Abstract: A new strategy for oriented immobilization of proteins was proposed. The strategy contains two steps. The first step is to search for a docking site away from the active site on the protein surface. The second step is trying to find a ligand that is able to grasp the targeted site of the protein. To avoid ligand binding to the active site of protein, the targeted docking site is selected to own opposite charges to those near the active site. To enhance the ligand-protein binding, both hydrophobic and electrostatic interactions need to be included. The targeted docking site should therefore contain hydrophobic amino acids. The ligand is then selected through the help of molecular docking simulations. The enzyme α-amylase derived from Aspergillus oryzae (TAKA) was taken as an example for oriented immobilization. The active site of TAKA is surrounded by negatively charged amino acids. All the possible hydrophobic sites on the surface of TAKA were evaluated by the free energy estimation through benzene docking. A hydrophobic site on the opposite side of TAKA-s active site was found to be positive in net charges. A possible ligand, 3,3-,4,4- – Biphenyltetra- carboxylic acid (BPTA), was found to catch TAKA by the designated docking site. Then, the BPTA molecules were grafted onto silica gels and measured the affinity of TAKA adsorption and the specific activity of thereby immobilized enzymes. It was found that TAKA had a dissociation constant as low as 7.0×10-6 M toward the ligand BPTA on silica gel. The increase in ionic strength has little effect on the adsorption of TAKA, which indicated the existence of hydrophobic interaction between ligands and proteins. The specific activity of the immobilized TAKA was compared with the randomly adsorbed TAKA on primary amine containing silica gel. It was found that the orderly immobilized TAKA owns a specific activity twice as high as the one randomly adsorbed by ionic interaction.
Abstract: Different agricultural waste peels were assessed for
their suitability to be used as primary substrates for the
bioremediation of free cyanide (CN-) by a cyanide-degrading fungus
Aspergillus awamori isolated from cyanide containing wastewater.
The bioremediated CN- concentration were in the range of 36 to 110
mg CN-/L, with Orange (C. sinensis) > Carrot (D. carota) > Onion
(A. cepa) > Apple (M. pumila), being chosen as suitable substrates
for large scale CN- degradation processes due to: 1) the high
concentration of bioremediated CN-, 2) total reduced sugars released
into solution to sustain the biocatalyst, and 3) minimal residual NH4-
N concentration after fermentation. The bioremediation rate constants
(k) were 0.017h-1 (0h < t < 24h), with improved bioremediation rates
(0.02189h-1) observed after 24h. The averaged nitrilase activity was
~10 U/L.
Abstract: Prickly pear juice has received renewed attention with regard to the effects of processing and preservation on its sensory qualities (colour, taste, flavour, aroma, astringency, visual browning and overall acceptability). Juice was prepared by homogenizing fruit and treating the pulp with pectinase (Aspergillus niger). Juice treatments applied were sugar addition, acidification, heat-treatment, refrigeration, and freezing and thawing. Prickly pear pulp and juice had unique properties (low pH 3.88, soluble solids 3.68 oBrix and high titratable acidity 0.47). Sensory profiling and descriptive analyses revealed that non-treated juice had a bitter taste with high astringency whereas treated prickly pear was significantly sweeter. All treated juices had a good sensory acceptance with values approximating or exceeding 7. Regression analysis of the consumer sensory attributes for non-treated prickly pear juice indicated an overwhelming rejection, while treated prickly pear juice received overall acceptability. Thus, educed favourable sensory responses and may have positive implications for consumer acceptability.
Abstract: Empty Fruit Bunches (EFB) and Palm Oil Mill
Effluent (POME) are two main wastes from oil palm industries which
contain rich lignocellulose. Degradation of EFB and POME by
microorganisms will produce hydrolytic enzyme which will degrade
cellulose and hemicellulose during composting process. However,
normal composting takes about four to six months to reach maturity.
