Study on EcoRI And HinDIII Immobilization Using Sodium Alginate and Their Restriction Activity on Lambda DNA

In general, enzyme technology refers to the synthesis, separation, purification, and application of enzymes (either soluble or immobilized) for the benefit of humankind. Enzyme technology also includes the use of protein engineering and recombinant DNA technologies to produce more efficient and useful enzymes. One of the main components of the biotechnology industry is the commercial manufacture and use of enzymes ^[1]. The commercialization of enzymes is restricted to a certain degree based on their cost, as more expensive enzymes are not cost-effective due to their poor reusability factor. Notwithstanding their high cost, immobilization of beneficial enzymes with higher repeatability and functional efficiency is seen to be more promising for overcoming these constraints. Enzyme immobilization may be defined as confining the enzyme molecules to a solid matrix or support that is distinct from the one that contains the products or the substrate. The enzymes are attached to or contained within an appropriate support material in order to do this ^[2]. Enzymes are frequently immobilized, primarily to reduce the amount of money that goes toward the process by allowing for multiple uses of the enzyme. This involves physically confining the enzyme so that it cannot dissolve into solution, usually in the form of beads or membranes made of polymer matrix. Because an immobilized enzyme can be easily removed through sieving, it also typically makes downstream processing easier. In contrast, extracting a soluble enzyme from a reactor stream would require significant time, money, and effort ^[3]. 

History of enzyme immobilization:

When Chibata and colleagues created the immobilization of Aspergillus oryzae aminoacylase for the resolution of synthetic racemic D-L amino acids, it was the first time that immobilized enzymes were used in industry. This was published in 1966. The initial stage was the empirical industrial use of immobilized microbes at the start of the 19^th century. Enzyme immobilization's contemporary history began in the late 1940s. Only immobilized single enzymes were employed in the second stage, but more sophisticated systems, such as two-enzyme processes including cofactor regeneration and live cells, were created by the 1970s ^[1]. The first international conference on enzyme engineering was held in 1971, greatly aided by the first practical use of immobilized enzymes. "Immobilized enzymes" was the conference's main focus. At the conference, the term "immobilized enzyme" was suggested. Prior to that, a number of terminologies were in use, including "matrix supported enzyme," "water-insoluble enzyme," "trapped enzyme," and "fix enzyme"^[4].

Components of enzyme immobilization:

The major components for an enzyme immobilization include an enzyme, a support matrix and mode of attachment of a catalyst to the carrier (Figure 1).