Abstract: Enzymatic modification of rice flour can produce highly functional derivatives use in food industries. This study aimed to evaluate the physical properties and resistant starch content of rice flour residues hydrolyzed by α-amylase. Rice flour hydrolyzed by α-amylase (60 and 300 u/g) for 1, 24 and 48 hours were investigated. Increasing enzyme concentration and hydrolysis time resulted in decreased rice flour residue’s lightness (L*) but increased redness (a*) and yellowness (b*) of rice flour residues. The resistant starch content and peak viscosity increased when hydrolysis time increased. Pasting temperature, trough viscosity, breakdown, final viscosity, setback and peak time of the hydrolyzed flours were not significantly different (p>0.05). The morphology of native flour was smooth without observable pores and polygonal with sharp angles and edges. However, after hydrolysis, granules with a slightly rough and porous surface were observed and a rough and porous surface was increased with increasing hydrolyzed time. The X-ray diffraction patterns of native flour showed A-type configuration, which hydrolyzed flour showed almost 0% crystallinity indicated that both amorphous and crystalline structures of starch were simultaneously hydrolyzed by α-amylase.
Abstract: The enzymatic hydrolysis of lignocellulosic biomass is one of the obstacles in the process of sugar production, due to the presence of lignin that protects the cellulose molecules against cellulases. Although the pretreatment of lignocellulose in ionic liquid (IL) system has been receiving a lot of interest; however, it requires IL removal with an anti-solvent in order to proceed with the enzymatic hydrolysis. At this point, introducing a compatible cellulase enzyme seems more efficient in this process. A cellulase enzyme that was produced by Trichoderma reesei on palm kernel cake (PKC) exhibited a promising stability in several ILs. The enzyme called PKC-Cel was tested for its optimum pH and temperature as well as its molecular weight. One among evaluated ILs, 1,3-diethylimidazolium dimethyl phosphate [DEMIM] DMP was applied in this study. Evaluation of six factors was executed in Stat-Ease Design Expert V.9, definitive screening design, which are IL/ buffer ratio, temperature, hydrolysis retention time, biomass loading, cellulase loading and empty fruit bunches (EFB) particle size. According to the obtained data, IL-enzyme system shows the highest sugar concentration at 70 °C, 27 hours, 10% IL-buffer, 35% biomass loading, 60 Units/g cellulase and 200 μm particle size. As concluded from the obtained data, not only the PKC-Cel was stable in the presence of the IL, also it was actually stable at a higher temperature than its optimum one. The reducing sugar obtained was 53.468±4.58 g/L which was equivalent to 0.3055 g reducing sugar/g EFB. This approach opens an insight for more studies in order to understand the actual effect of ILs on cellulases and their interactions in the aqueous system. It could also benefit in an efficient production of bioethanol from lignocellulosic biomass.
Abstract: Among agricultural residues, sugarcane bagasse is one of the most convincing raw materials for the production of bioethanol due to its availability, and low cost through enzymatic hydrolysis and yeast fermentation. A pretreatment step is needed to enhance the enzymatic step. In this study, sugarcane bagasse (SCB), one of the most abundant agricultural residues in Thailand, was pretreated biologically with various microorganisms of white-rot fungus—Phanerochaete sordid (SK 7), Cellulomonas sp. (TISTR 784), and strain A 002 (Bacillus subtilis isolated from Thai higher termites). All samples with various microbial pretreatments were further hydrolyzed enzymatically by a commercial enzyme obtained from Aspergillus niger. The results showed that the pretreatment with the white-rot fungus gave the highest glucose concentration around two-fold higher when compared with the others.
Abstract: The hydrolysis of lactose using β-galactosidase is one of the most promising biotechnological applications, which has wide range of potential applications in food processing industries. However, due to intracellular location of the yeast enzyme, and expensive extraction methods, the industrial applications of enzymatic hydrolysis processes are being hampered. The use of permeabilization technique can help to overcome the problems associated with enzyme extraction and purification of yeast cells and to develop the economically viable process for the utilization of whole cell biocatalysts in food industries. In the present investigation, standardization of permeabilization process of novel yeast isolate was carried out using a statistical model approach known as Response Surface Methodology (RSM) to achieve maximal b-galactosidase activity. The optimum operating conditions for permeabilization process for optimal β-galactosidase activity obtained by RSM were 1:1 ratio of toluene (25%, v/v) and ethanol (50%, v/v), 25.0 oC temperature and treatment time of 12 min, which displayed enzyme activity of 1.71 IU /mg DW.