Hence, application of fungi into compost can shorten the period of
composting. This study identifies the effect of xylanase and cellulase
produced by Aspergillus niger and Trichoderma virens on
composting process using EFB and POME. The degradation of EFB
and POME indicates the lignocellulolytic capacity of Aspergillus
niger and Trichoderma virens with more than 7% decrease in
hemicellulose and more than 25% decrease in cellulose for both
inoculated compost. Inoculation of Aspergillus niger and
Trichoderma virens also increased the enzyme activities during the
composting period compared to the control compost by 21% for both
xylanase and cellulase. Rapid rise in the activities of cellulase and
xylanase was observed by Aspergillus niger with the highest
activities of 14.41 FPU/mg and 3.89 IU/mg, respectively. Increased
activities of cellulase and xylanase also occurred in inoculation of
Trichoderma virens with the highest activities obtained at 13.21
FPU/mg and 4.43 IU/mg, respectively. Therefore, it is evident that
the inoculation of fungi can increase the enzyme activities hence
effectively degrading the EFB and POME.
Abstract: This study was conducted to evaluate the antifungal
activities of Cinnamomum zeylanicum and Origanum vulgare L.
essential oil against Aspergillus flavus in culture media and tomato
paste. 200 ppm of cinnamon and 500 ppm of oregano completely
inhibited A. flavus growth in culture media, while in tomato paste 300
ppm of cinnamon and 200 ppm of oregano had the same effect. Test
panel evaluations revealed that samples with 100 and 200 ppm
cinnamon were acceptable. The results may suggest the potential use
of Cinnamomum zeylanicum essential oil as natural preservative in
tomato paste.
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: The bioassay-guided isolation and purification of an
ethyl acetate extract of Aspergillus terreus MC751 led to the
characterization of butyrolactone I as an antidiabetic and antioxidant.
The antidiabetic activity of butyrolactone I was evaluated by α-
glucosidase and α-amylase inhibition assays. Butyrolactone I
demonstrated significant concentration-dependent, mixed-type
inhibitory activity against yeast α-glucosidase with an IC50 of 54μM.
However, the compound exhibited less activity against rat intestinal
α-glucosidase and α-amylase. This is the first report on α-glucosidase
inhibitory activity of butyrolactone I. The antioxidative activity of
butyrolactone I was evaluated based on scavenging effects on 1,1-
diphenyl-2-picrylhydrazyl (DPPH) (IC50 =51 μM) and hydrogen
peroxide (IC50= 141 μM) radicals as well as a reducing power assay.
The results suggest that butyrolactone I is a promising antidiabetic as
well as antioxidant and should be considered for clinical trials.
Abstract: A. niger XP isolated from Vietnam produces very low amount of acidic phytase with optimal pH at 2.5 and 5.5. The phytase production of this strain was successfully improved through gene cloning and expression. A 1.4 - kb DNA fragment containing the coding region of the phyA gene was amplified by PCR and inserted into the expression vector pPICZαA with a signal peptide α- factor, under the control of AOX1 promoter. The recombined plasmid was transformed into the host strain P. pastoris KM71H and X33 by electroporation. Both host strains could efficiently express and secret phytase. The multicopy strains were screened for over expression of phytase. All the selected multicopy strains of P. pastoris X33 were examined for phytase activity, the maximum phytase yield of 1329 IU/ml was obtained after 4 days of incubation in medium BMM. The recombinant protein with MW of 97.4 KW showed to be the only one protein secreted in the culture broth. Multicopy transformant P. pastoris X33 supposed to be potential candidate for producing the commercial preparation of phytase.
Abstract: In molecular biology, microarray technology is widely and successfully utilized to efficiently measure gene activity. If working with less studied organisms, methods to design custom-made microarray probes are available. One design criterion is to select probes with minimal melting temperature variances thus ensuring similar hybridization properties. If the microarray application focuses on the investigation of metabolic pathways, it is not necessary to cover the whole genome. It is more efficient to cover each metabolic pathway with a limited number of genes. Firstly, an approach is presented which minimizes the overall melting temperature variance of selected probes for all genes of interest. Secondly, the approach is extended to include the additional constraints of covering all pathways with a limited number of genes while minimizing the overall variance. The new optimization problem is solved by a bottom-up programming approach which reduces the complexity to make it computationally feasible. The new method is exemplary applied for the selection of microarray probes in order to cover all fungal secondary metabolite gene clusters for Aspergillus terreus.
