Institute of Industrial Fermentations of CSIC is offering licences recombinant bovine pepsin codified by a synthetic gene for bovine pepsinogen, expressible in various organisms. The resulting enzyme can be used for manufacturing cured cheese varieties, for the generation of bioactive peptides from various proteins or as a general purpose protease. Thus the use of animal tissues is avoided. A biotechnological company is required to licence or to collaborate to reach commercial production levels
DESCRIPTIONA complete cDNA version of the bovine pepsinogen gene has been constructed, with the exception of the original signal peptide. Two different cDNA clones, coming from different bovine cDNA libraries, as well a synthetic sequences added to the 5` end of some oligonucleotides, have been used for this construction. This gene, cloned in appropriate vectors has been expressed in both Saccharomyces cerevisiæ and Escherichia coli, thus allowing the production of pepsinogen. This pepsinogen can be activated by acidification of the medium, thus rendering active pepsin. The enzyme can be purified from the growth medium, in the case of the expression in Saccharomyces cerevisiæ or by breaking the cells, for the enzyme produced in Escherichia coli. The possible applications of recombinant bovine pepsinogen and pepsin include their use for the manufacture of cured cheese varieties in combination with recombinant chymosin. This could help to the acceleration and improvement of cheese maturation as compared to those made by using recombinant chymosin exclusively. However, in contrast to the use of natural bovine pepsin coming from the abomasum of cows, the use of the recombinant enzyme is safer than the use of some animal tissues, specially in relation to the possible presence of prions.On the other side it is known that the action of bovine pepsin on certain proteins, for example on caseins or casein macropeptide (resulting from the action of chymosin on casein), can result on the release of several peptides with biological activities potentially useful for the elaboration of functional foods. Similar to the above situation, the use of recombinant pepsin allows working with safer and pure enzymesRecombiant bovine pepsine can also be used as a general purpose protease.INNOVATIVE ASPECTSThe main advantage of this invention is that it allows the production of bovine pepsinogen and pepsin in microorganisms by the use of recombinant DNA technology. In this way, any risk related to infectious diseases transmitted by animal tissues is avoided. The enzyme can be used for the manufacture of cured cheese varieties, in combination with bovine chymosin, as well as for the production of bioactive peptides or as a general purpose protease.None of the cDNA clones available at the beginning of this work contained the complete bovine pepsinogen coding sequence. Site directed mutagenesis and synthetic sequences have been used in order to create a complete copy of the gene.COMPETITIVE ADVANTAGESIt would be possible to prepare mixtures of recombinant chymosin and pepsin; specially designed in order to accelerate maturation of some specific cured cheese varieties. In this way the advantages of chymosin and pepsin would be simultaneously exploited by avoiding the problems associated to the use of some animal tissues for food production. The facility of purification may constitute an advantae, not only for cheese-making, but also for the use of the enzyme for the production of bioactive peptides or as a general purpose protease.PATENTP200300179, applied for at 2003-01-24CONTACTEstrella Marotoemail: e.maroto@orgc.csic.esphone: +34 91 585 52 51FOR MORE CSIC TECHNOLOGY OFFERS CLICK HERE
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