反应 #1350567

反应方程式

bismuth nitrate

p-hydroxybenzoic acid hydrazide

potassium sodium tartrate

→

glucose

溶剂

glycine

NaOH

反应条件

后处理

1. 1
   workup.ADDITIONcontaining the agent
2. 2
   workup.ADDITIONare then added
3. 3
   温度after which the solution is heated for 10 minutes to 70° C
4. 4
   其他the absorption at 410 nm
5. 5
   其他is determined against a blank value at room temperature
6. 6
   其他is prepared in the same way as the measuring solution except that the glucan solution
7. 7
   workup.ADDITIONis added after the PAHBAH solution

实验过程

The detection and determination of the glucanolytic activity was based on modifications of the process described by M. Lever in Anal. Biochem. 47 (1972), 273-279 and Anal Biochem. 81 (1977), 21-27. A 0.5% by weight solution of β-glucan (Sigma No. G6513) in 50 mM glycine buffer (pH 9.0) was used for this purpose. 250 μl of this solution are added to 250 μl of a solution containing the agent to be tested for glucanolytic activity and incubated for 30 minutes at 40° C. 1.5 ml of a 1% by weight solution of p-hydroxybenzoic acid hydrazide (PAHBAH) in 0.5 M NaOH, which contains 1 mM bismuth nitrate and 1 mM potassium sodium tartrate, are then added, after which the solution is heated for 10 minutes to 70° C. After cooling (2 minutes/0° C.), the absorption at 410 nm is determined against a blank value at room temperature (for example with a Uvikon® 930 photometer) using a glucose calibration curve. The blank value is a solution which is prepared in the same way as the measuring solution except that the glucan solution is added after the PAHBAH solution. 1 U corresponds to the quantity of enzyme which produces 1 μmole of glucose per minute under these conditions.

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