2-2--azino-di-(3-ethylbenzothiazoline)-6-sulfonic-acid has been researched along with alcohol-oxidase* in 2 studies
2 other study(ies) available for 2-2--azino-di-(3-ethylbenzothiazoline)-6-sulfonic-acid and alcohol-oxidase
| Article | Year |
|---|---|
Simplified high-throughput screening of AOX1-expressed laccase enzyme in Pichia pastoris.
The heterologous protein expression in Pichia pastoris under the control of alcohol oxidase (AOX1)promoter comprises two steps, the growth and induction phases, which are time-consuming and technically demanding. Here, we describe an alternate method where expression is carried out directly in the methanol-containing medium. Using this method, we were successful in screening high-activity laccase clones from a library of laccase mutants generated by random mutagenesis. This simplified method not only saves time but also is highly efficient and can be used for screening a large number of clones. Topics: Alcohol Oxidoreductases; Benzothiazoles; Bleomycin; Clone Cells; Colorimetry; Coloring Agents; Enzyme Induction; Fungal Proteins; Gene Library; Kinetics; Laccase; Methanol; Mitochondrial Proteins; Mutagenesis; Mutant Proteins; Oxidoreductases; Pichia; Plant Proteins; Polymerase Chain Reaction; Promoter Regions, Genetic; Protein Engineering; Recombinant Proteins; Sulfonic Acids | 2015 |
A spectrophotometric assay for the enzymatic demethoxylation of pectins and the determination of pectinesterase activity.
A rapid spectrophotometric method for the determination of pectinesterase activity is presented. In this assay, methanol released from pectin by pectinesterase is oxidized with alcohol oxidase to form hydrogen peroxide and formaldehyde. Hydrogen peroxide is then quantitated with peroxidase and the chromogen 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid). Since both reactions exhibit the same pH optimum it was possible to couple the methanol assay directly to the action of pectinesterase for the real-time determination of this enzyme. The assay is reliable and sensitive, being capable of quantitating a minimum pectinesterase activity of 0.0625 unit (1 unit = 1 microM methanol released per minute). It is also capable of detecting the enzymatic demethoxylation of galactopyranosyl uronic acid methyl esters of pectin down to a minimum concentration of 1.56 nM of methanol per milliliter using a pectin substrate with a methoxy content of 10% (w/w) at a concentration of 0.5 microgram/ml. Topics: Alcohol Oxidoreductases; Benzothiazoles; Carboxylic Ester Hydrolases; Chromatography, Ion Exchange; Formaldehyde; Hydrogen Peroxide; Indicators and Reagents; Pectins; Peroxidase; Spectrophotometry; Sulfonic Acids | 1997 |