Search results for: Trichoderma viride

Authors: F. L. Motta, M. H. A. Santana

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Humic acids (HA) were produced by a Trichoderma viride strain under submerged fermentation in a medium based on the oil palm empty fruit bunch (EFB) and the main variables of the process were optimized by using response surface methodology. A temperature of 40°C and concentrations of 50g/L EFB, 5.7g/L potato peptone and 0.11g/L (NH4)2SO4 were the optimum levels of the variables that maximize the HA production, within the physicochemical and biological limits of the process. The optimized conditions led to an experimental HA concentration of 428.4±17.5 mg/L, which validated the prediction from the statistical model of 412.0mg/L. This optimization increased about 7–fold the HA production previously reported in the literature. Additionally, the time profiles of HA production and fungal growth confirmed our previous findings that HA production preferably occurs during fungal sporulation. The present study demonstrated that T. viride successfully produced HA via the submerged fermentation of EFB and the process parameters were successfully optimized using a statistics-based response surface model. To the best of our knowledge, the present work is the first report on the optimization of HA production from EFB by a biotechnological process, whose feasibility was only pointed out in previous works.

Keywords: Empty fruit bunch, humic acids, submerged fermentation, Trichoderma viride.

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Authors: R. Rajendiran, S. Gayathri devi, B.T. SureshKumar, V. Arul Priya

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Plackett-Burman statistical screening of media constituents and operational conditions for extracellular lipase production from isolate Trichoderma viride has been carried out in submerged fermentation. This statistical design is used in the early stages of experimentation to screen out unimportant factors from a large number of possible factors. This design involves screening of up to 'n-1' variables in just 'n' number of experiments. Regression coefficients and t-values were calculated by subjecting the experimental data to statistical analysis using Minitab version 15. The effects of nine process variables were studied in twelve experimental trials. Maximum lipase activity of 7.83 μmol /ml /min was obtained in the 6th trail. Pareto chart illustrates the order of significance of the variables affecting the lipase production. The present study concludes that the most significant variables affecting lipase production were found to be palm oil, yeast extract, K2HPO4, MgSO4 and CaCl2.

Keywords: lipase, submerged fermentation, statistical optimization, Trichoderma viride

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Authors: Olufunke O. Ezekiel, Ogugua C. Aworh

Abstract:

Solid state fermentation of cassava peel with emphasis on protein enrichment using Trichoderma viride was evaluated. The effect of five variables: moisture content, pH, particle size (p), nitrogen source and incubation temperature; on the true protein and total sugars of cassava peel was investigated. The optimum fermentation period was established to be 8 days. Total sugars were 5-fold higher at pH 6 relative to pH 4 and 7-fold higher when cassava peels were fermented at 30oC relative to 25oC as well as using ammonium sulfate as the nitrogen source relative to urea or a combination of both. Total sugars ranged between 123.21mg/g at 50% initial moisture content to 374mg/g at 60% and from 190.59mg/g with particle size range of 2.00>p>1.41mm to 310.10mg/g with 4.00>p>3.35mm.True protein ranged from 229.70 mg/g at pH 4 to 284.05 mg/g at pH 6; from 200.87 mg/g with urea as nitrogen source and to 254.50mg/g with ammonium sulfate; from 213.82mg/g at 50% initial moisture content to 254.50mg/g at 60% moisture content, from 205.75mg/g in cassava peel with 5.6>p> 4.75mm to 268.30 in cassava peel with particle size 4.00>p>3.35mm, from 207.57mg/g at 25oC to 254.50mg/g at 30oC Cassava peel with particle size 4.00>p>3.35 mm and initial moisture content of 60% at pH 6.0, 30oC incubation temperature with ammonium sulfate (10g N / kg substrate) was most suitable for protein enrichment with Trichoderma viride. Crude protein increased from 4.21 % in unfermented cassava peel samples to 10.43 % in fermented samples.

Keywords: Cassava peel, Solid state fermentation, Trichoderma viride, Total sugars, True protein.