Abstract: Twelve lactating Etawah Crossedbred goats were used
in this study. Goat feed consisted of Cally andra callothyrsus,
Pennisetum purpureum, wheat bran and dried fermented cassava
peel. The cassava peels were fermented with a traditional culture
called “ragi tape" (mixed culture of Saccharomyces cerevisae,
Aspergillus sp, Candida, Hasnula and Acetobacter). The goats were
divided into 2 groups (Control and Treated) of six does. The
experimental diet of the Control group consisted of 70% of roughage
(fresh Callyandra callothyrsus and Pennisetum purpureum 60:40)
and 30% of wheat bran on dry matter (DM) base. In the Treated
group 30% of wheat bran was replaced with dried fermented cassava
peels. Data were statistically analyzed using analysis of variance
followed SPSS program. The concentration of HCN in fermented
cassava peel decreased to non toxic level. Nutrient composition of
dried fermented cassava peel consisted of 85.75% dry matter;
5.80% crude protein and 82.51% total digestible nutrien (TDN).
Substitution of 30% of wheat bran with dried fermented cassava peel
in the diet had no effect on dry matter and organic matter intake but
significantly (P< 0.05) decreased crude protein and TDN
consumption as well as milk yields and milk composition. The study
recommended to reduced the level of substitution to less than 30% of
concentrates in the diet in order to avoid low nutrient intake and milk
production of goats.
Abstract: Antimicrobial resistant is becoming a major factor in
virtually all hospital acquired infection may soon untreatable is a
serious public health problem. These concerns have led to major
research effort to discover alternative strategies for the treatment of
bacterial infection. Nanobiotehnology is an upcoming and fast
developing field with potential application for human welfare. An
important area of nanotechnology for development of reliable and
environmental friendly process for synthesis of nanoscale particles
through biological systems In the present studies are reported on the
use of fungal strain Aspergillus species for the extracellular synthesis
of bionanoparticles from 1 mM silver nitrate (AgNO3) solution. The
report would be focused on the synthesis of metallic bionanoparticles
of silver using a reduction of aqueous Ag+ ion with the
culture supernatants of Microorganisms. The bio-reduction of the
Ag+ ions in the solution would be monitored in the aqueous
component and the spectrum of the solution would measure through
UV-visible spectrophotometer The bionanoscale particles were
further characterized by Atomic Force Microscopy (AFM), Fourier
Transform Infrared Spectroscopy (FTIR) and Thin layer
chromatography. The synthesized bionanoscale particle showed a
maximum absorption at 385 nm in the visible region. Atomic Force
Microscopy investigation of silver bionanoparticles identified that
they ranged in the size of 250 nm - 680 nm; the work analyzed the
antimicrobial efficacy of the silver bionanoparticles against various
multi drug resistant clinical isolates. The present Study would be
emphasizing on the applicability to synthesize the metallic
nanostructures and to understand the biochemical and molecular
mechanism of nanoparticles formation by the cell filtrate in order to
achieve better control over size and polydispersity of the
nanoparticles. This would help to develop nanomedicine against
various multi drug resistant human pathogens.
Abstract: In this study, the ability of Aspergillus niger and
Penicillium simplicissimum to extract heavy metals from a spent
refinery catalyst was investigated. For the first step, a spent
processing catalyst from one of the oil refineries in Iran was
physically and chemically characterized. Aspergillus niger and
Penicillium simplicissimum were used to mobilize Al/Co/Mo/Ni from
hazardous spent catalysts. The fungi were adapted to the mixture of
metals at 100-800 mg L-1 with increments in concentration of 100 mg
L-1. Bioleaching experiments were carried out in batch cultures. To
investigate the production of organic acids in sucrose medium,
analyses of the culture medium by HPLC were performed at specific
time intervals after inoculation. The results obtained from Inductive
coupled plasma-optical emission spectrometry (ICP-OES) showed
that after the one-step bioleaching process using Aspergillus niger,
maximum removal efficiencies of 27%, 66%, 62% and 38% were
achieved for Al, Co, Mo and Ni, respectively. However, the highest
removal efficiencies using Penicillium simplicissimum were of 32%,
67%, 65% and 38% for Al, Co, Mo and Ni, respectively
Abstract: To investigate the production of cellulases from Aspergillus heteromorphus, submerged fermentation was performed using wheat straw as substrate. Optimization of saccharification conditions like pH, temperature and time were studied. Highest reducing sugar was released on 5th day at 5 pH, 30° C temperature. When A. heteromorphous was grown on wheat straw in submerged fermentation after 5 days incubation at 30 ° C, 3.2 IU/ml and 83 IU/ml, filter paper activity and CMCase activity respectively.
