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Deciphering Proteinase K: Exploring Its Nonspecific Cleavage Sites

Explore the versatile enzyme, Proteinase K, and its broad spectrum of cleavage sites, offering insights into its applications in molecular biology research.

June 22, 2023

Proteinase K is a broad-spectrum serine protease commonly used in molecular biology research for its ability to digest proteins. It cleaves peptide bonds adjacent to the carboxylic group of aliphatic and aromatic amino acids with blocked alpha amino groups, such as phenylalanine, alanine, valine, leucine, and isoleucine. However, its activity is not limited to these amino acids, as it can also cleave other residues, albeit with lower efficiency.

Proteinase K does not recognize specific cleavage sites like restriction enzymes do in DNA. Instead, its cleavage activity is relatively nonspecific, leading to the digestion of proteins into smaller peptides and amino acids. This nonspecificity makes Proteinase K useful for various applications, such as removing proteins from nucleic acid preparations or digesting proteins in cell lysates for downstream analysis.

The lack of a specific recognition sequence means that Proteinase K cleaves proteins at multiple sites, resulting in a heterogeneous mixture of peptides. This broad specificity allows it to be effective in various experimental contexts but also requires careful optimization to achieve specific cleavage outcomes.

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.

 

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