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Authors: Hajji Lobna, Chattaoui Mayssa, Regaieg Hajer, M'Hamdi-Boughalleb Naima, Rhouma Ali, Horrigue-Raouani Najet

Abstract:

In this study, three local isolates of Trichoderma (Tr1: T. viride, Tr2: T. harzianum and Tr3: T. asperellum) were isolated and evaluated for their biocontrol effectiveness under in vitro conditions and in greenhouse. In vitro bioassay revealed a biopotential control against Fusarium oxysporum f. sp. radicis lycopersici and Meloidogyne javanica (RKN) separately. All species of Trichoderma exhibited biocontrol performance and (Tr1) Trichoderma viride was the most efficient. In fact, growth rate inhibition of Fusarium oxysporum f. sp. radicis lycopersici (FORL) was reached 75.5% with Tr1. Parasitism rate of root-knot nematode was 60% for juveniles and 75% for eggs with the same one. Pots experiment results showed that Tr1 and Tr2, compared to chemical treatment, enhanced the plant growth and exhibited better antagonism against root-knot nematode and root-rot fungi separated or combined. All Trichoderma isolates revealed a bioprotection potential against Fusarium oxysporum f. sp. radicis lycopersici. When pathogen fungi inoculated alone, Fusarium wilt index and browning vascular rate were reduced significantly with Tr1 (0.91, 2.38%) and Tr2 (1.5, 5.5%), respectively. In the case of combined infection with Fusarium and nematode, the same isolate of Trichoderma Tr1 and Tr2 decreased Fusarium wilt index at 1.1 and 0.83 and reduced the browning vascular rate at 6.5% and 6%, respectively. Similarly, the isolate Tr1 and Tr2 caused maximum inhibition of nematode multiplication. Multiplication rate was declined at 4% with both isolates either tomato infected by nematode separately or concomitantly with Fusarium. The chemical treatment was moderate in activity against Meloidogyne javanica and Fusarium oxysporum f. sp. radicis lycopersici alone and combined.

Keywords: Trichoderma spp., Meloidogyne javanica, Fusarium oxysporum f.sp. radicis lycopersici, biocontrol.

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Authors: Ishtiaq Ahmed, Muhammad Anjum Zia, Hafiz Muhammad Nasir Iqbal

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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.

Keywords: Cellulase, Lignocellulosic residue, Processoptimization, Proximal analysis, SSF, Trichoderma viride.

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Authors: Dayana Amira R., Roshanida A.R., Rosli M.I.

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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.

Keywords: EFB, cellulase, POME, xylanase

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Authors: M. D. S. Siti-Normah, S. Sabiha-Hanim, A. Noraishah

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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.

Keywords: Hemicellulose, oil palm fronds, Trichoderma longibrachiatum, xylooligosaccharides.

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Authors: Rashid S S, Alam M Z, Karim M I A, Salleh, M H

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 (<0.0011) and peptone (0.0021) indicated the significant effect on the production of cellulase with the determination coefficient (R2) of 0.995.

Keywords: Face centered central composite design (FCCCD), Liquid state bioconversion (LSB), Palm oil mill effluent, Trichoderma reesei RUT C-30.

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Authors: Khosro Mohammadi, Amir Ghalavand, Majid Aghaalikhani

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In order to Study the efficacy application of green manure as chickpea pre plant, field experiments were carried out in 2007 and 2008 growing seasons. In this research the effects of different strategies for soil fertilization were investigated on grain yield and yield component, minerals, organic compounds and cooking time of chickpea. Experimental units were arranged in splitsplit plots based on randomized complete blocks with three replications. Main plots consisted of (G1): establishing a mixed vegetation of Vicia panunica and Hordeum vulgare and (G2): control, as green manure levels. Also, five strategies for obtaining the base fertilizer requirement including (N1): 20 t.ha-1 farmyard manure; (N2): 10 t.ha-1 compost; (N3): 75 kg.ha-1 triple super phosphate; (N4): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost and (N5): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost + 50 kg.ha-1 triple super phosphate were considered in sub plots. Furthermoree four levels of biofertilizers consisted of (B1): Bacillus lentus + Pseudomonas putida; (B2): Trichoderma harzianum; (B3): Bacillus lentus + Pseudomonas putida + Trichoderma harzianum; and (B4): control (without biofertilizers) were arranged in sub-sub plots. Results showed that integrating biofertilizers (B3) and green manure (G1) produced the highest grain yield. The highest amounts of yield were obtained in G1×N5 interaction. Comparison of all 2-way and 3-way interactions showed that G1N5B3 was determined as the superior treatment. Significant increasing of N, P2O5, K2O, Fe and Mg content in leaves and grains emphasized on superiority of mentioned treatment because each one of these nutrients has an approved role in chlorophyll synthesis and photosynthesis abilities of the crops. The combined application of compost, farmyard manure and chemical phosphorus (N5) in addition to having the highest yield, had the best grain quality due to high protein, starch and total sugar contents, low crude fiber and reduced cooking time.