Abstract: Using an enzyme of known specificity the hydrolysis of protein was carried out in a controlled manner. The aim was to obtain oligopeptides being the so-called active peptides or their direct precursors. An original way of expression of the protein hydrolysis kinetics was introduced. Peptide bonds contained in the protein were recognized as a diverse-quality substrate for hydrolysis by the applied protease. This assumption was positively verified taking as an example the hydrolysis of albumin by thermolysin. Peptide linkages for this system should be divided into at least four groups. One of them is a group of bonds non-hydrolyzable by this enzyme. These that are broken are hydrolyzed at a rate that differs even by tens of thousands of times. Designated kinetic constants were k'F = 10991.4 L/g.h, k'M = 14.83L/g.h, k'S about 10-1 L/g.h for fast, medium and slow bonds, respectively. Moreover, a procedure for unfolding of the protein, conducive to the improved susceptibility to enzymatic hydrolysis (approximately three-fold increase in the rate) was proposed.
Abstract: Enzymatic hydrolysis is one of the major steps involved in the conversion from sugarcane bagasse to yield ethanol. This process offers potential for yields and selectivity higher, lower energy costs and milder operating conditions than chemical processes. However, the presence of some factors such as lignin content, crystallinity degree of the cellulose, and particle sizes, limits the digestibility of the cellulose present in the lignocellulosic biomasses. Pretreatment aims to improve the access of the enzyme to the substrate. In this study sugarcane bagasse was submitted chemical pretreatment that consisted of two consecutive steps, the first with dilute sulfuric acid (1 % (v/v) H2SO4), and the second with alkaline solutions with different concentrations of NaOH (1, 2, 3 and 4 % (w/v)). Thermal Analysis (TG/ DTG and DTA) was used to evaluate hemicellulose, cellulose and lignin contents in the samples. Scanning Electron Microscopy (SEM) was used to evaluate the morphological structures of the in natura and chemically treated samples. Results showed that pretreatments were effective in chemical degradation of lignocellulosic materials of the samples, and also was possible to observe the morphological changes occurring in the biomasses after pretreatments.
Abstract: Nowadays there is a growing interest in biofuel production in most countries because of the increasing concerns about hydrocarbon fuel shortage and global climate changes, also for enhancing agricultural economy and producing local needs for transportation fuel. Ethanol can be produced from biomass by the hydrolysis and sugar fermentation processes. In this study ethanol was produced without using expensive commercial enzymes from sugarcane bagasse. Alkali pretreatment was used to prepare biomass before enzymatic hydrolysis. The comparison between NaOH, KOH and Ca(OH)2 shows NaOH is more effective on bagasse. The required enzymes for biomass hydrolysis were produced from sugarcane solid state fermentation via two fungi: Trichoderma longibrachiatum and Aspergillus niger. The results show that the produced enzyme solution via A. niger has functioned better than T. longibrachiatum. Ethanol was produced by simultaneous saccharification and fermentation (SSF) with crude enzyme solution from T. longibrachiatum and Saccharomyces cerevisiae yeast. To evaluate this procedure, SSF of pretreated bagasse was also done using Celluclast 1.5L by Novozymes. The yield of ethanol production by commercial enzyme and produced enzyme solution via T. longibrachiatum was 81% and 50% respectively.
Abstract: Pretreatment is an essential step in the conversion of
lignocellulosic biomass to fermentable sugar that used for biobutanol
production. Among pretreatment processes, microwave is considered
to improve pretreatment efficiency due to its high heating efficiency,
easy operation, and easily to combine with chemical reaction. The
main objectives of this work are to investigate the feasibility of
microwave pretreatment to enhance enzymatic hydrolysis of
corncobs and to determine the optimal conditions using response
surface methodology. Corncobs were pretreated via two-stage
pretreatment in dilute sodium hydroxide (2 %) followed by dilute
sulfuric acid 1 %. Pretreated corncobs were subjected to enzymatic
hydrolysis to produce reducing sugar. Statistical experimental design
was used to optimize pretreatment parameters including temperature,
residence time and solid-to-liquid ratio to achieve the highest amount
of glucose. The results revealed that solid-to-liquid ratio and
temperature had a significant effect on the amount of glucose.
