tailieunhanh - Báo cáo Y học: The role of amino-acid residues Q39 and E451 in the determination of substrate specificity of the Spodoptera frugiperda b-glycosidase

The design of b-glycosidases with planed substrate specificity for biotechnological application has received little attention. This is mostly a consequence of the lack of data on the molecular basis of the b-glycosidase specificity, namely data on the energy of the noncovalent interactions in the enzymetransition state complex. In an attempt to fill this gap, sitedirected mutagenesis and enzyme steady-state kinetic experiments with different substrates were conducted, using as model a digestive b-glycosidase (glycoside hydrolase family 1) from Spodoptera frugiperda (Lepidoptera) (Sfbgly50). The active site of this enzyme was modeled based on its sequence and on crystallographic data of similar enzymes | Eur. J. Biochem. 269 3705-3714 2002 FEBS 2002 doi The role of amino-acid residues Q39 and E451 in the determination of substrate specificity of the Spodoptera frugiperda b-glycosidase Sandro R. Marana Walter R. Terra and Clelia Ferreira Departamento de Bioquimica Instituto de Quimica Universidade de Scio Paulo CP 26077 Sao Paulo 05513-970 Brazil The design of b-glycosidases with planed substrate specificity for biotechnological application has received little attention. This is mostly a consequence of the lack of data on the molecular basis of the b-glycosidase specificity namely data on the energy of the noncovalent interactions in the enzymetransition state complex. In an attempt to fill this gap site-directed mutagenesis and enzyme steady-state kinetic experiments with different substrates were conducted using as model a digestive b-glycosidase glycoside hydrolase family 1 from Spodoptera frugiperda Lepidoptera Sfbgly50 . The active site of this enzyme was modeled based on its sequence and on crystallographic data of similar enzymes. Energy of noncovalent interactions in transition state between Sfbgly50 amino acids and glycone hydroxyls was determined. Sfbgly50 residue E451 seems to be a key residue in determining b-glycosidase preference for glucosides vs. galactosides based on the following data a The energy of the noncovalent interaction between glycone equatorial hydroxyl 4 with E451 in the transition state is about 60 higher than its interaction with Q39. b The energy of the E451-hydroxyl 4 interaction decreases more than the Q39-hydroxyl 4 interaction when hydroxyl 4 is changed from equatorial to axial position. c A Sfbgly50 mutant where E451 was substituted by A hydrolyzes galactosides faster than glucosides. It was also shown that glycone hydroxyl 6 interacts favorably with Q39 but not with E451 probably due to steric hindrance. These interactions result in the b-glycosidase hydrolyzing fucosides 6-deoxygalactosides .

• Báo cáo khoa học
• Report medicine
• traditional medicine
• scientific reports
• genetics
• Structural biochemistry
• Structural Biology
• biology
• biological research
• applied biology
• modern biology
• biological engineering research
• metazoan systems
• Biochemistry cell and molecular biology test
• Molecular Biology Test
• Biochemistry Test
• Genetics test
• Cupramolecular complexes
• Campbell biology
• Biology systems
• Lecture Biology
• Biological sciences
• Molecular basis of inheritance
• Structural model of DNA
• scientific research
• molecular Biology
• structural information
• breast cancer
• BMC Bioinformatics
• Structural systems biology
• Oxidative stress
• Structural proteome
• Physicochemical properties
• Oxidative damage
• Metabolic model
• BMC Molecular and Cell Biology
• HIV 2 protease
• Drug resistance mutations
• Structural asymmetry
• Structural alphabet
• Structural variation
• Structural mutation
• 450 kb duplications
• Kidney cells
• Genome Biology
• Complex structural rearrangements
• Sequence analysis
• Copy number variant
• Tạp chí khoa học
• Structural basis of light energy
• Electron transfer in biology
• Models for energy and electron transfer
• Electron transfer in biological materials
• The blue multi copper
• Biological organization
• Systems biology
• Structural bioinformatics
• Integrative systems biology
• Bioinformatics methods
• Mass spectrometry
• medical reports
• thesis report
• health education
• medical report