Summary of the technology
The described process for collagenase production and purification is rapid, cheap and reproducible. The obtained product is highly active and stable for long period storage (in solution or lyophilised). The features of this novel product are, certainly related to a novel domain organization and architecture not yet described for other collagenases
Description of the technology
The present technology refers to a method for producing and purifying recombinant collagenase from microbial source. The collagenase is over-expressed as a recombinant protein in bacteria. The technology allows obtaining a novel collagenase that shows high level of collagenolytic activity and a remarkable storage stability. It is especially interesting for industrial applications or life-science and clinical research whereas collagen hydrolysis and tissues dissociation is required.
Collagenases have a key role in laboratorial practices involving cell and tissues manipulation in culture, namely for the establishment of primary cell cultures from collageneous organs or tissues. The main constraints associated with commercial collagenases are the high operational cost involved in their production and the low degree of storage stability. The most common strategies of production are based in methodologies involving multi-chromatography steps, with consequent lost of protein and enzymatic activity. The toxicity vs efficiency is also a limiting factor, especially for their application for the treatment of human pathologies associated to an excessive deposition of collagens, such as Dupuytren Syndrome or for the treatment of deep wounds or skin burns for the removal of necrotic tissue. For the reasons presented, several sectors may benefit from the use of this novel collagenase, both in industrial processes for product improving (such as in the food industry), and in clinical research by collagenase implementation on novel cell/tissue culture methodologies or in the development of new therapeutic strategies. Not least is its biotechnological potential for obtaining new formulations to create new cosmetic products.
Main advantages of its use
- It hydrolyses several collagen types: collagen type I (major component of connective tissues; i.e. bone, skin); collagen type II (cartilages); collagen type III (blood vessels).
- It presents high storage stability (lyophilized or in solution).
- Sub-toxic doses the collagenase have a direct specific proliferative effect on osteoblastic cells in vitro and a selective inhibitory effect on osteoclastic production of collagen
- The purification method is based in a single step of affinity chromatography leading to an efficient, fast and inexpensive process of collagenase production
- Healthcare sector
- Industry - cosmetic companies
- Industry - production and commercialization of bioactive compounds for different areas of research (cell biology, neuroscience, biotechnology, microbiology, clinical biology, etc).
- Industry - agro.food sector
- Industry - pharmaceutics