This was then used immediately to reconstitute enzyme reagent (R2c) which was protected from light and placed on board the instrument for analysis.
The reconstituted enzyme was found to crystallize using a replacement salt in the precipitant (KSCN), which afforded substrate binding, and three additional structures.
The reconstituted enzyme was used to determine pH and temperature optimum, pH and temperature stability, tolerance to chloride ions and several organic solvents using ABTS as the substrate.
All of the reconstituted enzyme mixtures were supplemented with GH3 B-glucosidase at a loading of 2.5 mg/g cellulose.
After adding triethanolamine solution and Assay Cocktail [a mixture of MES buffer (11.25 ml), reconstituted cofactor mixture (0.45 ml), reconstituted enzyme mixture (2.1 ml), water (2.3 ml), and reconstituted DTNB (0.45 ml)], total GSH in each of the deproteinated samples was measured at 405 nm.
Milligrams of extracted HMOs were then reduced, lyophilized, weighed using a microbalance, reconstituted in Nanopure water, and analyzed.
After three rounds of mutagenesis and screening, the triple mutant E163K/V193M/K170Q yielded a kcat > five times faster than wild type P450 1A2 in steady-state kinetic analysis using either isolated membrane fractions or purified, reconstituted enzymes.
CRAiLAR is turned into a fibre more like cotton when spun using a particular enzyme process.
To confirm that the functional R. castenholzii MPD (MDD-like protein sequence) catalyzes the transformation of MVAP (3) to IP (4) in a pathway specific manner with a measurable flux, we next reconstituted each pathway in vitro using a five-enzyme reaction mixture of homogenously purified enzymes and substrates.
In another reaction involving reconstituted enzymes, 80 nM CYP + 80 nM CPR mixture was used.
