Abstract: Nowadays, butyl acetate, a pineapple flavor has been applied widely in food, beverage, cosmetic and pharmaceutical industries. In this study, Butyl acetate, a flavor ester was successfully synthesized via green synthesis of enzymatic reaction route. Commercial immobilized lipase from Rhizomucor miehei (Lipozyme RMIM) was used as biocatalyst in the esterification reaction between acetic acid and butanol. Various reaction parameters such as reaction time (RT), temperature (T) and amount of enzyme (E) were chosen to optimize the reaction synthesis in solvent-free system. The optimum condition to produce butyl acetate was at reaction time (RT), 18 hours; temperature (T), 37°C and amount of enzyme, 25 % (w/w of total substrate). Analysis of yield showed that at optimum condition, >78 % of butyl acetate was produced. The product was confirmed as butyl acetate from FTIR analysis whereby the presence of an ester group was observed at wavenumber of 1742 cm-1.
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: Malting is usually carried out on intact barley seed,
while hull is still attached to it. In this study, oat grain with and
without hull was subjected to controlled germination to optimize its
enzymes activity, in such a way that lipase has the lowest and α-
amylase and proteinase the highest activities. Since pH has a great
impact on the activity of the enzymes, the pH of germination media
was set up to 3 to 8. In dehulled oats, lipase and α-amylase had the
lowest and highest activities in pHs 3 and 6, respectively whereas the
highest proteinase activity was evidenced at pH 7 and 4 in the oats
with and without hull respectively. While measurements indicated
that the effect of hull on the enzyme activities particularly in lipase
and amylase at each level of the pH are significantly different, the
best results were obtained in those samples in which their hull had
been removed. However, since the similar lipase activity in
germinated dehulled oat were recorded at the pHs 4 and 5, therefore
it was concluded that pH 5 in dehulled oat seed may provide the
optimum enzyme activity for all the enzymes.
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: Enzymatic saccharification of biomass for reducing
sugar production is one of the crucial processes in biofuel production
through biochemical conversion. In this study, enzymatic
saccharification of dilute potassium hydroxide (KOH) pre-treated
Tetraselmis suecica biomass was carried out by using cellulase
enzyme obtained from Trichoderma longibrachiatum. Initially, the
pre-treatment conditions were optimised by changing alkali reagent
concentration, retention time for reaction, and temperature. The T.
suecica biomass after pre-treatment was also characterized using
Fourier Transform Infrared Spectra and Scanning Electron
Microscope. These analyses revealed that the functional group such
as acetyl and hydroxyl groups, structure and surface of T. suecica
biomass were changed through pre-treatment, which is favourable for
enzymatic saccharification process. Comparison of enzymatic
saccharification of untreated and pre-treated microalgal biomass
indicated that higher level of reducing sugar can be obtained from
pre-treated T. suecica. Enzymatic saccharification of pre-treated T.
suecica biomass was optimised by changing temperature, pH, and
enzyme concentration to solid ratio ([E]/[S]). Highest conversion of
carbohydrate into reducing sugar of 95% amounted to reducing sugar
yield of 20 (wt%) from pre-treated T. suecica was obtained from
saccharification, at temperature: 40°C, pH: 4.5 and [E]/[S] of 0.1
after 72 h of incubation. Hydrolysate obtained from enzymatic
saccharification of pretreated T. suecica biomass was further
fermented into biobutanol using Clostridium saccharoperbutyliticum
as biocatalyst. The results from this study demonstrate a positive
prospect of application of dilute alkaline pre-treatment to enhance
enzymatic saccharification and biobutanol production from
microalgal biomass.
