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: 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: 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: 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: 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: 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: Rice bran has been abandoned as agricultural waste for million tonnes per year in Thailand, therefore they have been proposed to be utilized as a rich carbon source in the production of bioethanol. Many toxic compounds are possibly released during the pretreatment of rice bran prior the fermentation process. This study aims to analyze on the availability of toxic compounds and the amount of glucose obtained from 2 different pretreatments using sulfuric acid and mixed cellulase enzymes (without and with delignification/ activated charcoal). The concentration of furfural, 5- hydroxymethyl furfural (5-HMF), levulinic acid, vanillin, syringaldehyde and4-hydroxybenzaldehyde (4-HB) and the percent acetic acid were found to be 0.0517 ± 0.049 mg/L, 0.032 ± 0.06 mg/L, 21074 ± 1685.62 mg/L, 126.265 ± 6.005 mg/L, 2.89 ± 0.30 mg/L, 0.37 ± 0.031mg/L and 0.72% under the pretreatment process without delignification/ activated charcoal treatment and 384.47 ± 99.02 g/L, 0.068 mg/L, 142107.62 ± 8664.6 mg/L, 0.19 mg/L, 5.43 ± 3.29 mg/L, 4.80 ± 0.76 mg/L and 0.254% under the pretreatment process with delignification/ activated charcoal treatment respectively. The presence of high concentration of acetic acid was found to impede the growth of Zymomonas mobilis strain TISTR 551 despite the present of high concentration of levulinic acid. Z. mobilis strain TISTR 551 was found to produce 8.96 ± 4.06 g/L of ethanol under 4 days fementation period in biofilm stage in which represented 40% theoretical yield.