1. HYDROLYSIS OF PORCINE GLOBIN FOR FOOD USES C. Álvarez, M. Rendueles and M. Díaz Chemical Engineering and Environmental Technology. University of Oviedo. C/ Julián Clavería 8. 33071.Oviedo. Spain, E-mail: mrenduel@uniovi.esmrenduel@uniovi.es TBRTBR Technology of Bioprocesses and Reactors University of Oviedo HYDROLYSIS OF PORCINE GLOBIN FOR FOOD USES INTRODUCTION MATERIALS AND METHODS RESULTS AND DISCUSION CONCLUSIONS BIBLIOGRAPHY PERSPECTIVES Porcine blood from slaughtery house Refrigeration and centrifugation Plasma Red cells Fractionation Haemoglobin extraction with chloroform Lysis, osmotic shock Globin precipitation with acetone and wash with destilled water Hydrolysis ACID HYDROLYSIS The figures on the left shows how the size of hydrolysated evolves from whole proteins to small peptides, as the acid broke the peptidic bonds. The yellow line points the elution time of 10 Kda. The second gruop of peaks matches 1KDa size, and the third one to 0.1 Kda. The first peak decrease his height and the second one increase it. BASIC HYDROLYSIS This hydrolysis is less effective than acid is. The globin never is completly hydrolysated in experiment times, but the towards is clearly to produce small peptides. If the time of assay is extended all the peptides will be under 10 Kda. Some agregates are detected in the earlier stages. The isolated proteins contained in the waste blood could be recovered and used in food products. The aim goal of this work is analyze the hydrolysis of haemoglobin at different (acid or basis) concentration, and establish the optimum conditions to produce animal food according to the European normative (CE n° 1774/2002), which imposes 10 KDa as the maximum molecular weight of peptides. CONDITIONS OF HYDROLYSIS The effect of H 2 SO 4 and NaOH at different concentrations was studied allong the time.. MOLECULAR WEIGHT ANALYSIS Gel filtration was the method employed. The column (Superdex Peptide 10/300 GL) was calibrated with molecular weight markers. The buffer (230 ml of KCl 0.2M, 50 ml of HCl 1M, 30 ml of NaCl 5Mand 690 ml of Destilled water per litre) and the samples were gently mixed for 24 hours. SETUP OF HYDROLYSIS Globin Acid or alkali T = 70 ºC SAMPLING and neutralization with alkali or acid to stop the hydrolysis -.Radha, C., Kumar and P.R., Prakash V., 2007. Preparation and characterization of a protein hydrolysate from an oilseed flour mixture. Food Chemistry, 106, 1166-1174. - Landry, J. and Delhaye, S., 1996. A Simple and Rapid Procedure for Hydrolyzing minute Amounts of Proteins with Alkali. Analytical Biochemistry 243, 191–194. - Tsugita and Schefflere, 1982. A Rapid Method for Acid Hydrolysis of Protein with a Mixture of Trifluoroacetic Acid and Hydrochloric Acid. European Journal of Biochemistery, 124, 585-588. - Jiménez-Castaño, L., Villamiel, M. and López-Fandiño, R., 2006.Glycosylation of individual whey proteins by Maillard reaction using dextran of different molecular mass. Food Hydrocolloids 21, 433– 443. A)Chemical hydrolysis is a cheap and fast way to produce protein hydrolisated compare with enzymatic one. B)The acid hydrolysis is more effective than basic is, even at low molarity. C)The peptides produced are according to european normative. D)This procedure allows to re-use a very contaminant waste in the food industry. Functional properties (foam, solubility, gelling..) were measured The results were poor compared with patrons (BSA or egg albumin) How to improve the solubility and other properties?? Trough Maillard’s reaction, i.e. conjugation of dextran and proteins. Reducing carbohydrate Amino compound Freeze-dry N-Glycosilamine 80 ºC, 60-120 min. T1T2T3 17.85 10.66 11.14 20.68 19.03 17.98 21.99 11.13 17.97 20.64 19.08 19.04