Abstract: In this study, the designed dual stage membrane
bioreactor (MBR) system was conceptualized for the treatment of
cyanide and heavy metals in electroplating wastewater. The design
consisted of a primary treatment stage to reduce the impact of
fluctuations and the secondary treatment stage to remove the residual
cyanide and heavy metal contaminants in the wastewater under
alkaline pH conditions. The primary treatment stage contained
hydrolyzed Citrus sinensis (C. sinensis) pomace and the secondary
treatment stage contained active Aspergillus awamori (A. awamori)
biomass, supplemented solely with C. sinensis pomace extract from
the hydrolysis process. An average of 76.37%, 95.37%, 93.26 and
94.76% and 99.55%, 99.91%, 99.92% and 99.92% degradation
efficiency for total cyanide (T-CN), including the sorption of nickel
(Ni), zinc (Zn) and copper (Cu) were observed after the first and
second treatment stages, respectively. Furthermore, cyanide
conversion by-products degradation was 99.81% and 99.75 for both
formate (CHOO-) and ammonium (NH4
+) after the second treatment
stage. After the first, second and third regeneration cycles of the C.
sinensis pomace in the first treatment stage, Ni, Zn and Cu removal
achieved was 99.13%, 99.12% and 99.04% (first regeneration cycle),
98.94%, 98.92% and 98.41% (second regeneration cycle) and 98.46
%, 98.44% and 97.91% (third regeneration cycle), respectively.
There was relatively insignificant standard deviation detected in all
the measured parameters in the system which indicated
reproducibility of the remediation efficiency in this continuous
system.
Abstract: Studies on Simultaneous Saccharification and Fermentation (SSF) of corn flour, a major agricultural product as the substrate using starch digesting glucoamylase enzyme derived from Aspergillus niger and non starch digesting and sugar fermenting Saccharomyces cerevisiae in a batch fermentation. Experiments based on Central Composite Design (CCD) were conducted to study the effect of substrate concentration, pH, temperature, enzyme concentration on Ethanol Concentration and the above parameters were optimized using Response Surface Methodology (RSM). The optimum values of substrate concentration, pH, temperature and enzyme concentration were found to be 160 g/l, 5.5, 30°C and 50 IU respectively. The effect of inoculums age on ethanol concentration was also investigated. The corn flour solution equivalent to 16% initial starch concentration gave the highest ethanol concentration of 63.04 g/l after 48 h of fermentation at optimum conditions of pH and temperature. Monod model and Logistic model were used for growth kinetics and Leudeking – Piret model was used for product formation kinetics.
Abstract: The microbiological and physicochemical
characteristics of wetland soils in Eket Local Government Area were
studied between May 2001 and June 2003. Total heterotrophic
bacterial counts (THBC), total fungal counts (TFC), and total
actinomycetes counts (TAC) were determined from soil samples
taken from four locations at two depths in the wet and dry seasons.
Microbial isolates were characterized and identified. Particle size and
chemical parameters were also determined using standard methods.
THBC ranged from 5.2 (+0.17) x106 to 1.7 (+0.18) x107 cfu/g and
from 2.4 (+0.02) x106 to 1.4 (+0.04) x107cfu/g in the wet and dry
seasons, respectively. TFC ranged from 1.8 (+0.03) x106 to 6.6 (+
0.18) x106 cfu/g and from 1.0 (+0.04) x106 to 4.2 (+ 0.01) x106 cfu/g
in the wet and dry seasons, respectively .TAC ranged from 1.2
(+0.53) x106 to 6.0 (+0.05) x106 cfu/g and from 0.6 (+0.01) x106 to
3.2 (+ 0.12) x106 cfu/g in the wet and dry season, respectively.
Acinetobacter, Alcaligenes, Arthrobacter, Bacillus, Beijerinckja,
Enterobacter, Micrococcus, Flavobacterium, Serratia, Enterococcus,
and Pseudomonas species were predominant bacteria while
Aspergillus, Fusarium, Mucor, Penicillium, and Rhizopus were the
dominant fungal genera isolated. Streptomyces and Norcadia were
the actinomycetes genera isolated. The particle size analysis showed
high sand fraction but low silt and clay. The pH and % organic
matter were generally acidic and low, respectively at all locations.
Calcium dominated the exchangeable bases with low electrical
conductivity and micronutrients. These results provide the baseline
data of Eket wetland soils for its management for sustainable
agriculture.