Abstract: Enzymatic hydrolysis of starch from natural sources
finds potential application in commercial production of alcoholic
beverage and bioethanol. In this study the effect of starch
concentration, temperature, time and enzyme concentration were
studied and optimized for hydrolysis of Potato starch powder (of
mesh 80/120) into glucose syrup by immobilized (using Sodium
arginate) α-amylase using central composite design. The
experimental result on enzymatic hydrolysis of Potato starch was
subjected to multiple linear regression analysis using MINITAB 14
software. Positive linear effect of starch concentration, enzyme
concentration and time was observed on hydrolysis of Potato starch
by α-amylase. The statistical significance of the model was validated
by F-test for analysis of variance (p ≤ 0.01). The optimum value of
starch concentration, enzyme concentration, temperature, time and
were found to be 6% (w/v), 2% (w/v), 40°C and 80min respectively.
The maximum glucose yield at optimum condition was 2.34 mg/mL.
Abstract: Wheat has a bimodal starch granule population and the dependency of the rate of enzymatic hydrolysis on particle size has been investigated. Ungelatinised wheaten starch granules were separated into two populations by sedimentation and decantation. Particle size was analysed by laser diffraction and morphological characteristics were viewed using SEM. The sedimentation technique though lengthy, gave satisfactory separation of the granules. Samples (10μm and original) were digested with a-amylase using a dialysis model. Granules of 10μm (p10μm. Moreover, the digestion rate was dependent on particle size whereby smaller granules produced higher rate of release. The methodology and results reported here can be used as a basis for further evaluations designed to delay the release of glucose during the digestion of native starches.
Abstract: The present study addresses problems and solutions
related to new functional food production. Wheat (Triticum aestivum
L) bran obtained from industrial mill company “Dobeles
dzirnavieks”, was used to investigate them as raw material like
nutrients for Bifidobacterium lactis Bb-12. Enzymatic hydrolysis of
wheat bran starch was carried out by α-amylase from Bacillus
amyloliquefaciens (Sigma Aldrich). The Viscozyme L purchased
from (Sigma Aldrich) were used for reducing released sugar.
Bifidibacterium lactis Bb-12 purchased from (Probio-Tec® CHR
Hansen) was cultivated in enzymatically hydrolysed wheat bran
mash. All procedures ensured the number of active Bifidobacterium
lactis Bb-12 in the final product reached 105 CFUg-1. After enzymatic
and bacterial fermentations sample were freeze dried for analysis of
chemical compounds. All experiments were performed at Faculty of
Food Technology of Latvia University of Agriculture in January-
March 2013. The obtained results show that both types of wheat bran
(enzymatically treated and non-treated) influenced the fermentative
activity and number of Bifidibacterium lactis Bb-12 viable in wheat
bran mash. Amount of acidity strongly increase during the wheat
bran mash fermentation. The main objective of this work was to
create low-energy functional enzymatically and bacterially treated
food from wheat bran using enzymatic hydrolysis of carbohydrates
and following cultivation of Bifidobacterium lactis Bb-12.
Abstract: Agricultural residue such as oil palm fronds (OPF) is
cheap, widespread and available throughout the year. Hemicelluloses
extracted from OPF can be hydrolyzed to their monomers and used in
production of xylooligosaccharides (XOs). The objective of the
present study was to optimize the enzymatic hydrolysis process of
OPF hemicellulose by varying pH, temperature, enzyme and substrate
concentration for production of XOs. Hemicelluloses was extracted
from OPF by using 3 M potassium hydroxide (KOH) at temperature of
40°C for 4 hrs and stirred at 400 rpm. The hemicellulose was then
hydrolyzed using Trichoderma longibrachiatum xylanase at different
pH, temperature, enzyme and substrate concentration. XOs were
characterized based on reducing sugar determination. The optimum
conditions to produced XOs from OPF hemicellulose was obtained at
pH 4.6, temperature of 40°C , enzyme concentration of 2 U/mL and
2% substrate concentration. The results established the suitability of
oil palm fronds as raw material for production of XOs.
