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Journal ArticleDOI

Fermentative production and kinetics of cellulase protein on Trichoderma reesei using sugarcane bagasse and rice straw

01 Jan 2006-African Journal of Biotechnology (Academic Journals (Kenya))-Vol. 5, Iss: 20

TL;DR: This is the first attempt of combining the synthetic substrate (xylose, lactose) with natural substrate (sugarcane bagasse, rice straw) and the mixture of substrates produced the highest maximal enzyme activity on cellulose with xylose.

AbstractCellulase a multienzyme made up of several proteins finds extensive applications in food, fermentation and textile industries. Trichoderma reesei is an efficient producer of cellulase protein. The comparative study was made on various carbon sources on the production of cellulase using strains of T. reesei QM 9414, 97.177 and Tm3. Pretreatment of sugarcane bagasse and rice straw offers very digestible cellulose and potentially less inhibition. Cellulase production was enhanced by multiple carbon sources because of diauxic pattern of utilization of substrates. This is the first attempt of combining the synthetic substrate (xylose, lactose) with natural substrate (sugarcane bagasse, rice straw). The mixture of substrates produced the highest maximal enzyme activity on cellulose with xylose, cellulose with lactose, bagasse with xylose, bagasse with lactose, rice straw with xylose and rice straw with lactose. In addition Monod growth kinetics and Leudeking piret product formation kinetics were studied using T. reesei with optimized medium under optimized conditions of inoculum concentration, D.O. level, agitator speed, temperature and pH.

Topics: Cellulase (61%), Trichoderma reesei (59%), Bagasse (59%), Xylose (56%), Cellulose (52%)

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Journal ArticleDOI
Abstract: This study highlights the recent advances in the treatment and value addition of lignocellulosic wastes (LCW) with main focus on domestic and agro-industrial residues. Mechanical, physical and biological treatment systems are brought into perspective. The main value-added products from lignocellulosic wastes are summarized in a manner that pinpoints the most recent trends and the future directions. Physicochemical and biological treatment systems seem to be the most favored options while biofuels, biodegradable composites and biosorbents production paints a bright picture of the current and future bio-based products. Engineered microbes seem to tackle the problem of bioconversion of substrates that are otherwise non convertible by conventional wild strains. Although the main challenge facing LCW utilization is the high costs involved in treatment and production processes, some recent affordable processes with promising results have been proposed. Future trends are being directed to nanobiotechnology and genetic engineering for improved processes and products. The paper presents state of the art review of the dual advantage of handling LCW for cleaner environment and production of renewable bio-products.

219 citations


Cites background from "Fermentative production and kinetic..."

  • ...…production from LCW has been extensively studied (Jecu, 2000; Emtiazi and Nahvi, 2000; El-hawary and Mostafa, 2001; Ögel et al., 2001; Raj and Singh, 2001; Ojumu et al., 2003; Wen et al., 2005; Muthuvelayudham and Viruthagiri, 2006; Pothiraj et al., 2006; Daroit et al., 2007; Gao et al., 2008)....

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Journal ArticleDOI
TL;DR: Filamentous fungi Trichoderma reesei are considered to be one of the most efficient hyper producers of cellulase that is used in industry and were carried out using high concentration of cellulose to substitute glucose with the aim to improve cellulase production while trying to reduce production costs.
Abstract: Filamentous fungi Trichoderma reesei are considered to be one of the most efficient hyper producers of cellulase that is used in industry. Cellulase production by T. reesei was carried out using high concentration of cellulose to substitute glucose with the aim to improve cellulase production while trying to reduce production costs. The experiments were conducted separately as fed batch growth with T. reesei cultured using four media in a 7 L stirred tank bioreactor. A mixture of lactose and lactobionic acid was added into the bioreactor as cellulase inducers. The use of a cellulose–yeast extract culture medium yielded the highest enzyme and cell production with a volumetric enzyme activity of 69.8 U L−1 h−1, a filter paper activity of 5.02 U mL−1, a CMCase activity of 4.2 U mL−1, and a fungal biomass of 14.7 g L−1. The biomass concentration as a function of time was constant with relatively rapid, early growth on easily metabolized growth medium components (yeast extract), followed by a second slower growth phase due to hydrolysis of cellulose, which follow cellulase concentration augmentation. The costs to produce 1 L of production medium in laboratory-scale experiments were calculated to compare the tested media.

