARPOV, IGOR. Structure, computational and biochemical analysis of PcCel45A endoglucanase from Phanerochaete chrysosporium and catalytic mechanisms of GH45 subfamily C members. SCIENTIFIC REPORTS, v. 8, FEB 27 2018. Web of Science Citations: 2.
MELLO, BRUNO L.; ALESSI, ANNA M.; RIANO-PACHON, DIEGO M.; DEAZEVEDO, EDUARDO R.; GUIMARAES, FRANCISCO E. G.; ESPIRITO SANTO, MELISSA C.; MCQUEEN-MASON, SIMON; BRUCE, NEIL C.; POLIKARPOV, IGOR. Targeted metatranscriptomics of compost-derived consortia reveals a GH11 exerting an unusual exo-1,4-beta-xylanase activity. BIOTECHNOLOGY FOR BIOFUELS, v. 10, NOV 2 2017. Web of Science Citations: 6.
ALESSI, ANNA M.; BIRD, SUSANNAH M.; BENNETT, JOSEPH P.; OATES, NICOLA C.; LI, YI; DOWLE, ADAM A.; POLIKARPOV, IGOR; YOUNG, J. PETER W.; MCQUEEN-MASON, SIMON J.; BRUCE, NEIL C. Revealing the insoluble metasecretome of lignocellulose-degrading microbial communities. SCIENTIFIC REPORTS, v. 7, MAY 24 2017. Web of Science Citations: 8.
MELLO, BRUNO L.; ALESSI, ANNA M.; MCQUEEN-MASON, SIMON; BRUCE, NEIL C.; POLIKARPOV, IGOR. Nutrient availability shapes the microbial community structure in sugarcane bagasse compost-derived consortia. SCIENTIFIC REPORTS, v. 6, DEC 12 2016. Web of Science Citations: 11.
TOMAZINI JUNIOR, ATILIO; DOLCE, LUCIANO GRACIANI; DE OLIVEIRA NETO, MARIO; POLIKARPOV, IGOR. Xanthomonas campestris expansin-like X domain is a structurally disordered beta-sheet macromolecule capable of synergistically enhancing enzymatic efficiency of cellulose hydrolysis. Biotechnology Letters, v. 37, n. 12, p. 2419-2426, DEC 2015. Web of Science Citations: 4.
BERNARDINELLI, OIGRES DANIEL; LIMA, MARISA APARECIDA; REZENDE, CAMILA ALVES; POLIKARPOV, IGOR; DEAZEVEDO, EDUARDO RIBEIRO. Quantitative C-13 MultiCP solid-state NMR as a tool for evaluation of cellulose crystallinity index measured directly inside sugarcane biomass. BIOTECHNOLOGY FOR BIOFUELS, v. 8, AUG 5 2015. Web of Science Citations: 38.
BERNARDES, AMANDA; TEXTOR, LARISSA C.; SANTOS, JADEMILSON C.; CUADRADO, NAZARET HIDALGO; KOSTETSKY, EDUARD YA.; ROIG, MANUEL G.; BAVRO, VASSILIY N.; MUNIZ, JOAO R. C.; SHNYROV, VALERY L.; POLIKARPOV, IGOR. Crystal structure analysis of peroxidase from the palm tree Chamaerops excelsa. Biochimie, v. 111, p. 58-69, APR 2015. Web of Science Citations: 8.
MIOTTO, LIS SCHWARTZ; DE REZENDE, CAMILA ALVES; BERNARDES, AMANDA; SERPA, VIVIANE ISABEL; TSANG, ADRIAN; POLIKARPOV, IGOR. The Characterization of the Endoglucanase Cel12A from Gloeophyllum trabeum Reveals an Enzyme Highly Active on beta-Glucan. PLoS One, v. 9, n. 9 SEP 24 2014. Web of Science Citations: 13.
MELLO, BRUNO LUAN; POLIKARPOV, IGOR. Family 1 carbohydrate binding-modules enhance saccharification rates. AMB EXPRESS, v. 4, APR 25 2014. Web of Science Citations: 17.
LIMA, MARISA A.; GOMEZ, LEONARDO D.; STEELE-KING, CLARE G.; SIMISTER, RACHAEL; BERNARDINELLI, OIGRES D.; CARVALHO, MARCELO A.; REZENDE, CAMILA A.; LABATE, CARLOS A.; DEAZEVEDO, EDUARDO R.; MCQUEEN-MASON, SIMON J.; POLIKARPOV, IGOR. Evaluating the composition and processing potential of novel sources of Brazilian biomass for sustainable biorenewables production. BIOTECHNOLOGY FOR BIOFUELS, v. 7, JAN 18 2014. Web of Science Citations: 31.
DE ARAUJO, EVANDRO ARES; TOMAZINI, JR., ATILIO; SEIKI KADOWAKI, MARCO ANTONIO; MURAKAMI, MARIO TYAGO; POLIKARPOV, IGOR. Crystallization and preliminary X-ray diffraction analysis of a new xyloglucanase from Xanthomonas campestris pv. campestris. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 69, n. 6, p. 676-678, JUN 2013. Web of Science Citations: 0.
LIMA, MARISA A.; LAVORENTE, GABRIELA B.; DA SILVA, HANA K. P.; BRAGATTO, JULIANO; REZENDE, CAMILA A.; BERNARDINELLI, OIGRES D.; DEAZEVEDO, EDUARDO R.; GOMEZ, LEONARDO D.; MCQUEEN-MASON, SIMON J.; LABATE, CARLOS A.; POLIKARPOV, IGOR. Effects of pretreatment on morphology, chemical composition and enzymatic digestibility of eucalyptus bark: a potentially valuable source of fermentable sugars for biofuel production - part 1. BIOTECHNOLOGY FOR BIOFUELS, v. 6, MAY 9 2013. Web of Science Citations: 58.
ROSSETO, FLAVIO R.; PUHL, ANA C.; ANDRADE, MAXUEL O.; POLIKARPOV, IGOR. Crystallization and preliminary diffraction analysis of the catalytic domain of major extracellular endoglucanase from Xanthomonas campestris pv. campestris. ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS, v. 69, n. 2, p. 137-140, FEB 2013. Web of Science Citations: 2.

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