Purification and semienzymic synthesis of flavin adenine dinucleotide-3′-phosphate
| Autor: | Mark Fisher, Brian R. Rabin, Henny J. Eggelte, Stuart Harbron |
|---|---|
| Rok vydání: | 1994 |
| Předmět: |
D-Amino-Acid Oxidase
Ultrafiltration Bioengineering Flavin group Applied Microbiology and Biotechnology Biochemistry Cofactor Substrate Specificity Glucose Oxidase chemistry.chemical_compound Endoribonucleases Glucose oxidase Ribonuclease Flavin adenine dinucleotide Chromatography biology Hydrolysis Substrate (chemistry) Enzymes Immobilized chemistry Yield (chemistry) Ribonuclease T Flavin-Adenine Dinucleotide biology.protein Apoproteins Biotechnology |
| Zdroj: | Enzyme and Microbial Technology. 16:281-285 |
| ISSN: | 0141-0229 |
| DOI: | 10.1016/0141-0229(94)90167-8 |
| Popis: | Nonradioactive immunoassays incorporating an element of amplification in their detection system require the use of components that are highly purified. Flavin adenine dinucleotide-3′-phosphate (FADP) is the primary substrate used in such an amplification assay. For incorporation into a simple, single-pot assay system, the concentration of contaminating flavin adenine dinucleotide (a prosthetic group for the enzyme d -aminoacid oxidase used in the amplification cascade assay) in this primary substrate must be minimized to achieve maximum sensitivity. Production of the substrate to a high degree of purity has been achieved using apo-glucose oxidase to specifically remove contaminating flavin adenine dinucleotide from solution and hydrolysis of a cyclic intermediate as a final production protocol by ribonuclease T2 to give the product in high yield. The use of continuous ultrafiltration reactors at each stage is described and compared to a final production step utilizing immobilized ribonuclease T2. These reactors allow large volumes of material to be handled and assist in the scale-up of these processes. The suitability of each protocol is assessed for the commercial production of FADP. |
| Databáze: | OpenAIRE |
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