Abstract: Cassava bagasse is one of major biomass wastes in Thailand from starch processing industry, which contains high starch content of about 60%. The object of this study was to investigate the optimal condition for hydrothermally pretreating cassava baggasses with or without acid addition. The pretreated samples were measured reducing sugar yield directly or after enzymatic hydrolysis (alpha-amylase). In enzymatic hydrolysis, the highest reducing sugar content was obtained under hydrothermal conditions for at 125oC for 30 min. The result shows that pretreating cassava baggasses increased the efficiency of enzymatic hydrolysis. For acid hydrolysis, pretreating cassava baggasses with sulfuric acid at 120oC for 60 min gave a maximum reducing sugar yield. In this study, sulfuric acid had a greater capacity for hydrolyzing cassava baggasses than phosphoric acid. In comparison, dilute acid hydrolysis to provide a higher yield of reducing sugar than the enzymatic hydrolysis combined hydrothermal pretreatment. However, enzymatic hydrolysis in a combination with hydrothermal pretreatment was an alternative to enhance efficiency reducing sugar production from cassava bagasse.
Abstract: Lignocellulosic materials are new targeted source to
produce second generation biofuels like biobutanol. However, this
process is significantly resisted by the native structure of biomass.
Therefore, pretreatment process is always essential to remove
hemicelluloses and lignin prior to the enzymatic hydrolysis.
The goals of pretreatment are removing hemicelluloses and
lignin, increasing biomass porosity, and increasing the enzyme
accessibility. The main goal of this research is to study the important
variables such as pretreatment temperature and time, which can give
the highest total sugar yield in pretreatment step by using dilute
phosphoric acid. After pretreatment, the highest total sugar yield of
13.61 g/L was obtained under an optimal condition at 140°C for 10
min of pretreatment time by using 1.75% (w/w) H3PO4 and at 15:1
liquid to solid ratio. The total sugar yield of two-stage process
(pretreatment+enzymatic hydrolysis) of 27.38 g/L was obtained.
Abstract: Pretreatment of lignocellulosic biomass materials from
poplar, acacia, oak, and fir with different ionic liquids (ILs)
containing 1-alkyl-3-methyl-imidazolium cations and various anions
has been carried out. The dissolved cellulose from biomass was
precipitated by adding anti-solvents into the solution and vigorous
stirring. Commercial cellulases Celluclast 1.5L and Accelerase 1000
have been used for hydrolysis of untreated and pretreated
lignocellulosic biomass. Among the tested ILs, [Emim]COOCH3
showed the best efficiency, resulting in highest amount of liberated
reducing sugars. Pretreatment of lignocellulosic biomass using
glycerol-ionic liquids combined pretreatment and dilute acid-ionic
liquids combined pretreatment were evaluated and compared with
glycerol pretreatment, ionic liquids pretreatment and dilute acid
pretreatment.
Abstract: The complexity of lignocellulosic biomass requires
a pretreatment step to improve the yield of fermentable sugars. The
efficient pretreatment of corn cobs using microwave and potassium
hydroxide and enzymatic hydrolysis was investigated. The
objective of this work was to characterize the optimal condition of
pretreatment of corn cobs using microwave and potassium
hydroxide enhance enzymatic hydrolysis. Corn cobs were
submerged in different potassium hydroxide concentration at varies
temperature and resident time. The pretreated corn cobs were
hydrolyzed to produce the reducing sugar for analysis. The
morphology and microstructure of samples were investigated by
Thermal gravimetric analysis (TGA, scanning electron microscope
(SEM), X-ray diffraction (XRD). The results showed that lignin
and hemicellulose were removed by microwave/potassium
hydroxide pretreatment. The crystallinity of the pretreated corn
cobs was higher than the untreated. This method was compared
with autoclave and conventional heating method. The results
indicated that microwave-alkali treatment was an efficient way to
improve the enzymatic hydrolysis rate by increasing its
accessibility hydrolysis enzymes.
