Abstract: Telfairia occidentalis is a leafy vegetable commonly grown in the tropics for nutritional benefits. The use of enzymes and probiotics is becoming prominent due to the ban on antibiotics as growth promoters in many parts of the world. It is conceived that with enzymes and probiotics additives, fibrous leafy vegetables can be incorporated into poultry feeds as protein source. However, certain antinutrients were also found in the leaves of Telfairia occidentalis. Four broiler starter and finisher diets were formulated for the two phases of the broiler experiments. A mixture of fiber degrading enzymes, Roxazyme G2 (combination of cellulase, glucanase and xylanase) and probiotics (Turbotox), a growth promoter, were used in broiler diets at 1:1. The Roxazyme G2/Turbotox mixtures were used in diets containing four varying levels of Telfairia occidentalis leaf meal (TOLM) at 0, 10, 20 and 30%. Diets 1 were standard broiler diets without TOLM and Roxazyme G2 and Turbotox additives. Diets 2, 3 and 4 had enzymes and probiotics additives. Certain mineral elements such as Ca, P, K, Na, Mg, Fe, Mn, Cu and Zn were found in notable quantities viz. 2.6 g/100 g, 1.2 g/100 g, 6.2 g/100 g, 5.1 g/100 g, 4.7 g/100 g, 5875 ppm, 182 ppm, 136 ppm and 1036 ppm, respectively. Phytin, phytin-P, oxalate, tannin and HCN were also found in ample quantities viz. 189.2 mg/100 g, 120.1 mg/100 g, 80.7 mg/100 g, 43.1 mg/100 g and 61.2 mg/100 g, respectively. The average weight gain was highest at 46.3 g/bird/day for birds on 10% TOLM diet but similar (P > 0.05) to 46.2 g/bird/day for birds on 20% TOLM. The feed conversion ratio (FCR) of 2.27 was the lowest and optimum for birds on 10% TOLM although similar (P > 0.05) to 2.29 obtained for birds on 20% TOLM. FCR of 2.61 was the highest at 2.61 for birds on 30% TOLM diet. The lowest FCR of 2.27 was obtained for birds on 10% TOLM diet although similar (P > 0.05) to 2.29 for birds on 20% TOLM diet. Most carcass characteristics and organ weights were similar (P > 0.05) for the experimental birds on the different diets except for kidney, gizzard and intestinal length. The values for kidney, gizzard and intestinal length were significantly higher (P < 0.05) for birds on the TOLM diets. The nitrogen retention had the highest value of 72.37 ± 0.10% for birds on 10% TOLM diet although similar (P > 0.05) to 71.54 ± 1.89 obtained for birds on the control diet without TOLM and enzymes/probiotics mixture. There was evidence of a better utilization of TOLM as a plant protein source. The carcass characteristics and organ weights all showed evidence of uniform tissue buildup and muscles development particularly in diets containing 10% of TOLM level. There was also better nitrogen utilization in birds on the 10% TOLM diet. Considering the cheap cost of TOLM, it is envisaged that its introduction into poultry feeds as a plant protein source will ultimately reduce the cost of poultry feeds.
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: Soil enzyme activities in Kasuga-yama Hill Primeval Forest (Nara, Japan) were examined to determine levels of mineralization and metabolism. Samples were selected from the soil surrounding laurel-leaved (BB-1) and Carpinus japonica (BB-2 and Pw) trees for analysis. Cellulase, β-xylosidase, and protease activities were higher in BB-1 samples those in BB-2 samples. These activity levels corresponded to the distribution of cellulose and hemicellulose in the soil horizons. Cellulase, β-xylosidase, and chymotrypsin activities were higher in soil from the Pw forest than in that from the BB-2 forest. The relationships between the soil enzymes calculated by Spearman’s rank correlation indicate that the interactions between enzymes in BB-2 samples were more complex than those in Pw samples.
Abstract: Effect of enzyme on the yield and chemical composition of Artemisia campestris essential oil is reported in the present study. It was demonstrated that enzyme facilitated the extraction of essential oil with increase in oil yield and did not affect any noticeable change in flavour profile of the volatile oil. Essential oil was tested for antibacterial activity using Escherichia coli; which was extremely sensitive against control with the largest inhibition (29mm), whereas Staphylococcus aureus was the most sensitive against essential oil obtained from enzymatic pre-treatment with the largest inhibition zone (25mm). The antioxidant activity of the essential oil with hemicellulase pre-treatment (EO2) and control sample (EO1) was determined through reducing power. It was significantly lower than the standard drug (vitamin C) in this order: vitamin C˃EO2˃EO1.
