XXI SIMPÓSIO NACIONAL DE BIOPROCESSOS

XII SIMPÓSIO DE HIDRÓLISE ENZIMÁTICA DE BIOMASSA

3 a 6 de setembro de 2017
Aracaju, Sergipe, Brasil

OPTIMIZATION OF DILUTE ACID HYDROLYSIS OF MALT BAGASSE AIMING

FOR THE PRODUCTION OF FERMENTABLE SUGARS

A. N.R. Mota 1 , S.C.M. Silva 1 , Y. M. Anjos 1, B. Sarrouh2

1
Universidade Federal de São João del-Rei, Campus Alto Paraopeba - Departamento de Eng. Química,
2
Universidade Federal de São João del-Rei, Campus Alto Paraopeba - Departamento de Química,
Biotecnologia e Eng. Bioprocessos
E-mail: yunaanjos@gmail.com

ABSTRACT

The lignocellulosic materials are plentiful sources of organic compounds and present important
potential of use as raw material in industrial processes for the production of food, fuels, chemical
compounds and enzymes. These materials include agro-industrial residues such as coffee husk, rice
straws, sugarcane bagasse, corncob and malt bagasse, which have a limited used in industrial
processes. Several methods have been employed for the hydrolysis of the components of
lignocellulosic materials for the release of fermentable sugars. Acid hydrolysis stands out as an
example of these methods. The objective of this work was to assess the diluted acid hydrolysis of
malt bagasse under different conditions of reaction time and acid concentration in order to obtain a
hemicellulosic hydrolysate rich in sugars. According to the statistical analysis carried out, it was
possible to verify that the operational conditions to maximize the sugar concentration were at a
hydrolysis time of 10 minutes and a concentration of 150 mg.g-1 of H2SO4.

1. INTRODUCTION
The necessity to produce new fuels has led to the development of new technologies aiming for the
production second generation ethanol using agro-industrial waste. In this context, biomass
hydrolysis represents an efficient way to realize the breakage and consequent release of
fermentable sugars which will be used to produce alcohol (DA ROSA, 2009). Barley is a grain rich in
amide and protein, and it is produced in large scale in Brazil. It is known that malt bagasse, a
byproduct of brewery, is basically constituted by the husks of barley grain resulted from the
production of the wort. Since the brewing industry produces large quantities of malt, new
technologies have been sought for the processing of this residue into high value products. Several
studies have pointed to the use of acid hydrolysis of malt bagasse in order to release fermentable
 XXI SIMPÓSIO NACIONAL DE BIOPROCESSOS
XII SIMPÓSIO DE HIDRÓLISE ENZIMÁTICA DE BIOMASSA

3 a 6 de setembro de 2017
Aracaju, Sergipe, Brasil

sugars, mainly xylose and glucose, for the production of second generation ethanol as well as other
bioproducts of commercial interest (PELA, 2014).

In this work, dilute acid hydrolysis was studied, under different conditions, aiming for the
solubilization of the hemicellulosic fraction of malt bagasse and subsequent liberation of
fermentable sugars. In this process, protons were released from the acid, which were responsible
for breaking of the “1.4-Glycosidic” bonds in main chain; “-1.2-, “-1.3-, “-1.6-glycosidic bonds in side
chain of the hemicellulosic polymer. Several compounds such as xylose, arabinose and glucose were
liberated with the breakage of these bonds. (DUFF, 1996).

2. MATERIAL AND METHODS

2.1. Preparation and acid hydrolysis of malt bagasse

The malt bagasse used in the trials was supplied by a small-scale brewer. The bagasse was crushed
and sieved between 12 and 32 meshes, then it was dried in drying oven for later use in the
hydrolysis process. In each 125 mL Erlenmeyer, 5 g of dried malt bagasse were added to a volume of
50 mL of acid solution respecting a ratio of 1:10 (solid: liquid). The experiments were carried out
using factorial design 22, varying the reaction time (10 and 30 minutes) and sulfuric acid
concentration (50 and 150 mg.g -1 H2SO4) and a central point (PC) in triplicate (20 minutes and 100
mg.g -1 H2SO4). The acid hydrolysis trials were performed in an autoclave at 115 °C. The Erlenmeyers
were sealed to avoid possible losses of water mass.

