In the fields of biopharmaceuticals and molecular biology, Escherichia coli (E. coli) is widely used as a model organism. The efficiency of protein extraction significantly influences the success of downstream experiments. While traditional methods such as ultrasonic disruptors are commonly employed, they often result in sample heating and incomplete cell lysis. The CASPeter high-pressure homogenizer, with its full physical disruption mechanism, efficient cooling system, and GMP certification, has emerged as a preferred solution for laboratory cell lysis. This article presents a detailed guide on achieving optimal protein extraction from Escherichia coli using this advanced equipment.
I. Experimental Background: Why Choose High-Pressure Homogenization?
The cell wall of Escherichia coli consists primarily of peptidoglycan. Traditional methods, such as chemical lysis and ultrasonic disruption, may lead to protein denaturation or loss of biological activity. In contrast, high-pressure homogenization effectively disrupts cell walls at low temperatures through three key mechanisms: shear forces, impact, and cavitation, thereby preserving the native structure and function of proteins.
Key Features of the CASPeter High-Pressure Homogenizer
- Constructed from 316L stainless steel: Ensures corrosion resistance, ease of cleaning, and compliance with GMP standards.
- Integrated online cooling system: Allows temperature pre-setting within the range of 0–8°C to prevent protein denaturation.
- Processing capacity of 15 mL: Suitable for both small-scale and pilot-scale experimental applications.
II. Experimental Procedure: From Bacterial Culture to Protein Isolation
Materials Required
- Zhongke Peter High-Pressure Homogenizer (Model: PT-10)
- Chiller (preset to 3.5°C)
- Escherichia coli culture
- Lysis buffer (containing protease and phosphatase inhibitors)
Experimental Protocol
1. Equipment Pre-Cooling
Connect the chiller to the homogenizer’s discharge cooler. Fill with water and set the temperature to 3.5°C. Allow the system to pre-cool for 10 minutes.
(Rationale: Maintaining low temperatures helps inhibit protease activity and minimizes protein degradation.)
2. Sample Preparation
Add lysis buffer to the bacterial culture at a 1:1 volume ratio. Mix thoroughly and incubate on ice for 10 minutes.
(Rationale: The detergent components in the lysis buffer facilitate membrane disruption and promote the release of intracellular proteins.)
3. Homogenization Process
Ensure the pressure valve is fully open and flush the system with water to remove any air from the pipeline. Introduce the bacterial suspension into the feed port and adjust the operating pressure to 900–1200 bar. Perform 3–4 homogenization cycles.
(Outcome Assessment: Following homogenization, the suspension should appear uniformly turbid without visible sediment. Microscopic examination reveals cell fragments smaller than 1 μm.)
III. Experimental Conclusion
The CASPeter high-pressure homogenizer demonstrates superior performance over conventional methods in terms of lysis efficiency, protein integrity, and operational convenience. It is particularly well-suited for the extraction of temperature-sensitive proteins such as enzymes and antibodies. After use, fully depressurize the system, drain all residual liquid, rinse the internal components with clean water, and store with the feed inlet sealed using 75% alcohol.
For those interested in acquiring the CASPeter high-pressure homogenizer, complimentary sample testing and a one-year warranty are available. Click the link to schedule a demonstration and enhance your laboratory’s cell disruption capabilities → CASPeter Official Website
Post time: Aug-09-2025