203 citations


Journal Article
TL;DR: The production of cellulolytic enzymes by a local isolate of A. heteromorphus on lignocelluloses in submerged fermentation is reported, indicating potential to reduce use of fossil fuels and reduce environmental pollution in India.
Abstract: enzyme production and meeting the increase in awareness on energy conservation and recycling. The conversion of cellulosic mass to fermentable sugars through biocatalyst cellulase derived from cellulolytic organisms has been suggested as a feasible process and offers potential to reduce use of fossil fuels and reduce environmental pollution (5), (6). Cellulase production by different organisms in submerged state fermentation has received more attention and is found to be cost-prohibitive because of high cost of process engineering. India is an agricultural country and wheat is one of the most important agricultural crop in India. Every year nearly 78.4 Million Tones wheat is produced. With the processesing of wheat grains a large amount of straw is produced which can be used as a substrate for ethanol production. Currently a most important application of cellulases and hemicellulases in the pulp and paper industry is the biobleaching of pulp, the production of dissolving pulp, the treatment of wastewater and the deinking of recycled waste paper. The potential of enzymatic treatments has been assessed and the processes have proved successful (7), (8). In the present study the production of cellulolytic enzymes by a local isolate of A. heteromorphus on lignocelluloses in submerged fermentation is reported.

81 citations


Cites result from "Fermentative production and kinetic..."

  • ...And CMCase activity was similar as reported by Muthuvelayudham and Viruthagir [15] using Sugarcane baggase as substrate by T. reesei 94.144....

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  • ...And CMCase activity was similar as reported by Muthuvelayudham and Viruthagir [15] using Sugarcane baggase as substrate by T....

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Journal ArticleDOI
15 May 2014-Fuel
TL;DR: A mutant strain of Trichoderma asperellum RCK2011 was developed through UV-irradiation for enhanced cellulase production and lower catabolite repression and the production of FPase, CMCase and β-glucosidase was optimized under solid state fermentation.
Abstract: A mutant strain of Trichoderma asperellum RCK2011 was developed through UV-irradiation for enhanced cellulase production and lower catabolite repression. The production of FPase, CMCase and β-glucosidase was optimized under solid state fermentation; up to 20 mM of glucose did not inhibit cellulase production. The mutant strain T. asperellum SR1-7 produced FPase (2.2 IU/gds), CMCase (13.2 IU/gds), and β-glucosidase (9.2 IU/gds) under optimized conditions, which is, 1.4, 1.3, 1.5-fold higher than the wild type. The wild as well as mutant strain produced the cellulases at pH range, 4.0–10.0. Saccharification of pretreated corn cob, wheat straw, and sugarcane bagasse by cellulase from mutant strain SR1-7 resulted in release of reducing sugar at the rate of 530.0 mg/g, 290.0 mg/g, and 335.0 mg/g of substrate, respectively; this is 1.6-fold higher than the wild type strain.

72 citations


Journal ArticleDOI
TL;DR: Proteomic analyses and enzymology studies are combined to show that sequential and submerged cultivation methods differently influence both titers and secretion profile of key enzymes required for the hydrolysis of sugarcane bagasse.
Abstract: Cellulases and hemicellulases from Trichoderma reesei and Aspergillus niger have been shown to be powerful enzymes for biomass conversion to sugars, but the production costs are still relatively high for commercial application. The choice of an effective microbial cultivation process employed for enzyme production is important, since it may affect titers and the profile of protein secretion. We used proteomic analysis to characterize the secretome of T. reesei and A. niger cultivated in submerged and sequential fermentation processes. The information gained was key to understand differences in hydrolysis of steam exploded sugarcane bagasse for enzyme cocktails obtained from two different cultivation processes. The sequential process for cultivating A. niger gave xylanase and β-glucosidase activities 3- and 8-fold higher, respectively, than corresponding activities from the submerged process. A greater protein diversity of critical cellulolytic and hemicellulolytic enzymes were also observed through secretome analyses. These results helped to explain the 3-fold higher yield for hydrolysis of non-washed pretreated bagasse when combined T. reesei and A. niger enzyme extracts from sequential fermentation were used in place of enzymes obtained from submerged fermentation. An enzyme loading of 0.7 FPU cellulase activity/g glucan was surprisingly effective when compared to the 5-15 times more enzyme loadings commonly reported for other cellulose hydrolysis studies. Analyses showed that more than 80% consisted of proteins other than cellulases whose role is important to the hydrolysis of a lignocellulose substrate. Our work combined proteomic analyses and enzymology studies to show that sequential and submerged cultivation methods differently influence both titers and secretion profile of key enzymes required for the hydrolysis of sugarcane bagasse. The higher diversity of feruloyl esterases, xylanases and other auxiliary hemicellulolytic enzymes observed in the enzyme mixtures from the sequential fermentation could be one major reason for the more efficient enzyme hydrolysis that results when using the combined secretomes from A. niger and T. reesei.

