Protein 3D Structure



Protein Structure, Crystallization and Protein Modeling

Molecular Insights into Substrate Specificity and Thermal Stability of a Bacterial GH5-CBM27 endo-1,4-beta-D-mannanase









ARTICLE

Santos CR, Paiva JH, Meza AN, Cota J, Alvarez TMA, Ruller R, Prade RA, Squina FM and Murakami MT (2012) Molecular insights into substrate specificity and thermal stability of a bacterial GH5-CBM27 endo-1,4-β-d-mannanase. Journal of Structural Biology 177(2):469-476

The breakdown of bete-1,4-mannoside linkages in a variety of mannan-containing polysaccharides is of great importance in industrial processes such as kraft pulp delignification, food processing and production of second-generation biofuels, which puts a premium on studies regarding the prospection and engineering of beta-mannanases. In this work, a two-domain beta-mannanase from Thermotoga petrophila that encompasses a GH5 catalytic domain with a C-terminal CBM27 accessory domain, was functionally and structurally characterized. Kinetic and thermal denaturation experiments showed that the CBM27 domain provided thermo-protection to the catalytic domain, while no contribution on enzymatic activity was observed. The structure of the catalytic domain determined by SIRAS revealed a canonical (a/b)8-barrel scaffold surrounded by loops and short helices that form the catalytic interface. Several structurally related ligand molecules interacting with TpMan were solved at high-resolution and resulted in a wide-range representation of the subsites forming the active-site cleft with residues W134, E198, R200, E235, H283 and W284 directly involved in glucose binding 


MORE ARTICLES ABOUT PROTEIN STRUCTURE CRYSTALLIZATION AND PROTEIN MODELING

Souza TA, Santos CR, Souza AR, Oldiges DP, Ruller R, Prade RA, Squina FM, Murakami MT (2011) Structure of a novel thermostable GH51 alpha-L-arabinofuranosidase from Thermotoga petrophila RKU-1. Protein Science 20: 1632-1637

Dos Santos CR, Squina FM, Navarro AM, Oldiges DP, Leme AF, Ruller R, Mort AJ, Prade R, Murakami MT (2011) Functional and biophysical characterization of a hyperthermostable GH51 α-L arabinofuranosidase from Thermotoga petrophila.  Biotechnology Letters 33:131-137

Cota J, Alvarez TM, Citadini AP, Santos CR, de Oliveira Neto M, Oliveira RR, Pastore GM, Ruller R, Prade RA, Murakami MT, Squina FM. (2011) Mode of operation and low-resolution structure of a multi-domain and hyperthermophilic endo-β-1,3-glucanase from Thermotoga petrophila. Biochemical and Biophysical Research Communications 406:590-594

Squina FM, Santos CR, Ribeiro DA, Cota J, de Oliveira RR, Ruller R, Mort A, Murakami MT, Prade RA (2010) Substrate cleavage pattern, biophysical characterization and low-resolution structure of a novel hyperthermostable arabinanase from Thermotoga petrophila. Biochemical and Biophysical Research Communications 399:505-511

Santos CR, Meza AN, Hoffmam ZB, Silva JC, Alvarez TM, Ruller R, Giesel GM, Verli H, Squina FM, Prade RA, Murakami MT. (2010) Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1. Biochemical and Biophysical Research Communications 403:214-219

Santos CR, Squina FM, Navarro AM, Ruller R, Prade R, Murakami MT (2010) Cloning, expression, purification, crystallization and preliminary X-ray diffraction studies of the catalytic domain of a hyperthermostable endo-1,4-beta-D-mannanase from Thermotoga petrophila RKU-1. Acta Crystallographica. Section F, Structural Biology and Crystallization Communications 66(Pt 9):1078-1081

Santos CR, Meza AN, Hoffmam ZB, Silva JC, Alvarez TM, Ruller R, Giesel GM, Verli H, Squina FM, Prade RA, Murakami MT. (2010) Thermal-induced conformational changes in the product release area drive the enzymatic activity of xylanases 10B: Crystal structure, conformational stability and functional characterization of the xylanase 10B from Thermotoga petrophila RKU-1. Biochemical and Biophysical Research Communications 403:214-219

Squina FM, Prade RA, Wang H, Murakami MT (2009) Expression, purification, crystallization and preliminary crystallographic analysis of an endo-1,5-alpha-L-arabinanase from hyperthermophilic Thermotoga petrophila. Acta Crystallographica. Section F, Structural Biology and Crystallization Communications 65(Pt 9):902-905


http://www.sciencedirect.com/science/article/pii/S1047847711003376http://onlinelibrary.wiley.com/doi/10.1002/pro.693/fullhttp://www.springerlink.com/content/w27417057jr58l17/http://www.sciencedirect.com/science/article/pii/S0006291X11003032http://www.sciencedirect.com/science/article/pii/S0006291X11003032http://www.sciencedirect.com/science/article/pii/S0006291X10014178http://www.sciencedirect.com/science/article/pii/S0006291X10014178http://www.sciencedirect.com/science/article/pii/S0006291X10020589http://www.sciencedirect.com/science/article/pii/S0006291X10020589http://scripts.iucr.org/cgi-bin/paper?S1744309110029131http://scripts.iucr.org/cgi-bin/paper?S1744309110029131http://scripts.iucr.org/cgi-bin/paper?S1744309110029131http://www.sciencedirect.com/science/article/pii/S0006291X10020589http://www.sciencedirect.com/science/article/pii/S0006291X10020589http://journals.iucr.org/f/issues/2009/09/00/en5377/en5377bdy.htmlhttp://journals.iucr.org/f/issues/2009/09/00/en5377/en5377bdy.htmlhttp://lge.ibi.unicamp.br/lnbio/sbh.phpshapeimage_5_link_0shapeimage_5_link_1shapeimage_5_link_2shapeimage_5_link_3shapeimage_5_link_4shapeimage_5_link_5shapeimage_5_link_6shapeimage_5_link_7shapeimage_5_link_8shapeimage_5_link_9shapeimage_5_link_10shapeimage_5_link_11shapeimage_5_link_12shapeimage_5_link_13shapeimage_5_link_14shapeimage_5_link_15
Welcome

Instruction

Cell & Molecular Biology
ONLINE  Cell & Molec. Biol.
Medical Mycology
Eukaryotic Genetics
 
Research
Biomass Technology
Biomass Degradation
High Temp. Enzymes
Protein expression systems

Molecular Genetics
Gene Silencing
Stress Responses
Gene Regulation

Bioinformatics
DNA Bioinformatics
Structural Protein Modeling
Physical Mapping

Welcome.htmlInstruction.htmlMICR3033.htmlMICR3033ONL.htmlMICR3143.htmlMICR4263.htmlResearch.htmlHolocellulose.htmlThermophiles.htmlProtein_Expression.htmlGene_silencing.htmlStress.htmlRegulation.htmlDNA_Bioinfo.htmlPhysical_Maps.htmlshapeimage_6_link_0shapeimage_6_link_1shapeimage_6_link_2shapeimage_6_link_3shapeimage_6_link_4shapeimage_6_link_5shapeimage_6_link_6shapeimage_6_link_7shapeimage_6_link_8shapeimage_6_link_9shapeimage_6_link_10shapeimage_6_link_11shapeimage_6_link_12shapeimage_6_link_13shapeimage_6_link_14shapeimage_6_link_15