In the analysis of the gels was observed the presence of multiple bands in the samples of enzyme broths produced from different carbon sources used in the culture media.
76b When EPR measurements were carried out at 10 K during steady-state turnover, multiple [4Fe 4S] species were observed, and by comparison to samples of enzyme with and without SAM, two of these species were assigned as the unbound (g = 1.92, 2.04) and bound (g = 1.83, 1.99) forms of the [4Fe 4S]+ state of the enzyme (Table 2).
NaClO4 was added to a sample of enzyme and the DLS signal was monitored as a function of time.
The higher current in the ATR-IR cell reflects the larger sample of enzyme under electrochemical control on the high surface area electrode.
Experimental tests have been made on samples of industrial enzyme granules, which have a complex layered structure, and reproducible results have been obtained.
At intervals predetermined, samples of the enzyme were withdrawn, and their residual activities were measured under the standard assay condition.
Samples of purified enzyme (30 μL) were added to a 96-well microtiter plate with a 70 μL 1 mM solution (final concentration) of AlCl3, BaCl2, CaCl2, CdSO4, CoCl2, CuSO4, HgCl2, KCl, LiCl, MgCl2, MnCl2, PbCl2, and ZnSO4.
In order to evaluate the effect of pH on the stability of Spr Cel8A, samples of purified enzyme were incubated in a variety of buffers as mentioned previously, at pH values, ranging from 3.0 to 10.5, at 28 °C for 1 h.
By incubating samples of the enzymes at various temperatures and measuring the activity at regular intervals, the enzyme activity half-life was calculated at three different temperatures.
The samples of crude enzymes were centrifuged at 220000 g for 30 min and 0.15 M sodium chloride and 0.15 mM DTT (final concentrations) were added to the enzymes from Y. lipolytica and P. pastoris, and 0.15 M sodium chloride (final concentration) was added to the enzyme from P. angusta.
Purified enzymes were diluted in 0.5 mM HEPES, 1.5 mM NaCl pH 7.4 to a concentration of 40 ng μL 1, and then 10 μL samples of diluted enzymes were transferred to a 96-well PCR plate and mixed with either 10 μL of 40% (v/v) [C2mim][OAc]/water or 10 μL of 100 mM MES buffer pH 7.2 (MES buffer pH changed to 6.65 at 70 °C, matching the pH of the IL/water mixture).
