Enzovera Proteinase K is a broad-spectrum serine protease available in five catalogue variants: EV-NUC-001 (Lyophilized, ≥30 U/mg, 100 g/bottle), EV-NUC-001S (Sterile Lyophilized, ≥30 U/mg, 100 g/bottle), EV-NUC-002 (Solution 20 mg/ml, ≥900 U/ml, 10 ml and 1 L bottles), EV-NUC-003 (Solution 40 mg/ml, ≥1800 U/ml, 10 ml and 1 L bottles), and EV-NUC-004 (Cosmetic Grade, ≥150 U/g, 100 g/bottle). Originally isolated from the fungus Tritirachium album, Proteinase K is the preeminent protease for nucleic acid isolation workflows, where its capacity to digest proteins — including contaminating nucleases — in the presence of chaotropic agents, detergents, and chelating agents makes it indispensable at every scale of operation.

Mechanism. Proteinase K is a subtilisin-type serine endopeptidase (EC 3.4.21.64) that cleaves peptide bonds on the carboxyl side of aliphatic, aromatic, and other hydrophobic amino acid residues with low sequence specificity. The active-site catalytic triad (Asp-39, His-69, Ser-224) performs a nucleophilic attack on the carbonyl carbon of the scissile bond, generating an acyl-enzyme intermediate resolved by water. Critically, the enzyme retains full activity in 0.5–1% SDS, 0.5–1% Triton X-100, 8 M urea, and in the presence of EDTA — which chelates Mg²⁺ and inactivates contaminating DNases and RNases without affecting the serine active site. This detergent and chaotrope tolerance is the mechanistic basis of its dominance in DNA and RNA extraction: SDS denatures the protein matrix while Proteinase K digests it, freeing nucleic acids from protein associations without collateral nuclease activity on the target molecules.

Formulation variants. The lyophilized powder formats (EV-NUC-001, EV-NUC-001S) are the most economical choice for large-scale operations; EV-NUC-001S is sterile-filtered and suitable for clinical sample processing and applications requiring reduced bioburden. The 20 mg/ml solution (EV-NUC-002) provides ready-to-use convenience for genomic DNA and RNA extractions at standard scale. The 40 mg/ml solution (EV-NUC-003) is optimized for high-throughput workflows where maximizing protease activity in a minimum added volume is required. EV-NUC-004 is manufactured to cosmetic-grade standards with additional safety documentation for personal care product development.

Recommended applications:

• Proteinase K digestion step in genomic DNA extraction from tissue, cultured cells, and blood
• RNA isolation: digestion of ribonucleoprotein complexes and inactivation of RNases in cell lysates
• Deproteinization of nucleic acid samples prior to PCR, sequencing, and hybridization
• FFPE tissue DNA extraction — digestion of formalin cross-linked proteins
• Prion and amyloid protein digestion in protein conformation and aggregation studies
• ChIP-seq and ATAC-seq sample preparation: protein removal from chromatin lysates
• Biofilm disruption and extracellular matrix protein removal in microbiology workflows
• Cosmetic and personal care formulation (EV-NUC-004 only)