57 citations


Cites background from "Fermentative production and kinetic..."

  • ...21), which act synergistically in the conversion of cellulose into glucose [35, 36]....

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References
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Journal ArticleDOI
TL;DR: The conversion of both cellulose and hemicellulose for the production of fuel ethanol is being studied intensively, with a view to developing a technically and economically viable bioprocess.
Abstract: Lignocellulosic materials containing cellulose, hemicellulose, and lignin are the most abundant renewable organic resource on earth. The utilization of renewable resources for energy and chemicals is expected to increase in the near future. The conversion of both cellulose (glucose) and hemicellulose (hexose and pentose) for the production of fuel ethanol is being studied intensively, with a view to developing a technically and economically viable bioprocess. Whereas the fermentation of glucose can be carried out efficiently, the bioconversion of the pentose fraction (xylose and arabinose, the main pentose sugars obtained on hydrolysis of hemicellulose), presents a challenge. A lot of attention has therefore been focused on genetically engineering strains that can efficiently utilize both glucose and pentoses, and convert them to useful compounds, such as ethanol. Metabolic strategies seek to generate efficient biocatalysts (bacteria and yeast) for the bioconversion of most hemicellulosic sugars to products that can be derived from the primary metabolism, such as ethanol. The metabolic engineering objectives so far have focused on higher yields, productivities and expanding the substrate and product spectra.

481 citations


"Fermentative production and kinetic..." refers background in this paper

  • ...…growth rate (day)-1; µ, specific growth rate (day)–1; Ks, Monod constant; P, product concentration (g/l); α and β, Leudeking Piret constants; and X, substrate concentration (g/l). a number of applications including animal feed, pharmaceutical and textile industries (Aristidou and Penttilä, 2000)....

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  • ...a number of applications including animal feed, pharmaceutical and textile industries (Aristidou and Penttilä, 2000)....

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Journal ArticleDOI
TL;DR: RNA slot blot experiments showed that no expression could be observed on glucose-containing medium and that high glucose levels abolish the inducing effect of sophorose, showing that distinct and clear-cut mechanisms of induction and glucose repression regulate cellulase expression in an actively growing fungus.
Abstract: Basic features of regulation of expression of the genes encoding the cellulases of the filamentous fungus Trichoderma reesei QM9414, the genes cbh1 and cbh2 encoding cellobiohydrolases and the genes egl1, egl2 and egl5 encoding endoglucanases, were studied at the mRNA level. The cellulase genes were coordinately expressed under all conditions studied, with the steady-state mRNA levels of cbh1 being the highest. Solka floc cellulose and the disaccharide sophorose induced expression to almost the same level. Moderate expression was observed when cellobiose or lactose was used as the carbon source. It was found that glycerol and sorbitol do not promote expression but, unlike glucose, do not inhibit it either, because the addition of 1 to 2 mM sophorose to glycerol or sorbitol cultures provokes high cellulase expression levels. These carbon sources thus provide a useful means to study cellulase regulation without significantly affecting the growth of the fungus. RNA slot blot experiments showed that no expression could be observed on glucose-containing medium and that high glucose levels abolish the inducing effect of sophorose. The results clearly show that distinct and clear-cut mechanisms of induction and glucose repression regulate cellulase expression in an actively growing fungus. However, derepression of cellulase expression occurs without apparent addition of an inducer once glucose has been depleted from the medium. This expression seems not to arise simply from starvation, since the lack of carbon or nitrogen as such is not sufficient to trigger significant expression.

471 citations


"Fermentative production and kinetic..." refers background in this paper

  • ...Hypercelluloytic mutant strains secrete large amounts of cellulases (Ilmen et al., 1997)....

