DNA extraction is a fundamental process in molecular biology, biotechnology, and genetic research. It involves isolating DNA from various biological samples such as cells, tissues, or bodily fluids. However, the success of DNA extraction relies heavily on the elimination of contaminants and the preservation of DNA integrity. Proteinase K, an enzyme with potent proteolytic activity, plays a crucial role in this process by degrading proteins that could interfere with DNA isolation and analysis.
Proteinase K: An Overview
Proteinase K is a serine protease derived from the fungus Tritirachium album. It possesses a broad specificity, efficiently cleaving peptide bonds adjacent to the carboxyl group of aliphatic and aromatic amino acids. This robust enzymatic activity makes proteinase K invaluable in various molecular biology applications, including DNA extraction.
Role of Proteinase K in DNA Extraction
Protein Removal: Biological samples used for DNA extraction often contain proteins that can hinder the isolation and subsequent analysis of DNA. These proteins may include nucleases, which can degrade DNA molecules, as well as histones and other structural proteins. Proteinase K acts by hydrolyzing peptide bonds within these proteins, effectively degrading them and releasing DNA from their association.
- Inactivation of Nucleases: Nucleases are enzymes that degrade nucleic acids, including DNA and RNA. Their presence in biological samples can lead to the degradation of DNA during extraction and storage. Proteinase K helps inactivating these nucleases by cleaving their peptide bonds, rendering them non-functional. This action safeguards the integrity of the extracted DNA and prevents degradation.
- Enhanced DNA Yield: Proteinase K treatment enhances the yield of extracted DNA by facilitating the release of DNA from cellular structures and protein complexes. This is particularly crucial when isolating DNA from tough or recalcitrant samples such as plant tissues or microbial biofilms. Proteinase K effectively disrupts cell membranes and nuclear envelopes, releasing DNA into the extraction buffer for subsequent purification.
- Improved DNA Quality: The presence of intact DNA is essential for downstream applications such as PCR, sequencing, and cloning. Proteinase K aids in maintaining DNA integrity by minimizing shearing and fragmentation during extraction. By degrading proteins that could potentially cause physical damage to DNA molecules, proteinase K ensures that the extracted genetic material remains suitable for accurate analysis and manipulation.
In conclusion, proteinase K serves as a critical component in DNA extraction protocols, playing multiple essential roles in ensuring the success of the process. By effectively degrading proteins, inactivating nucleases, enhancing DNA yield, and preserving DNA integrity, proteinase K contributes to the isolation of high-quality DNA from diverse biological samples. Its versatility and efficacy make it an indispensable tool in molecular biology research, enabling scientists to unlock the secrets encoded within the genetic material of living organisms. At SBS Genetech, we proudly present a groundbreaking solution to this demand with its Mutant Proteinase K. Engineered through cutting-edge technology, this enzyme stands as a superior alternative to the wild-type Proteinase K, offering heightened performance and versatility across a myriad of applications.
Enhanced Properties
The hallmark feature of Mutant Proteinase K lies in its superior specific activity and enhanced stability, even at room temperature, setting it apart from its conventional counterpart. This non-specific serine proteinase boasts a broad substrate spectrum, ensuring efficacy in digesting both native and denatured proteins across diverse experimental setups.
Robust Characteristics
Mutant Proteinase K exhibits remarkable resilience, maintaining its activity over a wide pH range from 4 to 12. This adaptability renders it suitable for a multitude of experimental conditions, accommodating various research requirements with ease. Its compatibility with different substrates and experimental setups makes it an indispensable tool for molecular biologists and researchers alike.
Product Specifications
Designed to meet the exacting standards of the scientific community, Mutant Proteinase K is available in meticulously crafted formulations:
- Cat. No.: MPTK-10 (for 10g), MPTK-1000 (for 1kg)
- Form: White Powder, Lyophilized from purified solids or liquid enzyme
- Specific Activity: ≥30 U/mg dry weights
- Package: 100 mg/bottle (Standard) or customizable packing options ranging from 1 to 200 grams
- Source: Pichia Pastoris with a mutant cloned gene from Tritirachium album limber
- Molecular Weight: 29,300
Preparation and Storage
Mutant Proteinase K is conveniently soluble in water, yielding a clear, colorless solution at a concentration of 1 mg/ml. For optimal storage and longevity, it is recommended to store the enzyme below 4°C. With a shelf life of three years for powder and one year for liquid formulations, researchers can trust in its reliability for long-term experiments.
Quality Assurance
Stringent quality control assays validate the potency and purity of Mutant Proteinase K:
- Unit Definition: Defined as the amount of enzyme liberating 1 µmol of tyrosine per minute from casein at 37°C, pH 7.5.
- DNase and RNase Activity: Undetectable after prolonged incubation with DNA and RNA, respectively.
- Protein Purity: Exceeds 99% as confirmed by Native-PAGE and SDS-PAGE assays.
Versatile Applications
The versatility of Mutant Proteinase K is showcased through its wide-ranging applications, mirroring those of wild-type Proteinase K but with enhanced performance. From isolating mRNA and genomic DNA to modifying glycoproteins for structural studies, this enzyme proves indispensable in various molecular biology endeavors. Active in the presence of SDS, urea, and EDTA, and functional within a broad temperature range of 15°C to 75°C, its adaptability knows no bounds.
Scientific Validation
Backed by extensive research, Mutant Proteinase K has garnered recognition in esteemed scientific journals, including its inclusion in the New Products section of the Science Journal on March 8, 2019. This recognition underscores its significance and contribution to advancing molecular biology research.
Conclusion
SBS Genetech's Mutant Proteinase K stands as a testament to innovation and excellence in the field of molecular biology. With its unparalleled performance, enhanced stability, and broad applicability, it emerges as an indispensable tool for researchers worldwide. As a leading industry player in the Proteinase K market, SBS Genetech continues to spearhead advancements, empowering scientists in their quest for knowledge and discovery.
Related:
- Mutant Proteinase K: for common molecular biology applications
- Mutant Proteinase K (Sterile): for high-standard applications
- Mutant Proteinase K (Industrial Grade): for industrial applications