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: A large number of chemical, bio-chemical and pollution-control processes use heterogeneous fixed-bed reactors. The use of finite hollow cylindrical catalyst pellets can enhance conversion levels in such reactors. The absence of the pellet core can significantly lower the diffusional resistance associated with the solid phase. This leads to a better utilization of the catalytic material, which is reflected in the higher values for the effectiveness factor, leading ultimately to an enhanced conversion level in the reactor. It is however important to develop a rigorous heterogeneous model for the reactor incorporating the two-dimensional feature of the solid phase owing to the presence of the finite hollow cylindrical catalyst pellet. Presently, heterogeneous models reported in the literature invariably employ one-dimension solid phase models meant for spherical catalyst pellets. The objective of the paper is to present a rigorous model of the fixed-bed reactors containing finite hollow cylindrical catalyst pellets. The reaction kinetics considered here is the widely used Michaelis–Menten kinetics for the liquid-phase bio-chemical reactions. The reaction parameters used here are for the enzymatic degradation of urea. Results indicate that increasing the height to diameter ratio helps to improve the conversion level. On the other hand, decreasing the thickness is apparently not as effective. This could however be explained in terms of the higher void fraction of the bed that causes a smaller amount of the solid phase to be packed in the fixed-bed bio-chemical reactor.
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: Characterization and evaluation of the activity of Vespa basalis DPP-IV, which expressed in Spodoptera frugiperda 21 cells. The expression of rDPP-IV was confirmed by SDS–PAGE, Western blot analyses, LC-MS/MS and measurement of its peptidase specificity. One-step purification by Ni-NTA affinity chromatography and the total amount of rDPP-IV recovered was approximately 6.4mg per liter from infected culture medium; an equivalent amount would be produced by 1x109 infected Sf21 insect cells. Through the affinity purification led to highly stable rDPP-IV enzyme was recovered and with significant peptidase activity. The rDPP-IV exhibited classical Michaelis–Menten kinetics, with kcat/Km in the range of 10-500 mM-1×S-1 for the five synthetic substrates and optimum substrate is Ala-Pro-pNA. As expected in inhibition assay, the enzymatic activity of rDPP-IV was significantly reduced by 80 or 60% in the presence of sitagliptin (a DPP-IV inhibitor) or PMSF (a serine protease inhibitor), but was not apparently affected by iodoacetamide (a cysteine protease inhibitor).
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: It is believed that DNA damaging toxic metabolites contributes to the development of different pathological conditions. To prevent harmful influence of toxic agents, cells developed number of protecting mechanisms, such as enzymatic reaction of detoxification of reactive metabolites and repair of DNA damage. The aim of the study was to examine the association between polymorphism of GSTT1/GSTM1 and XRCC1/3 genes and coronary artery disease (CAD) incidence. To examine a polymorphism of these genes in CAD susceptibility in patients and controls, PCR based genotyping assay was performed. For GST genes, frequency of GSTM1 null genotype among CAD affected group was significantly increased than in control group (P0.1). We found that neither XRCC1 Arg399Gln nor XRCC3 Thr241Met were associated with CAD risk. Obtained data suggests that GSTM1 null genotype carriers are more susceptible to CAD development.
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.
Abstract: Prickly pear fruit (Opuntia ficus indica L. Miller) belongs to the Cactaceae family. This species is very sensitive to low storage temperatures (< 5°C) which cause damages. The fruits can be peeled, suitably packaged and successfully commercialized as a ready-to-eat product. The main limit to the extension of the shelf life is the production of off-flavors due to different factors, the growth of microorganisms and the action of endogenous enzymes. Lipoxygenase (LOX) and Pectinesterase (PE) are involved in fruit degradation. In particular, LOX pathway is directly responsible for lipid oxidation, and the subsequent production of off-flavours, while PE causes the softening of fruit during maturation. They act on the texture and shelf-life of post-harvest, packaged fruits, as a function of the the grown of microorganisms and packaging technologies used. The aim of this work is to compare the effect of different packaging technologies on the shelf life extension of ready-to-eat prickly pear fruits with regards for the enzymes activities.
Abstract: The dilute acid pretreatment and enzymatic
saccharification of lignocellulosic substrate, cogon grass (Imperata
cylindrical, L.) was optimized prior ethanol fermentation using
simultaneous saccharification and fermentation (SSF) method. The
optimum pretreatment conditions, temperature, sulfuric acid
concentration, and reaction time were evaluated by determining the
maximum sugar yield at constant enzyme loading. Cogon grass, at
10% w/v substrate loading, has optimum pretreatment conditions of
126°C, 0.6% v/v H2SO4, and 20min reaction time. These
pretreatment conditions were used to optimize enzymatic
saccharification using different enzyme combinations. The maximum
saccharification yield of 36.68mg/mL (71.29% reducing sugar) was
obtained using 25FPU/g-cellulose cellulase complex combined with
1.1% w/w of cellobiase, ß-glucosidase, and 0.225% w/w of
hemicellulase complex, after 96 hours of saccharification. Using the
optimum pretreatment and saccharification conditions, SSF of treated
substrates was done at 37°C for 120 hours using industrial yeast
strain HBY3, Saccharomyces cerevisiae. The ethanol yield for cogon
grass at 4% w/w loading was 9.11g/L with 5.74mg/mL total residual
sugar.