Abstract: This study was conducted for the investigation of
number of cellulolytic bacteria and their ability in decomposition.
Seven samples surface soil were collected on cellulose Zailiskii
Alatau slopes. Cellulolitic activity of new strains of Bacillus, isolated
from soil is determined. Isolated cellulose degrading bacteria were
screened for determination of the highest cellulose activity by
quantitative assay using Congo red, gravimetric assay and
colorimetric DNS method trough of the determination of the
parameters of sugar reduction. Strains are assigned to: B.subtilis,
B.licheniformis, B. cereus and, В. megaterium. Bacillus strains
consisting of several different types of cellulases have broad substrate
specificity of cellulase complexes formed by them. Cellulolitic
bacteria were recorded to have highest cellulase activity and selected
for optimization of cellulase enzyme production.
Abstract: Lignocellolusic material is a substance that is resistant to be degraded by microorganisms or hydrolysis enzymes. To be used as materials for biofuel production, it needs pretreatment process to improve efficiency of hydrolysis. In this work, chemical pretreatments on rice straw using three diluted organic acids, including acetic acid, citric acid, oxalic acid, were optimized. Using Response Surface Methodology (RSM), the effect of three pretreatment parameters, acid concentration, treatment time, and reaction temperature, on pretreatment efficiency were statistically evaluated. The results indicated that dilute oxalic acid pretreatment led to the highest enhancement of enzymatic saccharification by commercial cellulase and yielded sugar up to 10.67 mg/ml when using 5.04% oxalic acid at 137.11 oC for 30.01 min. Compared to other acid pretreatment by acetic acid, citric acid, and hydrochloric acid, the maximum sugar yields are 7.07, 6.30, and 8.53 mg/ml, respectively. Here, it was demonstrated that organic acids can be used for pretreatment of lignocellulosic materials to enhance of hydrolysis process, which could be integrated to other applications for various biorefinery processes.
Abstract: Rice straw is lignocellulosic biomass which can be utilized as substrate for the biogas production. However, due to the property and composition of rice straw, it is difficult to be degraded by hydrolysis enzymes. One of the pretreatment methods that modify such properties of lignocellulosic biomass is the application of lignocellulose-degrading microbial consortia. The aim of this study is to investigate the effect of microbial consortia to enhance biogas production. To select the high efficient consortium, cellulase enzymes were extracted and their activities were analyzed. The results suggested that microbial consortium culture obtained from cattle manure is the best candidate compared to decomposed wood and horse manure. A microbial consortium isolated from cattle manure was then mixed with anaerobic sludge and used as inoculum for biogas production. The optimal conditions for biogas production were investigated using response surface methodology (RSM). The tested parameters were the ratio of amount of microbial consortium isolated and amount of anaerobic sludge (MI:AS), substrate to inoculum ratio (S:I) and temperature. Here, the value of the regression coefficient R2 = 0.7661 could be explained by the model which is high to advocate the significance of the model. The highest cumulative biogas yield was 104.6 ml/g-rice straw at optimum ratio of MI:AS, ratio of S:I, and temperature of 2.5:1, 15:1 and 44°C respectively.
Abstract: Fungal mutant strains have produced cellulase and
xylanase enzymes, and have induced high hydrolysis with enhanced
of rice straw. The mutants were obtained by exposing Penicillium
strain to UV-light treatments. Screening and selection after treatment
with UV-light were carried out using cellulolytic and xylanolytic
clear zones method to select the hypercellulolytic and
hyperxylanolytic mutants. These mutants were evaluated for their
cellulase and xylanase enzyme production as well as their abilities for
biodegradation of rice straw. The mutant 12 UV/1 produced 306.21%
and 209.91% cellulase and xylanase, respectively, as compared with
the original wild type strain. This mutant showed high capacity of
rice straw degradation. The effectiveness of tested mutant strain and
that of wild strain was compared in relation to enhancing the
composting process of rice straw and animal manures mixture. The
results obtained showed that the compost product of inoculated
mixture with mutant strain (12 UV/1) was the best compared to the
wild strain and un-inoculated mixture. Analysis of the composted
materials showed that the characteristics of the produced compost
were close to those of the high quality standard compost. The results
obtained in the present work suggest that the combination between
rice straw and animal manure could be used for enhancing the
composting process of rice straw and particularly when applied with
fungal decomposer accelerating the composting process.
