Data on thermostable β-glucosidase immobilized by Zn2.

In this article, the methods for detection of enzyme activity and protein concentration are described. The data of the calibration curves can be used for a further understanding on the assays of enzyme activity measured with p-nitrophenyl-β-D-glucopyranoside (pNPG) or cellobiose as the substrate. In addition, the data presented provides an analytic method for measuring protein concentration in mixed samples.

Specifications table

Value of the data

The data makes available to the detection of enzyme activity with pNPG as the substrate.

The data will guide the assay of enzyme activity for immobilized enzyme with cellobiose as the substrate.

The data presented can provide a calibration curve for measuring protein concentration in mixed samples.

Data

The calibration curve about the assay of enzyme activity measured with pNPG as substrate is given in Fig. 1. Glucose Assay Kit purchased from Shanghai Rongsheng Biological Technology Co., Ltd provides the calibration curve about the assay of enzyme activity with cellobiose as substrate. Meanwhile, Fig. 2 showed a calibration curve for the detection of protein concentration in mixed samples.

Fig. 1

The calibration curve about assay of enzyme activity with pNPG as the substrate.

Fig. 2

The calibration curve about detection of protein concentration with bovine serum albumin (BSA) as the reference.

Experimental design, materials and methods

Enzyme activity of immobilized enzyme required was measured with pNPG as the substrate [1]. The pNP (0, 0.015, 0.030,0.0625, 0.125, 0.25, 0.50, 1.00 μmol/mL) was added to 1.5 mL tube containing 200 μL citric acid Na2HPO4 buffer (100 mM) and 600 μL Na2CO3 (1 M), The calibration curve was given by: y = 2.3764x + 0.0483 in Fig. 1, where x is the pNP concentration (μmol/mL) and , y is the absorbance of A405nm. Then the absorbance is calculated as enzyme activity from working curve. Enzyme activity of enzyme required was measured by cellobiose as the substrate. The calibration curve was provided by Glucose Assay Kit purchased from Shanghai Rongsheng Biological Technology Co., Ltd (Shanghai, China). And the calibration curve was given by: where y is the glucose concentration (mmol/L); A0 is the absorbance of A505nm from standard sample; A1 is the absorbance of A505nm from sample detected; B represented the concentration of standard sample (mmol/L). Then the absorbance is calculated as enzyme activity from working curve.

Protein concentration was detected by Bradford protein Assay Kit and the bovine serum albumin (BSA) was as the reference [2]. The mixture contained 200 μL Bradford protein Assay Kit and BSA (0, 25, 125, 250, 500, 750, and 1000 μg/mL) which was dissolved in deionized water. The calibration curve was given by: y = 0.0008x + 0.0081 in Fig. 2, where x is the BSA concentration (μg/mL), y is the absorbance of A595nm. Then the absorbance is calculated as protein concentration from working curve.

Subject area	Biology
More specific subject area	Enzyme Eng&Proteins
Type of data	Figure
How data was acquired	Using an absorbance microplate reader (SpectraMax190, Molecular Devices, LLC, Sunnyvale, CA)
Data format	Raw and analyzed data
Experimental factors	Assays of enzyme activity and protein concentration
Experimental features	The calibration curves of enzyme activity and protein concentration were offered
Data source location	Nanjing, China
Data accessibility	The data are available with this article