USE OF FORAGE PALM BIOMASS (OPUNTIA FICUS-INDICA MILL) TO OBTAIN CELLULASES AND

REDUCING SUGARS INTENDED FOR THE PRODUCTION OF BIOETHANOL

Fbio Batista Lima Filho1, Carlos Alberto Bispo de Sousa1, Lbia de Sousa Conrado2
1UFPB Universidade Federal da Paraba.
2UFCG Universidade Federal de Campina Grande.

INTRODUCTION 2- Saccharification assay

The demand for renewable energy sources has grown all over the world. The forage palm biomass and the fermentation residue were pretreated separately
Bioethanol obtained from the enzymatic hydrolysis of vegetable biomass is a promising by immersing in 0.1 mol L-1 NaOH for 1 hour in an autoclave at 120C. After cooling,
alternative. This process makes use of cellulase enzymes, whose production costs are the samples were washed and dried (65C ). One gram of each sample was transferred
still high. However, a considerable reduction in production costs can be obtained by to Erlenmeyer flasks containing 50 mL of crude enzyme extract, which were incubated
working with the concept of biorefinery, which is a productive unit that integrates the at 45 C for 40 hours at 100rpm. During the period of enzymatic hydrolysis, samples
generation of energy and value-added products high from biomass, with little waste and were collected to determine the reducing sugars released in the process.
emissions, bringing new opportunities for profit.
The forage palm (Opuntia ficus-indica Mill), is a cactus adapted to regions with semi-
arid climate. It is a raw material with potential application to provide lignocellulosic RESULTS AND DISCUSSION
biomass to obtain bioethanol in these regions. The objective of this research was to
Table 1. Cellulolytic activity of crude enzyme extract
obtain cellulase enzymes from the solid state fermentation of forage palm biomass and
verify the applicability of the enzymes obtained in the hydrolysis of the forage palm Enzyme Activity (U/mL)
biomass and the fermentation residue to obtain fermentable sugars. CMCase 0,181 0,006
FPase 0,101 0,005
Cellobiase 0,028 0,007
MATERIALS AND METHODS
1- Solid state fermentation
To obtain biomass of forage palm, cladodes were cut, dried (60 C) and milled. Part
of the biomass was used to obtain cellulases by solid state fermentation, which had the
following parameters: initial moisture of 90%, supplementation of 1% (NH4)2SO4,
temperature of 28 C and inoculation with 107 spores of Trichoderma reesei LCB 48 per
gram of moist biomass. The crude extract containing the cellulases was obtained by
leaching with 0.5 Mol.L-1 sodium citrate buffer, pH 4.8 (Figure 1)

Figure 1. Production of crude enzyme extract from the solid state fermentation of forage palm
Figure 3. Results of the enzymatic hydrolysis assay

Although the crude enzyme extract did not receive any additional treatment and
Biomass presented low values of cellulolytic activity, the saccharification study resulted in an
increase in the concentration of reducing sugars (glucose), which means that the
enzymes present in the extract were able to catalyze the cellulose cleavage of the palm
H2O and (NH4)2SO4 hydration biomass and the fermentation residue in reducing sugars. Maximum yields of 334.4
mg/L.h for saccharification of palm biomass and 308.2 mg/L.h were observed for the
sterilization fermentation residue in 4 hours, decreasing from that period by possible inhibitory
effects caused by the reducing sugars produced.
spore counts inoculation
(107 espores per gram)
CONCLUSION
Incubation

The forage palm biomass can be used as raw material to obtain cellulase enzymes
Leaching and also to provide reducing sugars for the production of bioethanol in biorefineries
installed in semi-arid regions. In addition, the residue generated from the fermentation
process can also be used for the enzymatic saccharification, representing a saving of
Crude enzyme extract biomass in the process. However, further studies are needed for process otimization.

Figure 2. Fluxogram of the fermentation process to obtain cellulases ACKNOWLEDGMENTS

CAPES for the opportunity to perform this research.