Novozymes requires novel enzymes from nature’s biodiversity for our discovery pipeline. Transferring the novel enzymes from their naturally occurring source to a recombinant host, or cloning, allows Novozymes to cleanly produce the enzyme quantities required with the desired level of purity.

Enzymes are produced by naturally occurring organisms along with many other proteins and bio products. The desired enzyme is often quite dilute and can be very difficult to purify in sufficient quantities for testing. In addition, other activities in the original organism are not desired in our assays or in the final product we sell. By transferring the enzyme gene into a new production host, Novozymes can eliminate both quantity and quality issues for the enzymes we evaluate. The transfer of the gene to a new host is facilitated by cloning.pYES2.0 expression plasmid. Genes can be cloned into the circular DNA and the resulting construct can be transferred to either yeast or E. coli.

In the past, enzyme products consisted solely of culture broths harvested directly from an enzyme producing organism. Increasingly our products have been cloned into our recombinant production hosts so that we can produce the single enzymes safely, reliably and in quantities needed for the industry.

In the laboratory, cloning facilitates identification of interesting new enzyme activities.Cloning is used at nearly every step from discovering new enzyme leads through to creation of a production microorganism capable of producing the enzyme in large quantities. The products Novozymes sells are the result of genes coding for protein product and it is these genes that the cloning technique involves. Cloning can be used to move the gene from the natural organism to a circular piece of DNA (called a plasmid). This plasmid can then be transferred to a recombinant host where the plasmid can be duplicated into many copies per cell. Such hosts used for cloning are the bacteria Escherichia coli or the yeast Saccharomyces cerevisiae.

Once the gene of interest has been cloned in a recombinant host, the gene itself can be easily analyzed and the protein it produces in small scale can often be studied. Once cloned, the gene is much easier to move around to other hosts that can produce more product for application trials and eventual products. Two such hosts used for producing more product from cloned genes are specially adapted fungus called Aspergillus oryzae and the bacterial host called Bacillus subtilis. B.subtilis is a common soil bacteria but laboratory strains have been selected to be good hosts of cloned genes..