Solid-β-Glucosidase(S-β-GC) Activity Assay Kit
Spectrophotometry
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description Product Description
The Solid-β-Glucosidase (S-β-GC) Activity Assay Kit is primarily used to measure the enzymatic activity of β-glucosidase in solid biological samples, such as soil, plant materials, or other solid matrices. This kit is essential in research focused on understanding the role of β-glucosidase in metabolic processes, particularly in the breakdown of glycosidic bonds in complex carbohydrates like cellulose into glucose. It is widely applied in studies related to plant biology, where β-glucosidase plays a critical role in cell wall metabolism and defense mechanisms. Additionally, the kit is utilized in the food and beverage industry to monitor and optimize enzymatic processes involved in fermentation, flavor development, and the production of bioactive compounds. In environmental science, it helps assess microbial activity in soil and water, contributing to studies on nutrient cycling and biodegradation. The assay kit is also valuable in medical research, particularly in investigating enzyme deficiencies and their implications in metabolic disorders. Its simplicity and accuracy make it a reliable tool for both academic and industrial applications.
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Soil β-Glucosidase (S-β-GC) Activity Assay Kit
Product Information
Product code: 67042
Method: Spectrophotometric method
S-β-GC catalyzes the hydrolysis of glycosidic bonds between aryl or alkyl groups and glycosyl moieties to generate glucose. It is an important component of the cellulolytic enzyme system and plays an important physiological role in soil microbial carbohydrate metabolism.
S-β-GC catalyzes p-nitrophenyl-β-D-glucopyranoside to generate p-nitrophenol, which has characteristic absorbance at 400 nm.
Package Contents and Storage
| Code | Item | Quantity | Storage |
|---|---|---|---|
| 67042.1 | Reagent 1 | 9 mg × 2 bottles | -20°C, protected from light |
| 67042.2 | Reagent 2 | 25 mL | 2-8°C |
| 67042.3 | Reagent 3 | 50 mL | 2-8°C |
| 67042.m | Instruction manual | 1 copy | / |
Toluene must be prepared separately.
Quality Standards and Safety Information
| Raw Material or Packaging Name | Quality Standard | Main Toxicity |
|---|---|---|
| Reagent 1 | -- | -- |
| Reagent 2 | -- | -- |
| Reagent 3 | -- | -- |
Transportation and Storage
Transportation: This product is transported with ice packs.
Storage: Store according to the instructions in this manual. Shelf life: 180 days.
Instructions for Use
Sample Preparation
- Air-dry fresh soil samples naturally or dry them in a 37°C oven.
- Pass the dried soil samples through a 30-50 mesh sieve.
Instrument Preparation
- Preheat the spectrophotometer for more than 30 minutes.
- Set the wavelength to 400 nm.
Reagent Preparation
Before use, add 6 mL double-distilled water to each bottle of Reagent 1. After use, store any unused solution at -20°C.
Assay Procedure
Use a 1 mL glass cuvette. Add the samples and reagents according to the table below.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Air-dried soil sample | 0.05 g | 0.05 g |
| Toluene | 25 μL | 25 μL |
Shake and mix thoroughly to moisten the soil sample, then place at room temperature for 15 minutes.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Reagent 1 | 400 μL | - |
| Reagent 2 | 500 μL | 500 μL |
| Distilled water | - | 400 μL |
Mix thoroughly and incubate in a 37°C water bath for 1 hour. Immediately incubate in a 90°C water bath for 5 minutes. Cap tightly to prevent water loss.
Cool under running water, then centrifuge at 10000 g at room temperature for 10 minutes. Collect the supernatant.
| Component | Assay Tube | Control Tube |
|---|---|---|
| Supernatant | 500 μL | 500 μL |
| Reagent 3 | 1000 μL | 1000 μL |
Mix thoroughly and let stand at room temperature for 2 minutes. Zero the spectrophotometer with the control tube, then measure the absorbance value A at 400 nm.
Calculation
The regression equation measured under standard conditions is:
y = 0.0032x - 0.0027
In this equation, x is the standard concentration in μmol/L, and y is the absorbance value.
One unit of enzyme activity is defined as the production of 1 μmol p-nitrophenol per g of soil sample in 24 hours.
Soil β-glucosidase activity:
Soil β-glucosidase activity (U/g soil sample) = (A + 0.0027) ÷ 0.0032 ÷ reaction time (1/24 h) × total reaction volume (0.925 mL) ÷ sample mass (0.05 g) ÷ 1000
Soil β-glucosidase activity (U/g soil sample) = 138.7 × (A + 0.0027)
Where:
- T: reaction time, 1 h = 1/24 d
- V total reaction: total volume of the reaction system, 9.25 × 10-4L
- W: sample mass, 0.05 g
Precautions
- Items to be prepared by the user: visible spectrophotometer, benchtop centrifuge, water bath, adjustable pipettes, 1 mL glass cuvette, toluene, and distilled water.
- Before the formal assay, select 2-3 samples with large expected differences for a preliminary assay.
- This 50T kit can test 24 samples.
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