2.2. Determination of reducing sugar concentration

The method used to determinate the amount of reducing sugars was the DNS method. It was
collected 50 μL of each hydrolyzed sample, which was previously filtered, and transferred to a test
tube. 3 ml of DNS solution was added to each trial tube. 50 μL of distilled water and 3 mL of DNS
solution were used for the blank sample. The trial tubes were placed in a water bath at
approximately 100 ° C for 5 minutes. The tubes were withdrawn and kept in an ice bath for 5 min.
Then, the absorbance of the samples was read using a spectrophotometer (Biospectro SP 220) at
540 nm.

3. RESULTS
3.1. Determination of reducing sugar concentration
 XXI SIMPÓSIO NACIONAL DE BIOPROCESSOS
XII SIMPÓSIO DE HIDRÓLISE ENZIMÁTICA DE BIOMASSA

3 a 6 de setembro de 2017
Aracaju, Sergipe, Brasil

The hydrolysates obtained under different acid hydrolysis conditions were analyzed to determine
the concentration of sugars from the hemicellulosic fraction during the reaction. The recovered
sugar values of the analyzed samples are shown in Table 1.

Table 1. Recovery of sugars from hydrolyzed samples

Time Conc. of H2SO4 Fermentable Sugars
Trial
(min) (mg g-1) (g L-1)
1 10 50 1,29
2 10 150 1,60
3 20 100 1,33
4 20 100 1,34
5 20 100 1,36
6 30 50 1,05
7 30 150 1,38

A variance analysis (ANOVA) was performed to determine the effects of variables and their
interactions. As shown in the Pareto chart (Figure 1.a), it was observed that the reaction time
variable (B) had a significant influence on the acid hydrolysis process of the malt bagasse. However,
this influence occurs in a negative way since the longer the hydrolysis time, the lower the amount of
sugars obtained. It could be noted through the Figure 1.b. that the increase in acid concentration
(positive slope effect line) favors the attaining of sugars, whereas shorter hydrolysis times (negative
slope effect) induce higher sugar levels.

Figure 1.a. The Pareto chart to estimate the effects of the variables: (A) H2SO4 concentration, (B) time, and the
interaction between them (AB) in the acid hydrolysis process of malt bagasse. 1.b. The Chart to analyze the influence of
reaction time and H2SO4 concentration on sugar production.

The Figure 2 represents the Response Surface of the total sugars obtained in the acid hydrolysis
trials of the malt baggace as a function of the variables of acid concentration and reaction time.
 XXI SIMPÓSIO NACIONAL DE BIOPROCESSOS
XII SIMPÓSIO DE HIDRÓLISE ENZIMÁTICA DE BIOMASSA

3 a 6 de setembro de 2017
Aracaju, Sergipe, Brasil

Figure 2. Response surface of the total sugars released as a function of the variables of acid concentration and reaction
time.
As shown in Figure 2, it was observed that the optimum conditions of hydrolysis to reach maximum
concentrations of sugars are at lower reaction time (10 min) and higher acid concentration (150
mg.g-1 of H2SO4). Within this work region, it is obtained the optimized concentration of 1.8 mg.L -1 of
total reducing sugars.

4. CONCLUSION
In this work, the diluted acid hydrolysis of malt bagasse was evaluated under different conditions of
reaction time and concentration of H 2SO4, aiming to obtain a hydrolysate rich in fermentable sugars.
It was noticed that, high acid concentrations provoked an increases in the recovery of fermentable
sugars. Regarding the reaction time of the hydrolysis it was observed that, an increase in this
variable promoted the degradation of sugars, possibly into phenolic compounds.

5. ACKNOWLEDGMENT

CNPq and FAPEMIG

6. REFERENCES
DA ROSA, Sergio Eduardo Silveira; GARCIA, Jorge Luiz Faria. O etanol de segunda geração: limites e
oportunidades. Revista do BNDES, v. 32, p. 118, 2009.
DUFF, S. J. B., MURRAY, W. D. Bioconversion of forest products industry waste cellulosics to fuel
etanol: a review. Bioresource Technology, 55:33, 1996.
PELÁ, A. L. B. Etanol de segunda geração a partir do bagaço de cana-de-açúcar: análise do ciclo de
vida com relação às emissões de CO2. 2014.