Keywords: chickpea, biofertilizer, nitrogen fixation.

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Authors: Khosro Mohammadi, Amir Ghalavand, Majid Aghaalikhani

Abstract:

In order to evaluation the effects of soil organic matter and biofertilizer on chickpea quality and biological nitrogen fixation, field experiments were carried out in 2007 and 2008 growing seasons. In this research the effects of different strategies for soil fertilization were investigated on grain yield and yield component, minerals, organic compounds and cooking time of chickpea. Experimental units were arranged in split-split plots based on randomized complete blocks with three replications. Main plots consisted of (G1): establishing a mixed vegetation of Vicia panunica and Hordeum vulgare and (G2): control, as green manure levels. Also, five strategies for obtaining the base fertilizer requirement including (N1): 20 t.ha-1 farmyard manure; (N2): 10 t.ha-1 compost; (N3): 75 kg.ha-1 triple super phosphate; (N4): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost and (N5): 10 t.ha-1 farmyard manure + 5 t.ha-1 compost + 50 kg.ha-1 triple super phosphate were considered in sub plots. Furthermoree four levels of biofertilizers consisted of (B1): Bacillus lentus + Pseudomonas putida; (B2): Trichoderma harzianum; (B3): Bacillus lentus + Pseudomonas putida + Trichoderma harzianum; and (B4): control (without biofertilizers) were arranged in sub-sub plots. Results showed that integrating biofertilizers (B3) and green manure (G1) produced the highest grain yield. The highest amounts of yield were obtained in G1×N5 interaction. Comparison of all 2-way and 3-way interactions showed that G1N5B3 was determined as the superior treatment. Significant increasing of N, P2O5, K2O, Fe and Mg content in leaves and grains emphasized on superiority of mentioned treatment because each one of these nutrients has an approved role in chlorophyll synthesis and photosynthesis abilities of the crops. The combined application of compost, farmyard manure and chemical phosphorus (N5) in addition to having the highest yield, had the best grain quality due to high protein, starch and total sugar contents, low crude fiber and reduced cooking time.

Keywords: chickpea, biofertilizer, nitrogen fixation.

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Authors: Snezana Rajkovic, Miroslava Markovic, Radoslav Rajkovic, Ljubinko Rakonjac

Abstract:

Grey mold on grape is caused by the fungus Botrytis cinerea Pers. Trichodex WP, a new biofungicide, that contains fungal spores of Trichoderma harzianum Rifai, was used for biological control of Grey mold on grape. The efficacy of Trichodex WP has been reported from many experiments. Experiments were carried out in the locality – Banatski Karlovac, on grapevine species – talijanski rizling. The trials were set according to instructions of methods PP1/152(2) and PP1/17(3) , according to a fully randomized block design. Phytotoxicity was estimated by PP methods 1/135(2), the intensity of infection according to Towsend Heuberger , the efficiency by Abbott, the analysis of variance with Duncan test and PP/181(2). Application of Trichodex WP is limited to the first two treatments. Other treatments are performed with the fungicides based on a.i. procymidone, vinclozoline and iprodione.

Keywords: Biofungicides, efficacy, grey mold, Trichodex WP.

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Authors: Mohamed F. Awad, M. Kraume

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The objective of this study is to evaluate the occurrence of fungi in aerobic and anoxic activated sludge from membrane bioreactors (MBRs). Thirty-six samples of both aerobic and anoxic activated sludge were taken from 2 MBR treating domestic wastewater. Over a period of eight months 2 samples from each plant were taken per month. The samples were prepared for count and definition of fungi. The obtained data show that, sixty species belonging to 27 genera were collected from activated sludge samples under aerobic and anoxic conditions. Regarding to the fungi definition, under aerobic condition the Geotrichum was found at (8.8%) followed by Penicillium (75.0%), Yeasts (65.7%) and Trichoderma (55.5%), while Yeasts (77.1%) Geotrichum candidumand Penicillium (61.1%) species were the most prevalent in anoxic activated sludge. The results indicate that activated sludge is habitat for growth and sporulation of different groups of fungi, both saprophytic and pathogenic.

