Alpha-glucosidase (Bacillus thermophilus) (recombinant)

reorganization - Thermal stability

E-ACTIVITIES

Item number:AS000007

EC: 3.2.1.20

Synonyms: alpha-glucosidaseα-D-Glucoside Glucohydrolase

CAZy Family: GH13

CAS: 9001-42-7

characteristic

Electrophoresis purity:

- Single band on SDS gel electrophoresis (MW~ 66,000)

- Single main band on isoelectric focusing (pI~ 6.7)

Enzyme activity detection method:

Defining enzyme activity: inFrom pH 6.5 and 40 ° C at 405 nm per minute4-Nitrophenyl α-D-glucopyranoside（pNPG）produceThe amount of enzyme required for 1 µmol p-nitrophenol (pNP) is a unit of enzyme activity. PNP has an absorption peak at 405 nm, and the absorbance measured at 405 nm by a microplate analyzer is used to calculate the content of the product pNP.

withPlot the standard curve for the standard solution with pNP, layer dilution: 250 umol/L with 0.1 mol/L PBS (pH 6.5)p-Nitrophenol, diluted into200, 150, 100, 50, 25, 5, 0 umol/L, take 100ul of standard solution for each layer, and then add 100ul 1mol/L Na₂CO₃Solution, assayOD₄₀₅Plot the enzyme activity standard curve.

Sample enzyme activity detection: take20UL 5mM pNPG and 60ul 0.1mol/L PBS (pH 6.5) and moderately diluted enzyme solution 20ul, mix well and react in a 40 ° C water bath for 10 min, then add 100ul 1mol/L Na immediately₂CO₃The solution terminates the reaction and develops color, which is then detectedOD₄₀₅。

Enzyme activity (U/mL）= C × 50 × N/10

C - the concentration of pNP on the curve corresponding to the absorption value of the enzyme solution;

50 - Convert the activity in 20ul diluted enzyme solution to the activity in 1ml enzyme solution;

N - the dilution multiple of the protoenzyme solution;

10 - Response time.