A Highly Stable Biocatalyst Obtained from Covalent Immobilization of a Non-Commercial Cysteine Phytoprotease

Walter David Florencia Ceccacci

doi:10.4172/2155-9821.1000211

Abstract

In this work, araujiain (enzymatic preparation obtained from the latex of Araujia hortorum fruits) was successfully immobilized on glyoxyl-agarose via multipoint covalent attachment. Thus, good efficiency of immobilization and high operational stability of immobilized enzyme were obtained. The activity of araujiain at alkaline pH was significantly improved after immobilization. In addition, immobilized araujiain also showed high activity and good stability, without significant loss in its activity, at temperatures between 37 and 60°C and in the presence of immiscible organic solvents. Immobilized araujiain also showed good performance in a mixture of 50% ethyl acetate in buffer, used for peptide synthesis, with better results than when the free enzyme was used as catalyst. These results indicate that immobilized araujiain via multipoint covalent attachment can be highly stabilized and this method might be used for practical applications of araujiain in hydrolytic and synthetic processes.

Highlights

Proteases are a family of enzymes that play a prominent role in plant physiology
The general conditions for the immobilization of araujiain on glyoxyl-agarose were selected in function of i) the immobilization of other enzymes on glyoxyl-agarose; ii) previous knowledge on the preparation and characterization of the proteolytic extract from A. hortorum (Asclepiadaceae) fruit latex, and iii) the non-deactivating conditions for the proteases present in the enzymatic extract: the use of high pH values does not affect the performance of araujiain [13,26,28,36]
The proteolytic extract obtained from latex of A. hortorum fruits, named araujiain, was characterized in terms of its protein content and specific activity

Summary

Introduction

Proteases are a family of enzymes that play a prominent role in plant physiology These enzymes are catalysts of processes such as storage of proteins during seed germination, activation of proenzymes, and degradation of defective proteins [1,2]. The recent application of proteases to the production of certain peptides and peptides derivatives has received great attention as a viable alternative to the chemical approach [9,10,11,12,13]. This wide range of applications insures proteases the first place in the world market of enzymes

Methods

Results

Conclusion