Abstract: The complex structure of lignocellulose leads to great
difficulties in converting it to fermentable sugars for the ethanol
production. The major hydrolysis impediments are the crystallinity of
cellulose and the lignin content. To improve the efficiency of
enzymatic hydrolysis, microbial pretreatment of corncob was
investigated using two bacterial strains of Bacillus subtilis A 002 and
Cellulomonas sp. TISTR 784 (expected to break open the crystalline
part of cellulose) and lignin-degrading fungus, Phanerochaete
sordida SK7 (expected to remove lignin from lignocellulose). The
microbial pretreatment was carried out with each strain under its
optimum conditions. The pretreated corncob samples were further
hydrolyzed to produce reducing glucose with low amounts of
commercial cellulase (25 U·g-1 corncob) from Aspergillus niger. The
corncob samples were determined for composition change by X-ray
diffraction (XRD), Fourier transform infrared spectroscopy (FTIR),
and scanning electron microscope (SEM). According to the results,
the microbial pretreatment with fungus, P. sordida SK7 was the most
effective for enhancing enzymatic hydrolysis, approximately, 40%
improvement.
Abstract: Enzymatic hydrolysis of starch from natural sources
finds potential application in commercial production of alcoholic
beverage and bioethanol. In this study the effect of starch
concentration, temperature, time and enzyme concentration were
studied and optimized for hydrolysis of cassava (Manihot esculenta)
starch powder (of mesh 80/120) into glucose syrup by immobilized
(using Polyacrylamide gel) a-amylase using central composite
design. The experimental result on enzymatic hydrolysis of cassava
starch was subjected to multiple linear regression analysis using
MINITAB 14 software. Positive linear effect of starch concentration,
enzyme concentration and time was observed on hydrolysis of
cassava starch by a-amylase. The statistical significance of the model
was validated by F-test for analysis of variance (p < 0.01). The
optimum value of starch concentration temperature, time and enzyme
concentration were found to be 4.5% (w/v), 45oC, 150 min, and 1%
(w/v) enzyme. The maximum glucose yield at optimum condition
was 5.17 mg/mL.
Abstract: Biofuels, like biobutanol, have been recognized for
being renewable and sustainable fuels which can be produced from
lignocellulosic biomass. To convert lignocellulosic biomass to
biofuel, pretreatment process is an important step to remove
hemicelluloses and lignin to improve enzymatic hydrolysis. Dilute
acid pretreatment has been successful developed for pretreatment of
corncobs and the optimum conditions of dilute sulfuric and
phosphoric acid pretreatment were obtained at 120 °C for 5 min with
15:1 liquid to solid ratio and 140 °C for 10 min with 10:1 liquid to
solid ratio, respectively. The result shows that both of acid
pretreatments gave the content of total sugar approximately 34–35
g/l. In case of inhibitor content (furfural), phosphoric acid
pretreatment gives higher than sulfuric acid pretreatment.
Characterizations of corncobs after pretreatment indicate that both of
acid pretreatments can improve enzymatic accessibility and the better
results present in corncobs pretreated with sulfuric acid in term of
surface area, crystallinity, and composition analysis.
Abstract: The present work represents an investigation of the
hydrolysis of hull-less pumpkin (Cucurbita Pepo L.) oil cake protein
isolate (PuOC PI) by pepsin. To examine the effectiveness and
suitability of pepsin towards PuOC PI the kinetic parameters for
pepsin on PuOC PI were determined and then, the hydrolysis process
was studied using Response Surface Methodology (RSM). The
hydrolysis was carried out at temperature of 30°C and pH 3.00. Time
and initial enzyme/substrate ratio (E/S) at three levels were selected
as the independent parameters. The degree of hydrolysis, DH, was
mesuared after 20, 30 and 40 minutes, at initial E/S of 0.7, 1 and 1.3
mA/mg proteins. Since the proposed second-order polynomial model
showed good fit with the experimental data (R2 = 0.9822), the
obtained mathematical model could be used for monitoring the
hydrolysis of PuOC PI by pepsin, under studied experimental
conditions, varying the time and initial E/S. To achieve the highest
value of DH (39.13 %), the obtained optimum conditions for time
and initial E/S were 30 min and 1.024 mA/mg proteins.