EXPECTED RESULTS

Research and characterise the products of commercial biotechnological importance such as cryophilic proteases (acting at low temperatures) obtained from a marine source: Antarctic krill, to be used as enzymes for detergents and wound-healing agents and chitin as adsorbent of toxic contaminating metals. This is achieved using advanced knowledge of biochemistry, biotechnology and characterisation, purification and chemistry of proteins including sequentiation and cloning of the proteases. The purified enzyme of the pool IV (Juanasa) has an important potential economic impact. Clearly, compared with the enzymes of standard detergents (subtilisina), it has a spectre of activities much more extended with high activity at 20ºC and also at higher temperatures. It shows high activity at alkaline pH, which is an expected characteristic of proteases in detergents. According to the results of the kinetic studies of adsorption of heavy metals it is possible to conclude that chitosan presents very fast adsorption kinetics, comparable to the observed adsorption kinetics in commercial activated carbon

SUMMARY

The use of enzymes at low temperatures has great potencial, both in the applications to be studied in this project (detergent proteases and wound debriding agents) and others in terms of lower energy costs and the possibility of therapeutic applications. In recent years there has been an interest in finding uses for chitin such as an adsorbant for contaminant metal ions.

Antarctic krill is extremely interesting as a source of enzymes with unique properties. particularly proteases which have high activities at low temperatures (cryophilic). These enzymes are ideal for use in detergents and as wound healing and debriding agents. There exists an important market for such products. In the first part of this project we have obtained samples of frozen krill (-80°C) that can be obtained internationally from different sources (e.g. Taiyo Fisheries, Japan, Taiwan fisheries Research Institute, Hiromatsu Fishery, Japan and the British Antarctic Survey, Cambridge). These samples were obtained by Bios Chile/Chiron and by Novo-Nordisk and were shipped frozen in dry ice to the lab of Dr. Asenjo where the enzymatic characterization has been carried out, this guarantees the supply of an appropriate raw material for the project.

The Center of Biochemical Engineering and Biotechnology of the Universidad de Chile has very well qualified academics of international standing with the necessary expertise to successfully carry out the proposed research. This centre is comparable with advanced research centres in Europe and the USA and. is fully equipped with the most modern equipment available thanks to a grant from the Fundación Andes, top level academics and researchers and a large number of Ph.D. candidates. The active participation of the company Bios Chile in the project will be important to assure its success. Bios Chile/Chiron will also carry out the work on cloning of the enzymes both for the fine characterization of the protein and to present a viable and economic alternative in case the best enzymes are in krill in relatively low concentrations or if its capture becomes difficult.

Marketing of the enzymes at an internationallevel will be done through Novo-Nordisk and Chiron who handle an important fraction of the world market of commercial proteases (ca. 30%) as well as 50% of that of recombinant insulin made in yeast. Both companies are already participating in the project.

This project involves the use of advanced biotecnological and biochemical techniques including the characterization, purification and chemistry of proteins, necessary to produce and characterize the cryophilic proteases and to study their action as ingredients of detergent and wound debriding preparations and to investigate the ability of chitin to adsorb toxic metal ions from waste streams.

MAIN PROJECT IMPACTS

SOCIO/ECONOMIC IMPACTS

SCIENTIFIC/TECHNOLOGICAL IMPACTS

The scientific-technological impact is very important because it integrates the use of advanced scientific knowledge in the techniques of purification, characterisation and the chemistry of sequentiation and cloning, applied to a product of commercial relevance as the Antarctic krill. The generation of new knowledge and technologies may now be transferred to industry. Also human resources in the area have been thoroughly trained so that new lines of research, development and application can be initiated.

INSTITUTIONAL IMPACTS

This project contributes to the development of the infrastructure, human resources and its recognition as a Centre of Excellence in Chile and Latin America. The areas under study in the project reflect the areas of interest of the Centre as well as those of Bios Chile whose collaboration has been fundamental

ENVIRONMENTAL IMPACTS

The development and use of chitin as adsorbent of toxic metals in effluents of the mineral industry represents a high environmental impact. Although the use of enzymes arising from krill has a neutral environmental effect, the development of cryophilic proteases (low temperatures) decreases the consumption of energy in these processes. The development of sequentiation and cloning of the proteases provides an important alternative for the production of these enzymes in fermentation reactors (for example the beer industry), that is why it is important to protect the krill population in the Antarctic seas.