Molecular characterization of a family 5 glycoside hydrolase suggests an induced-fit enzymatic mechanism.

Abstract Glycoside hydrolases (GHs) play fundamental roles in the decomposition of lignocellulosic biomaterials. Here, we report the full-length structure of a cellulase from Bacillus licheniformis (BlCel5B), a member of the GH5 subfamily 4 that is entirely dependent on its two ancillary modules (Ig-like module and CBM46) for catalytic activity. Using X-ray crystallography, small-angle X-ray scattering […]

Biochemical Characterization and Low-Resolution SAXS Molecular Envelope of GH1 β-Glycosidase from Saccharophagus degradans.

The marine bacteria Saccharophagus degradans (also known as Microbulbifer degradans), are rod-shaped and gram-negative motile γ-proteobacteria, capable of both degrading a variety of complex polysaccharides and fermenting monosaccharides into ethanol. In order to obtain insights into structure-function relationships of the enzymes, involved in these biochemical processes, we characterized a S. degradans β-glycosidase from glycoside hydrolase […]

Recombinant Trichoderma harzianum endoglucanase I (Cel7B) is a highly acidic and promiscuous carbohydrate-active enzyme

Trichoderma filamentous fungi have been investigated due to their ability to secrete cellulases which find various biotechnological applications such as biomass hydrolysis and cellulosic ethanol production. Previous studies demonstrated that Trichoderma harzianum IOC-3844 has a high degree of cellulolytic activity and potential for biomass hydrolysis. However, enzymatic, biochemical, and structural studies of cellulases from T. […]

HTP-OligoDesigner: An Online Primer Design Tool for High-Throughput Gene Cloning and Site-Directed Mutagenesis

Following burgeoning genomic and transcriptomic sequencing data, biochemical and molecular biology groups worldwide are implementing high-throughput cloning and mutagenesis facilities in order to obtain a large number of soluble proteins for structural and functional characterization. Since manual primer design can be a time-consuming and error-generating step, particularly when working with hundreds of targets, the automation […]

Nuclear receptor full-length architectures: confronting myth and illusion with high resolution

The crystal structures of three nuclear receptor (NR) complexes have emerged to reveal their multidomain architectures on DNA. These pictures provide unprecedented views of interfacial couplings between the DNA-binding domains (DBDs) and ligand-binding domains (LBDs). The detailed pictures contrast with previous interpretations of low-resolution electron microscopy (EM) and small angle X-ray scattering (SAXS) data, which […]

RXR agonist modulates TR: corepressor dissociation upon 9-cis retinoic acid treatment

Transcriptional regulation controlled by thyroid hormone receptor (TR) drives events such as development, differentiation and metabolism. TRs may act either as homodimers, or as heterodimers with retinoid X receptor (RXR). Thyroid hormone T3 (3, 3`,5 triiodo-L-thyronine) preferentially binds TR:RXR heterodimers, which activate transcription through coactivator recruitment. However, it is unclear if TR:RXR heterodimers may also […]

Mapping the lignin distribution in pretreated sugarcane bagasse by confocal and fluorescence lifetime imaging microscopy.

Mapping the lignin distribution in pretreated sugarcane bagasse by confocal and fluorescence lifetime imaging microscopy. Coletta VC, Rezende CA, da Conceição FR, Polikarpov I, Guimarães FE.   BACKGROUND: Delignification pretreatments of biomass and methods to assess their efficacy are crucial for biomass-to-biofuels research and technology. Here, we applied confocal and fluorescence lifetime imaging microscopy (FLIM) […]

X-ray Structure and Molecular Dynamics Simulations of Endoglucanase 3 from Trichoderma harzianum: Structural Organization and Substrate Recognition by Endoglucanases That Lack Cellulose Binding Module

X-ray Structure and Molecular Dynamics Simulations of Endoglucanase 3 from Trichoderma harzianum: Structural Organization and Substrate Recognition by Endoglucanases That Lack Cellulose Binding Module Érica T. Prates, Ivana Stankovic, Rodrigo L. Silveira, Marcelo V. Liberato, Flávio Henrique-Silva, Nei Pereira Jr, Igor Polikarpov, Munir S. Skaf   Plant biomass holds a promise for the production of […]

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

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. Lima MA, Lavorente GB, da Silva HK, Bragatto J, Rezende CA, Bernardinelli OD, de Azevedo ER, Gomez LD, McQueen-Mason SJ, Labate CA, Polikarpov I.   BACKGROUND: In recent years, the growing demand for biofuels has encouraged the search for […]

Joint x-ray crystallographic and molecular dynamics study of cellobiohydrolase I from trichoderma harzianum: deciphering the structural features of cellobiohydrolase catalytic activity

Textor LC, Colussi F, Silveira RL, Serpa V, de Mello BL, Muniz JR, Squina FM, Pereira N Jr, Skaf MS, Polikarpov I.   Aiming to contribute towards the characterization of new, biotechnologically relevant cellulolytic enzymes, we report here the first crystal structure of the catalytic core domain of Cel7A (CBHI) from the filamentous fungus Trichoderma harzianum IOC 3844. Our structural studies and molecular dynamics […]