Hidrólisis enzimática del mucilago de cacao ccn-51 (theobroma cacao l.) para incrementar la concentración de azúcares fermentables

Abstract

The cocoa beans are surrounded by an aromatic pulp rich in fermentable sugars, this pulp is called cocoa mucilage is discarded in the form of exudate during fermentation, the cacao producers, due to lack of knowledge, allow the sea to decompose in its entirety, which causes an environmental contamination, being the means of diverse microorganisms and parasites that sometimes can cause diseases and damages to the health of the inhabitants of these plantations. The objective of the present research was to hydrolyze the carbohydrates present in the cocoa mucilage CCN-51 to increase the concentration of fermentable sugars, using hydrolytic enzymes. The CCN-51 cocoa harvest was carried out in the agricultural farm of the Technical University of Machala, carrying out a process of selection of the ears in optimum state of maturity, proceeded to carry out the harvest, after being harvested, to each ear proceeded to make a cross section to extract the mucilage-coated seeds, said seeds were wrapped in a mesh, so that by squeezing, the cocoa mucilage is obtained in the form of exudate. The physical-chemical characterization of cocoa mucilage was carried out by high performance liquid chromatography, using a SUPERCOGEL C-610H column at 50 ° C, for the measurement of total sugars such as; glucose, sucrose, fructose and cellobiose, moisture and ash by thermogravimetry, pH by potentiometry, total acidity by titration expressed as a percentage of the dominant acid (citric acid), elemental analysis to determine the content of Carbon, Hydrogen, Nitrogen and Sulfur, which it was carried out using a Flas EA1112 elemental analyzer (Thermo Finnigan) within a range of 323-1173 ° K and the measurement of reducing sugars by the 3.5 Dinitro salicylic acid (DNS) method. The process of enzymatic hydrolysis was carried out at a constant concentration of sugars and varying the type of enzyme (α Amylase, β Glucosidase and CTEC2). The results of the characterization of the cocoa mucilage CCN-51 were: 45.03% Carbon, 5.40% Hydrogen, ˂0.05 Sulfur and 0.80% Nitrogen; 17.60 ° Brix, 3.40 pH, 82.02% humidity, 0.94% ash, 1.5% total acidity of which 0.94 belongs to the predominant acid of the mucilage (citric acid) and 15% of reducing sugars and total sugars: 1.78 g / l of Celobiosa, 108.43 g / l of sucrose, 79.78 g / l of fructose and of glucose 67.18 g / l and the results of the elemental analysis carried out were the following: 45.03% of Carbon (C), 5.40% Hydrogen (H), ˂0.05 Sulfur (S), and 0.80% Nitrogen (N). The enzymatic hydrolysis process was carried out for 1 hour at room temperature, for each treatment and samples were taken every 15 min, to perform pH, temperature (° C) and reducing sugar (%) analyzes. After enzymatic hydrolysis, the best conditions corresponded to treatment 3, in which the enzyme CTEC2 (cellulase enzyme blend) was used, whose optimal conditions were pH = 3.50 and temperature of 22 ° C, giving an increase of 25% of sugars reducers. Since the cocoa mucilage is a product rich in fermentable sugars, it can be used in its entirety to obtain derivatives such as; jellies, syrups, jams, nectars and alcohol, improving its process through enzymatic hydrolysis.