Biomaterial Database

A novel prokaryotic expression system for biosynthesis of recombinant human membrane-bound catechol-O-methyltransferase. 2011

A Q Pedro, and M J Bonifácio, and J A Queiroz, and C J Maia, and L A Passarinha

CICS - Centro de Investigação em Ciências da Saúde, Universidade da Beira Interior, 6201-001 Covilhã, Portugal.

Membrane proteins constitute 20-30% of all proteins encoded by the genome of various organisms. While large amounts of purified proteins are required for pharmaceutical and crystallization attempts, there is an unmet need for the development of novel heterologous membrane protein overexpression systems. Specifically, we tested the application of Brevibacillus choshinensis cells for the biosynthesis of human membrane bound catechol-O-methyltransferase (hMBCOMT). In terms of the upstream stage moderate to high expression was obtained for complex media formulation with a value near 45 nmol/h/mg for hMBCOMT specific activity achieved at 20 h culture with 37°C and 250 rpm. Subsequently, the efficiency for reconstitution of hMBCOMT is markedly null in the presence of ionic detergents, such as sodium dodecyl sulphate (SDS). In general, for non-ionic and zwiterionic detergents, until a detergent critic micellar concentration (CMC) of 1.0 mM, hMBCOMT shows more biological activity at lower detergent concentrations while for detergent CMC higher than 1 mM, higher detergent concentrations seem to be ideal for hMBCOMT solubilization. Indeed, from the detergents tested, the non-ionic digitonin at 0.5% (w/v) appears to be the most suitable for hMBCOMT solubilization.

UI	MeSH Term	Description	Entries
D008565	Membrane Proteins	Proteins which are found in membranes including cellular and intracellular membranes. They consist of two types, peripheral and integral proteins. They include most membrane-associated enzymes, antigenic proteins, transport proteins, and drug, hormone, and lectin receptors.	Cell Membrane Protein,Cell Membrane Proteins,Cell Surface Protein,Cell Surface Proteins,Integral Membrane Proteins,Membrane-Associated Protein,Surface Protein,Surface Proteins,Integral Membrane Protein,Membrane Protein,Membrane-Associated Proteins,Membrane Associated Protein,Membrane Associated Proteins,Membrane Protein, Cell,Membrane Protein, Integral,Membrane Proteins, Integral,Protein, Cell Membrane,Protein, Cell Surface,Protein, Integral Membrane,Protein, Membrane,Protein, Membrane-Associated,Protein, Surface,Proteins, Cell Membrane,Proteins, Cell Surface,Proteins, Integral Membrane,Proteins, Membrane,Proteins, Membrane-Associated,Proteins, Surface,Surface Protein, Cell
D011485	Protein Binding	The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments.	Plasma Protein Binding Capacity,Binding, Protein
D011994	Recombinant Proteins	Proteins prepared by recombinant DNA technology.	Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D002394	Catechol O-Methyltransferase	Enzyme that catalyzes the movement of a methyl group from S-adenosylmethionone to a catechol or a catecholamine.	Catechol Methyltransferase,Catechol-O-Methyltransferase,Catechol O Methyltransferase,Methyltransferase, Catechol,O-Methyltransferase, Catechol
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D058675	Brevibacillus	A genus of GRAM-POSITIVE ENDOSPORE-FORMING RODS in the family Paenibacillaceae. Most strains have been isolated from the natural environment, particularly soils.
D061249	Batch Cell Culture Techniques	Methods for cultivation of cells, usually on a large-scale, in a closed system for the purpose of producing cells or cellular products to harvest.	Batch Culture Techniques,Batch Culture,Continuous Batch Culture,Fed Batch Culture Techniques,Fed-Batch Culture,Fed-Batch Culture Techniques,Shake-Flask Culture,Batch Culture Technique,Batch Culture, Continuous,Batch Cultures,Batch Cultures, Continuous,Continuous Batch Cultures,Culture Technique, Batch,Culture Technique, Fed-Batch,Culture Techniques, Batch,Culture Techniques, Fed-Batch,Culture, Batch,Culture, Continuous Batch,Culture, Fed-Batch,Culture, Shake-Flask,Cultures, Batch,Cultures, Continuous Batch,Cultures, Fed-Batch,Cultures, Shake-Flask,Fed Batch Culture,Fed-Batch Culture Technique,Fed-Batch Cultures,Shake Flask Culture,Shake-Flask Cultures,Technique, Batch Culture,Technique, Fed-Batch Culture,Techniques, Batch Culture,Techniques, Fed-Batch Culture