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Journal ArticleDOI
TL;DR: This mini-review summarises the considerable amount of data accumulated over the past three decades regarding promoters of genes encoding hydrolytic enzymes, inducer molecules, and models for the recognition of the insoluble substrates cellulose and xylan.
Abstract: The genus Trichoderma comprises a group of filamentous ascomycetes that are now widely used in industrial applications because of their ability to produce extracellular hydrolases in large amounts. In addition, strong inducible promoters together with high secretory capacity have made Trichoderma an attractive host for heterologous protein production. Several promoters of genes encoding hydrolytic enzymes have been investigated in detail regarding their cis-acting elements and trans-acting factors. Potent inducer molecules, for both xylanolytic and cellulolytic enzyme systems, have been identified and characterized. Furthermore, models for the recognition of the insoluble substrates cellulose and xylan have been developed based on a large set of experiments. This mini-review summarises the considerable amount of data accumulated over the past three decades.

201 citations


"Fermentative production and kinetic..." refers background in this paper

  • ...The genus Trichoderma, filamentous ascomycetes are widely used in industrial applications because of high secretory capacity and inducible promoting characteristics (Mach and Zeilinger, 2003)....

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Journal ArticleDOI
TL;DR: The growth and enzyme production by Trichoderma reesei Rut C-30 using different lignocellulosic materials as carbon source were investigated and it was found that endoglucanase and endoxylanse activities were produced throughout the cultivations, whereas α-arabinosidase was induced late during the cultivation.
Abstract: The growth and enzyme production by Trichoderma reesei Rut C-30 using different lignocellulosic materials as carbon source were investigated Cellulose, sugar beet pulp and alkaline extracted sugar beet pulp (resulting in partial removal of hemicellulose, lignin and pectin) or mixtures thereof were used as carbon sources It was found that endoglucanase and endoxylanse activities were produced throughout the cultivations, whereas α-arabinosidase was induced late during the cultivation The highest amounts of endoglucanse, could be measured when T reesei Rut C-30 was grown on cellulose or cellulose containing mixtures Endoxylanase was produced on all substrates, but the presence of cellulose was favourable for the production Polygalacturonase activity could be measured at high varying levels throughout the cultivations, except during growth on cellulose The varying levels might originate from the production of different isoenzymes of polygalacturonase

150 citations


"Fermentative production and kinetic..." refers background in this paper

  • ...Biosynthesis of cellulase was also made on lignocellulo-sic materials such as sugar beet pulp and alkaline extracted sugar beet pulp and cellulose (Olsson et al., 2003)....

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  • ...The general trend is that more cellulose in the mixture results in higher levels of endoglucanase (Olsson et al., 2003)....

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Journal ArticleDOI
TL;DR: The production of cellulose and hemicellulose-degrading enzymes by cultivation of Aspergillus niger, Botrytis cinerea, Penicillium brasilianum, Schizophyllum commune, and Trichoderma reesei Rut-C30 was studied to obtain an enzyme mixture optimal for enzymatic hydrolysis of wet-oxidised wheat straw.
Abstract: The production of cellulose and hemicellulose-degrading enzymes by cultivation of Aspergillus niger ATCC 9029, Botrytis cinerea ATCC 28466, Penicillium brasilianum IBT 20888, Schizophyllum commune ATCC 38548, and Trichoderma reesei Rut-C30 was studied. Wet-oxidised wheat straw suspension supplemented with NH4NO3, MgSO4, and KH2PO4 was used as cultivation medium aiming to obtain an enzyme mixture optimal for enzymatic hydrolysis of wet-oxidised wheat straw. The cultivations with B. cinerea and P. brasilianum gave the highest endoglucanase (EC 3.2.1.4) and β-glucosidase (EC 3.2.1.21) activities, in contrast to the other fungi where lower activities were found. The culture filtrates were concentrated by ammonium sulphate precipitation. After enzyme concentration, the highest enzyme activities (1.34 FPU/ml) were found in the culture broth originating from P. brasilianum. Enzymatic hydrolysis of filter cake from wet-oxidised wheat straw for 48 h with an enzyme loading of 5 FPU/g biomass resulted in glucose yields from cellulose of 58% (w/w) and 39% (w/w) using enzymes produced by P. brasilianum and a commercial enzyme mixture, respectively. At higher enzyme loading (25 FPU/g biomass) using either enzyme mixtures the glucose yield from cellulose was in the range of 77–79% (w/w).

124 citations


"Fermentative production and kinetic..." refers background or methods in this paper

  • ...The formation of reducing sugar was measured by DNS method (Thygesen et al., 2003)....

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  • ...Cellulose and hemi cellu-lose-degrading enzyme was produced on wet-oxidized wheat straw using various filamentous fungi (Thygesen et al., 2003)....

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