Abstract: Composting is the process in which municipal solid
waste (MSW) and other organic waste materials such as biosolids
and manures are decomposed through the action of bacteria and other
microorganisms into a stable granular material which, applied to
land, as soil conditioner. Microorganisms, especially those that are
able to degrade polymeric organic material have a key role in speed
up this process. The aim of this study has been established to
isolation of microorganisms with high ability to production
extracellular enzymes for degradation of natural polymers that are
exists in MSW for decreasing time of degradation phase. Our
experimental study for isolation designed in two phases: in first
phase we isolated degrading microorganism with selected media that
consist a special natural polymer such as cellulose, starch, lipids and
etc as sole source of carbon. In second phase we selected
microorganism that had high degrading enzyme production with
enzymatic assay for seed production. However, our findings in pilot
scale have indicated that usage of this microbial consortium had high
efficiency for decreasing degradation phase.
Abstract: Sunflower stalks were analysed for chemical
compositions: pentosan 15.84%, holocellulose 70.69%,
alphacellulose 45.74%, glucose 27.10% and xylose 7.69% based on
dry weight of 100-g raw material. The most optimum condition for
steam explosion pretreatment was as follows. Sunflower stalks were
cut into small pieces and soaked in 0.02 M H2SO4 for overnight.
After that, they were steam exploded at 207 C and 21 kg/cm2 for 3
minutes to fractionate cellulose, hemicellulose and lignin. The
resulting hydrolysate, containing hemicellulose, and cellulose pulp
contained xylose sugar at 2.53% and 7.00%, respectively.The pulp
was further subjected to enzymatic saccharification at 50 C, pH 4.8 citrate buffer) with pulp/buffer 6% (w/w)and Celluclast 1.5L/pulp
2.67% (w/w) to obtain single glucose with maximum yield 11.97%.
After fixed-bed fermentation under optimum condition using
conventional yeast mixtures to produce bioethanol, it indicated
maximum ethanol yield of 0.028 g/100 g sunflower stalk.
Abstract: The use of bioindicators plants (lichens, bryophytes
and Sphagnum....) in monitoring pollution by heavy metals has been
the subject of several works. However, few studies have addressed
the impact of specific type-s pollutants (fertilizers, pesticides.) on
these organisms.
We propose in this work to make the highlighting effect of NPKs
(NPK: nitrogen-phosphate-potassium-sulfate (NP2O5K2O) (15,15,15),
at concentrations of 10, 20, 30 , 40 and 50mM/L) on the activity of
detoxification enzymes (GSH/GST, CAT, APX and MDA) of plant
bioindicators (mosses and lichens) after treatment for 3 and 7 days.
This study shows the important role of the defense system in the
accumulation and tolerance to chemical pollutants through the
activation of enzymatic (GST (glutathione-S-transferase, APX
(ascorbat peroxidase), CAT (catalase)) and nonenzymatic biomarkers
(GSH (glutathione), MDA (malondialdehyde)) against oxidative
stress generated by the NPKs.
Abstract: The possibility of using cassava residue containing
49.66% starch, 21.47% cellulose, 12.97% hemicellulose, and 21.86%
lignin as a raw material to produce glucose using enzymatic
hydrolysis was investigated. In the experiment, each reactor
contained the cassava residue, bacteria cells, and production medium.
The effects of particles size (40 mesh and 60 mesh) and strains of
bacteria (A002 and M015) isolated from Thai higher termites,
Microcerotermes sp., on the glucose concentration at 37°C were
focused. High performance liquid chromatography (HPLC) with a
refractive index detector was used to determine the quantity of
glucose. The maximum glucose concentration obtained at 37°C using
strain A002 and 60 mesh of the cassava residue was 1.51 g/L at 10 h.