Keywords: Aerobic conditions, Anoxic conditions, Activated sludge, Membrane bioreactor, Fungi.

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Authors: Rashid S. S., Alam M. Z.

Abstract:

A statistical optimization of the saccharification process of EFB was studied. The statistical analysis was done by applying faced centered central composite design (FCCCD) under response surface methodology (RSM). In this investigation, EFB dose, enzyme dose and saccharification period was examined, and the maximum 53.45% (w/w) yield of reducing sugar was found with 4% (w/v) of EFB, 10% (v/v) of enzyme after 120 hours of incubation. It can be calculated that the conversion rate of cellulose content of the substrate is more than 75% (w/w) which can be considered as a remarkable achievement. All the variables, linear, quadratic and interaction coefficient, were found to be highly significant, other than two coefficients, one quadratic and another interaction coefficient. The coefficient of determination (R2) is 0.9898 that confirms a satisfactory data and indicated that approximately 98.98% of the variability in the dependent variable, saccharification of EFB, could be explained by this model.

Keywords: Face centered central composite design (FCCCD), Liquid state bioconversion (LSB), Palm oil mill effluent, Trichoderma reesei RUT C-30.

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Authors: Mohamed F. Awad, M. Kraume

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Thirty six samples from each (aerobic and anoxic) activated sludge were collected from two wastewater treatment plants with MBRs in Berlin, Germany. The samples were prepared for count and definition of fungal isolates; these isolates were purified by conventional techniques and identified by microscopic examination. Sixty tow species belonging to 28 genera were isolated from activated sludge samples under aerobic conditions (28 genera and 58 species) and anoxic conditions (26 genera and 52 species). The obtained data show that, Aspergillus was found at 94.4% followed by Penicillium 61.1 %, Fusarium (61.1 %), Trichoderma (44.4 %) and Geotrichum candidum (41.6 %) species were the most prevalent in all activated sludge samples. The study confirmed that fungi can thrive in activated sludge and sporulation, but isolated in different numbers depending on the effect of aeration system. Some fungal species in our study are saprophytic, and other a pathogenic to plants and animals.

Keywords: Activated sludge, membrane bioreactors, aerobic, anoxic conditions, fungi

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Authors: L. V. Ignatova, Y. V. Brazhnikova, T. D. Mukasheva, A. A. Omirbekova, R. Zh. Berzhanova, R. K. Sydykbekova, T. A. Karpenyuk, A. V. Goncharova

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The comparative analysis of different taxonomic groups of microorganisms isolated from dark chernozem soils under different agricultures (alfalfa, melilot, sainfoin, soybean, rapeseed) at Almaty region of Kazakhstan was conducted. It was shown that the greatest number of micromycetes was typical to the soil planted with alfalfa and canola. Species diversity of micromycetes markedly decreases as it approaches the surface of the root, so that the species composition in the rhizosphere is much more uniform than in the virgin soil. Promising strains of microscopic fungi and yeast with plant growth-promoting activity to agricultures were selected. Among the selected fungi there are representatives of Penicillium bilaiae, Trichoderma koningii, Fusarium equiseti, Aspergillus ustus. The highest rates of growth and development of seedlings of plants observed under the influence of yeasts Aureobasidium pullulans, Rhodotorula mucilaginosa, Metschnikovia pulcherrima. Using molecular - genetic techniques confirmation of the identification results of selected micromycetes was conducted.

Keywords: Agricultures, biodiversity, micromycetes, plant growth-promoting microorganisms.

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Authors: R.Muthuvelayudham, T.Viruthagiri

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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.

Keywords: Banana fiber, Cellulase, Optimization, Rice straw

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Authors: Nasim Shaibani, Saba Ghazvini, Mohammad R. Andalibi, Soheila Yaghmaei

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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.

Keywords: Alkali pretreatment, bioethanol, cellulase, simultaneous saccharification and fermentation, solid statefermentation, sugarcane bagasse

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Authors: M. A. Kassim, R. Potumarthi, A. Tanksale, S. C. Srivatsa, S. Bhattacharya

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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.

Keywords: Microalgal biomass, enzymatic saccharification, biobutanol, fermentation.

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Authors: Md. Z. Alam, Amal A. Elgharbawy, Muhammad Moniruzzaman, Nassereldeen A. Kabbashi, Parveen Jamal

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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.

Keywords: Cellulase, hydrolysis, lignocellulose, pretreatment, stability.

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