Proteinase K has received more attention from the medical and pharmaceutical industries since the COVID-19 outbreak in late 2019. In fact, proteinase K has always been active in the scientific research field, while the global COVID-19 pandemic further promoted the growth of the proteinase K market. In this article, we will introduce proteinase K from three aspects: the general definition, the applications, and market development.
Proteinase K was initially isolated and purified from the fungus Engyodontium album culture medium in 1974 and named proteinase K because of its ability to digest keratin. It is a highly active serine Proteinase that shows a preference for cleaving ester and peptide bonds connected to the carboxy-termini of aromatic and aliphatic amino acids. Few proteins are resistant to proteinase K due to the enzyme's potency, wide pH optimum (4-12.5), and broad substrate specificity.
Compared to other proteinases, proteinase K displays high activity and stability. Denaturants such as urea, SDS and EDTA, can not inactivate proteinase K. Proteinase K still maintains high activity even under the conditions of high temperature, high salt, or high pH. As a biochemical reagent, proteinase K usually provides products in lyophilized powder form and liquid form. Lyophilized form dominates the commercially available proteinase K due to its easy store and sustained integrity for a long time. The cofactor, calcium ion provides stability to the enzyme, however, it cannot increase the activity of the reaction.
Proteinase K remains stable in denaturants such as SDS or urea, and it can degrade native proteins. These two properties of proteinase K contribute to it be used in various fields, including biological scientific research, medical ＆ pharmaceutical industry, environment protection, food processing, washing industry, cosmetic industry, and leather industry. And the Proteinase K used in these applications is recombinant Proteinase K.
The isolation and purification of nucleic acids is a basic technique in biochemistry and molecular biology. Proteinase K can degrade proteins (such as histon) bound with nucleic acid and facilitate the separation of nucleic acid. Proteinase K is also often used to inhibit and degrade RNase during RNA extraction. RNA extracted by proteinase K method creates an RNase-free environment and get high-quality RNA, which is critical to the success of Northern blotting and hybridization analysis, cDNA synthesis, and in vitro translation.
Proteinase K is commonly used in situ hybridization for the treatment of prehybridization specimens. It can degrade the protein surrounding the target DNA to facilitate the penetration of the probe and improve the sensitivity of detection.
Proteinase K can determine the enzyme localization on the membrane by modifying proteins and glycoproteins on cell surfaces.
Proteinase K also promotes cell lysis by activating a bacterial autolytic factor and is used to remove cellular debris during the preparation of colony lifts.
Proteinase K is useful for the separation of mitochondria from the tissue.
In protein fingerprinting (an analytical technique for protein identification), the proteinase K catalyzes the hydrolysis of the unknown protein into smaller peptides, allowing for accurate measurement of their absolute masses with a mass spectrometer such as MALDI-TOF or ESI-TOF.
SARS-Cov-2, the causative agent for COVID-2019, is an RNA virus that mainly attacks the lung, intestine, liver, kidney, and other human organs after entering the body. The RT-PCR of SARS-CoV-2 is the “gold standard” for viral diagnosis and confirmation. Proteinase K is the primary component during SARS-CoV-2 extraction and purification. Besides, proteinase K is used for the digestion of proteins from brain tissue samples for prions in Transmissible Spongiform Encephalopathies (TSE) research. It is also applied in the forensic gene identification for comparison.
Proteinase K is used to isolate and extract DNA or RNA of disease-causing viruses without DNase and RNase. These pure DNA or RNA is injected into the animals or human bodies and is synthesized into the viral antigen protein, inducing an immune response that achieves a preventative or therapeutic role.
Proteinase K can also be used as a disinfectant in sewage treatment by inactivating bacteria in domestic and industrial water. The capsid protein in the outer layer of the virus protects the virus from environmental factors and determines the specificity of the virus infection. Proteinase K can destroy the viral capsid protein, thus interfering with the virus's binding to host cells. Compared with chlorine-based disinfection, Proteinase K-mediated virus inactivation is not only efficient but also environmentally friendly because Proteinase K itself is harmless and the disinfection process does not produce harmful disinfection products.
Proteinase K can improve meat quality by decomposing muscle fiber of meat, the protein of connective tissue, even including elastin and collagen. Proteinase K helps to clarify the wine during the brewing process.
Adding Proteinase K to laundry detergents or detergents helps to better clean protein-based stains from clothing.
The proteinase K can digest keratin (a hair protein), which allows for its contamination-preventing role in the cosmetics industry, as well as in the food and beverage sector.
Proteinase K is also involved in leather softening and refinement, depilation and softening, and silk degumming.
In recent years, the global proteinase K market has gradually increased in size due to its broad applicability.
The global proteinase K market is mainly distributed in North America, Latin America, Europe, Asia Pacific, Middle East, and Africa. According to its applications, the end-users of proteinase K include contract research organizations, academic institutions, biotechnology companies, and diagnostic centers. And the demand for proteinase K is dramatically increased as the COVID-19 outbreak persistently spread around the world.
The growth of the global proteinase K market is predominantly driven mainly by the growing prevalence of chronic diseases, increased government investment in research, increasing global demand for gene therapy, and increasing consumer awareness. Growing research and technological progress, including some positive developments such as rising demand for nucleic acid purification, improved cloning efficiency, and tissue degradation, dominate the proteinase K market growth. Besides, good product stability and specificity, low production costs, and extensive applications are expected to drive market growth during the forecast period.
The COVID-19, which has attracted worldwide attention, has significantly increased the market demand for Proteinase K.
The purity of a protein reflects its quality to a certain extent. Therefore, the purity of existing Proteinase K in the market and its doping with other proteins and nucleases are the main factors constraining the growth of the Proteinase K market. The low thermal stability of proteinase K limits hydrolysis and aminolysis reactions during industrial production, which also hinder the market growth in the forecast period.
Additionally, the good availability of other Proteinases and the introduction of newer advanced technologies for DNA purification might impede market growth over the forecast period.
Driven by positive trends such as rising manufacturing activity, increasing demand for purification, growing awareness about food contamination, widespread product adoption, and increased investment in research and development by the government, the global proteinase K market is expected to achieve strong growth in the near future.
In recent years, various chronic diseases such as cancer and infectious diseases such as emerging COVID-2019 have continued to spring up, which enhances proteinase K wider applicability in DNA & RNA extraction for in vitro diagnosis. In the medical and pharmaceutical industry, proteinase K plays an important role in the production of Klenow fragments, peptide synthesis, and the hydrolysis of unnecessary proteins in the process of nucleic acid purification or cell culture. Besides, the proteinase K market is likely to cater to the growing demand in new areas such as agriculture, where proteinase K is expected to help purify and effectively use basic mixtures like urea. In the regenerative tissue engineering industry, proteinase K also shows a distinguished promise.