Abstract: Biofuels production has come forth as a future
technology to combat the problem of depleting fossil fuels. Bio-based
ethanol production from enzymatic lignocellulosic biomass
degradation serves an efficient method and catching the eye of
scientific community. High cost of the enzyme is the major obstacle
in preventing the commercialization of this process. Thus main
objective of the present study was to optimize composition of
medium components for enhancing cellulase production by newly
isolated strain of Bacillus tequilensis. Nineteen factors were taken
into account using statistical Plackett-Burman Design. The significant
variables influencing the cellulose production were further employed
in statistical Response Surface Methodology using Central
Composite Design for maximizing cellulase production. The
optimum medium composition for cellulase production was: peptone
(4.94 g/L), ammonium chloride (4.99 g/L), yeast extract (2.00 g/L),
Tween-20 (0.53 g/L), calcium chloride (0.20 g/L) and cobalt chloride
(0.60 g/L) with pH 7, agitation speed 150 rpm and 72 h incubation at
37oC. Analysis of variance (ANOVA) revealed high coefficient of
determination (R2) of 0.99. Maximum cellulase productivity of 11.5
IU/ml was observed against the model predicted value of 13 IU/ml.
This was found to be optimally active at 60oC and pH 5.5.
Abstract: In this study, six bacterial isolates of a slightly
thermophilic organism from the Sg. Klah hot spring, Malaysia were
successfully isolated and designated as M7T55D1, M7T55D2,
M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The
bacterial isolates were screened for their cellulose hydrolytic ability
on Carboxymethlycellulose agar medium. The isolated bacterial
strains were identified morphologically, biochemically and
molecularly with the aid of 16S rDNA sequencing. All of the bacteria
showed their optimum growth at a slightly alkaline pH of 7.5 with a
temperature of 55°C. All strains were Gram-negative, non-spore
forming type, strictly aerobic, catalase-positive and oxidase-positive
with the ability to produce thermostable cellulase. Based on BLASTn
results, bacterial isolates of M7T55D2 and M7T53D1 gave the
highest homology (97%) with similarity to Tepidimonas ignava while
isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed
their closest homology (97%-98%) with Tepidimonas thermarum.
These cellulolytic thermophiles might have a commercial potential to
produce valuable thermostable cellulase.
Abstract: Due to a high ethanol demand, the approach for effective ethanol production is important and has been developed rapidly worldwide. Several agricultural wastes are highly abundant in celluloses and the effective cellulase enzymes do exist widely among microorganisms. Accordingly, the cellulose degradation using microbial cellulase to produce a low-cost substrate for ethanol production has attracted more attention. In this study, the cellulase producing bacterial strain has been isolated from rich straw and identified by 16S rDNA sequence analysis as Acinetobacter sp. KKU44. This strain is able to grow and exhibit the cellulase activity. The optimal temperature for its growth and cellulase production is 37°C. The optimal temperature of bacterial cellulase activity is 60°C. The cellulase enzyme from Acinetobacter sp. KKU44 is heat-tolerant enzyme. The bacterial culture of 36h. showed highest cellulase activity at 120U/mL when grown in LB medium containing 2% (w/v). The capability of Acinetobacter sp. KKU44 to grow in cellulosic agricultural wastes as a sole carbon source and exhibiting the high cellulase activity at high temperature suggested that this strain could be potentially developed further as a cellulose degrading strain for a production of low-cost substrate used in ethanol production.
Abstract: Conversion of lignocellulosic biomass is the basis process for production of fuels, chemicals and materials in the sustainable biorefinery industry. Saccharification of lignocellulosic biomass is an essential step which produces sugars for further conversion to target value-added products e.g. bio-ethanol, bio-plastic, g-valerolactone (GVL), 5-hydroxymethylfuroic acid (HMF), levulinic acid, etc. The goal of this work was to develop an efficient enzyme for conversion of biomass to reducing sugar based on crude fungal enzyme from Chaetomium globosum BCC5776 produced by submerged fermentation and evaluate its activity comparing to a commercial Acremonium cellulase. Five local biomasses in Thailand: rice straw, sugarcane bagasse, corncobs, corn stovers, and palm empty fruit bunches were pretreated and hydrolyzed with varying enzyme loadings. Saccharification of the biomass led to different reducing sugar levels from 115 mg/g to 720 mg/g from different types of biomass using cellulase dosage of 9 FPU/g. The reducing sugar will be further employed as sugar feedstock for production of ethanol or commodity chemicals. This work demonstrated the use of promising enzyme candidate for conversion of local lignocellulosic biomass in biorefinery industry.
Abstract: A statistical optimization was studied to design a media composition to produce optimum cellulolytic enzyme where palm oil mill effluent (POME) as a basal medium and filamentous fungus, Trichoderma reesei RUT-C30 were used in the liquid state bioconversion(LSB). 2% (w/v) total suspended solid, TSS, of the POME supplemented with 1% (w/v) cellulose, 0.5%(w/v) peptone and 0.02% (v/v) Tween 80 was estimated to produce the optimum CMCase activity of 18.53 U/ml through the statistical analysis followed by the faced centered central composite design(FCCCD). The probability values of cellulose (
Abstract: The purpose of the present work was to study the
production and process parameters optimization for the synthesis of
cellulase from Trichoderma viride in solid state fermentation (SSF)
using an agricultural wheat straw as substrates; as fungal conversion
of lignocellulosic biomass for cellulase production is one among the
major increasing demand for various biotechnological applications.
An optimization of process parameters is a necessary step to get
higher yield of product. Several kinetic parameters like pretreatment,
extraction solvent, substrate concentration, initial moisture content,
pH, incubation temperature and inoculum size were optimized for
enhanced production of third most demanded industrially important
cellulase. The maximum cellulase enzyme activity 398.10±2.43
μM/mL/min was achieved when proximally analyzed lignocellulosic
substrate wheat straw inocubated at 2% HCl as pretreatment tool
along with distilled water as extraction solvent, 3% substrate
concentration 40% moisture content with optimum pH 5.5 at 45°C
incubation temperature and 10% inoculum size.
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: Response Surface Methodology (RSM) is a powerful
and efficient mathematical approach widely applied in the
optimization of cultivation process. Cellulase enzyme production by
Trichoderma reesei RutC30 using agricultural waste rice straw and
banana fiber as carbon source were investigated. In this work,
sequential optimization strategy based statistical design was
employed to enhance the production of cellulase enzyme through
submerged cultivation. A fractional factorial design (26-2) was applied
to elucidate the process parameters that significantly affect cellulase
production. Temperature, Substrate concentration, Inducer
concentration, pH, inoculum age and agitation speed were identified
as important process parameters effecting cellulase enzyme synthesis.
The concentration of lignocelluloses and lactose (inducer) in the
cultivation medium were found to be most significant factors. The
steepest ascent method was used to locate the optimal domain and a
Central Composite Design (CCD) was used to estimate the quadratic
response surface from which the factor levels for maximum
production of cellulase were determined.
Abstract: the objective of this study is to measure the levels of
cellulas activity of ostrich GI microorganisms, and comparing it with
the levels of cellulas activity of rumen-s microorganisms, and also to
estimate the probability of increasing enzyme activity with injecting
different dosages (30%, 50% and 70%) of pure anaerobic goat rumen
fungi. The experiment was conducted in laboratory and under a
complete anaerobic condition (in vitro condition). 40 ml of
“CaldWell" medium and 1.4g wheat straw were placed in incubator
for an hour. The cellulase activity of ostrich microorganisms was
compared with other treatments, and then different dosages (30%,
50% and 70%) of pure anaerobic goat rumen fungi were injected to
ostrich microorganism-s media. Due to the results, cattle and goat
with 2.13 and 2.08 I.U (international units) respectively showed the
highest activity and ostrich with 0.91 (I.U) had the lowest cellulose
activity (p < 0.05). Injecting 30% and 50% of anaerobic fungi had no
significant incensement in enzyme activity, but with injecting 70% of
rumen fungi to ostrich microorganisms culture a significant increase
was observed 1.48 I.U. (p < 0.05).
Abstract: Simultaneous Saccharification and Fermentation (SSF) of sugarcane bagasse by cellulase and Pachysolen tannophilus MTCC *1077 were investigated in the present study. Important process variables for ethanol production form pretreated bagasse were optimized using Response Surface Methodology (RSM) based on central composite design (CCD) experiments. A 23 five level CCD experiments with central and axial points was used to develop a statistical model for the optimization of process variables such as incubation temperature (25–45°) X1, pH (5.0–7.0) X2 and fermentation time (24–120 h) X3. Data obtained from RSM on ethanol production were subjected to the analysis of variance (ANOVA) and analyzed using a second order polynomial equation and contour plots were used to study the interactions among three relevant variables of the fermentation process. The fermentation experiments were carried out using an online monitored modular fermenter 2L capacity. The processing parameters setup for reaching a maximum response for ethanol production was obtained when applying the optimum values for temperature (32°C), pH (5.6) and fermentation time (110 h). Maximum ethanol concentration (3.36 g/l) was obtained from 50 g/l pretreated sugarcane bagasse at the optimized process conditions in aerobic batch fermentation. Kinetic models such as Monod, Modified Logistic model, Modified Logistic incorporated Leudeking – Piret model and Modified Logistic incorporated Modified Leudeking – Piret model have been evaluated and the constants were predicted.
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
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.